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للكيمياء العربية

The Official Journal For the Arab Union of Chemists

Published By Saudi Chemical Society

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Arabian Journal of Chemistry

ArabianJournal of Chemistry

Editor -in- Chief Prof. Abdulrahman A . AlwarthanChemistry DepartmentKing Saud University,Riyadh, Saudi ArabiaE-mail: [email protected]

Prof. Sultan T. Abu-OrabiPresidentYarmouk UniversityIrbid -JordanJordanian Chemical SocietyArab Union of Chemists/Secretary GeneralE-mail: [email protected]

Prof. Yousry M. IssaChemistry DepartmentCairo University, CairoEgyptE-mail: [email protected]

Vice-editors -in- Chief

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EDITORIAL BOARDProf. Belkheir HammoutiDirector, Laboratory of Applied Chemsitry and Envi-ronmentFaculty of ScienceUniversity of Mohammed PremierMoroccoE-mail: [email protected]

Prof. Mamia El RhaziChemistry DepartmentFaculty of Science and TechnologyHassan II Univeristy-MohammediaMoroccoE-mail: [email protected]

Prof. Hassan M. Al-HazimiChemistry DepartmentScience College, King Saud UniversitySaudi Arabia E-mail: [email protected]

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Prof. El Sayed H. El AshryChemistry DepartmentFaculty of ScienceUniversity of AlexandriaAlexandria, EgyptE-mail: [email protected]

Dr. Lassaad BakloutiLaboratoire de Chimie des Interactions MoleculairesFaculte de Sciences de Bizerte7021 Zarzouna, TunisieE-mail: [email protected]

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Mohammad HouraniDeartment of Chemistry, Al Balqa Applied University, Al-Salt, Jordan.E-mail:[email protected]

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Prof. Nouria A. Al-AwadiProfessor of Organic ChemsitryDepartment of Chemistry Faculty of Science P.O.Box: 5969 Safat-13060Kuwait UniversityE-mail: [email protected]

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Dr. Abdulaziz A. AlNajjarApplied Science, College of Technological Studies, Public Authority of Applied Education and TrainingP.O. Box 34484 Adeilia, 73255, Kuwait e-mail: [email protected]

Dr. Ameera Saeed Al-HaddadUniversity of Bahrain, Kingdom of Bahrain , Department of Chemistry, P.O. Box 32028, Isa Town, Kingdom of Bahrain E=mail: [email protected]

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INTERNATIONAL ADVISORY BOARDProf. Issa YavariChemistry DepartmentTarbiat Modarres UniversityTehran, IranE-mail: [email protected]

Dr. Paul S. FrancisSchool of Life and Environmental SciencesDeakin University, Geelong Victoria 3217, AustraliaE-mail: [email protected]

Prof. Jose Martinez CalatayudChemistry DepartmentUniversity of ValenciaValencia, SpainE-mail: [email protected]

Prof. Robert G. MichelDepartment of ChemistryUniversity of Connecticut55 North Eagleville RoadStorrs, CT 06269-3060E-mail: [email protected]

Prof. Motaza M. KhaterChemistry DepartmentCairo UniversityCairo, EgyptE-mail: [email protected]

Prof. Jacques VicensDirecteur de recherche at CNRSUMR 7178-CNRSInstitut Pluridisciplinaire Hubert CurienUniversite Louis Pasteur de StrasbourgLaboratoire de Conception Moleculaire`ECPM25, rue BecquerelF-67087FranceE-mail: [email protected]

Prof. Jean-Michel KAUFFMANNFree University of BrusselsLab, Instrumental Analysis and BioelectrochemistryPharmaceutical Institute, ULB 205/6Campus PlaineB-1050 Brussels, BelgiumE-mail: [email protected]

Prof. Essam Khamis Al-HanashVice-Dean for Graduate Studies & Research.

Faculty of Science, Mohram Bey, Alexandria University.Alexandria, EgyptE-mail: [email protected]

Prof. Bryan R. HenryUniversity of GuelphDepartment of ChemistryGuelph, Ontario NIG 2W1CanadaE-mail: [email protected]

Prof. Samy El-ShallDepartment of Chemistry, Virginia Commonwealth University Richmond,Virginia 23284-2006USAE-mail: [email protected].

Prof. JIN, JUNG-IL(M)35-41Ku-Ui 2-DongKwang-Jin Ku, Seoul 133-202, KoreaChemistry Department and Center for Electro-and Photo-Responsive Molecules, College of Sciences, Korea University5-1 Anam-Dong, Seoul 136-701, KoreaE-mail: [email protected], [email protected]

Prof. Alan TownshendThe University of HullDepartment of ChemistryHull, HU6 7RXUnited KingdomE-mail: [email protected]

Dr. Danielle M. Cleveland18347 woodland Ridge Drive # 14 Spring Lake, MI 49456E-mail: [email protected]

Prof. Yuhan SunInstitute of Coal ChemistryChinese Academy of SciencesP.O.Box: 165,Taiyuan, Shanxi, 030001, PR. ChinaE-mail: [email protected]

Prof. Ishaque KhanDirector, Materials and Chemical Synthesis ProgramDepartment of Biological Chemical and Physical SciencesCollege of Science and LettersRoom 125, E110W, 32nd StreetChicago, IL 60616-3793E-mail: [email protected]

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INTERNATIONAL ADVISORY BOARDProf. Mikhail M. KrayushkinHead of Laboratory of Heterocyclic CompoundsN.D.Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences119991, Moscow, Leninsky Propspect 47, RussiaE-mail: [email protected]

Prof. Volker Schurig,Institute of Organic chemsitry,University of Tubingen, Auf der Morgenstelle 18,72076 Tubingen, GermanyE-mail:[email protected]

Prof. Angel Rios CastroDepartment of Analytical Chemistry and food Tech., Faculty of ChemistryUniversity of Castilla- La ManchaAV. Camilo Jose Cela. 10, E--13004 Ciudad Real, Spain.Phone: +34 926 295232 /Fax: +34 926 295318E-mail: [email protected]

Prof. Faiza M. Al-KharafiDepartment of ChemsitryFaculty of ScienceKuait UniversityKuwaitE-mail: [email protected]

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Scope and DescriptionArabian Journal of Chemistry (AJC) is an international quarterly

peer-reviewed research journal issued by the Arab Union of Chemists, and published by the Saudi Chemical Society, Riyadh, Saudi Arabia. The Journal publishes new and original Research Articles, Short Communications, Technical Notes, Feature Articles and Review Articles encompassing all fields of chemistry,experimental and theoretical, written either in English or Arabic.

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Manuscript Submission1- Hardcopy: The Original and three copies of the manuscript, together with a covering letter from the corresponding author, should submitted to the:

Editor-in-chief:Prof. Abdulrahman. A. AlwarthanEditor-in-ChiefArabian Journal of ChemistryChemistry Department, Faculty of ScienceKing Saud UniversityP.O.Box: 2455, Riyadh-11451Saudi ArabiaTel: 00966 1 4676005Fax: 00966 1 4675888E-mail:[email protected]

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Table of Contents

Volume 2, Number 1

Page

Physicochemical Studies on Cross-Linked Thorium(IV)-Alginate Complex Especially the Electrical Conductivity and Chemical Equilibrium Related to the Coordination Geometry. Ishaq A. Zaafarany Khalid S. Khairou , Refat M. Hassan and Yasuhisa Ikeda

1-10

Fluorescence Spectrometric Study of Eosin Yellow Dye-Surfactant Interactions Seema Acharya And Babulal Rebery 11-19

Photochromic Properties of 1,3,3-Trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-b][1,4]oxazine]

Doped in PMMA and Epoxy Resin Thin Films

Abdullah M. Asiri , Abood A. Bahaja, Abdullah G. Al-Sehem

21-30

The Use of Kinetic Methods For the Determination of Ultra-Trace Amount of Iodide in Water

F. Z. Shtewi, R. A. Mokhtar*, A. Al-Zawik and S. Karshman 31-42

Simultaneous Determination of Metal Ions as Complexes of Pentamethylene Dithiocarbamate IN Indus River Water , Pakistan Muhammad Amir Arain, Feroza Hamid Wattoo, Muhammad Hamid Sarwar Wattoo, Allah Bux Ghanghro, Syed Ahmad Tirmizi, Javed Iqbal and Shahnila Amir Arain

43-48

New Ceramic Microfiltration Membranes From Mineral Coal Fly Ash Ilyes Jedidi, Sami Saïdi, Sabeur Khmakem, André Larbot , Najwa Elloumi-Ammar, Amine Fourati, Aboulhassen Charfi and *Raja Ben Amar

49-62

Flow Injection Potentiometric Sensor for Determination of Phenylpropanolamine Hydrochloride Y. M. Issa, M. M. Khalil, S. I. M. Zayed and Ahmed Hussein 63-72

Experimental Study on Effect of Different Parameters on Size and Shape of Triangular Silver Nanoparticles Prepared by a Simple and Rapid Method in Aqueous Solution Seyed Soheil Mansouri, Sattar Ghader

73-88

Microwave and Ultrasound Promoted Synthesis of Substituted New Arylhydrazono Pyridinones Khadijah M. Al-Zaydi 89-94

Utility of Oxidation-Reduction Reaction for the Spectrophotometric Determination of AmlodipineBesylate Sayed A. Shama, Alaa S. Amin, El Sayed M. Mabrouk and Hany A. Omara

95-102

Synthesis and Characterization of New Poly(ester-amide)s containing Diarylidenecyclohexanone in the Main Chain. Part: II Khalid S. Khairou, Mohamed A. Abdullah, Kamal I. Aly*, Nariman M. Nahas

103-112

Study of Effect of Energy Drinks on Biochemical and Histological Markers in Rats Amani A. Al-Rasheedi 113-126

Arabian J. Chem. Vol. 2, No. 1, 1-10(2009)

Arabian J. Chem. Vol. 2, No. 1, (2009)

Physicochemical Studies on Cross-Linked Thorium(IV)-Alginate Complex Especially the Electrical Conductivity and Chemical Equilibrium Related to the Coordination Geometry

Ishaq A. Zaafarany1.* Khalid S. Khairou1 , Refat M. Hassan2 and Yasuhisa Ikeda3

1 Chemistry Department, Faculty of Applied Sciences, Umm Al-Qura University, Makkah

Al-Mukarramah- 13401, Saudi Arabia 2 Chemistry Department, Faculty of Science, Assiut University, Assiut- 71516, Egypt

3Research Laboratory for Nuclear Reactors, Tokyo Institute of Technology, Tokyo 152-8550, Japan

E-mail: [email protected]

Abstract

The electrical conductivity of cross-linked thorium(IV)-alginate complex in the form of circular disc has been

investigated as a function of temperature. The Arrhenius plot of log σ vs. 1/T showed a simple parabolic

shape at the early stages, followed by a sharply increase in ơ values with raising the temperature at the final

stages. This behaviour was interpreted by the formation of free-radicals at the initial stages, followed by the

degradation of the complex at elevated temperatures to give rise to thorium oxide product. The heterogeneous

chemical equilibrium for exchange of Th4+ counter ions in the complex by H+ ions has been investigated by

titrimetric and complexometric techniques. The thermodynamic equilibrium constant was found to be 26 ±

0.25 dm9 mol-3 at 25o C. The X-ray diffraction pattern indicated that thorium(IV)-alginate complex is

amorphous in nature. Infrared absorption spectra indicated that Th4+ is chelated to alginate macromolecular

chains and displayed υs OCO- and υas OCO- in the ranges of 1419 and 1635 cm-1, respectively. A geometrical

structure for chelation of thorium(IV) to the functional groups of alginate macromolecule is suggested and

discussed in terms of complex stability.

Keywords: Thorium(IV); Alginate; Complex; Electrical Conductivity; Chemical Equilibria.

1. Introduction

Alginic acid is a polyuronide comprising D-mannuronic

and L- guluronic acids linked through β(1→4) positions in

a linear block copolymer structure [1-4]. It is well known

that alginate has a high affinity for chelation with

polyvalent metal ions to form the corresponding cross-

linked complexes in either gel or granule forms depending

on the method of preparation [5,6]. A kind of chelation

occurs between the interdiffused metal ions and the

carboxylate and hydroxyl groups of the alginate

macromolecular chains [7-10].

Although, the electrical conductivity of synthetic

polymer complexes [11] has attracted many investigators

from both theoretical and practical points of view, a little

attention has been focused to that of natural polymers such

as metal alginate complexes. Indeed, Hassan and

coworkers studied the electrical conductivity of these

natural polymer derivative complexes under the influence

of high frequencies for the acid, divalent and trivalent

metal alginate complexes in either gel [12,13] or granule

[14] forms. On the other hand, analogous studies of the

Ishaq A. Zaafarany Khalid S. Khairou1 and Refat M. Hassan


- 2 -

change of conductance as a function of temperature for

monovalent [15], divalent [16], trivalent [17] and

hexavalent [18] metal alginate complexes in the form of

granules have been reported earlier.

In view of the above aspects, the present study seems

to be of interest to gain some information on the electrical

properties and chemical equilibrium of alginate complexes

containing cross-linked tetravalent metal ions. In addition,

the results obtained may shed some light on the stability of

these complexes in terms of their coordination geometry.

2. Experimental

Materials

The sodium alginate used was Cica-Reagent (Kanto Chem.

Co.). All other materials used were of analytical grade.

Doubly distilled conductively water was used in all

preparations.

Preparation of thorium (IV) alginate granules

Thorium(IV)-alginate complex in the form of granules was

prepared by the replacement of Na+ counter ions of alginate

macromolecule by Th4+ cations. This process was

performed by stepwise addition of the alginate powder to

an electrolyte of thorium(IV) ions while rapidly stirring the

solution to avoid the formation of lumps, which swell with

difficulty. After completion of the exchange process, the

grains formed were washed with deionized water until the

resultant water became free of Th4+ ions and then dried

under vacuum as described elsewhere [15,16].

Samples in the form of circular discs of diameter 13

mm and thickness 2-3 mm were obtained using an infrared

disc press at a constant pressure of 1500 p.s.i. (103 p.s.i. =

6.89 Nm-2).

X-ray diffraction

The X-ray diffraction patterns was obtained using a Philip

1710 diffractometer, with copper as target and nickel as a

filter (λ = 1.54178 Ă) at 40 kV and 30 mA. The scanning

speed was 3.6 min-1 in the range of 2θ = 2-60 (298 K) as

described elsewhere [15,16].

Infrared spectrum

The IR spectra were scanned on a Pye Unicam Sp3100

spectrophotometer using the KBr disc technique (4000-400

cm-1). The method include mixing few mgs of a fine

powder of the sample with KBr powder in agate mortar.

The mixture was then pressed by means of a hydraulic

press. The transmittance was automatically registered

against wavenumber (cm-1). Relevant IR bands which

provide considerably structural evidence for the mode of

attachment of alginate functional groups to thorium(IV)

were obtained.

Conductance measurements

The dc conductance was measured over the temperature

range 290-560o K using a Keithely 610 C electrometer as

described previously [15-18]. The thorium(IV)-alginate

complex was sandwiched between two standard electrodes

(graphite, copper or silver paste) mounted into a specially

designed temperature-controlled electric furnace provided

with a special copper-constantan thermocouple. The

sample was kept for about 5 h to make it ready for the

experiment. The electrical resistance of the sample was

measured, and from this the electrical conductivity (ơ) was

calculated as follows

σ= (1/R)(L/a) (1)

where R is the Ohmic resistance (Ω), a is the area of the

sample (cm2) and L is the thickness of the specimen (cm).

Equilibrium measurements

Aqueous solutions containing mixtures of thorium(IV)-

alginate complex grains and hydrogen ions (HClO4) of

known concentrations were thermally equilibrated in a

constant temperature water-bath maintained at the desired

temperature within ±0.05o C with continuous stirring using

a magnetic stirrer. After equilibrium had been attained (24

Physicochemical Studies on Cross-Linked Thorium(IV)-Alginate Complex Especially the Electrical ..


- 3 -

h), clear solutions containing both reactants were syringed

out and the concentrations of H+ and Th4+ were determined

titrimetrically and compelxometrically [19,20],

respectively.

The ionic strength of the mixture was maintained

constant at 0.1 mol dm-3 by adding NaClO4 as an inert

electrolyte.

3. Results and Discussion

The replacement of Na+ counter ions of alginate

macromolecule by a polyvalent metal ions is an inherent

stoichiometric exchange process [4,21] which leads to the

formation of the corresponding cross-linked metal alginate

complexes as follows :

Mz + z (Na-Alg)n → (M-Algz)n + z Na+ (2)

where M denotes the polyvalent metal ion and z stands to

its valency. The interdiffused metal ions chelate the

carboxylate and hydroxyl functional groups of alginate

macromolecular chains by partially ionic and partially

coordinate bonds [5,11], respectively.

In general, there are two types of chelation in

these cross-linked metal alginate complexes [10]. The first

type in which the interdiffused metal ion cross-links the

functional groups of two different chains and the plane

containing the chelated metal ion is perpendicular to the

plane of alginate chains. This type of chelation corresponds

to the intermolecular association or non-planar geometry.

The second type of chelation represents the intermolecular

association or planar geometry in which the metal ion

cross-links the functional groups of the same chain and the

plane containing the metal ion is parallel to the plane of

alginate chains. The type and nature of chelation depends

on the valency and coordination number of the

interdiffused metal ion, respectively.

It is well known that most of the divalent metal

cations are of octahedral six coordination geometry in their

complexes [22]. Therefore, these metal ions have the

choice to chelate the functional groups of alginate via

either inter- or intramolecular association in order to attain

the octahedral geometry.

However, in tri- and tetravalent metal cations, the

octahedral geometry can be attained only via an

intermolecular association. This fact is owing to the

difficulty of stretching the chemical bonds to involve three

or four neighbor monomers of the same chain in case of

intramolecular association. The chelates of metal ions in

case of intrarmolecular association mechanism, resemble

an egg-carton like structure [7,8].

The X–ray diffraction patterns indicate that

thorium(IV)–alginate complex is amorphous in nature and,

hence, the stacking alginate chains blocks are mediated by

thorium(IV) ions. Relevant infrared bands which provide

considerable structural evidence for the mode of

attachment of alginate functional groups to Th4+ ion are

shown in Fig. 1. The appearance of a band at 890 cm-1

indicates the presence of chelated thorium(IV) ions [23].

The bands of υs CO2- and υas CO2

- are shifted from 1400

and 1600 cm-1 in alginate to 1419 and 1635 cm-1 in the

complex, respectively, indicating the complexation of Th4+

ion and the functional groups of alginate chains. The broad

band observed at 3461 cm-1 is due to υOH of water or (OH-

free functional groups) [23]. The displacement of this band

to 1749 cm-1 of the spectrum of thorium(IV)-alginate

complex (Fig. 1) may indicate the coordination of the

carboxylate group with the appearance of both symmetric

(υs) and asymmetric (υas) vibrations of COO- groups.

Again, the location of υs OCO is diagnostic of a bridging

carboxylate groups.



- 4 -

a

b

Wavenumber (cm-1)

Figure 1. Infrared spectrum of (a) alginate (b) cross linked thorium(IV)- Alginate complex.

The values of electrical conductivity which were

measured using different electrodes were found to be in

good agreement with each other confirming the

reproducibility of the conductance measurements. The plot

of log σ vs. 1/T displayed a simple parabolic shape at the

early stages, followed by a slight increase in σ values on

raising temperatures. Then, a sharply increase in the

electrical conductivity is observed at elevated temperatures

of measurements as shown in Fig. 2.

1 . 6 2 . 0 2 . 4 2 . 8 3 . 2

9 . 0

8 . 5

8 . 0

7 . 5

( v )

( i v )

( i i i )

( i i )

( i )

1 0 3 ( 1 / T ) , K - 1

Figure 2. The electrical conductivity as a function of temperature for cross- linked thorium(IV)-alginate complex

-logσ

% T

rans

mitt

ance



- 5 -

It has been previously reported [10] that the

metal-alginate complexes of planar geometry show

electrical properties similar to those of insulators, whereas

those of non–planar structure possess electrical

conductivity values in the range of semiconductors. This

fact can be explained by the charge carriers which tend to

gain maximum speed in case of a perpendicular geometry

owing to the presence of multi-channels around the planes.

These channels facilitate the migration of charge carriers

and, hence, an increase in the electrical conductivity

occurs.

Conductance mechanism

In general, the electrical conductivity of polymeric

materials is usually attributed to the presence of low

molecular mass impurities of free-ions not connected

chemically with the macromolecules [24-26]. Therefore, a

suitable conductance mechanism for the electrical

properties of Th4+- alginate complex may be suggested.

The small increase of σ values observed at the initial stages

(i) may be attributed to the slight density of charge carriers

(intrinsic conductance). The subsequent appreciable

increase of σ values may be due to either the dehydration

process of the coordinated water molecules in the complex

sphere or the formation of free- radicals.

Since the alginate complexes of divalent metal

ions contain similar coordinated water molecules in their

atmospheric region [27,28] and there is no any prabolic

behaviour [16],then the suggestion based on increasing the

electrical conductivity by the dehydration process would be

excluded. Hence, the formation of free-radicals is the more

favorable explanation for the increase of σ values of the

prabolic shape.

Similar prabolic behaviour was observed with respect to

cross-linked trivalent [17] and hexavalent [18] metal

alginate complexes. The increase of σ values was

interpreted by the formation of free-radical complexes as a

result of electron transfer from alginate macromolecule to

the chelated metal ions to form metal ions of lower

oxidation states. However, there is no evidence for

existence of any lower oxidation states for thorium cation

in solution [29], the existence of Th3+ and Th2+ is possible

in the solid state [29,30].

Consequently, the increase of ơ values in the prabolic

shape at the early stages (ii) can be interpreted by the

formation of free-radical complex with lower oxidation

state of thorium ion as follows :

(RCOO- zMz+)n1 → (RCOO.

(z-1) .M(z-1)+)n2 (3)

where RCOO. represents the alginate macromolecule,

RCOO- is the formed radical, M is the thorium metal ion

and z stands for its valency. Again, the dimerization of the

free-radicals should be accompanied by a decrease in the

electrical conductivity as it is experimentally observed in

region (iii).

(RCOO(z-1) .M(z-1)+)n2 + (RCOO.(z-1) .M(z-1)+)n2 →

2(RCOO(z-1) .M(z-1))n2 (4)

The thermal decomposition of thorium(IV)–alginate

complex [31] indicated that the dehydration of the

coordinated water molecules occurs at the initial stage

(< 375 K). This dehydration is followed by a degradation

process to form the corresponding metal oxalate (< 450 K).

This intermediate is subsequently decomposed to give the

metal oxide product at the final stage. Accordingly, the

region at which a slightly increase in σ values (iv) can be

explained by the formation of oxalate intermediate

resulting from the decomposition of the complex formed

(Eq. 4)

2 (RCOO(z-1) . M(z-1))n2 → M(C2O4)2 . 6H2O + CO2 +

H2O (5)

Again, the sharp increase in σ values (v)

observed at high temperatures (> 450 K) can be attributed

to the decomposition of the oxalate intermediate to give

rise to thorium oxide product at the elevated temperatures

as follows:

M (C2O4)2 . 6H2O + O2 → MO2 + 4 CO2 + 6H2O (6)

Furthermore, the change in colour for cross-

linked thorium(IV)-alginate complex before and after



- 6 -

temperature treatment may confirm the formation of

various oxidation states of thorium ion and, hence, supports

the suggested mechanism. Typical photos are shown in Fig.

3.

Ion exchange equilibrium has been attained when the

Th4+ counter ions in the alginate complex grains are

replaced by other different counter cations of the same

sign. Hydrogen ions were selected for replacement owing

to the easiness and simplicity of exchange [32]. The

equilibrium of ion exchange between Th4+ and H+ ions can

be expressed by the following stoichiometric equation

(Th4+-Alg4)n(s) + 4 H+(aq) → 4 (H-Alg)n(s) + Th4+

(aq) (7)

Applying the mass action-law for such a heterogeneous

system and assuming that the activities of the solid phase

are always unity [33] and the ratio of the activity

coefficient in the solid phase is constant [34], the following

relationship is obtained

Ka = Kc (γTh4+/ γ4

H+) (8)

(a) (b)

Figure 3. Optical images in cross-linked thorium(IV)-alginate complex: (a) before and (b) after treatment.

where γ is the activity coefficient of the respective ions, Ka

is the thermodynamic equilibrium constant and may be

vary with the composition of the solid phase and Kc is the

equilibrium constant and can be defined as

Kc = [Th4+]/ [H+]4 (9)



- 7 -

The values of Ka were found to be 25.75 and 15.40

dm9. mol–3 at 25o C and 40o C, respectively. The values of

the thermodynamic parameters were calculated from the

temperature dependence of the equilibrium constant and

found to be ∆Ho = -4.76 kJ mol-1, ∆So = +10.92 J K-1 mol-1

and ∆Go = -8.05 kJ mol-1, respectively.

In view of these interpretation and the experimental

observation, thorium(IV) should be chelated to the

functional groups of alginate macromolecular chains via

intermolecular association mechanism of non-planar

geometry (Scheme I). This configuration maybe

considered as an indirect evidence to explain the high

electrical conductivity of the complex, which lies in the

magnitude of semi conductors, compared to that of other

complexes of planar structures and low electrical

conductivities [16] which lie in the region of insulators as

shown in (Table 1).

Scheme I Table 1. The electrical conductivity of some cross-linked metal-alginate complexes at 290 K.

Metal-alginate complexes σ (Ω-1 cm-1) Reference

UVI-alginate 1.69x10-12 18

ThIV-alginate 2.01x10-9 This work

CrIII-alginate 1.0x10-10 17

FeIII-alginate 2.0x10-9 17

CaIII-alginate 1.3x10-13 16

CuII-alginate 1.82x10-12 16

CdII-alginate 2.40x10-12 16

PbII-alginate 1.84x10-12 16

AgI-alginate 1.78x10-8 15

OH OH

O

OH OH

O

O OO

O

OHOHO O

O

OHOH

C = O

O

O = C

C = OO = C

OO

O O

H2O.................... ThIV...................OH2

n



- 8 -

The conductance of polymeric compounds is

usually occurred by two conductance mechanisms, ionic

and electronic, depending on the nature of the charge

carriers existing within the network of the macromolecular

chains [16]. The formation of free-radicals demonstrates

the electronic conduction mechanism. Therefore, the

gradual increase in the conductance at the initial stage of

ThIV–alginate can be explained by the increase of charge

carriers within the solid, whereas the sharp increase in σ

values at the final stage may be interpreted by the

formation of thorium oxide, respectively.

The activation energy may reflect the mechanism of

conductance. The activation energy is evaluated from the

slope of log σ–1/T plot using the Arrhenies equation as

following:

σ= σθ exp (-Ea/RT ) (10)

where ơ is the electrical conductivity, σθ is a constant and

Ea is the activation energy of the charge carriers. This value

was evaluated by using the least-squares method and is

summarized along with the values of other cross-linked

metal alginate complexes in Table 2. The lower activation

energy ≤ 1.0 eV corresponds to the electronic structure,

whereas the higher values refer to the ionic conduction

mechanism.

The magnitude of the equilibrium constant for

exchange obtained, may indicate the high stability of the

thorium(IV)-alginate complex. The negative value of ∆Ho

indicates that the exchange process is an exothermic

process. Whereas, the negative value of ∆Go reflects the

spontaneity of a such exchange process [35].

Table 2. The activation energies in eV for some cross-linked metal-alginate complexes.

Metal-alginate complexes Ea (initial stage) Ea (final stage) Reference UVI-alginate 0.37 - 18

ThIV-alginate 0.86 1.20 This work

CrIII-alginate 0.16 2.74 17

FeIII-alginate 0.22 1.41 17

CaIII-alginate - 2.12 16

CuII-alginate - 5.21 16

CdII-alginate - 1.75 16

PbII-alginate - 0.18 16

AgI-alginate 0.21 3.15 15

References

[1] Specker, H.; Kuchner, M.; Hortkamp, H., Z.

Anal. Chem. 1954, 33, 141.

[2] Thiele, H.; Anderson, G., Kolloid Z. 1955,

76, 140; Thiele, H.; Hallich, K., Kolloid Z.

1957, 1, 151.

[3] Haug, A.; Smidsrod, O., Acta Chem. Scand.

1965, 19, 341.

[4] Muzzarelli, R. A. A., Natural Chelating

Polymers. 1st ed., Pergamon Press, Oxford.

1972.



- 9 -

[5] Awad, A.; El-Cheikh, F.; Hassan, R. M., Rev.

Roum. Chim. 1979, 211, 563; Awad, A.; El-

Cheikh, F., J. Coll. Interf. Sci. 1981, 80, 107.

[6] Hassan, R. M.; El-Shatoury, S. A.; Makhlouf, M. Th.,

Coll. Polym. Sci. 1992, 12, 1237; Hassan, R. M.;

Wahdan, M. H.; Hassan, A., Eur. Polym. J. 1988, 24,

281.

[7] Rees, D. A., Biochem. J. 1972, 126, 257; Hirst, E.;

Rees, D. A., J. Chem. Soc. 1965, 1182; Rees, D. A.;

Scott, W. E., J. Chem. Soc. B, 1971, 469.

[8] Schweiger, R. G., J. Org. Chem. 1962, 27, 1786;

Schweiger, R. G., Kolloid Z. 1964, 196, 47.

[9] Hassan, R. M.; Awad, A.; Hassan, A., J. Polym. Sci.

1991, 29, 1645.

[10] Hassan, R. M., Polym. Inter. 1993, 31, 81.

[11] Khan, A. A.; Khan, A., Talanta 2007, 73, 50; Khan,

A. A.; Khan, A.; Talanta 2007, 72, 699.

[12] Hassan, R. M., High Perform. Polym. 1989, 1, 275.

[13] Hassan, R. M.; Makhlouf, M. Th.; Summan, A. M.;

Awad, A., Eur. Polym. J. 1989, 25, 993.

[14] Abdel-Wahab, S. A.; Ahmed, M. A.;

Radwan, F. A.; Hassan, R. M.; El-Refae, A.

M., Mater. Lett. 1997, 20, 183; Ahmed, M.

A.; Radwan, F. A.; El-Refae, A. M.; Abdel-

Wahab, S. A.; Hassan, R. M., Ind. J. Phys.

1997, 71A, 395.

[15] Hassan, R. M., Coll. Surf. 1991, 60, 203.

[16] Khairou, K. S.; Hassan, R. M., High

Perform. Polym. 2002, 14, 93.

[17] Zaafarany, I. A.;Khairou, K. S. Hassan, R.

M.: High Perf. Polym.(in press 2009).

[18] Hassan, R. M., Ekeda, Y.; Tomiyasu, H., J.

Mater. Sci. 1993, 28, 5143.

[19] Hassan, R. M., J. Mater. Sci. 1991, 26, 5806.

[20] Vogel, A. I., Textbook of Quantitative

Inorganic Chemistry. 4th ed., Longman,

New York 1978.

[21] Hellferich, H.: Ion exchange. McGraw-Hill,

New York 1962.

[22] Martell, A. E.: Coordination Chemistry.

New York, Van Nastrand-Rainhold 1972.

[23] Cozzi, D.; Desider, P. G.; Leppri, L.;

Cinatelli, G., Alginic acid, J. Chromatogr.

1965, 35, 369; Bellamy, L. J., The Infrared

Spectra of Complex Molecules. vol. 1

Chapman and Hall, London 1975.

[24] Seanor, D. A., J. Polym. Sci. A, 1968, 2,

463.

[25] Miyoshi, Y.; Saito, N., J. Phys. Soc. Jpn.

1968, 24, 1007.

[26] Baird, M. E., J. Polym. Sci. A, 1970, 2, 739.

[27] Said, A. A.; Hassan, R. M., Polym. Degrad.

Stabil. 1993, 93, 393.

[28] El-Gahami, M. A.; Khairou, K. S.; Hassan,

R. M., Bull. Polish Acad. Sci. 2003, 51, 105.

[29] Cotton, A. F.; Wilkinson, G., Advanced

Inorganic Chemistry. 3rd ed., John Wiley,

New York 1972.



- 10 -

[30] Moller, T.: Inorganic Chemistry. 1st ed.,

John Wiley, New York, 1967.

[31] Hassan, R. M.: Zaafarany, I. A.; Ikeda

Y.(Submitted for publication 2008).

[32] Hassan, R. M., J. Mater. Sci. 1993, 28, 384.

[33] Glasstone, S., Lewis, D., Elements of

Physical Chemistry, 2nd ed., Van Nostrand

1960.

[34] Baugh, P. J., Lawton, J. B., Philips, G. O., J.

Phys. Chem. 1972, 76, 658; Eisenman, G.

Biophys. J. 1962, 2, 2590.

[35] Hassan, R. M., J. Coord. Chem. Rev. 1992,

27, 255.

Arabian J. Chem. Vol. 2, No. 1,11-19(2009)

Arabian J. Chem. Vol. 2, No. 1(2009)

- 11 -

Fluorescence Spectrometric Study of Eosin Yellow Dye-Surfactant Interactions

Seema Acharya1 And Babulal Rebery*

*Central salt and marine chemical research institute (CSIR) Bhavnagar, Gujarat-364002 (INDIA)

1Spectroanalytical laboratory, Department of Chemistry J.N.Vyas University Jodhpur –342005 (INDIA)

Email: [email protected], [email protected]

Abstract

The spectrofluorimetric behavior of an analytically important molecule eosin yellow was studied in

the presence of various surfactant solutions. The relatively weak fluorescence of eosin yellow was

significantly enhanced in micellar media formed by cationic and anionic (DBSS) surfactants. The

influence of the surfactant structures, concentrations and working experimental conditions on the

fluorescence spectra of eosin yellow was thoroughly evaluated and discussed. The solubilizing action

of the surfactant has been supplemented by the theoretically calculated spectral parameters like,

empirical fluorescence coefficient, quantum yield, molar extinction coefficient and Stokes' shift.

Keywords: Eosin yellow, Fluorescence, Absorption and Solubilization.

1. Introduction Analytical methods which rely on the use of surfactants are

becoming more and more numerous, since addition of

surfactants provides an increase in selectivity and

sensitivity[1,2]. Eosin (yellowish)-Tetrabromo fluorescein

sodium salt is an acid xanthene (natural anionic) dye. A

comparative photophysical study of rose bengal, eosin

yellow and their monomethyl and dimethyl derivatives

shows that aggregates of these dyes are probably non

emissive[3]. Dye sensitized chemiluminescence of luminol

and related cyclic hydrazides shows that this emission can

be initiated by triplet states of methylene blue and eosin

yeollw[4]. Chao Lu et al.[5] found that fluorescein, eosin

yellow and uranine have evidence of a chemiluminescence

enhancing of the CuII(H2O2) and CoII(H2O2) systems. Eosin

yellow provides example of direct measurement of

elementary processes like singlet excited state absorption

of the excited singlet state[6]. Color removal from effluent

is one of the most difficult requirements faced by the textile

finishing, dye manufacturing, and pulp and paper

industries. These industries are major consumers of water

and, therefore, cause water pollution. Most of these dyes

are harmful when brought in contact with living tissues for

a long time. The discharge of such dyes to the river stream

without proper treatment causes irreparable damage to the

crops and living beings, both aquatic and terrestrial[7].

Separation of Congo red by surfactant mediated

cloud point extraction, removal of dye from wastewater

using micellar enhanced ultrafiltration and regeneration of

surfactant and resistance in series model for micellar

enhanced ultrafiltration of eosin dye have been studied[8-

10]. The photophysical and photo catalytic parameters of

sulfo and tetrabromo sulfo derivatives of fluorescein have

also been studied[11]. Seret et al.[12] have studied

Seema Acharya And Babulal Rebery


- 12 -

solubility properties of eosin yellow and rose bengal triplet

state in sodium dodecyl sulfate micellar solutions.

From an analytical view point, the use of

surfactants increases the solubility of organic substances in

water, through shallow or deep penetration of the micelles

or simply by surface adsorption[13], and can also catalyze

specific reactions by modification of the micro-

environment in which these reactions take place[14].

Surfactants at concentrations higher than the critical

micelle concentration (cmc) has been extensively used in

the application of spectroscospic (ultra-violet, fluorescence,

phosphorescence, atomic spectroscopy), electroanalytical

and separation methods to sparingly soluble

analytes[15,16].

Eosin has been used as a groundwater migration

tracer by capillary electrophoresis/laser-induced

fluorescence using a multi wavelength laser[17]. The

decomposition of eosin (yellow) under UV–visible light

irradiation in the presence of CeO2–CeTi2O6 films shows

the presence of photoactivity in these films[18].

Modification of the properties of NaDS micellar solutions

by adding electrolytes and nonelectrolytes: investigations

with decyl eosin as a pKa probed by Loginova et al.[19].

The staining of eosin with haematoxylin have

been used in structure determination of grasshopper and

mammalian testis as well as supporting structure

determination of destruction of dental tissues[20].

This paper includes study of the influence of

various nonionic, anionic and cationic surfactants on the

fluorescence and absorption spectra of eosin yellow. The

optimum solubilization showing dye-surfactant interaction

can be utilized as separation of dyes from waste dye-stuffs

of different textile, paper and pulp industries. The results

have been interpreted from the calculation of molar

extinction coefficient, empirical fluorescence coefficient

and quantum yield of eosin yellow fluorescence in various

micellar media. Stokes' shift calculation at various

concentration of eosin yellow is also supportive.

2. Experimental Materials and Method

Fluorimetric studies were carried out with a

Perkin Elmer spectrophotometer 204 A. The slit width was

kept at 10 nm throughout for excitation as well as emission

spectra. Absorption spectra of eosin yellow were taken on a

chemito UV-VIS 2600 double beam spectrophotometer.

The stock solution of analytically pure eosin

yellow (Sd fine chemicals) was prepared in double distilled

water. All the experiments were made at room temperature

(23-25 °C) and were performed in aqueous medium

keeping the final concentration of eosin yellow at 10-6 M.

All the surfactants used were either of sigma (USA) or

BDH products.

(A) Nonionic

Polyoxyethylene 23 lauryl ether (Brij-35)

Polyoxyethylene sorbitan monopalmitate

(Tween-40). Polyoxyethylene tertoctyl phenol

(Eq-10) (Tx-100)

(B) Anionic Dodecylbenzene sodium sulphonate (DBSS)

Sodiumlauryl sulphate (SLS)

Dioctylsodium sulphosuccinate (DSSS)

(C) Cationic

Cetyltrimethyl ammonium Bromide (CTAB)

Cetylpyridinium chloride (CPC)

Myrstyltrimethyl ammonium bromide (MTAB)

The purity of surfactants was checked by

determining their CMC values with the help of surface

tension measurements, employing drop weight method.

The absolute fluorescence quantum yield of the compound

was calculated relative to anthracene solution used as a

standard. Each time the total intensity of fluorescence

emission was measured for the standard and the sample

from the area of fluorescence spectrum recorded over the

whole range of emission under identical conditions.



- 13 -


The aqueous solution of eosin yellow showed

maximum excitation peak at 510 nm while the emission

spectrum showed a peak at 535 nm. The cationic

surfactants caused an enhancement in the fluorescence

intensity with 15–20 nm gradual red shifts. Among these

surfactants CTAB exerted maximum effect. The changes in

fluorescence intensity of eosin yellow on addition of

CTAB are shown in Fig. I. On addition of a cationic

surfactant red shift occurs at maximum. This may be

attributed to the difference in solvation energy of the solute

in the ground state and the excited state.

Fig. I: The changes in the fluorescence intensity of eosin

yellow on adding different concentrations of CTAB

are given;

(a) 1 x 10–6 M eosin yellow

(b) 1 x 10–6 M eosin yellow + 0.05% CTAB

(c) 1 x 10–6 M eosin yellow + 0.3% CTAB

(d) 1 x 10–6 M eosin yellow + 0.5% CTAB On addition of the nonionic surfactants like Brij-

35 and Tween-40, fluorescence intensity decreased with 5-

10 nm blue shift while for TX-100 fluorescence intensity

reached maximum initially and then it decreased with the

increase in concentration of the surfactant accompanied by

red shift of 15-20 nm was observed while on addition of

anionic surfactants like DBSS, fluorescence intensity

increased with a red shift of 5 nm. For SLS and DSSS,

initially fluorescence intensity reduced to a very low value

accompanied by 15 nm blue shift and then it was gradually

increased with the concentration of the surfactant. The

changes in fluorescence intensity of eosin yellow on

addition of DBSS are shown in Fig. II.

Fig. II: The changes in the fluorescence intensity of eosin

yellow on adding different concentration of

DBSS are given

(a) 1 x 10–6 M eosin yellow

(b) 1 x 10–6 M eosin yellow + 0.003% DBSS

(c) 1 x 10–6 M eosin yellow + 0.005% DBSS

(d) 1 x 10–6 M eosin yellow + 0.07% DBSS

(e) 1 x 10–6 M eosin yellow + 0.1% DBSS



- 14 -

The changes observed in fluorescence emission intensity in

presence of surfactants are as given in table 1, 2 and 3.

Table 1: Effect of nonionic surfactants on the fluorescence intensity (F.I.) of eosin yellow

exλ = 510 nm emλ = 535 nm P.M. Gain = 2 Sensitivity = 0.3

S. No. % of

Brij-35 F.I.

emλ (nm)

% of Tween-

40 (w/v)

F.I. emλ

(nm)

% of TX-100

(w/v)

F.I. emλ

(nm)

1.

2..

3.

4.

5.

6.

7.

8.

9.

10.

11.

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

29

22

11

10

10

9

8

7

7

6

5

535

535

535

520

520

520

520

520

520

515

515

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

31

29

29

26

24

14

12

12

11

8

4

535

540

538

538

538

538

535

535

530

515

515

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

30

64

41

35

34

33

25

18

16

10

8

535

535

535

535

535

535

535

555

555

550

550

Table 2: Effect of anionic surfactants on the fluorescence intensity (F.I.) of Eosin Yellow

exλ = 510 nm emλ = 535 nm P.M. Gain = 2 Sensitivity = 0.3

S. No.

% of DBSS (w/v)

F.I. emλ (nm)

% of SLS (w/v) F.I. emλ

(nm)

% of DSSS (w/v)

F.I. emλ (nm)

1.

2..

3.

4.

5.

6.

7.

8.

9.

10.

11.

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

32

34

38

40

41

42

44

46

53

53

54

535

535

540

535

535

535

535

535

535

535

535

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

31

9

9

11

12

12

13

15

17

35

47

535

520

520

520

520

520

520

520

520

520

520

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

31

9

8

8

10

10

12

12

14

19

21

535

520

520

520

520

520

520

520

520

520

515



- 15 -

Table 3: Effect of cationic surfactants on the fluorescence intensity (F.I.) of Eosin Yellow

exλ = 510 nm emλ = 535nm P.M. Gain = 2 Sensitivity = 0.3

S.

No.

% of CTAB (w/v)

F.I. emλ (nm)

% of CPC (w/v)

F.I. emλ (nm)

% of MTAB (w/v)

F.I. emλ (nm)

1.

2..

3.

4.

5.

6.

7.

8.

9.

10.

11.

12.

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

31

31

32

32

32

32

33

43

41

53

68

535

520

520

520

520

520

545

545

545

545

545

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

31

31

31

25

19

18

40

59

68

80

83

535

520

520

520

520

550

550

550

550

545

545

0.000

0.003

0.005

0.007

0.01

0.03

0.05

0.07

0.1

0.3

0.5

0.7

31

31

31

32

33

34

34

34

35

36

38

43

535

520

520

520

520

520

520

550

550

550

550

550

The absorption spectrum gave peak at 505 nm. On addition

of any of the nonionic surfactants, a continuous decrease in

absorbance was observed with 5 nm blue shift in peak

position. Among anionic surfactants, DBSS showed a

gradual enhancement in the absorbance without any shift

while for SLS and DSSS, initially absorbance reached a

lowest value with 15 nm blue shift and then gradually

increased.

For cationics absorbance spectra enhanced

without any shift in peak position. Molar extinction

coefficient (log ε) calculations showed a gradual increase

in log ε values with the increase in cationic surfactant

concentration. It was due to the strong π → π* transitions,

while on increasing concentration of cationic surfactant

n → π* transition decreased. With nonionic surfactants as

the concentration increased, the (log ε) values decreased

gradually, with anionic surfactants, the (log ε) values

initially decreased and then increased.

The molar extinction coefficient log ε values of

the solubilizate molecule in different micellar media follow

the same trend as their emission intensity. Hence it proves

the well known fact that fluorescence intensity of a

fluorophore is directly related to its molar extinction

coefficient (log ε) [22]. The empirical fluorescence

coefficient (kf) values showed a similar trend to the

fluorescence emission intensity.

The value of kf confirms this observation and

attributes to the increased sensitivity of fluorimetric

analysis of the organic molecule by solubilization. This

was attributed to the fact that surfactants offer protective

microenvironment, leading to enhanced fluorescence of the

guest molecule (solubilizate) by shielding the excited state

from non-radiative decay that normally occurs in bulk

aqueous solution. The empirical fluorescence coefficient

(kf) is the ratio of fluorescence intensity and the

concentration of the fluorescent molecule and it was

determined by the formula given below[23].



- 16 -

If Kf = ———

C

Where :

If = Fluorescence intensity C = Concentration in

moles/litre.

The fluorescence quantum yield ( fφ ) values of

eosin yellow have been determined in aqueous medium at

different concentrations of aqueous surfactant solution

added to it. For nonionic surfactants added solutions the

quantum yield fφ ) values decreased. With anionic

surfactants like DBSS, the fφ values increased while for

SLS and DSSS, the fφ values were initially decreased and

then increased. For cationic surfactants, fφ values were

increased. These spectral parameters (fluorescence

coefficient and quantum yield) are shown in table no 4.

Stokes' shift value continuously increased as the

concentration of eosin yellow increased. The magnitude of

Stokes' shift depends on several factors. The large Stokes'

shift values for eosin yellow are due to hydrogen bond

formation between the solute and the solvent in the ground

state. This bond breaks following excitation to S1 but

reforms following proton transfer[24]. When photons from

molecules in an excited state are emitted by fluorescence,

one of the most important observations was that they are

emitted at longer wavelengths (lower frequency) and

consequently are less energetic than the photons

responsible for the excitation. This difference between the

excitation and emission maxima is termed the Stokes' shift.

Table 4 :Empirical Fluorescence Coefficient (kf) and quantum yield ( fφ )for CTAB

S. No. Concentration of CTAB

(w/v) in %

Empirical Fluorescence Coefficient

(kf) x 104 per mole Quantum yield ( fφ )

1.

2.

3.

4.

0.000

0.05

0.3

0.5

3100

3300

5300

6800

0.427

0.460

0.488

0.582

Stokes' shift is a physical constant of luminescent

molecules. It indicates the energy dissipated in bringing

about ionization during the lifetime of excited state before

return to the ground state.

Stokes' shift 7 1 110

ex emλ λ

= −

Where λex and λem are corrected maximum excitation and

emission wavelength and are expressed in nanometers. The

Stokes' shift is of interest to analytical chemists since the

emission wavelength can be greatly shifted by varying the

form of the molecule being excited. Electrolytic

dissociation in the excited state can also give rise to

apparently large Stokes' shift. Several factors influence the

magnitude of the Stokes' shift. If the environment is rigid



- 17 -

so that little rearrangement is possible then the Stokes shift

is expected to be small. The magnitude of the shift depends

on factors such as solvent polarity, viscosity and

polarisability. It also depends on whether the excited state

can undergo any specific interactions such as proton

transfer or charge transfer to other molecules or

(sometimes) within the same molecule. Where fluorescent

materials are used as detectable labels a large Stokes shift

is highly desirable because it makes life easier when optical

filters are used to separate exciting light and fluorescence

emission. The changes in stokes’ shift on increasing

concentration of eosin yellow is given in table no. 5.

Table 5: Stokes' shift data of eosin yellow at room temperature

S.No. Concentration of

compound (M)

F.I. exλ (nm)

emλ (nm)

P.M. Gain Sensitivity Stokes'

Shift (cm-1)

1.

2.

3.

4.

5.

1 x 10-6

3 x 10-6

5 x 10-6

7 x 10-6

1 x 10-5

18

20

22

37

56

515

515

515

515

518

540

540

540

540

545

3

3

3

3

2

0.1

0.1

0.1

0.1

0.1

898

898

898

898

956

Fluorescence intensity of the compound on

adding surfactants can be attributed to the increase in the

quantum yield. The fluorophore is the fluorescein in the

dye molecule, which is disodium salt of dibromo

fluorescein. The fluorophore exists in two forms, one is

more stable quinoid structure (A) which is coloured and

gives intense fluorescence while the other one is colourless

lactone form (B) which is non-fluorescent as shown in Fig.

III.

C

OBr Br

O

BrBr

NaO

COONa

OBr Br

OH

BrBr

NaO

O

O

(A) (B) Coloured Colourless

Fig. III: The different form of fluorophore (eosin yellow)



- 18 -

The initial enhancement in the fluorescent

intensity of dye eosin yellow on adding TX-100 surfactant

was due to the interaction of hydrophilic part of the

surfactant with the polymeric part of dye molecules which

results in breaking them into monomeric form. This causes

an increase in emission intensity initially but at its higher

concentration the geometry of the fluorophore in eosin

yellow changes to the lactone form which is non-

fluorescent. The decrease in emission intensity of eosin

yellow on addition of Tween-40 and Brij-35 with a blue

shift in emλ may be due to the increase hydrophobicity of

the surfactants. The dye being anionic in nature so there

should not be any interaction with anionic surfactants.

However the DBSS with bulky size was able to cause a

change in geometry of dye molecules. Wherein they make

the dye more coplanar hence enhance the emission

intensity. The interaction between the anionic dye and

cationic surfactants leads to an initial charged

neutralization; i.e. dye-surfactant ion-pair formation which

further induces the protonation of the system. The

preferential interaction of cationic surfactants with anionic

dye resulted into inactivation of fluorescing sites. Now in

its changed conformation it appears to be susceptible to

disaggregation on further adding the surfactant. Thus, it

causes subsequent micellization and further solubilization

hence increase the fluorescence emission intensity.

In micellar media many characteristics of organic

molecules e.g. absorption and fluorescence spectra are

changed drastically. Thus the above observations can be

explained by the solubilizing action of surfactant micelles.

This process is expected to be most pronounced in the

region of critical micelle concentration (CMC) of particular

surfactant. During the experiment it was observed that a

sudden increase in the fluorescence intensity occurred at

particular concentration range of each surfactant, which

was in the CMC range of the respective surfactant. In case

of ionic surfactant the changes observed were below CMC

and this was probably due to the premicellar aggregation in

the surfactant micelle.

On adding the surfactants to the aqueous solution

of the compound, the surfactant micelles get adsorbed at

the interfaces and remove the hydrophobic groups from

contact with water, thereby reducing the free energy of the

system. But in transferring the hydrophobic groups from

solution, to the micelle in the solvent, may experience

some loss of freedom confined to the micelle and, in the

case of ionic surfactants, from electrostatic repulsion from

other similarly charged surfactant molecules in the micelle.

These forces increase the free energy of the system and

thus oppose micellization.

Whether micellization occurs in a particular case

and, if so, at what concentration of monomeric surfactant,

therefore depends on the balance between the factors

promoting micellization and opposing it. Thus the increase

in quantum yield suggests that the surfactants have

solubilized the suspended solubilizate molecules (eosin

yellow). The higher fφ values in cationic micellar media

are because of the lesser effect of other deactivation

processes, which compete with fluorescence. Sufficiently

large values of log ε is assigned to the π-π* transitions and

also confirms the increasing trend of Stokes' shift values.

The red shift in the peak wavelength of eosin yellow in

micellar media is attributed to the hydrogen bonding

capacity of the solubilizate molecule.

4. Conclusion The present analysis and interpretation suggest that

experimental results observed and the theoretically

calculated spectral data are found to be in good agreement.

This proves the validity of the investigation made. Hence

the process of micellization followed by solubilization of

the eosin yellow substrate would catalyze its activities

which may serve better results in pollution removal in

analytical fields and color stabilization in textile industries.



- 19 -

Thus in analytical chemistry, surfactants have been

recognized as being very useful for improving analytical

methodology, e.g. in chromatography and luminescence

spectroscopy.

Acknowledgement The council of scientific and industrial research New Delhi

is highly acknowledged for financial support.

References

[1] Peris-Cardells, E.; Guardia, M. L.; Pramau, E.;

Savarino P.; Viscardi, G.,Quím. Anal. 1993, 12, 38.

[2] Beltrán, J. L.; Prat M. D.; Codony, R., Talanta

1995, 42, 1989.

[3] Delvalle, J. C.; Catalan, J.; Amat-Guerri, F., J.

Photochem. Photobiol. A:Chem. 1993, 72, 49.

[4] Klimov, A. D.; Lebedkin, S. F.; Emokhonov, V. N.,

J. Photochem. Photobiol. A:Chem. 1992, 68, 191.

[5] Lu, C.; Lin, J. M.; Hule, C.W.; Yamada, M.,

Analytical Sciences 2003, 19, 557.

[6] Penzkofer, A.; Beidoum, A.; Speiser, S., Chem.

Phys. 1993, 170, 139.

[7] Purkait, M. K.; DasGupta, S.; De, S., J. of Environ.

Manage. 2005, 76, 135.

[8] Purkait, M. K.; Vijay, S. S.; DasGupta, S.; De, S.,

Dyes Pigments 2004, 63 (2), 151.

[9] Purkait, M. K.; DasGupta, S.; De, S., Sep. Purif.

Technol. 2004, 37 (1), 81.

[10] Purkait, M. K.; DasGupta, S.; De, S., J. Colloid

Interface Sci. 2004, 270, 496.

[11] Ponyaev, A. I.; Martynova, V. P.; El’tsov, A.V.,

Russian J. of Gen. Chem. 2001, 71(11), 1744.

[12] Seret, A.; Vorst, A.V. D., J. Phys. Chem. 1990,

94, 5293.

[13] Pal, T.; Jana, N. R., Talanta 1994, 41, 1291.

[14] Sicilia, D.; Rubio, S.; Pérez-Bendito, D., Anal.

Chim. Acta 1994, 297, 453.

[15] Neal, S. L.; Villegas, M. M., Anal. Chim. Acta

1995, 307, 419.

[16] Khaledi, M. G.; Rodgers, A. H., Anal. Chim. Acta

1990, 239, 121.

[17] Brumley, W. C.; Farley, J. W., Electrophoresis

2003, 24, 2335.

[18] Verma, A.; Srivastava, A.K.; Sood, K. N., Solid

State Ionics 2007, 178, 1288.

[19] Loginova, L. P.; Samokhina, L. V.; Mchedlov-

Petrossyan, N. O.;Alekseeva, V. I.; Savvina, L. P.,

Colloids Surfaces A 2001, 193, 207.

[20] Espada, J.; Valverde P.; Stockert, J. C.,

Histochemistry 1993, 99,385.

[21] Rossi, A. D.; Rocha, L. B.; Rossi, M. A.,J. Oral

Pathol. Med. 2007, 36, 377.

[22] Practical Fluorescence : Theory, Methods and

Techniques; Guilbault, G.,Ed.;Marcel

Dekker:New York, 1993.

[23] Aithal, K. S.; Sreenivasan, K. K.; Udupa, N., J.

Indian Chem. Soc. 2005, 82, 575.

[24] Solntsev, K. M.; Huppert D.; Agmon, N., J. Phys.

Chem. 1998, 102, 9599.



- 20 -

Arabian J. Chem. Vol. 2 , No. 1, 21-30 ( 2009)

Arabian J. Chem. Vol. 2 , No. 1, ( 2009)

- 21 -

Photochromic Properties of 1,3,3-Trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-b][1,4]oxazine] Doped in

PMMA and Epoxy Resin Thin Films

Abdullah M. Asiria, , Abood A. Bahajajb, Abdullah G. Al-Sehemic

and Amerah M. Alsoliemya

a Chemistry Department, Faculty of Science, King Abdul Aziz University, Jeddah- 21413, P.O. Box 80203, Saudi Arabia.

b Chemistry Department, Faculty of Science, Hadhramout University of Science & Technology, P.O. Box 50512, Mukalla, Republic of Yemen.

cChemistry Department, Faculty of Science, King Khalid University, Abha, Saudi Arabia. aE-mail: [email protected]

Abstract Irradiation of colorless 1,3,3-trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-b][1,4]oxazine] SO doped in

PMMA and epoxy resin with UV light (at 366 nm) results in the formation of the intense colored

zwitterionic photomerocyanine PMC. The reverse reaction was photochemically induced by irradiation

with white light. Photocoloration and photobleaching reactions follow a first-order rate equation. It was

found that photocoloration rate constant of SO in PMMA film is greater than that in epoxy resin. On the

other hand, the photobleaching rate constant is almost identical in both matrices. Spirooxazine doped in

epoxy resin shows much better fatigue resistance than that doped in PMMA.

Keywords: Spirooxazine; Photochromism; Polymer film; Epoxy resin; Photobleaching, Photocoloration,

kinentics, Fatigue resistance.

1. Introduction

The IUPAC definition of photochromism is “a reversible

transformation of a chemical species induced in one or both

directions by absorption of electromagnetic radiation

between two forms, A and B, having different absorption

spectra” [1]. This transformation is usually induced by

electromagnetic radiation in the ultraviolet wavelengths

range, where a deeply colored specie B is generated from

the uncolored or weakly colored specie A. The reverse

reaction (bleaching) could be accomplished by thermal

and/or photochemical effects. Various organic systems

were found to show potential thermal and photochromic

properties. The widest and most important groups of such

systems are those based on the reversible light-induced

hexatriene/cyclohexadiene pericyclic reactions such as in

fulgides [2], diarylethenes [3], spiropyrans [4] and

spirooxazines [5].

Spiropyrans and spirooxazines are much closed

compounds in their photochromism. The photochromism of

these colorless or weakly colored spiro compounds SO

arises from the photo cleavage of the C–O spiro bond upon

the UV irradiation. Such cleavage results in the formation

of an intense colored zwitterionic open form known as

photomerocyanine PMC which absorbs in the visible

region. The reverse reaction (fading) proceeds thermally or

under irradiation with white light. The immediate cleavage

of the spiro carbon-oxygen bond upon UV irradiation

results in the formation of a highly unstable intermediate

Abdullah M. Asiri, , Abood A. Bahajaj, Abdullah G. Al-Sehemi and Amerah M. Alsoliemy


- 22 -

cis-cisoid isomer [6]. This intermediate isomerizes to the

more stable zwitterionic open form which in turn

undergoes subsequent geometrical isomerism to give

several quinoidal forms (Scheme 1).

O

N

N

N

N O

N

N

O

N

NN N

N N

O

O

O

+ -UV

Vis or heat

(TTT)

(TTC)(CTT)

(CTC)

colorless spirooxazineColored photomerocyanine

(zwitterionic form)

(quinoidal forms of photomerocyanine)

(SO) (PMC)

Scheme 1: Photochemical reactions of spiroindolinonphthooxazin

Time-resolved absorption spectroscopy [7] and

NMR studies [8] reveal the coexistence of four geometrical

isomers (TTC, CTT, CTC, and TTT) which are in thermal

equilibrium at room temperature. Takahashi [9] have

found that TTC isomer (trans-trans-cis isomer) is more

stable in aliphatic hydrocarbons, while CTT and CTC are

more stable in polar solvents. The recent interest in the

photochromism of spiropyrans and spirooxazines is due to

their fast coloration rate under UV irradiation, fast thermal

fading, and excellent fatigue resistance. These criteria are

indispensable for applications to optoelectronic devices,

such as memories and switches, and nonlinear optics

[5],.Spiropyrans and spirooxazines have been recently used

as nucleic acid hybridization probes [10]

2. Experimental

1,3,3-trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-

b][1,4]oxazine] was prepared according to general

procedure previously reported [11]. The films were

prepared as follow: 2.23 g of PMMA (Aldrich product)

was dissolved in 20 mL chloroform and warmed to ensure

complete dissolution. The solution was then cooled. Two

blank films were prepared by taking 0.3 mL of the above

solution and spread over a quartz plate. These plates were

covered and left overnight in the dark. To the remaining

Photochromic Properties of 1,3,3-Trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-b][1,4]oxazine]…..


- 23 -

solution was added about 1mg of spirooxazine SO and

mixed well. Then 0.3mL of the mixture was added to each

of the quartz plates which then covered and left overnight

in the dark. The epoxy resin film was prepared as follow:

A 10 mL of epoxy resin (diglycidyl ether of bisphenol A)

and 1 mL of diamine hardener were added and mixed

thoroughly to ensure complete dissolution. Two blank

films were prepared by taking 0.3 mL of the above solution

and spread over each quartz plate. To the remaining

mixture, a solution of 2 mg of SO dissolved in 15 mL

chloroform was added and mixed. Then 0.3mL of the

mixture was added to each of the quartz plates which then

covered and left overnight in the dark.

Ultraviolet and visible spectra were measured

using Perkin-Elmer lambda EZ210 spectrophotometer.

Photocoloration (at 366 nm) was carried out using Blak-

Ray lamp model UVL-56 and photobleaching was obtained

using a tungsten filament lamp. Two fresh films of SO in

PMMA and epoxy resin are annealed for three hours at 75

°C. The fatigue resistant of the annealed SO doped PMMA

and epoxy resin films was carried out by photocoloring and

photobleaching SO, consecutively for 9 cycles. In each

cycle, the film was irradiated with UV lamp for 30 min and

photobleached with white light for 20 min.


3.1 Photocoloration 1,3,3-Trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-

b][1,4]oxazine] SO doped in PMMA polymer film was

irradiated with mercury lamp (366 nm) and the

photocoloration process was followed

spectrophotometrically by monitoring the absorption of the

intense purple colored open form photomerocyanine PMC

at its λmax (555 nm) at intervals of time as shown in Fig. 1.

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

250 300 350 400 450 500 550 600 650 700Wavelength (nm)

Abs

orba

nce

Fig. 1: Photocoloration of SO doped in PMMA polymer film. The arrow direction indicates the increase of absorbance with increasing exposure time (sec.), 0; 20; 40; 100; 120; 140; 200; 240; 280; 340; 520; 560; 600.



- 24 -

The film of SO doped in an epoxy resin was

similarly treated. The resulting PMC has its λmax at 560

nm (Fig. 2). The observed red shift of the absorption band

of PMC in epoxy resin compared to that in PMMA film is

a result of increasing the polarity of the medium. It is well

known that this absorption band shows a bathochromic

shift as the solvent polarity increase [12-15] which

represents a positive solvatochromism.

0.1

0.3

0.5

0.7

0.9

1.1

1.3

1.5

250 300 350 400 450 500 550 600 650 700Wavelength (nm)

Abs

orba

nce

Fig. 2: Photocoloration of SO doped in epoxy resin polymer film. The arrow direction indicates the increase of absorbance with increasing exposure time (sec.), 0; 20; 40; 60; 100; 140; 180; 220; 300; 480; 540.

Kinetics of spiropyrans and spirooxazines have

been studied in solutions [12-15], in phospholipid bilayers

[16], in solid polymer matrices [17, 18 ] and in the

crystalline state [19]. It shows, in general, first-order rate

dependence, especially in solutions [13, 14]. However, in

polymer matrices deviation from simple first-order reaction

was observed. Such nonlinearity of the first-order plots was

attributed to several factors such as the effects of the

polymer matrix on the photochromic compound, the

presence of more than one conformers or the dye might be

confined in the solid polymer matrix [20]

The integrated form of the first-order rate law for

the photocoloration process is:

( )( ) kt

AAAA

t

=−−

∞

∞ 0ln

where k is the rate constant, A∞, At, and , A0 are the

absorbance of the PMC at infinite time, at time t, and zero

time, respectively. Plot of [ln(A∞ −A0)/(A∞ −At)] against

time, gives a straight line with slope equals as shown in k

(Fig. 3). From the graph the apparent first-order rate

constant (k) for the photocoloration of spirooxazine SO

doped PMMA and epoxy resin films was found to be

0.0040 s-1 and 0.0021 s-1, respectively. It is clear that the

photocoloration of SO in PMMA is almost twice faster

than that in epoxy resin. This could be attributed to the

larger free volume of the PMMA compared to that of

epoxy resin which offer large space for the dye to undergo

isomerization. Another factor could be the possible

hydrogen bonding between the epoxy resin and the

spirooxazine which stabilize the later.



- 25 -

0

0.5

1

1.5

2

2.5

0 100 200 300 400 500 600Time (sec.)

Photocoloration in PMMA polymerPhotocoloration in epoxy resin

Fig. 3: First-order plot of the photocoloration of SO doped in PMMA and epoxy resin

3.2 Photobleaching Spiropyrans and spirooxazines are known [15] to be

thermal reversible photochromic compounds. This thermal

reversibility is due to the sigma bond cleavage of the spiro

C-O single bond. The open form photomerocyanine reverts

thermally to the closed form spiro compound. The rate of

thermal fading of photomerocyanine to spiro form was

found to decrease with increasing solvent polarity [15] and

metal complexation [14, 21]. This trend was attributed to

the stabilization of the polar zwitterionic

photomerocyanine through complexation and hydrogen

bonding.

Photobleaching and thermal fading of PMC to

SO follow simple first-order rate law. When the open form

PMC was irradiated with white light, it is converted to the

colorless spirooxazine SO. Thus, the purple color is

gradually disappeared with time. Figures 4 and 5 shows

the absorption spectrum of the photobleaching reaction of

photomerocyanine PMC doped in PMMA and epoxy resin

polymer films, respectively.

ln (A

∞ -

A0)

- ln

(A∞ -

At)



- 26 -

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

250 300 350 400 450 500 550 600 650 700Wavelength (nm)

Abs

orba

nce

Fig. 4: Photobleaching reaction of PMC in PMMA polymer film. The arrow direction indicates the decrease of absorbance with increasing exposure time (sec.), 0; 20; 100; 120; 200; 240; 280; 340; 560.

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

250 300 350 400 450 500 550 600 650 700Wavelength (nm)

Abs

orba

nce

Fig. 5: Photobleaching reaction of PMC in epoxy resin film. The arrow direction indicates the decrease of absorbance with increasing exposure time (sec.), 0; 20; 40; 60; 80; 140; 180; 320; 400; 440; 500; 540; 680; 720.



- 27 -

The integrated form of the first-order rate law for

the photobleaching process is

ktAAt −=

0

ln

Where k is the rate constant, A0 is the absorbance of the

PMC at zero time and At is its absorbance at time t. Plot of

[ln(At) – ln(A0)] against time, gives a straight line with a

slope equals (-k). Fig. 6 shows the simple first-order plots

of the photobleaching reaction of PMC doped in PMMA

and epoxy resin polymer films, respectively. The apparent

first-order rate constant (k) for the photobleaching of

spirooxazine SO doped in PMMA equals 0.0012 s-1 and

that for SO doped in epoxy resin equals 0.0013 s-1. The

rate of photobleaching reaction PMC in both films is

almost the same. We found that the reaction rate for the

photobleaching process is slower than that of the

photocoloration process in both matrices. This is expected

because PMMA and epoxy resin are both polar and could

form hydrogen bond with photomerocyanine and thus

stabilize the later which results in retardation of the

photobleaching rate.

-1.2

-1

-0.8

-0.6

-0.4

-0.2

0

0 100 200 300 400 500 600 700 800Time (sec.)

Photobleaching in PMMA polymerPhotobleaching in epoxy resin

Fig. 6: First-order plot of the photobleaching reaction of PMC in PMMA and epoxy resin films.

ln A

t / A

0



- 28 -

3.3 Photochemical fatigue resistance of spirooxazine in

PMMA and epoxy resin polymer films

One of the indispensable properties that should be fulfilled

by a photochrome to be used as a data-storage medium is

its high resistance to photochemical degradation. The

fatigue resistance of spirooxazine SO doped in PMMA and

epoxy resin polymer films is reported as the changes in

An/A0 with UV/visible irradiation cycles numbers, A0 and

An are the absorbance of the open form PMC at its λmax

obtained on the first and nth cycles, respectively (Fig. 7).

As it is clear from Fig. 7, the fatigue resistance of

spirooxazine doped in epoxy resin is much better than that

of spirooxazine doped in PMMA. Similar results were

reported with fulgides [22-25]. This was attributed to the

decrease of available polymer free volume due to

increasing cross-linkage in the epoxy resin [25].

0

0.2

0.4

0.6

0.8

1

1.2

1 2 3 4 5 6 7 8 9Number of irradiation cycles

A(n

) / A

(0)

SO doped in PMMA polymerSO doped in epoxy resin

Fig.7: Photochemical fatigue resistance of (SO) doped in PMMA and epoxy resin polymer films.

4. Conclusion The photochromic properties of 1,3,3-

trimethylspiro[indoline-2,3′-[3H]naphtho[2,1-b][1,4]

oxazine] SO doped in epoxy resin and PMMA films are

investigated. Kinetics of photocoloration and

photobleaching reactions were followed spectrophoto-

metrically. Irradiation of colorless spirooxazine SO doped

in PMMA and epoxy resin with UV light (366 nm) results

in the formation of the intense colored zwitterionic

photomerocyanine PMC.

The reverse reaction was photochemically

induced by irradiation with white light. It was found that

the visible absorption band is red shifted of the PMC

doped in epoxy resin compared to that doped in PMMA.

Such bathchromic shift was attributed to the higher polarity

of epoxy resin compared to PMMA. Photocoloration and

photobleaching reactions follow a first-order rate equation.



- 29 -

It was found that photocoloration rate constant of SO in

PMMA film is greater than that in epoxy resin. On the

other hand, photobleaching rate constant is almost identical

in both matrices. This was attributed to the higher stability

of the zwitterionic photomerocyanine PMC in both

matrices compared to the coloreless closed spirooxazine

SO. Spirooxazine doped in epoxy resin shows much better

fatigue resistance than that doped in PMMA.

Acknowledgment The authors wish to thank King Abdul Aziz City for

Science and Technology (KACST) For funding this

research work via grant no. At – 27 - 68.

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(2005) 21893.

[17] S. Fua, W. Hub, M. Xieb, G. Wanga, Y. Lia, Q.

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139.



- 30 -

Arabian J. Chem. Vol. 2, No. 1,31-42 (2009)


- 31 -


F. Z. Shtewi, R. A. Mokhtar*, A. Al-Zawik and S. Karshman

Chemistry Department, Faculty of Science, Zawia-Libya *Chemistry Department ,Faculty of Education, 7th April University, Zawia-Libya

[email protected]

Abstract A new method for the determination of ultra-trace amounts of iodide ion w as developed. The proposed

method employs ABTS, (2.2`-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)), as the chromogenic

reagent, and made full use of the advantages of stopped-flow methodology. This method was found to be

selective and sensitive. The method was based on the monitoring of the alteration in the rate of formation of

the cation radical of ABTS by oxidation with chloramine-T using a stopped-flow system. Traces of iodide

markedly increases the rate of the reaction. The alteration is proportional to the concentration of the iodide

which can be determined over the range 0-50 ppb with an RSD of less than 0.5% over this range.

1. Introduction

Iodine, as iodide is one of the trace elements present in

some foodstuffs at below the 50 mg/kg level. It is generally

regarded as one of the essentially nutritive elements. The

analysis of iodine at trace and ultra-trace level is becoming

increasingly important in the food industry [1, 2] and in the

analysis of environmental samples such as natural waters.

Attention has been growing to the role of iodide and

bromide in the formation of trihalomethanes, which are

regarded as possible carcinogens. Such trihalomethanes are

formed during the oxidative treatment of drinking water

[3]. Several methods have been reported for the

determination of iodide at ultra-trace levels, i.e. at ppb

concentrations, viz:10-7M and below this concentration.

Several different techniques have been employed.

A kinetic method [4] for the determination of

iodide in iodinated salt, based on the catalytic effect of

iodide on the chlorpromazine-bromate reaction reported the

limits of determination to be between 5 and 75 ppb. Other

methods [5, 6] involved the effect of iodide on the cerium

(IV)- arsenic(III) reaction. A spectrophotometric method

[7] has been reported for the determination of iodide in

river water over the range 20-100 ppb.

A catalytic reduction method [8] was chosen in

1992 as a standard method for the range 20-60 ppb. Mitic,

et.al[9]. reported that a kinetic method for the

determination of trace amounts of iodide by a catalytic

effect on the oxidation of sodium pyrogallol-5-sulfonate by

hydrogen peroxide. The reaction is followed

spectrophotochemically at 436.8 nm, the calibration graph

was found linear in the range 10-200ppb [9].

The inhibitory effect of iodide on the Pd(II)

catalyzed reaction between the EDTA-Co(II) complex and

hypophosphite has been used in spectrophotometry,

measuring the decrease in the absorbance at 540 nm. The

range of determination reported was 2-28 ppb [10]. Iodide

and thiosulphate have been determined using ion-pair,

response for the iodide was over the range 10-100 ppb [12].

Flow analysis technique was used for the determination of

iodide ion at a nanogram level in water by Chandrawanshi

et al. [13]. Iodide ion was also determined in urine and

water samples using isotope dilution analysis. The range of

determination reported in urine was 0.22-124.22 ppb [14].



- 32 -

Iodide has been separated from other species by

oxidation to iodine element, extraction into carbon

tetrachloride, re-conversion to iodide ion, reaction with

methylene blue to form an ion-pair. Extraction of this ion-

pair complex into 1,2-dicloroethane and

spectrophotometric determination of the complex. The

range of determination was 7.5×10-8 M to 3×10-6 M [15].

The use of a stopped-flow technique coupled

with a fixed optics system capable of monitoring changes

of 0.001 absorbance units has been previously reported

[16]. It was considered that using this, coupled with a

kinetic method based on a selective reaction and involving

the formation of a color, would have potential for the

development of a method capable for the determination of

ultra-trace amounts of iodide.

Consideration of potential chromogens indicated

that the choice should be governed by the commercial

availability of the chromogen in an analytical acceptable

state of purity as well as using a substance that gave a

product with a high molar absorptivity. The compound

2,2`-azinobis (3-ehylbenzothiazoline)-6-sulphonic acid

(ABTS) is used as an indicator for redox titrations

involving glucose [17]. It is readily available in the

required state of analytical purity and the product, the

cation radical, is highly colored in dilute solutions. This

compound was chosen for further study and use in the

proposed reaction sequences.

2. Experimental A block diagram of the experimental layout is given in Fig.

1. Light from a 50 watt tungsten-lamp passes through

approximately 1cm length of the water in the thermostatted

water bath (to act as a heat filter). Then through the optical

cell and an optical filter (to select the required wavelength),

mounted on the side of the optical cell and then directly on

to a focusing lens in front of a photodiode.

Signal output

The progress of the reaction is then followed by measuring

the change in absorbance of the beam of monochromatic

light. A simple electrical circuit allows the signal to be

amplified (sensitivity control) and zero-end on the scale by

appropriate “backing off”. The electronically amplified

signal from the detector is then recorded on a millivolt

potentiometric recorder. If required, the signal can be

further amplified using the recorder’s sensitivity control.

From the trace, suitable reaction parameters such

as the initial rate of reaction can be calculated. Using the

apparatus as presently designed it is possible to obtain

800 % “back off” and this is equivalent to using a recorder

with a chart width of 2 meters.



- 33 -

Figure 1. Block diagram of the stopped-flow system Dispensing of reagents Glass syringes fitted with Hamilton stainless steel/Teflon

3-way valves were used. The pistons of both syringes are

connected to a block which allows them to be operated

singly or simultaneously. In practice, it was found that a

volume of ca. 0.5 cm3, for each syringe was an acceptable

volume for all experiments.

The reactants pass through separate coils of thin walled

polyethylene delivery tubing (1mm; length 80 cm; nominal

volume 0.625 cm3) immersed in the thermostatted bath

(±0.5 oC) and into a mixing chamber which causes

homogeneous mixing by tangential action. From the

mixing chamber the solutions pass immediately into an

optical cell of 1.5 cm optical bath length, with an internal

volume of approximately 0.14 microliters, permanently

fixed in position. The solutions then pass to waste.

Reagents and Solutions Solution A Chloramine-T solution (1.0x10-3 M)

Solution B

Solution B contains a mixture of fixed amounts of

potassium iodide solution (i.e. 2.0 cm3; 1.0×10-6 M) and

ABTS solution (i.e. 2.0 cm3; 1.0×10-3 M). To this mixture

various amounts of 0.1 M HCl were added in order to give

different solutions with different ranges (i. e. in the range

of 7-9).

Each solution was made up to 25cm3 with de-

ionized water. The results are given in table 1. A pH of 7.0

was chosen for further studies. (see discussion). A stock

solution of pH 7 buffer, with a molarity of 0.02M with

respect to TRIS was prepared by dissolving 24.23g of the

solid in water, adding HCl (430cm3; 0.02 M) and diluting

with water to 1 liter.


The effects of variations of the experimental parameters

were investigated. The parameters which were individually

varied and included:- (i) pH of the system, (ii)

concentration of the TRIS buffer used, (iii) concentration



- 34 -

of the ABTS, (iv) concentration of the chloramines-T, (v)

temperature of the reaction cell. In each set of experiments

equal volumes (0.5 cm3) of two solutions (A and B) were

mixed in the stopped-flow apparatus and the absorbance of

the solution in the reaction cell was monitored at 625 nm.

The relative rates of formation of the green color were

calculated from the traces.

(i) Effect of varying the pH of the system The effect of variations in the buffer concentrations showing in table 1.

Table 1. shows the variety of buffer solutions regarding the relative rate.

pH 7.0 7.2 7.4 7.9 8.4 9.0 Relative rate 4.5 3.2 1.7 0.02 0.01 0.0

(ii) Effect of variations in the concentration of the buffer The reaction cell thermostatted at 25± 0.1oC. The results are shown in table 2. A buffer concentration of 4×10-2 M was

chosen for further work.

Table 2 shows the variety of TRIS buffer solutions regarding the relative rate.

TRIS buffer conc (10-3 M) 8 20 32 44 60 Relative rate 16.0 13.4 12.0 11.7 11.5

(iii) Effect of variations in the ABTS concentration

A series of 25 cm3 of solutions was prepared each

containing potassium iodide solution (0.50 cm3 of 1×10-5

M) and TRIS buffer (10 cm3 of 0.1 M; pH 7.0). To each

solution of the series 2 cm3 of ABTS solution was added to

have solutions of concentration ranging from 10-3 M to

2x10-4 M. The final volume was completed to the mark

with de-ionized water.

A sample of each solution was mixed with

Chloramine-T (1×10-3 M) in the thermostatted reaction cell.

The initial slopes of the reactions were calculated and

corrected for the blank. The results are given in table 3.

Table 3 shows the ABTS concentrations regarding the Initial rate

Final conc. ABTS 10-5M 2 4 8 12 16 20 24

Initial rate 1.3 2.5 5.0 7.3 9.7 11.0 11.0 The results obtained indicate an increase in the rate with

increase in the concentration of the ABTS until a

concentration of 2×10-4 M is reached. This concentration

was selected for further work.

(iv) Effect of variations in the temperature of the system Each of a series of aliquots of an iodide solution containing

10 ppb was mixed with aliquots of the buffered ABTS

reagent and then with aliquots of Chloramine-T to form a

mixture. The initial rate of the reacting compounds of each

mixture was monitored at different temperatures. The

results obtained are shown in table 4. From the data shown



- 35 -

in table 4, it appears that it is necessary to control the

temperature of the reaction. A temperature of 25oC was

chosen.

Table 4 shows the effect of temperature of the system

Temp. (oC) 19 21 25 34

Initial Rate 61.5 61.5 70 78

An aliquot (0.5 ml) of a standard iodide solution (solution

C) and an aliquot (0.5 ml) of the oxidant were

simultaneously injected into the system and the course of

the reaction was monitored at 625 nm for at least 60

seconds.

Calculations from the Recorder output

It may be seen from typical traces (Figure 2) that

the slope of the “blank” reaction, i.e. the non-catalysed

oxidation of the ABTS, depends upon the sensitivity

employed to monitor the reaction. The initial rate of the

reaction is calculated by measuring the slope and

calculating the tangent of the angle to the horizontal. It is

necessary to correct for any blank value. In practice it is

found to be more convenient to allow the trace to continue

for at least 60 seconds after the onset of the oxidation

reaction. And then measure the intercept of the trace on the

vertical axis of the chart at 60 seconds (or some other fixed

time) after the onset of the oxidation reaction. This

intercept is designated the I(0) intercept, or I(t) intercept.

From the recorder trace the initial rate of reaction and the

I(60) values were calculated as indicated. The process was

repeated for the series of solutions C and the rates of the

reactions and the I(60) values were plotted against the

concentration of iodide. The results are given in table 6.

Table 5 shows I(60) values were plotted against the concentration of iodide.

Conc. of iodide (10-8 M) 0 2.0 4.0 8.0 12.0 20.0 28.0 Relative Rate 0 1.05 2.24 4.44 6.66 11.0 15.5 I60, (mm) intercept 0 30.5 67.0 133 200 330 461

When the graph of the relative rate is plotted

against iodide concentration a straight line plot with a

linear correlation coefficient of 0.999 is obtained. A similar

linear correlation exists between the intercept60 and the

concentration of the analyte.

Interferences From the prepared calibration graph the initial rate of the

reaction , (corrected for any blank value) Or: intercept on

the vertical axis of the trace at 60 secs. (or t secs.) after the

start of the reaction. [I(60) or I(t)] (corrected for any blank

value), can be calculated. If very low concentrations are to

be monitored it may be necessary to allow the intervals of

time for obtaining the intercept to be up to 100 secs. This

allows the difference between the “blank” intercept and the

analyte intercept to be seen. Table 6 shows the

concentrations of the foreign species that can be tolerated

without significant effect (less than 5% interference).



- 36 -

Table 6 shows the concentrations (ppm) of the foreign species that can be tolerated without significant

effect

Anions Iodate (K)

Bromide (K)

Bromate (K)

Chloride (Na)*

Sulphate (Na)

Nitrite (Na)

Nitrate (Na)

EDTA (Na)

Conc. 1000 300 1000 2000 1400 300 1000 170 Cations Ca(II) Mg(II) Fe(II) Fe(III) Cu(II) Zn(II) Cd(II) Hg(II) Pb(II) Mn(II) Conc. 2000 2000 5 20 5 500 400 1 200 900

Removal of interferences

The interference of most of the cations is readily removed

using a suitable cation exchange. However, the cation

exchanger did not completely remove the interference

caused by the addition of mercury. (see discussion). Removal of the effect of mercury ions present

The above results indicate that the system can tolerate the

presence of up to 2000 ppm of chloride ion without

showing any effect on the determination of iodide.

Attempts were made to eliminate the effects of mercury by

addition of chloride to a sample. It was found that it is

necessary to remove residual iodide from a commercial

sample of A.R. sodium chloride by repeated

recrystallisation. (the results reported above are using

iodide-free sodium chloride).

Using sodium chloride which had been twice

recrystallised from water to remove any residual iodide.

The following results shown in table 7 were obtained in the

analysis of water sample which containing 25 ppb of iodide

and a set of samples which contained 25 ppb of iodide and

250 ppb of mercury(II).

Table 7 shows the effect of addition of sodium chloride to a solution containing Hg(II) 250 ppb

and 25 ppb of potassium iodide.

Conc. of Cl-(ppm) 0 400 1200 2000 2800

Initial Rate 9.0 12.0 15.0 19.9 16.0

The initial rate for the solution without mercury(II) present was 16.0. Determination of an unknown sample 1- use the same experimental conditions used in the

calibration exercise. As shown in table 8. Switch on the

electrical systems and allow warming up for at least 5

minutes. Ensure that all controls are locked into the

positions previously determined in the calibration

sequence. Ensure that the interference filter, for monitoring

the absorbance of the cell solution, is monitored at 625 nm.

2 (i)- If no cationic interferences are present, pipette 5 cm3

of the buffered ABTS reagent solution into a 25 cm3 flask.

Make up to the mark with the sample. Shake the mixture to

achieve homogeneity and place in the thermostat. Connect

to the syringe B and flush out the syringe and coil by

ejecting 2 aliquots (0.5 cm3) of the buffered ABTS and

sample mixture through the system.

2 (ii)- If cationic interferences other than Hg(II) are

present, pipette 5 cm3 of the buffered ABTS reagent

solution into a 25 cm3 flask. Connect a 25 cm3 syringe



- 37 -

filled with the sample to the ion-exchange column. Slowly

eject the solution through the ion-exchanger into the flask,

making the volume to the mark. Shake the mixture to

achieve homogeneity and place the flask into the main

thermostat. Connect to the syringe B and flush out the

syringe and coil by ejecting 2 aliquots (0.5 cm3) of the

buffered ABTS reagent and sample mixture through the

system.

2 (iii)- If cationic interferences including mercury(II) are

present, to 100 cm3 of the unknown sample add

approximately 0.1g of bromide free sodium chloride. Shake

to dissolve the solid and to achieve homogeneity, then

proceed as in 2 (ii).

3- Switch on the recorder. 4- With both syringes full, inject aliquots of the two

solutions into the coils and hence to the mixing cell.

Allow recording of the trace to continue for at least 60

secs.

5- From the trace obtained, calculate: either, the initial rate

of the reaction; or, the intercept on the vertical axis of

the trace at 60 secs. after the start of the reaction, (I(60)).

6- Calculate the concentration if iodide in the sample using

one of the previously prepared calibration curves.

Table 8 Conditions for the determination of iodide, and for calibration purposes, using the stopped-flow

apparatus. Parameter Instrument condition

Wavelength 625 nm Chart speed 60 mm per minute

Recorder sensitivity (mv for full scale deflection)

20 mv

Offset 800 % Temperature In range 20-25 oC ± 0.1 oC

solution Concentration Chloramine-T 1x10-3 M

Buffered reagent ABTS (2x10-4 M) in pH 7.0 TRIS buffer

4. Discussion

The main aim of the investigation was to design a selective

and sensitive method for the determination of ultra-trace

amounts of iodide present in water. An associated aim was

to ensure that the method was simple and required both

relatively low cost reagents and equipment. The latter

being such that on economic ground, it was suitable to

become a dedicated instrumental system in general

analytical laboratory used for routine or semi-routine assay

of the chosen analyte.

The reason for the choice of the stopped-flow

technique and a system involving the measurement of

initial rates of the analyte reaction has been previously

established. The fixed optical and physical geometries of

the system ensure that the system is optically stable and

capable of reproducing minute changes in the optical

absorbance of the solution under investigation. Using the

optical “backing off” system, with a back-off” of 800 %

gives the equivalent of a chart width of approximately 2

meters for a full scale deflection an ability to reproduce a

single to 1 millimeter. Thus, the present apparatus is

capable of reproducibly detecting and measuring



- 38 -

absorbance changes of the order of 0.0001 absorbance

units. This factor ensures that a sensitivity physical system

is of relatively low-cost, easy to use and service and

sufficiently robust in design to be used in a general

laboratory by skilled or semi-skilled workers after the

various reagents have been prepared.

When choosing a substance to be used as a

selective and sensitive reagent for ultra-trace amounts of a

particular analyte in aqueous media, the factors governing

selectivity and sensitivity should not be separately

considered. The choice of a chromogenic reagent which is

selective towards the changes in the system is primarily

governed by the type of analyte reactions available. In any

selective determination of iodide, use may be made of the

redox properties of the iodide/iodine system. This is

especially so when other ions present may also undergo

redox reactions. The use of the hypochlorite ion for the

oxidation of iodide is well established. Its use in systems

which have bromide present requires the control of the pH

of the medium to decrease the probability of interference

by the competing bromide/bromine system.

The choice of the pH was governed partly by the

fact that any bromide present is less likely to be oxidized at

this pH that is the iodide and also by the fact that mixing

equal volumes of 0.1 M TRIS and 0.1 M HCl and diluting

with water gives a buffer with a pH of approximately 7.0.

Thus, a usable buffer is easily made by even unskilled

labor and it is possible to dispense with the need to check

the pH if all calibrations are done using such a mixture. A

further consideration for choosing this pH is that at the

chosen temperature (25 °C) the pKa of TRIS is

approximately 8.0 and thus alteration in the ionic strength

of the solution when sodium chloride is added to sequester

Hg(II) will not have a significant effect on the pH of the

medium.

A result of any oxidative reaction involving the

chromogen will be either two forms of the reagent (a leuco

and a coloured form) or a new compound. In either case it

is essential that there should be a fairly large difference in

the molar extinction coefficients of the two compounds or

forms so that small changes in the amount of the compound

measured in the system are manifested as significant

changes in absorbance.

An oxidative is ABTS, which is an established

chromogenic reagent, readily available in an acceptable

state of purity, capable of being stored in normal laboratory

conditions for months without deterioration. It gives a

product with an acceptable high difference in its molar

extinction coefficient to that of the parent compound.

When a solution of Chloramine-T is mixed with

an iodide solution, buffered at pH 7.0 and containing

ABTS, the almost colourless solution first becomes yellow

and then quite rapidly turns a blue-green colour. The rapid

formation of the yellow colour is explained by the

formation of iodine from the iodide by its oxidation by

Chloramine-T. The disappearance of the yellow colour and

the formation of the green colour of the cation radical of

ABTS are explained as following:-

−+ +=+ I2ABST2ABSTI2

In this way the iodide is regenerated to be re-

oxidized by the HOCl. Thus, assuming the two reactions

are rapid, there is practically little decrease in the original

concentration of the iodide which can be regarded as acting

as a “catalyst”. The rate of formation of the ABTS radical

will be governed by the rate of formation of the iodine

molecules and thus is indirectly governed by the initial

concentration of the iodide ion.

Substances which remove iodide by

complexation, such as mercury(II) or remove iodine by

other redox reaction are potential sources of interference.

However, the latter type of potential interferences is

removed by the presence of the excess of Chloramine-T.

Possible cationic interferences, other than mercury, may be

removed by a suitable ion-exchange system. Mercury may

be sequestered by taking advantage of the ability to form



- 39 -

chloro-complexes of mercury which, although having

lower stability constants than the iodo-complex, are formed

because of the large excess of chloride used.

The proposed method may thus be used for a

wide variety of samples of the types generally found in

initial waters used for the generation of steam for boilers

…etc or for potable purposes. A comparison of the method

with others reported for the determination of iodide at the

ppb level is given below in table 9.

Table 9 Comparison of methods for the determination of iodide in the concentration range 0-50 ppb.

Technique Range (ppb) % RSD Spectrophotometry 2-28 2.2 Spectrophotometry 5-70 3.9 HPLC 3-1600 1.43 Colorimetry 20-80 NL Potentiometry 10-400 2.7 Voltammetry 0.3-17 5.0 Flow Injection 50-150 1.0 Proposed Method 0.5-5.0 1.0 2.5-36 0.3

When selecting a method for general industrial

use in routine or semi-routine analysis, the range of any

method may not be the only consideration. The overall cost

per analysis is also important. This is governed by the cost

of the apparatus and materials used and often more

importantly by the cost of the labor involved. A method

requiring many operational steps between receiving of

sample and dispatching of the results may require the use

of skilled (and thence expensive) labor. In many of modern

industrial work analytical results are needed as fast as

possible and often in a matter of minutes from receiving of

sample. Apparatus which is expensive may not always be

able to be dedicated to a particular analysis and thus time

will be required setting up a method for semi-routine

analysis, and overall time taking to obtain a result after

receiving of the sample may be relatively long.

As stated, one of the aims of the present study

was to design a method and procedure which is simple,

robust and relatively low-cost. Consideration of the above

methods indicates that this has been achieved. The

voltammetric method requires a pre-concentration step

before analyzing for the iodide and the concentration of the

analyte in the material undergoing analysis is greater than

50 ppb. The colorimetric method uses 10 different reagents

in the procedure. The cost of HPLC equipment is much

higher than that required for other methods. The flow

injection method is reported to be able to deal with

approximately 50 analyses per hour but no indication is

given if these are analyses (done in at least duplicates) or

are single results. In use as a routine assays (in duplicate) to

be obtained per hour, (If interferences are present, the rate

is reduced to about 20 per hour). The method of

calculation, involving the use of fixed time intercepts, is

both simple and rapid.

The proposed method uses low cost apparatus,

only a few reagents and dose not require any pre-

concentration steps before the analyte is determined. The

ability to vary the sensitivity of the system ensures that it

may be used over various ranges of concentrations

appropriate to the particular industrial problem.

After preparation of the solutions and calibration graphs,

the method dose not requires the use of highly skilled labor

and thus overall costs are reduced.

Thus, from various considerations, including ease

of operating procedure, initial cost of equipment, running

cost and the speed of analyses, the proposed method



- 40 -

appears to have industrial potential. Figure 2 shows the

typical recorder traces of iodide in solutions regarding the

following concentrations.

1-Blank (no iodide ion). 2-2.0×10-8 M, 3-4.0×10-8 M, 4-

1.0×10-7 M, 5-1.4×10-7 M.

1 cm of original chart corresponds to 10 secs.

T60 is time ordinate 60 secs. after onset of oxidation

reaction.

I60 is the intercept 60 secs. after the onset of the oxidation

reaction.

Note:

(i) Absorbance: 1 cm of original chart corresponds

to 0.001 a.u.

(ii) For a concentration of 1.4x10-7 M, I60

corresponds to 0.017 a.u. with the

parameters as indicated.

(iii) (iv) (v) Figure 2. Typical recorder traces of concentration of iodide in solutions (1) blank, (2) 2.0x10-8M, (3)

4.0x10-8M, (4) 1.0x10-7M and (5) 1.4x10-7M.

References

[1] Holak, W., Anal. Chem., (1987), 59, 2218.

[2] Van staden, J. F., Anal Lett., (1986), 19, 1407.

[3] Verma, K. K., Jain. A. and Verma. A., Anal.

Chem., (1992), 64, 1484.

[4] Vinas, P., Cordoba. M. H. and Sanchez-Pedreno.

C., Talanta, (1987), 34, 351.

[5] Rubio, S., and Perez-Bendito. D., Anal Chim Acta.,

(1989), 224, 185.

[6] Kenney, R. O., Bator. J. and Reading C., Anal

Biochem. (1989), 179, 139.

[7] Truesdale, V. W., and smith P., analyst. (2003),

100, 111.

[8] Greenberg, A. E., Clesceri L. S. and Eaton A. S.,

“Standard Methods for the Examination of Water

and Wastewater.” American Public Health

association. Washington. 18th Edn (1992) pp 4-72.

[9] Mitic, S. S., Miletic G. Z. and Kostic D. A. Anal.

Sci., vol,19(2005), No 6, 913.

[10] Garcia, M. S., Sanchez-Pedreno C., Albero M.I.

and Sanchez C., analyst (1991), 116, 653.



- 41 -

[11] Myashita, M. and Yamashita S., J. Chromatog.

1(990), 498, 137.

[12] Lookabaugh, M., Krull I. S., and Lacourse. W.R.,

J. Chromatog. (1987), 387, 301.

[13] Chandrawanshi, S. K., Chandrawanshi, S. K., and

Patel K. S., Journal of automated method and

management in chemistry, volume 18 (2005)

Issue 5, 181.

[14] Unak, P., Darcan S., Yurt F., Biber Z, and Coker

M., Boil Trace Elem Res. 1999 Winter; 71-72:

463-70.

[15] KOH, T., Ono, M. and Makino, I., Analyst,

(1988), 113, 945.

[16] Mokhtar, R. A., Shtewi, F.Z., Al-Zawik, A.,

Karshman, S., Jordan Journal of Chemistry,

(2008), V.3, 305.



- 42 -

Arabian J. Chem. Vol. 2, No. 1,43-48 (2009)


- 43 -

Simultaneous Determination of Metal Ions as Complexes of Pentamethylene Dithiocarbamate IN Indus River Water , Pakistan

1Muhammad Amir Arain, 2Feroza Hamid Wattoo*, 3Muhammad Hamid Sarwar Wattoo,

2Allah Bux Ghanghro,3Syed Ahmad Tirmizi, 4Javed Iqbal and 5Shahnila Amir Arain

1Dr. A.Q. Khan Research Laboratories, P.O. Box 502, Rawalpindi, Pakistan 2Institute of Biochemistry, University of Sindh, Jamshoro-76080, Pakistan

3Department of Chemistry, Quaid-i-Azam University, Islamabad-45320, Pakistan 4Institute of Chemistry, University of the Punjab, Lahore-54590, Pakistan

5 M.A. Kazi Institute of Chemistry, University of Sindh, Jamshoro-76080, Pakistan

*E-mail: [email protected]

Abstract River water samples before and after mixing with industrial effluents were collected at an interval of 4

weeks for one year and analyzed for simultaneous determination of Fe3+, Cr3+, Mn2+, Cu2+, Ni2+and Co2+

after preconcentration using pentamethylene dithiocarbamate (PMDTC) as derivatizing reagent and

subsequent solvent extraction by high performance liquid chromatography (HPLC). The average levels

(n = 12) of metal ions were found in the range of 14.2 to 542 µg/L. The results were then compared with

a standard flame atomic absorption spectrophotometric method revealed no significant differences.

Keywords: Liquid Chromatography, Pentamethylene Dithiocarbamate Complexes, Metals, Fresh Water

1. Introduction Determination of trace metals in water [1-5] is often made

possible by the addition of complexing agent and analyzing

the sample by spectrophotometry or by liquid

chromatography. Most separation methods in use are based

on the formation of metal dithiocarbamate especially

ammonium pyrrolidine dithiocarbamate and sodium diethyl

dithiocarbamate as ligands to stabilize high oxidation states

which allow monitoring of the oxidation rather than the

reduction of metal dithiocarbamate complex formed in situ

in the liquid chromatographic system [6-7]. Sodium diethyl

dithiocarbamate has been used as a derivatizing reagent for

gas chromatography as well as liquid chromatography [3,

7, 8] using electrochemical and spectrophotometric

detections [9].

Mostly chromatographic separations of metal

dithiocarbamates were achived using normal phase

chromatography with UV-Visible spectrophotometric

detection [10-15]. Babu and Naidu [16] reported the use of

pentamethylene dithiocarbamate for the complexation,

solvent extraction and AAS determination of Fe, Ni, Cr and

Mn from water. Asolkar et al.[17] used the same reagent

for the determination of Cd2+, Cu2+, Fe3+ and Pb2+ on thin

layer chromatography. Arain et al. [5] separated the series

of six metal ions as chelates of pentamethylene

dithiocarbamate by capillary gas chromatography (CGC)

and high performance liquid chromatography (HPLC); see

figure 1.

N C

S

S

M

S

C

S

N

Figure 1. Structural diagram of PMDTC–metal

complex

Muhammad Amir Arain, Feroza Hamid Wattoo , Muhammad Hamid Sarwar Wattoo


- 44 -

In the present work, we have investigated the determination

of Fe3+, Cr3+, Mn2+, Cu2+, Ni2+, Co2+ ions from fresh water

samples collected from river Indus at Ghulam Muhammad

barrage. The metal contents were preconcentrated as

complexes of pentamethylene dithiocarbamte, extracted in

organic solvent and simultaneously determined by HPLC.

The seasonal variations in the metal contents of river Indus

water were also evaluated.

2. Experimental

2.1 Instrumentation

A liquid chromatograph (Perkin-Elmer 8700) equipped

with LiChrosorb ODS column (150 x 4.6 mm, i.d., 5 µm),

UV-detector, Rheodyne 7125 injector and D-2500

chromato-integrator and an atomic absorption spectrometer

(Hitachi-18050) were used in present work.

Electrochemical measurements were made with Pye

Unicame model 292 pH meter. Single channel transfer

Pipettes using 100 µL (0.1 ml tip) were used to deliver the

metal ion solution.

2.3 Reagents and Solutions

Stock metal ion solutions containing 1 mg/ml of each metal

ion were prepared from their nitrate. Methanol, sulfuric

acid, nitric acid, hydrochloric acid, acetic acid, hydrogen

peroxide and sodium acetate were all purchased from E.

Merck Germany. All chemicals used were of AR grade

purity. Deareated high purity double distilled

demineralized water was used for mobile phase and

solution preparation.

2.4 Synthesis of Pentamethylene Dithiocarbamate

Reagent (PMDTC)

Carbon disulfide (76 g/mol) was slowly added to 80 g

freshly vacuum distilled pipridine (80 g/mol) in 25 ml of

water at temperature > 5 °C with a constant stirring

followed by the addition of 40 g sodium hydroxide

dissolved in 20 ml water [16]. The reagent solution was

prepared by dissolving 1 g of the reagent in 100 ml of

water.

2.5 Analytical Procedure

250 ml of aqueous solution containing chromium, cobalt

and manganese (0-20 µg), iron (0-25 µg), nickel and

copper (0-30 µg) was transferred to a 500 ml separating

funnel. Then the reagent solution of PMDTC (5 ml, 0.1%

w/v in water) and acetate buffer (pH 5, 5 ml) were added.

pH was adjusted to 5. Chloroform (5 ml) was added and

the contents were mixed well for 3 minutes and aqueous

layer was allowed to separate from organic layer, which

was transferred to a volumetric flask. The extraction was

repeated with chloroform (5 ml). The chloroform layers

were combined and volume was made up to 10 ml. 20 µL

of this extract was injected into RP-HPLC connected with

ODS column (150 x 4.6 mm. i.d., 5 µm), with a mobile

phase consisting of methanol: 1% 0.1M acetate (30: 70,

v/v), with a flow rate of 1.2 ml/min. and detection was at

260 nm by UV detector [3, 5].

2.6 Determination of Cr, Mn, Fe, Co, Ni and Cu in

River Indus Water Samples

Indus river water samples (n = 12) were collected from

Ghulam Muhammad barrage (before mixing of industrial

effluents) and near Kotri SITE area (after mixing with

industrial waste water), with the interval of one month in

2.5 L glass bottles. Subsurface water samples were

collected at the depth of one foot. All samples were

preserved as per standard procedure [1, 2]. The samples

were analyzed for the metal contents next day using the

above mentioned analytical procedure.

3. Results and Discussions

The reagent reacts with iron, chromium, manganese,

copper, nickel and cobalt to form color complexes [5, 18,

19]. Maximum color development occurs in neutral to

slightly acidic media. The metal chelates are easily

Simultaneous Determination of Metal Ions as Complexes of Pentamethylene Dithiocarbamate……..


- 45 -

extractable in chloroform. The reagent was examined for

preconcentration, extraction and simultaneously

determination of Fe3+, Cr3+, Mn2+, Cu2+, Ni2+ and Co2+.

Metal pentamethylene dithiocarbamate chelates (Figure 1)

were separated as reported [5] on HPLC column (150 x 4.6

mm. id, 5 µm). HPLC was calibrated with six standards

and extraction efficiency was evaluated by adding 250 ml

distilled water. The instrument was recalibrated after five

samples; it was observed that percentage recovery was 94-

100% with a coefficient of variation (C.V) up to 3.9%.

This method was applied for the determination of

metal ions in water samples collected from Indus river and

were examined quantitatively from April to March (n =

12). The percentage recovery (Table 1) of each of the metal

ions was examined using analytical procedure and the

average recovery (n = 5) was observed within 92-99% with

coefficient of variation within 1.2-3.9%. The average

concentration of Cr3+, Mn2+, Fe3+, Co2+, Ni2+ and Cu2+

observed were 78, 61, 542, 23, 14.2 and 42.6 µg/L,

respectively as shown in (Table 2). The concentration of

metal ions exhibited the Fe3+>Cr3+>Mn2+>Cu2+>

Ni2+>Co2+, decreasing sequence.

Table 1. Percentage recovery of metal ions by preconcentration (n = 5)

Metal Ions Metal added (µg/ml)

Metal found By HPLC*

Metal found By AAS** % Recovery

0.25 0.248±0.013 0.249 99.20 0.50 0.496±0.025 0.496 99.20 1.00 0.994±0.182 0.999 99.40 1.50 1.488±0.028 1.492 99.20 2.00 1.982±0.013 1.995 99.10

Fe3+

Mean % Recovery 99.22 0.25 0.240±0.012 0.244 96.00 0.50 0.480±0.024 0.495 96.00 1.00 0.956±0.052 0.980 95.60 1.50 1.470±0.035 1.473 98.00 2.00 1.928±0.072 1.990 96.40

Cr3+

Mean % Recovery 96.40 0.25 0.234±0.011 0.242 93.60 0.50 0.468±0.023 0.489 93.60 1.00 0.960±0.037 0.983 96.00 1.50 1.390±0.030 1.486 92.70 2.00 1.932±0.024 1.964 96.60

Mn2+

Mean % Recovery 94.50 0.25 0.240±0.011 0.246 96.00 0.50 0.480±0.015 0.493 96.00 1.00 0.961±0.016 0.983 96.00 1.50 1.446±0.016 1.486 96.40 2.00 1.937±0.005 1.970 96.90

Cu2+

Mean % Recovery 96.26 0.25 0.246±0.130 0.249 98.40 0.50 0.492±0.010 0.498 98.40 1.00 0.980±0.016 0.992 98.00 1.50 1.476±0.016 1.486 98.40 2.00 1.962±0.022 1.983 98.10

Ni2+

Mean % Recovery 98.26 0.25 0.234±0.011 0.243 93.60 0.50 0.468±0.016 0.488 93.60 1.00 0.928±0.015 0.983 92.80 1.50 1.376±0.052 1.486 91.70 2.00 1.858±0.040 1.940 92.90

Co2+

Mean % Recovery 92.92 HPLC* = High performance liquid chromatography, AAS** = Atomic absorption spectroscopy, Average values, n = 5, Confidence interval at 95%.



- 46 -

Table 2. Average concentration of metal ions in µg/l (n = 12) with confidence interval at 95%.

Metals Ions Fe3+ Cr3+ Mn2+ Cu2+ Ni2+ Co2+

Minimum 15.60 238.00 11.20 22.00 2.14 8.40 Maximum 82.80 960.00 198.00 106.00 31.20 37.50

G.M

. Bar

rage

(n

= 1

2)

Mean 42.6±17 542.0±188 78.0±21.2 61.0±17 14.2±5.5 23.0±8

Minimum 21.20 292 47.40 36.50 4.60 9.20 Maximum 164.00 1383.00 418.00 285.00 72.00 47.80

Kot

ri In

dust

rial A

rea

(n =

12)

Mean 56.0±21 766.0±212 96.0±29 72.0±22 21.0±7.3 29.0±11.2

Ghulam Muhammad barrage = Actual Indus river water, Kotri industrial area = River water after mixing with industrial effluents

The total metal ions concentration fluctuated between 2.12

to 960 µg/L at Ghulam Muhammad barrage and 4.6 to

1383 µg/L at Kotri SITE area. The seasonal variation of

metal ions (Figure 2) was uniform and depended upon

water flow. High flow occurs in summer, when snow

melts extensively and dominant monsoon rains augment

many fold. Metal contents were diluted in peak flow season

June to September and concentration level was high in

winter due to the shortage of water especially during

December to February (Figure 2). Figure 2 also indicates

maximum concentration of metal ions in the month of

January and minimum in the July, which is due the water

discharge in river Indus. The results also indicate highest

concentration of iron through out the study.

Figure 2. Seasonal variation of metal ions

Simultaneous Determination of Metal Ions as Complexes of Pentamethylene Dithiocarbamate……..


- 47 -

4. Conclusion This method have been used for the determination of

chromium, manganese, iron, cobalt, nickel and copper ion

as pentamethylene dithiocarbamate chelates in Indus river

water and effluent water samples (after mixing industrial

effluents from SITE area) and good correlation was

observed with that of atomic absorption spectrometry. The

metal ions contents were observed in a safe limit but

concentration of iron and copper contents were slightly on

the higher side. This is might be due to the extraction from

sediments at acidic pH adjusted for the preservation of

water samples.

Acknowledgments

The authors acknowledge continuous support of the

research laboratories of National Center of Excellence in

Analytical Chemistry, University of Sindh, Jamshoro,

Pakistan. Mr. Saad Iqbal, Senior Scientist, Pakistan Atomic

Energy Commission, Islamabad, is acknowledged for his

help in necessary computational facilities about this

project.

References [1] M. H. S. Wattoo, F. H. Wattoo, S. A. Tirmizi, T. G.

Kazi, M. I. Bhanger and J. Iqbal, “Pollution of Phulali

canal water in the city premises of Hyderabad: Metal

monitoring”, J. Chem. Soc. Pak., 28 (2006) 136.

[2] M. H. S. Wattoo, F. H. Wattoo, T. G. Kazi, S. A.

Tirmizi, M. I. Bhanger, R. B. Mahar and J. Iqbal,

“Quality characterization of Phulali canal water for

agricultural purposes”, The Nucleus, 41 (2004) 69.

[3] M.A. Arain, M.I. Bhanger and M.Y. Khuhawar,

“Capillary gas chromatography of metal chelates of

diethyl dithiocarbamates, Chromatographia, 55,

(2001) 349.

[4] P. Wang, H.K. Lee, “Recent applications of high-

performance liquid chromatography to the analysis of

metal complexes”, J. Chromat. A, 789, (1997) 437.

[5] M.A. Arain, M.I. Bhanger and M.Y. Khuhawar, “Gas

and liquid chromatography of metal chelates of

pentamethylene dithiocarbamate”, J. Chromatogr. A.,

973, (2002) 235.

[6] A. M. Bond and G. G. Wallace, “Determination of

copper as a dithiocarbamate complex by reverse-phase

liquid chromatography with electrochemical

detection”, Anal. Chem., 53 (1981) 1209.

[7] A. M. Bond and G. G. Wallace, “Simultaneous

determination of copper, nickel, cobalt,

chromium(VI), and chromium(III) by liquid

chromatography with electrochemical detection”,

Anal. Chem., 54 (1982) 1706.

[8] T. J. Cardwell, D. J. Desarro and P. C. Uden, “Gas

chromatography of some volatile metal diethyldi-

thiocarbamates”, Analytica Chimica Acta, 85 (1976)

415.

[9] A. M. Bond and G. G. Wallace, “Automated

determination of nickel and copper by liquid

chromatography with electrochemical and

spectrophotometric detection”, Anal. Chem., 55,

(1983)718.

[10] O. Li ka, J. Lehotay, E. Brand teterová, G. Guiochon

and H. Colin, “Liquid chromatography of metal

complex of N-disubstituted dithiocarbamic acids: IV.

Separation of mixtures of Zn(II), Cu(II), Mn(II),

Ni(II), Pb(II), Cr(III), Co(II), Cd(II) and Fe(II)

diethyldithiocarbamate complexes by high-

performance liquid chromatography”, J. Chromatogr.,

172 (1979) 384.

[11] M. Moriyasu and Y. Hashimoto, “Microdetermination

of Heavy Metal Chelates by High-Performance Liquid

Chromatography”, Anal. Lett., 11 (1978) 593.

[12] R. R. Brooks, B. J. Presley and I. R. Kaplan, “APDC-

MIBK extraction system for the determination of trace



- 48 -

elements in saline waters by atomic-absorption

spectrophotometry”, Talanta, 14 (1967) 809.

[13] T. K. Jan and D. R. Young, “Determination of

microgram amounts of some transition metals in sea

water by methyl isobuthyl ketone-nitric acid

successive extraction and flameless atomic absorption

spectrophotometry”, Anal. Chem., 50 (1978) 1250.

[14] L. Danielsson, B. Magnusson and S. Westerlund, “An

improved metal extraction procedure for the

determination of trace metals in sea water by atomic

absorption spectrometry with electrothermal

atomization”, Anal. Chim Acta, 98 (1978) 47.

[15] R. E. Sturgeon, S. S. Berman, A. Desaulniers and D.

S. Russell, “Pre-concentration of trace metals from

sea-water for determination by graphite-furnace

atomic-absorption spectrometry”, Talanta, 27 (1980)

85.

[16] D. R. Babu and P.R. Naidu, “A solvent extraction-

atomic absorption technique for the simultaneous

determination of low concentrations of iron, nickel,

chromium and manganese in drinking water”,

Talanta, 38 (1991) 175.

[17] A. Asolkar, A. Kumar, P. Pandey and R. Bhardwaj,

“TLC resolution studies of some metal-

piperidinedithiocarbamate complexes on various

surfactant impregnated silica gel-g plates”, J. Liq.

Chromatogr., 15 (1992) 1689.

[18] V. V.Ramana, K. Santha, M. Dasaratha Ramaiah, K.

Saraswathi, “Determination of molybdenum in soil

using sodium pentamethylene dithiocarbamate”, J.

Indian. Chem. Soc., 68 (1991) 178.

[19] P. Kuba´nˇ, R. Guchardi and P. C. Hauser, “Trace-

metal analysis with separation methods”, Trends

Analyt. Chem., 24 (2005) 192.

New Ceramic Microfiltration Membranes From Mineral Coal Fly Ash


- 49 -


Ilyes Jedidi1,2,3, Sami Saïdi1, Sabeur Khmakem1, André Larbot 2, Najwa Elloumi-Ammar3, Amine Fourati3, Aboulhassen Charfi3 and *Raja Ben Amar1

1 Laboratoire Sciences des Matériaux et Environnement, Faculté des sciences de Sfax, Rte. de Soukra Km 4, 3018, Sfax, Tunisia.

2 Institut Européen des Membranes, UMR 5635 (CNRS, ENSCM, UM II), 1919 Route de Mende, 34293, Montpellier, CEDEX 5, France.

3 Groupe Chimique Tunisien, Centre de Recherche de Sfax, B.P : S, Route de Gabes km 4,5, Sfax, 3018, Tunisia. * Email: [email protected]

Abstract This work aims to develop a new mineral porous tubular membrane based on mineral coal fly ash.

Finely ground mineral coal powder was calcinated at 700°C for about 3 hours. The elaboration of the

mesoporous layer was performed by the slip-casting method using a suspension made of the mixture

of fly ash powder, water and PVA. The obtained membrane was submitted to a thermal treatment

which consists in drying at room temperature for 24 hours then a sintering at 800 °C. SEM

photographs indicated that the membrane surface was homogeneous and did not present any macro

defects (cracks, etc…). The average pore diameter of the active layer was 0.25 µm and the thickness

was around 20µm. The membrane permeability was 475 L/h.m2.bar.

This membrane was applied to the treatment of the dying effluents generated by the washing baths in

the textile industry. The performances in term of permeate flux and efficiency were determined and

compared to those obtained using a commercial alumina microfiltration membrane. Almost the same

stabilised permeate flux was obtained (about 100 L.h-1.m-2). The quality of permeate was almost the

same with the two membranes: the COD and color removal was 75% and 90%, respectively.

Keywords: Mineral coal fly ash; Ceramic microfiltration membrane; Slip-casting process; dying

effluents.

1.Introduction

Ceramic membranes are used in the crossflow filtration

mode, which allows maintaining a high filtration rate

compared with the direct-flow filtration mode used in

conventional filtration process.

Thermal, chemical and mechanical properties of

ceramic membranes give them significant advantages over

polymeric ones [1]. Conventionally, alumina, zirconia,

titania and silica are considered as the main materials of

commercialized ceramic membranes [2]. Unfortunately,

these membranes are too expensive for a technico-

economic point of view. For example, in the

environmental field, great volumes of wastewater are

generally treated. So, the use of membrane separation

techniques requires a great membrane area. Recently, the

development of low cost ceramic membranes based on

natural materials such as clays and apatite appeared as an

efficient solution to treat waste water at a low cost [3-7].

Mineral coal fly ash obtained from coal-fired

power stations could be also a good material to make low

cost membranes. Indeed, this way allows a good

management of this subproduct which represents a major

Ilyes Jedidi, Sami Saïdi, Sabeur Khmakem, André Larbot , Najwa Elloumi-Ammar, Amine Fourati, …

Arabian J. Chem. Vol. 2, No. 1 (2009)

- 50 -

problem in many parts of the world due to the resulting

pollution. It is noticed, by the same way, that significant

quantities are being used in some range of applications like

in construction and other civil engineering applications [8]

where fly ash is used as a substitute for cement in concrete

[9]. During the past years, some researches have been

performed concerning the integration of fly ash in the

manufacture of bricks and tiles which use a large volume

of silicate-based raw materials [10,11]. Conventional

porous ceramic products prepared using only fly ash have

been also investigated [12,13]. Little research work has

focused on upgrading this material in the membrane

preparation field like the preparation of stainless-steel / fly

ash membrane suitable for hot gas cleaning [14].

This work describes the elaboration worth on

ceramic fly ash microfiltration membrane applied to the

clarification and the decolouration of the effluents coming

from the dying industry.

2. Experimental

2.1 Materials and methods

2.1.1 Characterisation of the fly ash powder

The fly ash powder used was obtained by calcination at

700 °C of a finely ground mineral coal. The particle size

analysis of the powder was determined using the Particle

Sizing System AccuSizer Model 770 (Inc. Santa Barbara,

Calif., USA). The grinding of the mineral coal was

performed using a planetary crusher at 300

revolutions/min.

A Hitashi scanning electron microscope (SEM)

was used to study the powder morphology as well as the

microstructure formed in the sintered material. The

chemical composition of the powder was determined by

spectroscopic techniques: X-ray fluorescence for metals

and atomic absorption for alkaline earth metals. Phases

present in the powder composition were analysed using an

X-ray diffractometer (Siemens, Germany) with Cu Kα

radiation (λ = 0.154 nm).

The thermogravimetric analysis (TGA) and differential

scanning calorimetry (DSC) of the fly ash powder were

carried out at temperature ranging between 0 and 1000 °C

at a rate of 5°C/min under air.

2.2 Membrane elaboration

The slip-casting process was applied to form a

microfiltration layer based on mineral coal fly ash, coated

on a macroporous support, previously elaborated in our

laboratory from the same material with the following

characteristics: a mean pore diameter of 4.5 µm and a

porosity of 51%.

2.3 Slip casting process

The active microfiltration layer from fly ash was prepared

by a slip casting process on fly ash support (closed-end

tubes of 150 mm in length, with an inner diameter of 5

mm) in dip solution containing the powder and an aqueous

solution of polyvinyl alcohol (PVA) (Rhodoviol 25/140

(Prolabo)), used as a binder.Figure 1 describes the slip

casting process. It consists of:

• Putting in suspension the mineral powder in

water.

• Adding a binder (12-wt % aqueous solution of

PVA) and homogenisation by a magnetic stirring.

• Coating the support for a few minutes at room

temperature. In the case of the tubular

membranes, the tube was closed at one end and

filled with the solution.

• Drying is carried out for 24 h at room

temperature.



- 51 -

Figure 1: showing scheme of a slip casting process.

2.3.1 Composition and characterisation of the slip

In order to make a slurry solution suitable for the slip

casting, empirical study was performed to select the

optimum composition. The optimised slip composition was

based on a rheological study using a viscosimeter LAMY

model TVe-05 (five shear speeds were used) and the SEM

observation of the sintered layer obtained according to a

fixed temperature-time schedule previously determined

using clay material [15].

The investigation was focused on the uniformity

of the coating deposited on the inner surface of the

macroporous support. The optimum composition was

shownin table 1.

Table 1 : Composition of the slurry solution:

Component Conditions Proportion (wt %) Water Deionised 66

Polyvinyl alcohol (aqueous solution) 12 % aqueous solution 33 Fly ash Particle size < 5 µm 4

2.4 Sintering Program

The firing temperature, fixed at 800 °C, is reached

following the program shown in figure 2. A temperature

plate at 250 °C for 1 h is necessary in order to completely

eliminate the PVA, which is in great quantities in the slip.

A relatively slow temperature increasing rate (2°C/min)

was needed in order to avoid the formation of cracks on the

layer.

.



- 52 -

Figure 2: shown the temperature-time Schedule used in the active layer sintering.

2.5 Membrane characterisation

The average pore diameter of the active layer was

determined by mercury porosimetry on a Micrometrics

Autopore II9220 V3.05. The membrane texture was

characterised by Scanning Electron Microscopy (SEM).

2.6 Filtration tests

Crossflow microfiltration tests were performed using a

home-made pilot plant (Fig.3) at a temperature of 25°C and

transmembrane pressure (TMP) range between 1 and 4

bars. The transmembrane pressure was controlled by an

adjustable valve at the concentrate side. The flow rate was

fixed at 1.76 m s-1. Before the tests, the membrane had

been conditioned by immersion in pure deionised water for

at least 24 h. The duration of each test ranged from 1 to 3

hours.

Figure 3: shown the scheme of the pilot plant.

2.7 Effluents characterisation

The microfiltration membranes have been applied to

wastewater treatment coming from the dying industry.

Conductivity, absorption (using an “OPTIMA SP-3000”

UV-VIS spectrophotometer at a λ = 600nm, since the raw



- 53 -

effluent colour is blue) and pH measurements were performed.

3. Results

a. Fly ash characterization

i. Chemical composition and particle size distribution

The chemical composition of the fly ash is given in Table

2. The majority of the used fly ash (82.4%) consists of

SiO2, Al2O3 and Fe2O3. The other percentage is a mixture

of different alkali metals.

The fly ash powder obtained by calcination of the

finely ground mineral coal at 800°C showed a particle size

diameter less than 2 µm (figure 4). Figure 5 shows that the

particles size distribution of the powder used for the

elaboration of the microfiltration layer is homogeneous

within the interval 0 to 5 µm. However, it appears that a

majority of the particles are sized between 0 and 1µm

which is in accordance with the particle size distribution

diagram of figure 4.

Table 2: Chemical composition of the used fly ash.

Elements Proportion (wt %) SiO2 49.09 Al2O3 24.34 Fe2O3 8.93 CaO 4.88 MgO 3.15 K2O 1.74 SO3 2.15 LOIa 1.07

a Loss on ignition.

Figure 4: Fly ash particle size distribution.



- 54 -

Fig. 5: SEM picture of fly ash powder after calcination of a finely ground mineral coal

at 700°C.

ii. Thermal analysis

The DSC-TGA data shows that the mass loss is around

1.5% (figure 6) which is due to some impurities and the

small percentage of unburned mineral coal powder, since

the phenomenon started at 250°C and lasted until it reached

800°C.

iii. Phase identification

XRD data for a sample sintered at 800°C are shown in

figure 7. The major crystalline phases identified were

quartz (SiO2), anorthite (CaAl2Si2O8), gehlenite

(Ca2Al2SiO7), hematite (Fe2O3) and mullite

(3Al2O3·2SiO2). A minority of anhydrite (CaSO4) can be

seen on the spectrum.

Figure 6: DSC-TGA data of the fly ash powder.



- 55 -

Figure 7: XRD pattern of fly ash powder fired at 800°C (Q = quartz, M = mullite,

An = anhydrite, Ge = Gehlenite, H = Hematite).

b. Membrane characterisation

i. Slip characterisation

Three slips with a percentage of 4% of fly ash and three

different rates of PVA (30%, 39% and 45%) were

prepared. The rheological data of the three compositions

are given in figure 8 which represents the curve of shear

stress (τ) versus shear rate (D). Fly-ash slip was found to

exhibit a rheo-thickener behaviour, controlled by the

presence of PVA. The decrease of PVA percentage leads to

the maintenance of particles in a stable suspension.

ii. Scanning Electron Microscopy

Slips S1 and S2 were used to prepare an active layer on the

macroporous support. The same casting time was used

during the slip casting operation. The sintering conditions,

previously mentioned, were respected. Figures 9 and 10,

which show SEM pictures for surface and cross section of

respectively S1 and S2 elaborated layers, give information

about the thickness and texture. For S1 slip, a defect-free

microfiltration membrane was obtained with a layer

thickness of around 20 µm. However, as regards to S2 slip,

a multi-defect layer was noticed (a thick layer with many

cracks).

iii. Determination of the porosity

Porosity and pore size distribution were measured by

mercury porosimetry. This technique is based on the

penetration of mercury into a membrane’s pores under

pressure. The intrusion volume is recorded as a function of

the applied pressure and then the pore size was determined.

The pore diameters measured were centred near 0.25µm for

the deposited microfiltration layer (figure 11).

iv. Determination of membrane permeability

The membrane permeability (Lp) can be determined using

the variation of the water flux (Jw) with the transmembrane

pressure (∆P) following the Darcy’s law:

Jw = Lp . ∆P, where ∆P = [(Pinlet + Poutlet) / 2 – Pf]

Pinlet = inlet pressure ; Poutlet = outlet pressure ; Pf = filtrate

pressure.



- 56 -

It can be seen that the water flux increases linearly with

increasing applied pressure (figure 12). The membrane

permeability (Lp) was found to be equal to 475 l/ h.m2.bar.

Figure 8: Evolution of the shear stress (Tau, τ) versus shear rate (D) for different

PVA percentages in fly ash slip: (S1: 4% fly ash / 66% water / 30%

PVA), (S2: 4% fly ash / 57% water / 39% PVA), (S3: 4%

fly ash / 51% water / 45% PVA).

Figure 9: SEM micrographs of the optimised active layer obtained with the slip

composition S1 (4 % fly ash / 66 % water / 30% PVA) and sintered at

800 °Ca) cross-section, b) surface.



- 57 -

Figure 10: SEM micrographs of the multi-defects active layer obtained with the

slip composition S2 (4 % fly ash / 57% water / 39 % PVA) and sintered

at800°C. a) and b) surface views with different magnitudes, c) Cross-

section.



- 58 -

Figure 11: Pore diameters of the Fly ash membrane.

Microfiltration Layer

Support

0.25



- 59 -

Figure 12: Water fluxes vs. working pressure.

c. Application to the treatment of textile dye waste

water

The treatment of industrial waste water can be achieved by

membrane process. Thus, microfiltration was used in this

study to the clarification of textile dye waste water. Two

ceramic membranes are used: on 0.2 µm Alumina

membrane and the fly ash elaborated membrane. Figure 13

shows typical microfiltration experiments for the two

membranes. For fly ash membrane, the flux drops fast in

the first 15 minutes from 410 l/h.m2 to 135 l/h.m2 then

stabilises at a permeate flux (Jf) of about 90 l/h.m2. The

same behaviour

was obtained with alumina membrane which then was

made to be stabilised at a permeate flux of 110 l/h.m2.

The average effluent quality (before and after

microfiltration treatment) is illustrated in the table (3).

Microfiltration using fly ash membrane proved to be

effective in removing the COD, turbidity and color with

almost the same efficiencies as that obtained with alumina

membrane: 75% for COD, 90 % for color. A very low

turbidity value of the two permeates was also obtained (0.5

NTU).

Table 3 : Characteristics of the effluent before and after microfiltration at 1 bar with the fly ash and the

alumina membranes.

Sample Conductivity Turbidity (NTU) COD (mg.L-1) Absorbance at 600 nmRaw effluent 6.16 45.5 3440 0.104

Fly ash membrane 5.38 0.58 880 0.010 Alumina membrane 5.6 0.62 834 0.013



- 60 -

Figure 13: Variation of permeate flux with time (T = 25°C, TMP = 1 bar).

Figure 14: A photograph of the dying effluent before and after MF treatment (T = 25 °C,

TMP = 1 bar).



- 61 -

4. Conclusion

New ceramic microfiltration membranes made of mineral

coal fly ash have been prepared and characterised. The fly-

ash powder characterisation was performed. It was found

that the crystalline phases composition is changing with the

increase of the calcinations temperature and that the weight

loss is very slight. The optimised composition of the slip

was determined: 30% PVA, 66% water and 4% fly-ash

powder. The obtained membrane was defect free and has

the following characteristic: thickness of about 20 µm,

mean pore diameter of 0.25 µm and porosity of 51 %.

The performances of the fly-ash microfiltration

membrane for the treatment of the textile dye waste water

was determined and compared with those obtained using

commercial 0.2 µm alumina membrane. Almost the same

stabilised permeate flux was obtained (about 100 l.h-1.m-2).

The quality of permeate was almost the same with the two

membranes: the COD and color removal was 75% and

90%, respectively.

These experimental results show that mineral

coal fly-ash is an appropriate material for the development

of microfiltration membranes which could be applied to the

industrial wastewater treatment.

Acknowledgement

This work was supported in part by The Tunisian Chemical

Group Company.

List of symbols

abbreviations

Nomenclature

Pinlet : inlet pressure (bar)

Poutlet : outlet pressure (bar)

Pf : filtrate pressure (bar)

Jw : Water permeate flux (l/h.m2)

Lp : Water permeability (l/h.m2.bar)

COD : Chemical Oxygen Demand (mg/l)

Jf : Permeate flux (l/h.m2)

Jw : Water flux (l/h.m2)

TMP : Trans-membrane pressure (bar)

τ : shear stress (mPa)

D : shear rate (s-1)

References

[1] K.K. Chan and A.M. Brownstein, Ceramic membranes:

growth prospects and opportunities,Ceramic Bulletin,

70 (1991) 703.

[2] T. Tsuru, Inorganic porous membranes for liquid phase

separation, Sep. Purif. Meth. 30 (2001) 191–220.

[3] M.R. Weir, E. Rutinduka, C. Detellier, C.Y. Feng, Q.

Wang, T. Matsuura, R. Le VanMao, Fabrication,

characterization and preliminary testing of all-

inorganic ultra-filtration membranes composed

entirely of a naturally occurring sepiolite clay mineral,

J. Membr. Sci. 182 (2001) 41.



- 62 -

[4] S. Masmoudi, R. Ben Amar, A. Larbot, H. El Feki, A.

Ben Salah, L. Cot, Elaboration of inorganic

microfiltration membranes with hydroxyapatite

applied to the treatment of wastewater from sea

product industry, J. Membr. Sci. 247 (2005) 1.

[5] J. Bentama, K. Ouazzani, Z. Lakhliai, M. Ayadi,

Inorganic membranes made of sintered clay for the

treatment of biologically modified water, Desalination

168 (2004) 295.

[6] J. Bentama, K. Ouazzania, P. Schmitz, Mineral

membranes made of sintered clay: application to

crossflow microfiltration, Desalination 146 (2002) 57.

[7] S. Khemakhem, R. Ben Amar, A. Larbot., Synthesis

and characterization of a new inorganic ultrafiltration

membrane composed entirely of Tunisian natural illite

clay, Desalination, 206 (2007) 210.

[8] E. Mulder, A mixture of fly ashes as road base

construction material, Waste Mgmt., 16 (1996) 15.

[9] R. asserman, A. Bentur, Effect of lightweight fly ash

aggregate microstructure on the strength of concretes.

Cem. Concr. Res., 27 (1997) 525.

[10] WM. Carty, U. Senapati, Porcelain raw materials

processing, phase evolution and mechanical

behaviour. J. Am. Ceram. Soc., 81 (1998) 3–20

[11] Palmonari C, Nassetti G. Evolution and future trends

of traditional ceramics, Am. Ceram. Soc. Bull., 73

(1994) 42–6.

[12] M. Ilic, C. Cheeseman, C. Sollars, J. Knight,

Mineralogy and microstructure of sintered lignite coal

fly ash, Fuel, 82 (2003) 331.

[13] L Barbieri, I. Lancellotti, T. Manfredini, I. Queralt,

JM. Rincon, M. Romero, Design obtainment and

properties of glasses and glass–ceramics from coal fly

ash, Fuel, 78 (1999) 271–276.

[14] Y. M. Jo, R. Huchinson, J.A. Raper, Preparation of

ceramic membrane filters, from waste fly ash, suitable

for gas cleaning, Waste Mgmt. Res., 14 (1996) 281-

295.

[15] S. Khemakhem, A. Larbot, R. Ben Amar, New

ceramic microfiltration membranes from Tunisian

natural materials: Application for the cuttlefish

effluents treatment, Ceramics International, Online in

2007.

Arabian J. Chem. Vol. 2, No. 1, 63-72(2009)


- 63 -

Flow Injection Potentiometric Sensor for Determination of Phenylpropanolamine Hydrochloride

Y. M. Issa1, M. M. Khalil2, S. I. M. Zayed3* and Ahmed Hussein2

1Faculty of Science, Cairo University, Giza, Egypt, 2 Faculty of Science, Beni Suef University, Beni Suef, Egypt 3Faculty of Industrial Education, Beni Suef University, Beni Suef, Egypt

*E-mail: [email protected]

Abstract A new polymeric membrane electrode has been constructed for the determination of

phenylpropanolamine hydrochloride. The electrode was prepared by solubilizing the

phenylpropanolamine phosphomolybdate ion associate into a polyvinyl chloride matrix plasticized by

dibutylphthalate as a solvent mediator. The electrode showed near-Nernstian response over the

concentration range of 1x10-5 - 1x10-2 M with low detection limit of 6.3x10-6 M. The electrode displays

a good selectivity for phenylpropanolamine with respect to a number of common inorganic and organic

species. The electrode was successfully applied to the potentiometric determination of

phenylpropanolamine ion in its pure state and its pharmaceutical preparation in batch and flow injection

conditions.

Keywords: Phenylpropanolamine hydrochloride; Ion selective electrodes; Flow injection analysis;

Potentiometry

1. Introduction

Phenylpropanolamine hydrochloride (PPACl),

Benzenemethanol, α-(1-aminoethyl) hydrochloride, (R*,

S*) (±) [154-41-6], belongs to the sympathomimetic amine

class of drugs and is structurally related to ephedrine

hydrochloride [1] (scheme I).

H

OH

C

CH3

C

H

NH3 .HCl

(Scheme I)

A number of analytical methods have been

reported for the determination of PPACl. Among these are

HPLC [2-7] , gas chromatographic [8,9], capillary

electrophoresis [10-12], conductimetric [13] and

spectrophotometric methods [14-19]. Potentiometric ion-

selective electrodes based on phenylpropanolamine-

tetraphenylborate or phenylpropanolamine-

phosphotungstate have been reported [20]. Ion-selective

membrane electrodes play an increasing role in

pharmaceutical analysis with further use in FI [21-23]

offering advantages of simplicity, rapidity and accuracy.

Liquid membrane electrodes using phosphotungestic and

phosphomolybdic acids were previously described [24]

The present work describes the construction and

potentiometric characterization of new potentiometric

sensor for PPA. The electrode is based on the incorporation

of phenylpropanolamine-phosphomolybdate (PPA)3-PM

ion associate in a polyvinyl chloride (PVC) membrane

plasticized with dibutylphthalate (DBP). Applications of

the electrode for the determination of PPACl in

Y. M. Issa, M. M. Khalil, S. I. M. Zayed and Ahmed Hussein


- 64 -

pharmaceutical preparation for batch and FI analysis

system were also described.

2. Experimental 2.1. Reagents and materials

All chemicals were of analytical grade. Double distilled

water was used throughout all experiments. Pure grade

phenylpropanolamine hydrochloride (PPACl) and the

pharmaceutical preparation Contac 12 capsules were

provided by Kahira pharmaceutical and Chemical

Industries Co., Egypt, and Egyptian International

Pharmaceutical Industries Co., (EIPICO), respectively.

Phosphomolybdic acid (PMA), dioctyl sebacate (DOS),

and tricresyl phosphate (TCP) were from Fluka, dibutyl

phthalate (DBP), and dioctyl phthalate (DOP) from Merck.

PVC of relatively high molecular weight was from Aldrich.

2.2. Apparatus

Potentiometric and pH measurements were carried out

using a Seibold G-103 digital pH/mV meter (Vienna,

Austria). A techne circulator thermostat Model C-100 was

used to control the temperature of the test solutions. A

saturated calomel electrode (SCE) was used as the external

reference, while an Ag/AgCl wire as an internal electrode.

The flow injection setup as previously reported

[24]. Fig. 1 represents the schematic diagram of the flow

injection system used in the measurements.

to waste

Fig. 1 Schematic diagram of the flow injection system used in the measurements

2.3. Preparation of the ion associate

The ion associate (PPA)3-PM, was prepared by mixing 150

ml of 10-2 M PPACl solution with 50 ml of 10-2 M

phosphomolybdic acid. The formed precipitate was

filtered, washed thoroughly with bidistilled water until

chloride free and dried at room temperature. The

composition of the ion-associate was found to be 3:1 as

confirmed by elemental analysis data done at

microanalytical research laboratory in National Research

Centre (Dokki, Cairo, Egypt). The percentages values

ISE + SCE

Recorder

mV-meter

50 cm 0.5 mm

Water Water



- 65 -

found are 14.20, 1.91 and 1.82 and the calculated values

are 14.24, 1.73 and 1.84 for C, H and N, respectively.

2.4. Electrode preparation

The electrode was constructed as previously described

[24]. The membranes were prepared by dissolving the

required amount of the ion associate, PVC and DBP, in

about 10 ml of THF. The solution mixture was poured into

a 6.0 cm Petri dish and left to dry in air. To obtain a

uniform membrane thickness, the amount of THF was kept

constant, and its evaporation was fixed for 24 h. The

thickness of the membrane was about 0.2 mm.

A 12 mm diameter disk was cut out from the

prepared membrane and glued using PVC-THF paste to the

polished end of a plastic cap attached to a glass tube. The

electrode body was filled with a solution of 1x10-1 M NaCl

and 1x10-2 M PPACl. The electrode was pre-conditioned

before use by soaking in a 1x10-3 M PPACl solution.

2.5. Potentiometric determination of PPACl

The standard addition method [25] was applied, in which

small increments of the standard solution (10-1 M) of

PPACl were added to 50 ml aliquot samples of various

concentrations from pure drug or pharmaceutical

preparations. The change in millivolt reading was recorded

for each increment and used to calculate the concentration

of PPACl sample solution using the following equation: 1

)/(10−

∆

+

−

+

=VsV

VVV

VCCx

xSEn

sx

ssx

where xC and xV are the concentration and the volume of

the unknown, respectively, sC and sV the concentration

and the volume of the standard solution, respectively, s

the slope of the calibration graph and E∆ is the change in

millivolt due to the addition of the standard solution.

2.6. Determination of phenylpropanolamine

hydrochloride in Contac 12 capsules

Twenty capsules were accurately weighed and powdered in

a mortar, the required amount from the capsules powder

was dissolved in chloroform to separate

phenylpropanolamine hydrochloride from the capsules

matrix (chloroform dissolves isopropamide iodide only).

The separated phenylpropanolamine

hydrochloride was dried and then dissolved in about 30 ml

bidistilled water and filtered in a 50 ml measuring

flask.The residue was washed three times with bidistilled

water, the volume was completed to the mark by the same

solvent, the contents of the measuring flask were

transferred into a 100 ml beaker and subjected to a

potentiometric determination of PPACl.

3. Results and discussion 3.1. Optimization of the ISE response in batch

conditions

Four membrane compositions were prepared by varying the

percentages of the ion associate, while keeping the

percentages of the PVC and the plasticizer equal 1:1 (Table

1).



- 66 -

Table 1 composition of the membrane and the slope of the calibration graphs at 25 + 1 oC and 30 min. of soaking in 10-3 M PPACl

Composition% (W/W) Membrane

Ion associate PVC DBP

Slope mV/decade RDS%

I 3.0 48.5 48.5 53.53 0.62 II 5.0 47.5 47.5 54.88 0.68 III 7.0 46.5 46.5 54.31 0.61 VI 10.0 45.0 45.0 50.78 0.54 RSD: relative standard deviation (four determinations)

Table 2 Effect of plasticizers on PPA responsive membranes and slopes of the corresponding calibration

graphs at 25+1 oC and 30 min. of soaking in 10-3 M PPACl

Plasticizer Slope mV/decade

Usable concentration range (M)

Detection limit[26]

DBP 54.9 1.00x10-5-1.00x10-2 6.31x10-6 DOP 44.0 1.00x10-5-1.00x10-2 7.94x10-6

DOS 50.1 3.98x10-5-1.00x10-2 1.12x10-5

TCP 50.2 3.98x10-5-1.00x10-2 1.41x10-5

The results showed that the electrode made of

membrane with 5% PPA-PM ion associate exhibits the best

performance characteristics[slope 54.88 mV concentration

decade-1 at 25 oC, usable concentration range 1x10-5 –

1x10-2 M and detection limit [26] 6.31x10-6 M PPACl]

(Table 2). The role of the membrane liquid is significant

because the nature of the selected organic solvent

determines the extraction parameters of the ion associates

and consequently, the electrode selectivity towards the ion

of interest [21].

Four plasticizers were tested to evaluate the

effect of the plasticizer on the response of PPA electrode

(Table 2). The results indicate that DBP is the best

plasticizer tested. Poor sensitivities for the electrodes

plasticized using DOP, DOS and TCP are due to low

solubilities or low distributions of (PPA)3-PM ion

associates in these solvents [27]. The electrode using DBP

as a plasticizer provides not only higher Nernstian slope

but also a wide response range more stable potential

reading and lower detection limit. It seems that DBP, as a

low polarity compound, provides more appropriate

conditions for incorporation of the highly lipophilc PPA+

ion into the membrane prior to its exchange with the soft

ion exchanger. Therefore, we used DBP as a suitable

plasticizer for further studies.

The effect of temperature on the electrode

response was studied at different temperatures. The

electrode gave good Nernstian response over the

temperature range 25-60oC. The standard electrode

potentials, Eo, were determined at different temperatures

and used to calculate the thermal coefficient of the

electrode [28], which were found to be -0.0020 V/oC and of

the cell to be 0.0014 V/oC. These values indicate fairly

good thermal stability of the electrodes.



- 67 -

The life time of the electrode was investigated by

performing the calibration graphs after the electrode was

soaked continuously in 10-3 M PPACl periodically till 28

days and calculating the response slopes. The results

indicate that during the first day the slope remains constant

at about 55.0 mV/concentration decade, then slightly

decreased reaching 54.0, 53.0 and 52.0 mV/concentration

decade after 12, 18 and 22 days, respectively. A further

decrease reaching 49.0, 46.0 and 43.0 mV/concentration

decade was observed after 24, 26 and 28 days, respectively.

This decrease in the slope of the electrode may be due to

the leaching of the lipophilic salts from the gel layer at the

electrode surface.

3.2. Optimization of FI parameters

FI parameters were optimized in order to obtain the best

signal sensitivity and sampling rate under low dispersion.

The dispersion coefficient was 1.23, i.e., limited dispersion

that aids optimum sensitivity and fast response of the

electrode [29]. The influence of the injected volume was

assessed for sample volumes from (4.7-500.0 µl). In

general, the higher the sample volume, the higher the peak

heights and residence time of the sample at the electrode

surface, requiring a longer time to reach a steady state and

greater consumption of sample [30]. A sample loop of size

150 µl was used throughout this work, giving maximum

peak height, less consumption of reagents, and a short time

to reach the base line.

The effect of the flow rate was evaluated using

different flow rates (4.15, 5.35, 7.50, 9.70, 12.50, 17.85,

23.25, 25.00, 27.00 and 30.00 ml/min.) at a constant

sample loop of size 150 µl . It was found that, as the flow

rate increased, the peaks became higher and narrower until

a flow rate of 7.50 ml/min. Then the peaks obtained above

this flow rate was nearly the same. Therefore this flow rate

was used throughout this work providing the maximum

peak height, a shorter time to reach line and less

consumption of the carrier solution. Under these conditions

the electrode presented detection limit of 1.12x10-5 and a

linear range of 5.0x10-5-1x10-2 M PPACl. Fig. 2a

represents the recorded peaks and Fig. 2b, shows the

calibration graph for the electrode at the optimum

conditions.

Fig. 2 Recording (a) and its corresponding calibration graph (b) for PPA-PM electrode under FI conditions



- 68 -

The effect of the pH of the test solution on the

electrode potentials was studied in batch and FI conditions.

In batch measurements the effect of pH of the test solution

(10-4, 10-3 and 10-2 M PPACl) on the electrode potential

was investigated by following the variation in potential

with the change in pH by adding of very small volumes of

hydrochloric acid and sodium hydroxide (each 0.1-1.0 M).

The results indicated that the electrode did not respond to

the pH change in the range 2.8-6.8 (Fig. 3). In this pH

range, the electrode can be used safely for the respective

determination of PPACl in the pharmaceutical

preparations. The increase in mV reading at pH less than

2.8 may be due to the penetration of H+ into the membrane

surface. While the decrease in the potential reading after

pH 6.8 most probably attributed to the formation of the free

phenylpropanolamine base in the solution, leading to a

decrease in the concentration of phenylpropanolamine

cation. In FI a series of solutions of concentration that are

10-3 M PPACl and have pH values ranging from 1 to 12

were injected in the flow stream adjusted to the same pH,

then the peak heights representing the variation of potential

with pH were measured. No variation in the peak height

was observed in the same pH ranges registered in the

steady state mode for the electrode. This indicates that the

electrode do not respond to pH changes in these ranges

under FI conditions

.

pH0 2 4 6 8 10 12 14

E, m

V

-120

-100

-80

-60

-40

-20

0

20

40

60

80

a

b

c

Fig. 3. Effect of the test solution on the potential response of the PPAFM electrode (a) 1×10-4 , (b) 1×103 and (c) 1×102



- 69 -

3.3. Selectivity of the electrode

The effect of some inorganic cations, sugars, amino acids

and vitamins on the response of phenylpropanolamine ion

selective electrode was investigated. The selectivity

coefficients were determined using two methods, the

separate solution method (SSM)[31] and the matched

potential method (MPM) [32,33]. In the separate solution

method, the Nicolsky -Eisenman equation was used:

log potJPPA ZK +, = (E2 - E1)/S + log [PPACl] – log [Jz+]1/z

where, E1 and E2 are the electrode potentials in a 1x10-3 M

solutions of PPACl and interfering ions Jz+, respectively,

and S is the slope of the calibration graphs in mV

concentration decade-1.

In matched potential method, the potentiometric

selectivity coefficient is defined as the activity ratio of

primary ions and interfering ions that give the same

potential change under identical conditions. At first, a

known activity of phenylpropanolamine ion solution is

added into a reference solution that contains a fixed activity

of phenylpropanolamine αPPA, (α'PPA-αPPA is the change in

activity), and the corresponding potential change ∆E is

recorded. The change in potential produced at the constant

background of the primary ion must be the same in both

cases.

J

PPAPPApotJPPA a

aaK Z

−=+

',

Where, Ja is the activity of the added interferent.

In FI, a series of standard PPACl solutions

between 5x10-6 and 1x10-2 M was prepared; their

corresponding heights were measured and used to

determine the slope of the calibration graph. Solutions that

are 1x10-3 M of interfering ions were prepared; and their

corresponding peak heights were also measured. The

selectivity coefficients were calculated using the the

separate solution method. The selectivity coefficients

values potJPPA ZK +, of the electrode listed in Table 3 reflect a

high selectivity of this electrode towards

phenylpropanolamine cation. The inorganic cations do not

interfere owing to the differences in ionic size and

consequently in their mobilities and permeabilities as

compared with PPA+. In case of non ionic species, the high

selectivity is due to the difference in polarity and to the

lipophilic nature of their molecules relative to PPA cation.

3.4. Analytical applications

In order to assess the applicability of the proposed selective

electrode, the method was applied for the determination of

PPACl in pure solutions and in the pharmaceutical

preparation Contac 12 Capsules (phenylpropanolamine

HCl, 50 mg and isopropamide, 3.4 mg under batch and FI

conditions. The mean recovery and the relative standard

deviation values are summarized in Table 4. The

interference resulted from the other drug, isopropamide

was prevented by dissolving the capsules powder in

chloroform, that dissolve only isopropamide, the data

indicated that there is no interference from the other

excipients used in the formulations of the capsules.

The results obtained were compared with the

official method [34] (Table 5) and found to be in good

agreement with those obtained from the official method.

Student’s t- and F-tests (at 95% confidence level) were

applied [35]and the results show that the calculated t- and

F-values did not exceed the theoretical values.



- 70 -

Table 3. Selectivity coefficients for PPA-PM responsive electrode

Interferent potJPPA ZK +,

Batch FI

SSM MPM ----

Na+ 2.41x10-3 7.44x10-4 1.90x10-2

K+ 1.53x10-2 6.53x10-4 1.83x10-2

Mg2+ 9.78x10-4 4.87x10-4 2.24x10-4

Ca2+ 1.19x10-4 7.75x10-4 4.69x10-4

Ba2+ 1.99x10-4 7.57x10-4 1.47x10-4

Sr2+ 4.88x10-4 6.45x10-4 7.93x10-4

Co2+ 2.13x10-4 7.24x10-4 1.32x10-4

Zn2+ 1.82x10-4 8.77x10-4 3.42x10-4

Cu2+ 2.23x10-5 6.80x10-4 2.24x10-4

Vitamine B1 1.05x10-2 1.44x10-3 2.51x10-2

Vitamine B6 8.97x10-2 3.28x10-3 4.72x10-2

Glucose -- 2.85x10-4 ---

Fructose -- 2.69x10-4 --

Maltose -- 3.03x10-4 --

Lactose -- 2.92x10-4 --

Alanine -- 2.40x10-4 --

Glycine -- 2.45x10-4 --

Table 4. Determination of PPACl in pure form and in pharmaceutical preparation by applying standard additions method and under FI conditions (n = 4). Taken, mg Mean recovery, % RSD, %

1.0x10-4 99.73 0.732 2.0x10-4 99.39 1.475 3.0x10-4 98.72 0.650

Pure solution Standard additions method

5.0x10-4 98.48 1.156 Capsules (Contac 12)

Standard additions method 1.0x10-4 100.52 0.422

2.0x10-4 100.78 0.537

3.0x10-4 100.50 0.717

5.0x10-4 101.46 0.742

FI 5.0x10-5 100.86 0.984

1.0x10-4 100.68 0.836

5.0x10-4 100.72 0.477

1.0x10-3 100.62 0.802



- 71 -

Table 5. Statistical comparison between the results of determination of the pharmaceutical preparation Contac 12 capsules applying the proposed and official methods (n = 4)

Parameter Standard additions FI Official method [34] Mean recovery, % 100.81 100.72 101.72 SD 0.609 0.780 0.655 RSD 0.604 0.774 0.644 F-ratio (9.28)a 1.157 1.418 t-test (2.447)b 2.036 1.964 SD: standard deviation RSD: relative standard deviation a: tabulated F-value at 95% confidence level b: tabulated t-value at 95% confidence level and six degrees of freedom

4. Conclusion The proposed sensor based on (PPA)3-PM ion associate as

the electroactive compounds might be a useful detector for

the determination of PPACl in pharmaceutical

preparations, in batch and FI system. The inherent

advantages of the proposed techniques are their high

selectivity, rapid response, simple operation, precise results

and low cost.

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Arabian J. Chem. Vol. 2, No.1, 73-88 ( 2009)


- 73 -

Experimental Study on Effect of Different Parameters on Size and Shape of Triangular Silver

Nanoparticles Prepared by a Simple and Rapid Method in Aqueous Solution

Seyed Soheil Mansouri, Sattar Ghader*

Department of Chemical Engineering, College of Engineering,

Shahid Bahonar University of Kerman P. O. Box 76175-133, Kerman, Iran


Abstract

This paper continues our previous work on preparation of truncated triangular silver

nanoparticles. The method proceeds with reaction of silver nitrate with hydrazine in the

presence of sodium citrate in aqueous solution, in which triangular nanoparticles are formed

in a few minutes with some spherical ones. In particular range of reactants, especially high

reductant concentration, only spherical nanoparticles are formed. In further investigation we

observed that spherical nanoparticles shape could change to triangular by aging. This means

that controlled growth of nanoparticles could lead to the formation of triangular ones.

Therefore, a method was devised to slow down the rate of reduction by adding Fe3+ to the

reaction solution. The results show that in this case more triangular nanoparticles are formed

compared to the original one. This result also confirms that with the increasing hydrazine

concentration, growth becomes less important compared to nucleation and smaller triangles

are formed.

Keywords: triangular silver nanoparticles, aqueous solution, citrate, hydrazine.

1. Introduction Research on metal nanoparticles has increased extensively

in recent years due to their size and shape dependent

optical [1], physical [2] and chemical properties [3].

Developing methods for tailoring metal nanoparticle size

and shape enable us easy and large scale production and

correlating the optimal properties to structure. Different

shapes of silver nanoparticles have size and shape sensitive

surface plasmon resonance bands and applications in optics

[1], electronics [2], sensors [4], surface enhanced Raman

spectroscopy (SERS) [5], catalysts [6], biological detection

and drug delivery [7].

Synthesis of triangular silver nanoparticles has

become important since pioneering work of Jin et al. [8] in

photo induced conversion of silver nanospheres to

nanoprism in several hours. Subsequently, many methods

for preparation of silver nanoprism have been reported.

Callegari et al. [9] could adjust size of triangular silver

nanoplates by choosing the wavelength of light used to

transform spherical nanoparticles to triangular nanoplates.

Seyed Soheil Mansouri, Sattar Ghader


- 74 -

Bastys et al. [10] also modified Jin et al. [8] method to

form silver nanoprisms with surface plasmons at

communication wavelengths. Jia et al. [11] described

synthesis of triangular silver nanoparticles by

photoreducing silver ions by citrate. All photo induced

methods need several hours to prepare nanotriangles.

Triangular silver nanoplates were also formed by reverse

micelles of di(2-ethyl-hexyl)sulfosuccinate (AOT) [12] and

seed-mediated growth in the presence of

cetyltrimethylammonium bromide (CTAB) micelles [13].

Silver nanoprisms are also synthesized by boiling

silver nitrate in dimethyl formamide (DMF) in the presence

of poly(vinyl pyrrolidone) (PVP) [14] and shape

transformation by refluxing spherical silver nanoparticles

[15]. Metraux and Mirkin [16] developed thermal synthesis

of silver nanoprism in 20-30 min with controllable

thickness.

These methods for synthesis of triangular silver

nanoparticles used photo, template, seed and thermal

processing which were either time consuming or needed

some steps and processings after initial synthesis of silver

nanoparticles. No method is reported to produce silver

nanotriangles directly as a product of a reaction in aqueous

solution. Some methods are also reported in non-aqueous

solutions i.e. reducing silver perchlorate in formamide in

the presence of polyethylene glycol (PEG) at room

temperature [17] and shape transformation to triangular

nanoplates by aging spherical silver nanoparticles prepared

in pyridine in the presence of poly(vinyl pyrrolidone)

(PVP) [18].

In this paper we completed our previous work

[19] on synthesis of truncated triangular silver

nanoparticles by improving the yield of triangular

nanoparticles. In previous study, spherical silver

nanoparticles were formed in especial range of

concentrations. In this study, we observed that these

particles turned to triangular ones after two months. This

means that after nucleation, when growth becomes a

controlling factor the kinetic effects favors triangular

nanoparticles formatiom. This hypothesis was confirmed

by slowing down the rate of original synthesis method by

introducing Fe3+ to the reaction solution. Shape change of

triangular nanoparticles to smaller one with increasing

concentration of hydrazine could also be justified by

considering kinetics effects in shape evolution of particles.

2. Experimental Silver nitrate, tri-sodium citrate dihydrate, hydrazine

hydrate and ammonium iron sulfate were supplied by

Merck. A solution of 100 mL silver nitrate 0.1 mM and 5

mL of sodium citrate 34 mM (1 wt%) is made. This

solution is stirred by a magnet and 5 mL of 2 mM

hydrazine is added to the solution drop by drop. Then, the

sample exhibited two color changes. After about 3 min the

solution color changed to yellow which turned to green in

about 2 min indicating formation of truncated triangular

silver triangular nanoparticles. The colloid was stable for

months. A Zeiss transmission electron microscope (TEM)

operating at 80 kV was used to observe the nanoparticles.

The samples were prepared by dropping 10 µL of solution

on the copper grid covered with amorphous carbon and let

to dry in air. UV-vis absorption spectra were recorded with

a Varian Cary 50 Conc spectrophotometer with 1-cm

length optical cell.

Experimental Study on Effect of Different Parameters on Size and Shape of Triangular Silver……


- 75 -


TEM Images

Silver nanotriangles are the product of a reaction,

which includes reducing silver nitrate with hydrazine

in the presence of sodium citrate as a stabilizing

agent. TEM images were taken from the product of

the reaction and as Fig. 1 shows triangular

nanoparticles are truncated in shape. TEM images

indicate that the size of truncated triangular

nanoparticles (maximum length of the base as

defined by Brioude and Pileni [20]) is 94± 8 nm.

TEM images show a mixture of circular, hexagonal

and triangular particles.

Fig. 1. TEM images of truncated triangular silver nanoparticles synthesized by reduction of silver nitrate with

hydrazine in the presence of sodium citrate (at different areas of TEM grid).

UV-vis Absorption Spectroscopy The changing in the shape of the silver nanoparticles

prepared by adding hydrazine was investigated by UV-vis

spectra. Based on the theoretical calculations by Brioude

and Pileni [20] four peaks in absorption spectra of silver

nanotriangles can be attributed to in-plane dipolar, in-plane

quadrupolar, out-of-plane dipolar, and out-of-plane

quadrupolar resonances. In-plane dipolar resonance, at the

longest wavelength, is sensitive to the size of the triangle

and red shifts with size. The out-of-plane quadrupolar peak

appears at the lowest wavelength and is located around 340

nm [20]. Fig. 2 shows the experimental UV-vis spectra of

silver nanoparticles during the formation of nanotriangles.



- 76 -

Fig. 2. Time change of UV-vis absorption spectra during synthesis of truncated triangular nanoparticles

Two peaks at the longest wavelengths and the

lowest wavelength peak increase as the reaction proceeds,

which implies that the amount of nanotriangles increases.

Peaks gradually increase and the final absorption peak is

observed after 5 min. Final UV-vis spectra exhibit four

distinct peaks at 334, 404, 672 and 740 nm. The peak at

334 nm is assigned to out-of-plane quadrupolar resonance.

The peaks at 740 and 672 nm are attributed to in-plane

dipolar and in-plane quadrupolar resonances, respectively.

An intense peak is observed at 404 nm. Since spherical

nanoparticles have their absorption peak in this region it

shows the existence of spherical nanoparticles in the

solution – as observed in TEM images – which are more

than triangular nanoparticles.

To examine the reproducibility of the

nanotriangles preparation, usually more than 20 samples

were prepared and the absorbance was recorded for each

sample. Usually only two or three of these samples

exhibited more than 5% deviation in the spectrum.

Comparing the absorption of nanotriangles to nanospheres

at 404 nm 22% of product is nanotriangles. Nanotriangles

can be separated from spherical nanoparticles by

centrifugation. This method is reported earlier in many

references, for example Maillard et al. [12], Deivaraj et al.

[18] and Chen et al. [21]. The original solution was

centrifuged at 6000 rpm for 20 min. Removing the

supernatant, the precipitate was dispersed in water. This

procedure was repeated once more and the UV-vis

spectrum of triangular nanoparticles was obtained. The

UV-vis spectrum after centrifugation is shown in Fig. 3

which is more similar to the theoretical calculations by

Brioude and Pileni [20]. By collapsing the intense peak of

spherical nanoparticles, a peak at 475 nm appears which is

attributed to out-of-plane dipolar resonance. The weak

peak at 420 nm relates to residual spherical nanoparticles.

A TEM image (Fig. 4) was taken from nanoparticles after



- 77 -

centrifugation which shows successful separation of triangular nanoparticles.

Fig. 3. UV-vis spectrum of truncated triangular silver nanoparticles after centrifugation.

Fig. 4. TEM image of truncated triangular silver nanoparticles after centrifugation.

Effect of Silver Nitrate Concentration and Aging Silver nitrate concentration was also an important

parameter in triangles synthesis. When silver nitrate

concentration was increased to 0.5 mM triangles did not

form. The solution color was yellow and the UV-vis

absorption spectra of colloid completely changed and a



- 78 -

peak at 408 nm was observed (Fig. 5). TEM images of

resulting nanoparticles show that spherical nanoparticles

are produced (Fig. 6). Similar results on the changing of

the shape with concentration of silver nitrate was reported

by Pastoriza and Marzan [14]. Very small amount of tiny

nanotriangles can be seen in the TEM image as shown in

Fig. 6. When this solution was remained unstirred for two

months solution color changed from yellow to green. In

this case nanotriangles were formed in the solution by

aging – as UV-vis spectrum (Fig. 7) and TEM (Fig. 8)

shows – indicating when the amount of silver nitrate is

increased; hydrazine is not at sufficient amount to

influence synthesis of nanotriangles. Effect of hydrazine is

more discussed in subsequent sections. Aging small

spheres increased the yield of nanotriangles. Based on the

UV-vis spectrum (Fig. 7) the yield of triangles after ageing

increased to 44%.

In another experiment, the nanospheres were kept

unstirred in darkness after formation. The nanotriangles

were formed in this system too by aging and color of

solution also changed to green. UV-vis spectrum (Fig. 9)

was taken from this solution after aging it for 45 days

which indicates formation of triangles. So it can be

concluded that nanotriangles are also formed in darkness

and the process is not affected by photo.

In summary, the reduction could be significantly

slowed to induce anisotropic growth in the solution by

aging. The initial product of such a synthesis was Ag

nanoparticles. Once the supersaturation had been reduced

to a certain level, the growth of Ag atoms would be

switched to a highly anisotropic mode to form Ag

nanotriangles. Since the Ag+ existed in the solution at a low

concentration for a long period of time, the Ag atoms could

grow into triangles. Increasing yield of triangles and

influence of slowing down reduction rate is more discussed

in subsequent sections.

Fig. 5. UV-vis spectrum of resulting nanoparticles when silver nitrate concentration increased to 0.5 mM.



- 79 -

Fig. 6. TEM image of resulting nanoparticles when silver nitrate concentration increased to 0.5 mM.

Fig. 7. UV-vis spectrum of resulting nanoparticles after aging solution of Fig. 5 for two months (silver nitrate

concentration 0.5 mM).



- 80 -

Fig. 8. TEM image of resulting nanoparticles after aging solution of Fig. 5 for two months (silver nitrate concentration 0.5 mM).

Fig. 9. UV-vis spectrum of resulting nanoparticles after ageing solution of Fig. 5 for 45 days in darkness (silver nitrate

concentration 0.5 mM).



- 81 -

Effect of Sodium Citrate Concentration

Some other experiments were carried out to attain

citrate effect on nanoparticles. It was found that

triangular nanoparticles were formed at any citrate

concentration. Fig. 10 shows, the UV-vis absorption

spectrum of on the product colloid at low citrate

concentration (0.34 mM) as well as TEM image (Fig.

11).

Fig. 10. UV-vis spectrum of obtained truncated triangular nanoparticles with citrate concentration 0.3 mM.

Fig. 11. TEM image of obtained truncated triangular nanoparticles with citrate concentration 0.3 mM.

Citrate likely effects face-selective growth by

adsorbing more strongly to the Ag(111) surface to direct

the final shape to be a triangle. Many studies have



- 82 -

mentioned importance of citrate for preparation of silver

triangular nanoparticles [8-11, 15, 16].

Effect of pH

We also studied the effect of hydrazine solution pH on the

synthesis of the triangular nanoparticles since it influences

reducing power of hydrazine and formation of triangles.

The pH of original hydrazine solution 9.2. Nitric acid and

sodium hydroxide were added to hydrazine solution for pH

2 – 8 and 10 – 12, respectively. We tried to reveal the role

of hydrazine by varying its effect; i.e. changing pH. At pH

2 almost a clear solution obtained and UV-vis absorption

spectra (Fig. 12a) shows weak peaks. indicating very small

amount of triangular nanoparticles is formed. This result

also emphasizes critical role of hydrazine in formation of

triangular nanoparticles. Since at low pH condition most of

hydrazine is removed, nanotriangles are decreased

dramatically. Thus, hydrazine may not just providing

reducing power, but participate in shape control. Maillard

et al. [12] also indicated important role of hydrazine in

nanotriangles formation. At pH = 3.5 solution becomes

green and four peaks in absorption spectra grow and

become evident. As Fig. 12 represents from pH = 4 to 8

plasmon resonance bands increase indicating formation and

increase of triangular nanoparticles

Fig. 12. UV-vis spectra of resulting nanoparticles at different pH of hydrazine solution (a) pH = 2, 3.5, 4, 6 and (b) pH

= 8, 10, 12.

Nevertheless, further increasing pH results in blue shift of

in-plane dipolar resonance band and at pH = 12 in-plane

dipolar resonance almost disappear and spherical

nanoparticles are formed in a yellow color solution as TEM

image of Fig. 13 shows. Similar change in absorption

spectra with pH was observed by Chen et al. [21]. With the

pH being 12 the stabilizing agent is most likely OH- which

adsorbs on the nanoparticle surface. Nickel et al. [22] could

also produce spherical nanoparticles by hydrazine pH

above 10.

The reaction of hydrazine with silver nitrate

produces Ag, N2 and proton. With increasing pH protons



- 83 -

are neutralized with sodium hydroxide and hydrazine does

not remain partially protonated. Because of the high redox

potential of protonated hydrazine, with increasing pH, the

actual redox potential of reducing agent decreases.

Therefore, the initial pH should be higher than 10 to see

this effect. Furthermore, adsorbed OH- on nanoparticles

prevents triangles production. The peak for spheres also

increases a little as triangles disappear. This set of

experiments show that choosing proper pH is important for

triangles synthesis and it can affect product shape. pH also

accelerated the reduction time and even at pH = 12 spheres

are formed at 30 sec.

Effect of Hydrazine Concentration and Controlling

Rate of Reduction

A study concerned the effect of hydrazine concentration on

nanoparticles synthesis is being investigated. Interestingly,

with increasing hydrazine concentration in-plane dipolar

resonance blue shifts (Fig. 14) was occured. This effect is

in accordance with the result observed with varying pH, as

increasing pH has unfavorable effect on the production of

truncated triangular nanoparticles and ultimately prevent

their formation. As the theoretical calculations of Brioude

and Pileni [20] show for nanotriangles smaller than 60 nm

two intermediate peaks almost disappear; only in-plane

dipolar and out-of-plane quadrupolar resonances remains

while smaller nanotriangles are formed as in-plane dipolar

resonance blue shifts. TEM image shows that smaller

nanotriangles are formed (Fig. 15 for the case c in Fig. 14)

after increasing hydrazine. In order to justify why

increasing hydrazine concentration leads to smaller

triangles, an experimental study was conducted which is

explained below.

Fig. 13. TEM image of resulting nanoparticles at pH of hydrazine solution pH = 12.



- 84 -

Fig. 14. UV-vis spectra of nanoparticles at different hydrazine concentration: (a) 2 mM (b) 3 mM (c) 5 mM (d) 10

mM.

Fig. 15. TEM image of truncated triangular nanoparticles at hydrazine concentration 5 mM.



- 85 -

Since a quasi-spherical nanoparticle has the

lowest possible surface energy and is therefore favored by

thermodynamics, the growth kinetics of a particle must be

carefully controlled to obtain a shape that does not

represent an energy minimum. Factors that influence the

growth kinetics of a solution-phase synthesis include (i) the

concentration of metal precursor, (ii) the rate of reduction

(the concentration and power of the reductant); (iii) the

presence of a soft template or capping agent; and (iv) the

specific adsorption of a capping agent to a particular

crystallographic plane. Several research groups have

employed such kinetic controls to generate triangular and

circular nanoplates of silver in a number of different

solvent systems [8, 13, 14].

Therefore, we explored a way of controlling the

rate of reduction to understand the effect of slower growth

rate and thus a kinetically favored shape. Since reduction

potential of Fe3+ is very close to Ag+, the key strategy is the

introduction of either Fe3+ or Fe2+ species to change the

growth rate by slowing down the reduction reaction or

change the level of supersaturation of Ag atoms,

respectively. The addition of Fe3+ (0.1 mM) to the

triangular silver synthesis slowed down the rate of

reduction. This protocol produced results that were

concentration dependent and with Fe3+ (0.01 M)

nanoparticles did not form in the solution, unless excess

hydrazine was added to the solution. The addition of Fe3+

(0.1 mM) increased the yield of triangular nanoparticles as

UV-vis spectrum shows (Fig. 16) with increasing about 1

min the time previously required. The peak of nanospheres

is almost reduced to one third of the original solution. The

yield of triangular silver nanoparticles in this case was 74%

based on absorptions in UV-vis spectrum.

Furthermore, the function of Fe2+ (0.1 mM) was

not similar to Fe3+ and Fe2+ accelerated the rate of

reduction. With high reducing potential of hydrazine it may

be concluded that Fe3+ is reduced to Fe2+ after addition

which in turn changed the kinetics of the system to a

slower growth which has led to nanotriangles synthesis

which are to considered more favorable to be formed at

slower growth. Nevertheless, adding Fe2+ to the synthesis

has an effect similar to hydrazine since it could reduce

more silver ions to Ag atoms. This procedure accelerates

the formation of nanotriangles, in spite of Fe3+, leading to

increasing the growth rate and favors a shape that is not

kinetically favored.

As a result smaller triangular nanoparticles are

formed which are more similar to quasi-spherical

nanoparticles (thermodynamically favored) as UV-vis

spectrum shows (Fig. 17). With the increasing the

hydrazine concentration in the original procedure,

acceleration in reduction was observed. Considering the

results of this experimental study, kinetics of the system at

accelerated growth favors a shape more similar to quasi-

spherical nanoparticles; i.e. smaller nanotriangles.



- 86 -

Fig. 16. UV-vis spectrum of nanoparticles when adding +3Fe (0.1mM) to the solution (before adding hydrazine).

Fig. 17. UV-vis spectrum of nanoparticles when adding +2Fe (0.1mM) to the solution (before adding hydrazine).



- 87 -

4. Conclusions In this study we have continued our work on the synthesis

of triangular silver nanoparticles. In previous work a

simple method for the synthesis of truncated triangles was

described at room temperature. The reaction occurs in

aqueous solution between silver nitrate and hydrazine.

Citrate is used to stabilize the nanoparticle formed.

Concentration of reactants are important in formation of

nanotriangles. When concentration of silver nitrate is

increased to 0.5 mM triangular nanoparticles are not

formed and the product is dominated by spherical

nanoparticles.

Nevertheless, we observed that the shape of these

spherical nanoparticles changed to a triangular after aging

for two months. In other words, when growth becomes a

controlling, factor low supersaturation for a long time

provides space for shape change to a triangular in the

presence of citrate and hydrazine. To confirm this

hypothesis, Fe3+ was added to solution to slow down

reduction rate. In this case more triangular nanoparticles

were formed with a delay in formation time. Based on

these observations, high concentration of hydrazine must

favors smaller triangles, because it favors nucleation rather

growth - which was observed experimentally.

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[8] Jin, R. C.; CaO, Y. W.; Mirkin., C. A.; Kelly, K. L.;

Schatz, G. C.; Zheng, J. G., Science 2001, 294,

1901.

[9] Callegari., A.; Tonti, D.; Chergui, M., Nano Lett.

2003, 3, 1565.

[10] Bastys, V.; Pastoriza-Santos, I.; Rodriguez-Gonzalez,

B.; Vaisnoras, R.; Liz-Marzan, L. M., Adv. Funct.

Mater. 2006, 16, 766.

[11] Jia, H.; Xu, W.; An, J.; Li, D.; Zhao, B.,

Spectrochimica Acta Part A 2006, 64, 956.

[12] Maillard, M.; Giorgio, S.; Pileni, M. P., J. Phys.

Chem. B 2003, 107, 2466.

[13] Chen, S.; Carroll, D. L., Nano Lett. 2002, 2, 1003.

[14] Pastoriza-Santos, I.; Liz-Marzan, L. M., Nano Lett.

2002, 2, 903.

[15] Sun, Y.; Mayers, B.; Xia, Y., Nano Lett. 2003, 3,

675.

[16] Metraux, G.; Mirkin, C. A., Adv. Mater. 2005, 17,

412.

[17] Sarkar, A.; Kapoor, S.; Mukherjee, T., J. Colloid

Interf. Sci. 2005, 287, 496.

[18] Deivaraj, T. C.; Lala, N. L.; Lee, J. Y., J. Colloid

Interf. Sci. 2005, 289, 402.

[19] Ghader, S.; Manteghian, M.; Kokabi, M.; Sarraf

Mamoori, R., Polish J. Chem. 2007, 81, 1555.

[20] Brioude, A.; Pileni, M. P., J. Phys. Chem. B 2005,

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Langmuir 2000, 16, 9087.



- 88 -

Arabian J. Chem. Vol. 2, No. 1, 89-94(2009) - 89 -

Microwave and Ultrasound Promoted Synthesis of Substituted New Arylhydrazono Pyridinones

Khadijah M. Al-Zaydi

Department of Chemistry, Girls' College of Education, King Abdul-Aziz University, Jeddah, P. O. Box

50918, Jeddah 21533, Kingdom of Saudi Arabia.


Abstract:

A variety of arylhydrazonopyridinones 6a,b were prepared via heating cyanoacetamide derivative with

ethyl acetoacetate in absence of solvent under reflux conventionally or ultrasound irradiation or in a

microwave oven then coupling with heteroaromatic diazonium salts. Several attempts were attained to

synthesize corresponding aminothienopyridinones 7a,b from 6a,b. Also, attempts to add electron poor

olefins to 6a,b have failed and only arylhydrazonopyridinones recovered. Keywords: Green Chemistry, Microwave Irradiation, Ultrasound Irradiation, heteroaromatic

hydrazonopyridinones.

1. Introduction Arylhydrazonopyridinones are now rapidly replacing

arylazopyrazolones in classical dye industry. Moreover,

reasonable solubility of these derivatives in lipophilic

solvents gives these dyes high potential for utility in D2T2

(Dye Diffusion Thermal Transfer) printing. Although

almost all commercial arylhydrazonopyridinones have an

alkyl function utility of these pyridinones, synthesis of

arylhydrazone condensed pyridinones have not received

interest.

Moreover, to our knowledge, modern green

synthetic methodologies have not yet been adopted for the

synthesis of these pyridinones. Hence, there remains a

demand for more efficious and safer green technologies [1-

8] for synthesis of alkyl azinylcarbonitriles as precursors to

condensed azines.

We report here about an adoptation of green

methodologies for the synthesis of heteroaromatic

hydrazonopyridinones [9-16].

2. Experimental

General All melting points were measured on a Gallenkamp

electrothermal melting point apparatus and are uncorrected.

The IR absorption spectra were measured on a Nicolet

Magna 520FT IR spectrophotometer. 1H NMR, 13C NMR

spectra were recorded in deuterated dimethylsulfoxide

[DMSO] or deutrated chloroform (CDCl3) at 200 MHz on

a Varian Gemini NMR spectrometer and a Bruker DPX

400 MHZ spectrometer using tetramethylsilane (TMS) as

an internal reference. Mass spectra were performed on a

Shimadzu GCMS-QP 1000 EX mass spectrometer at 70

eV. Microwave irradiation was carried out using the

commercial microwave oven (SGO 1000 W), a

thermocouple used to monitor the temperature inside the

vessel, it was found that ≈ 105-110 0C.

Ultrasound, microprocessor controlled-2004,

high intensity ultrasonic processor with temperature

controller (750 W), the ultrasonic frequency of the cleaning

bath used equal 25 KHz. The reaction temperature

Khadijah M. Al-Zaydi - 90 -

stabilized at 35-40 0C even after more than one hour by

addition or removal of water in ultrasonic bath to keep the

required temperature. Elemental analyses have been done

using Perkin Elmer 2400 CHN Elemental analyzer

flowchart

General Procedure for the preparation of N-

benzyl-2-cyano-acetamide 3[6]

Method I (thermal) Equimolar amounts (0.1 mol) of ethyl cyanoacetate and

benzyl amine were stirred at room temperature for 60 min..

The resulting solid product was recrystallized from ethanol.

Method II (microwave) A mixture of ethyl cyanoacetate (0.1 mol) and benzyl

amine (0.1 mol) were placed in the microwave oven and

irradiated at 400 W for 1 min. Then left to cool to room

temperature. The solid so formed was filtered and

recrystallized from ethanol.

Method III (ultrasound) Equimolar amounts (0.1 mol) of ethyl cyanoacetate and the

benzyl amine were mixed and the reaction mixture was

heated under ultrasound irradiation at 40 ºC for 2 min, and

then left to cool to room temperature. The solid so-

formed was filtered and recrystallized from ethanol.

Preparation of 1-Benzyl-4-methyl-2,6-dioxo-1,2,5,6-

tetrahydropyridine-3-carbonitrile (4);

Method I (thermal)[6] Ethyl acetoacetate (0.1 mol) was added to N-Benzyl-2-

cyano-acetamide (0.1 mol) (3). The reaction mixture was

refluxed for 13 h. The reaction mixture was poured into

ice-cold water and acidified with dilute HCl and then left to

cool to room temperature. The solid so- formed was

filtered and recrystallized from ethanol.

Method II (microwave) A mixture of ethyl acetoacetate (0.1 mol) and N-Benzyl-2-

cyano-acetamide(0.1 mol) (3)., was placed in the

microwave oven and irradiated at 400 W for 20 min. The

reaction mixture was poured into ice-cold water and

acidified with dilute HCl and then left to cool to room

temperature. The solid product so formed was filtered and


Method III (ultrasound) Ethyl acetoacetate (0.1 mol) was added to a mixture of

amine derivative (0.1 mol) and ethyl cyanoacetate (0.1

mol) and the reaction mixture was catalyzed by 0.1 mol of

ceric ammonium nitrate under ultrasound irradiation at 40

ºC for 7 hours. The reaction mixture was poured into ice-

cold water and acidified with dilute HCl and then left to

cool to room temperature. The solid product so formed was

filtered and recrystallized from ethanol.

Preparation of heterohydrazone compounds ( 6a,b)

[17,18]: A cold solution of arenediazonium salt (10 mmol),

[prepared by adding a solution of sodium nitrite (1g in 10

ml H2O) to a cold solution of aryl amine hydrochloride or

aryl amine nitrate (10 mmol) with stirring as described

earlier]. The resulting solution of the arenediazonium was

then added to a cold solution of 4 (0.1 mol) in ethanol (50

ml) containing sodium acetate (1g in 10 ml H2O). The

mixture was stirred at room temperature for 1 h and the

solid product so formed was collected by filtration and


1-Benzyl-4-methyl-2,6-dioxo-5[(2H-[1,2,4]triazol-3-yl)-hydrazono]-1,2,5,6-tetrahydro-pyridine-3-carbonitrile(6a) m. p. 255 ºC. IR (KBr): υ = 3333(2NH), 3032(CH

aromatic), 2924 (CH aliphatic), 2229(CN) and 1685,

1639(2C=O ring) cm-1. 1H NMR (400 MHz, DMSO-d6, 25

ºC, TMS): δ = 2.61(s, 3H, CH3), 5.02(s, 2H, CH2ph), 7.24-

7.37(m, 5H, ph-H), 8.63(s, 1H, CH triazole ring), 14.30(s,

1H, NH triazole ring) and 14.57(s, 1H, NH) ppm; 13CNMR

(100 MHz, DMSO-d6, 25 ºC, TMS): δ =16.93(CH3),

43.50(CH2Ph), 102.90 (C-3), 115.23(CN), 125.59, 127.87,

128.25, 128.93 (phenyl carbons), 136.69(C-4), 146.20,

151.22(triazole ring carbons), 159.40(C-5) and 160.28,

160.48(2C=O) ppm; MS: m/z = 335. Anal. For C16H13N7O2



- 91 -

(335.33) calcd .C 57.31, H 3.91, N 29.24; Found C 57.40,

H 3.82, N 29.30.

2[N-(1-Benzyl-5-Cyano-4-methyl-2,6-dioxo-1,6-dihydro-

2H-pyridine-3-ylidene)-hydrazino]-4,5,6,7-tetrahydro-

benzo [b]thiophene-3-carboxylic acid ethyl ester (6b)

m. p. 247 ºC. IR (KBr): υ = 3349(br NH due to H-bond

between O and NH), 3091(CH aromatic), 2947(CH

aliphatic), 2223(CN), 1670(C=O ester) and 1624(2C=O

ring) cm-1. 1H NMR (400 MHz, DMSO-d6, 25 ºC, TMS): δ

= 1.28(t, 3H, COOCH2CH3, J= 7Hz), 1.64-2.59(m, 8H,

cyclohexene-H), 2.65(s, 3H, CH3), 4.21-(q, 2H,

COOCH2CH3, J= 7Hz), 5.03(s, 2H, CH2ph), 7.06-7.14(m,

5H, ph) and 14.21(s, 1H, NH) ppm; MS: m/z = 476. Anal.

For C25H24N4O4S (476.56) calcd .C 63.01, H 5.08, N

11.76; Found C 63.10, H 5.13, N 11.83.

3. Result and Discussion The standard route to arylhydrazonopyridinones is

coupling of 4, prepared from 1 and benzyl amine 2, with

heteroaromatic diazonium salts. In our laboratory several

cyanoacetamides 3 have been prepared via treatment of 1

with primary amines either at room temperature for a

longer time or via irradiation with microwave for 1 minute

at 100 W or with ultrasound (US) for 2 mine at 40 ºC.

Compound 3 was reacted with an ethyl

acetoacetate also either via a longer time using reflux of

neat reagents and by a short time microwave or by US to

afford product 4 which may be exist in another tautmeric

form 5 (Scheme 1). In Table 1 yields as well as reaction

times by the three methodologies are compared.

O

OEtNC

OHN

NC

MeO

CO2Et

N OOCH2Ph

CH3CN

N OCH2Ph

HO

CH3CN

12 3

4

CH2+

CH2NH2

5

Scheme 1


Table 1. Time and yield of compounds 3, 4 by (∆ = thermal, MW = microwave irradiation and US = ultrasound )

Yield % Time/min.

US MW ∆ US MW ∆

No.

90 93 89 2 1 60 3 88 93 72 420 20 780 4

Coupling of 4 with heteroaromatic diazonium salts

afforded the corresponding heteroaromatic hydrazones

6a,b (scheme 2).

The isolated products 6a,b gave satisfactory

elemental analyses and spectroscopic data (IR, 1HNMR,

13CNMR, MS) consistent with their assigned structures.

Their IR spectra of the products showed presence of imino

group (NH) absorption band. The mass spectra of the

isolated product such as 6a showed, a peak corresponding

to the molecular ion at 335 (cf. experimental part).

NCH2Ph

OO

CH3CN

Het N N ClNCH2Ph

OO

CH3CNN

NH

Het

HNN

N

S

CO2Et

-+

4 6 a,bHet; a =

b =

Scheme 2

As anticipated the heteroaromatic hydrazones

6a,b reacted with elemental sulfur either by heating with

microwave or by US and by conventional heating to the

corresponding aminothienopyridinones 7a,b. But Several

attempts were attained to synthesize corresponding

aminothienopyridinones in presence of elemental sulfur

using different conditions (changing Temperature and

Time) under microwave irradiation, ultrasonic irradiation,

and by conventional heating, the reaction did not takeplace

(monitoring reaction by TLC).

Also, reaction of heteroaromatic hydrazones

6a,b with acrylonitrile and methyl acrylate to afford

isoquinoline derivatives did not occur using different

conditions under microwave irradiation, ultrasonic

irradiation, and by conventional heating as examined by

TLC. In our opinion this may be due to a steric factor.

4.Conclusion

We have synthesized under a variety of

arylhydrazonopyridinors microwave, sonication and

classical conditions. In general, improvements in rates and

yield of reactions are observed when reactions were carried



- 93 -

out under microwave and sonication compared with

classical condition.

It should be noted, however, that activation

occurs at different temperatures with these techniques and,

therefore strict comparisons will require a balance between

effectiveness and energy costs.

References

[1] D. L. Hjeresen, D. L. Schutt, J. M. Boese, J. Chem.

Ed. 2000, 77, 1543.

[2] P. Tundo, P. Anastas, D. S. Black, J. Breen, T.

Colline, S. Memoli, J. Miyamoto, M. Polyakoff, W.

Tumas, Pure Appl. Chem. 2000, 72, 1207.

[3] P. Lidstrom, J. Tierney, B. Wathey, J. Westman,

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[4] M. Poliakoff, J. M. Fitzpatrick, T.R. Farren, P.T.

Anastas, Science 2002, 297, 807.

[5] W. Bonrath, Ultrasonics Sonochemistry 2004, 11,

1-4.

[6] a- K. M. Al-Zaydi, R. M. Borik, M. H. Elnagdi,

Ultrasonics Sonochemistry, 2009, 16(6),660.

b-K. M. Al-Zaydi, Heterocycles, 2009,

78(8),2003.

[7] K. M. Al-Zaydi, Ultrasonics Sonochemistry, 2009,

inpress

[8] K. M. Al-Zaydi, R. M. Borik, M. H. Elnagdi,

J.Heterocyclic Chem., 2007, 44, 1187.

[9] K. Bougrin, A. Loupy, M. Soufiaoui, J. Photochem.

Photobiol. 2005, 139.

[10] G. Cravotto, P. Cintas, Chem. Soc. Rev. 2006, 35,

180.

[11] J. N. Heo, Y.S. Song, B. T. Kim, Tetrahedron Lett.

2005, 46, 4621.

[12 G. Cravotto, L. Boffa, M. Turello, M. Parenti, A.

Barge, Steroids 2005, 70, 77.

[13] R. S. Disselkamp, T. R. Hart, A. M. Williams, J. F.

White, C. H. F. Peden, Ultrasonics Sonochemistry

2005, 12, 319.

[14] F. Priego-Capote, M. D. Luque de Castro, J.

Biochem. Biophys. Methods 2007, 70, 299.

[15] M. H. Elnagdi, A. M. Negm, A.W. Erian, Liebigs

Ann. Chem. 1989, 1255.

[16] M. H. Elnagdi, A. W. Erian, Liebigs Ann. Chem.

1990, 1215.

[17] K. M. Al-Zaydi, R. M. Borik, M. H. Elnagdi,

Molecules 2003, 9, 910.

[18] K. M. Al-Zaydi, R. M. Borik., Molecules 2007, 12,

2061.


Arabian J. Chem. Vol. 2, No. 1, 95-102 (2009)


- 95 -

Utility of Oxidation-Reduction Reaction for the Spectrophotometric Determination of Amlodipine Besylate

Sayed A. Shama, Alaa S. Amin*, El Sayed M. Mabrouk and Hany A. Omara

Chemistry Department, Faculty of Science, Benha University, Benha, Egypt. *E-mail: [email protected]

Abstract A simple, rapid, accurate, precise and sensitive spectrophotometric method for the determination of

amlodipine besylate (ADB) in bulk sample and in dosage forms is described. The method is based on

oxidation of the drug by potassium permanganate in acidic medium and determine the unreacted oxidant by

measuring the decrease in absorbance for five different dyes; methylene blue (MB), acid blue 74 (AB), acid

red 73 (AR), amaranth dye (AM) and acid orange 7 (AO) at a suitable λmax 663, 609, 511, 520, and 484 nm,

respectively. Regression analysis of Beer's law plots showed good correlation in the concentration ranges 1.0-

24, 0.9-22, 1.2-26, 0.9-12.8 and 1.0-14 µg ml-1, respectively. The apparent molar absorptivity, Sandell

sensitivity, detection and quantitation limits were calculated. For more accurate results, Ringbom optimum

concentration ranges were 1.2-22.4, 1.1-20, 1.4-24.5, 1.0-12.3 and 1.3-13.2 µg ml-1, respectively. Statistical

treatment of the results reflects that the proposed procedures are precise, accurate and easily applicable for

the determination of amlodipine besylate in pure form and in pharmaceutical preparations.

Keywords: Amlodipine besylate; spectrophotometry; redox reaction; potassium permanganate; pharmaceutical

analysis.

1. Introduction Amlodipine besylate is 2-[(2-Aminoethoxy) methyl]-4-(2-

chloro-phenyl)-1,4-dihydro-6-methyl-3,5-pyridindicarboxy-

late-3-ethyl-5-methyl ester mono-benzene sulphonate. It is a

new calcium channel-blocking agent with vasodilator

activity similar to that of nifedipine [1]. It is mainly used for

its antianginal, antihypertensive and antiarrhythic activity.

The drug in pure form and its formulations are not official in

USP pharmacopoeia, and therefore require much more

investigation. Different analytical methods that have been

reported for its determination including, high-performance

liquid chromatography [2-9], liquid chromatography coupled

with tandem mass spectrometry [10], gas liquid

chromatography [11], gas chromatography coupled with

mass spectrometry [12], high performance thin layer liquid

chromatography [13-15] high-performance capillary

electrophoresis [16] and fluorimetry [17]. Visible

spectrophotometric methods are commonly used in industrial

laboratories because of their simplicity, selectivity and

sensitivity. The amlodipine besylate in pharmaceutical

preparations was determined by the spectrophotometric

method [18,19] involving oxidation of the drug,

voltammetrically [20]. A number of other extractive

spectrophotometric methods [21-27] have been also

reported. However, some of these methods are somewhat

tedious and time consuming. Therefore, the need for a fast,

low cost, accurate, precise and sensitive method is obvious,

especially for a routine quality control analysis of

pharmaceutical products containing ADB.

All five dyes, methylene blue (MB), Basic blue 9

[122965-43-9]; acid blue 74 (AB 74), indigocarmine,indigo-

5,5`-disulfononic acid disodium salt [860-22-0]; acid red 73

(AR 73), Brilliant crocein MOO C.I. 27290; amaranth (AM),



- 96 -

acid red 27, azorubin S, [915-67-3] and acid orange 7, (AO),

orange II sodium salt [633-96-5] are well known for their

high absorptivity and have been utilized for estimation of

excess oxidant. The work aims to demonstrate a simple,

rapid, accurate, precise and sensitive spectrophotometric

method suitable and convenient for the determination of

amlodipine besylate in pure and in dosage forms.

2. Experimental

Apparatus All the absorption spectral measurement were made using

JASCO V-530 (UV-VIS) spectrophotometer (Japan), with

scanning speed 400 nm min-1 and band width 2.0 nm,

equipped with 10 mm matched quartz cells.

Reagents All chemicals used were of analytical or pharmacopoeial

grade purity and bidistilled water was used. Standard

amlodipine besylate was obtained from Egyptian

International Pharmaceutical Industries Co. (EIPICO) 10th

of Ramadan City, Egypt. Stock amlodipine besylate solution

(100 µg ml-1) was prepared by dissolving 0.01 g in

bidistilled water and adjusted to 100 ml with bidistilled

water in 100 ml measuring flask. Working solutions of lower

concentration were prepared by serial dilutions.

Aqueous solutions of 10-3 M AB (Merck), and AO, AM

and AR (Aldrich), or 10-4 M for MB (Merck) were prepared

by dissolving an appropriate weight in 100 ml bidistilled

water. A stock (5.0 x 10-4 M) solution of KMnO4 (Aldrich),

was freshly prepared by dissolving an accurate weight in

bidistilled water, and standardized as recommended [28].

A solution of 0.2 M H2SO4, was prepared by adding

exact volume from stock (98%) concentrated acid to

bidistilled water, cooled to room temperature, transferred to

500 ml measuring flask, diluted to the mark and standardized

as recorded [29].

General procedure The method depends on oxidation of amlodipine besylate by

addition of 0.1-2.6 ml ADB (100 µg ml-1) to 1.0 ml of 5.0 x

10-4 M KMnO4 and 1.0 ml of 0.2 M H2SO4. The solution

was heated in a water bath at 50 ± 1 oC for 10 min, the

mixture was cooled and 2.0 ml (10-4 M) of MB, 0.8, 0.35,

0.8 and 0.7 ml (10-3 M) of AB, AR, AM and AO,

respectively was added. The volume was completed to 10 ml

with bidistilled water. The decrease in color intensity of dyes

were measured spectrophotometrically against a blank

solution containing the same constituent except drug treated

similarly, at their corresponding λmax 663, 609, 511, 520 or

484 nm, respectively. The concentration range was

determined in each case by plotting the concentration of

amlodipine besylate against absorbance at the corresponding

maximum wavelengths.

Procedure for determination of dosage forms At least ten tablets of ADB were weighed into a small dish,

powdered and mixed well. A portion equivalent to 10 mg

was weighed and dissolved in 100 ml bidistilled water,

mixed well for 15 min using a magnetic stirrer and filtered

through a sintered glass crucible G4. A 1.0 ml aliquot of the

test solution (100 µg ml-1 of ADB) was treated as described

above in the general procedure.



- 97 -

3. Results and discussion An analytical procedure based on the specific reactivity of an

amino group was investigated. The method involves two

steps namely:

1- Oxidation of amlodipine besylate with KMnO4 in acidic

medium by heating in water bath of 50 ± 1 oC.

2- Determination of unreacted oxidant by measuring the

decrease in absorbance of dyes at a suitable λmax.

Optimization The influence of each of the following variables on the

reaction was tested.

Effect of permanganate concentration The influence of KMnO4 concentration was studied in the

range from 10-5 - 10-4 M, as final concentration. The

optimum results were obtained with 5.0 x 10-5 M; higher

concentration of KMnO4 caused the color to disturbed.

Effect of acid concentration Different types of acid were examined (HCl, H2SO4, H3PO4,

CH3COOH and HNO3). The most suitable acid to achieve

maximum yield of redox reaction was found to be sulphuric

acid. Moreover, various volumes of H2SO4 were tested and

found to be 1.0 ml of 0.2 M as shown in Fig. 1.

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1.0

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2

ml added of sulphoric acid (0.2 M)

Abs

orba

nce

Methylene blueAcid blue 74Acid red 73Acid red 27Acid orange 7

Fig. 1. Effect of ml added of sulphuric acid (0.2 M) on absorbance of 10 µg ml-1 of amlodipine besylate

with KMnO4 (5.0 x 10-4 M) and dyes (1.0 x 10-3 M) except on using methylene blue (1.0 x 10-4 M)

Effect of temperature and time The oxidation process of amlodipine besylate is catalyzed by

heating in water bath of 50 ± 1 oC. The time required to

complete the reaction is 10 min. After oxidation process, the

solution must be cooled at least for 3.0 min before addition

of dye.

Effect of dye concentration The optimum volume of dye used for production of

maximum color intensity is 2.0 ml of 10-4 M MB, or 0.8,

0.35, 0.8 and 0.7 ml of 10-3 M AB, AR, AM and AO,

respectively. The effect of time after the addition of dye

indicated that shaking for 1.0 min is sufficient to give

reliable results for all dyes. The color remains constant for at

least 48 h.

Analytical data Beer’s law limits, molar absorptivities, Sandell sensitivities,

regression equations and correlation coefficients were

calculated and recorded in Table 1. The limits of detection

(K=3) and quantitation (K=10) were established according to

IUPAC definitions [30] and recorded in Table 1. In order to

determine the accuracy and precision of the methods,



- 98 -

solution containing three different concentrations of ADB

were prepared and analyzed in six replicates. The analytical

results obtained from this investigation were summarized in

Table 2.

Table 1. Optical and regression characteristics of amlodipine besylate with five different dyes.

Parameters MB AB AR AM AO

λmax (nm) 663 609 511 520 484

Beer’s law limits, (µg ml-

1)

1.0-24 0.9-22 1.2-26 0.9-12.8 1.0-14

Ringbom limits, (µg ml-1) 1.2-22.4 1.1-20 1.4-24.5 1.0-12.3 1.3-13.2

Molar absorptivity, (L

mol-1 cm-1)

2.25 x 104 3.12 x 104 2.01 x 104 22 x 104 3.42 x 104

Sandell sensitivity, (ng

cm-2)

25.19 18.15 28.17 13.44 16.58

Detection limits, (µg ml-1) 0.277 0.249 0.329 0.239 0.272

Quantitation limits, (µg

ml-1)

0.923 0.831 1.096 0.798 0.907

Regression

equation*:Slope (b)

0.0397 0.0551 0.0355 0.0744 0.0603

Intercept (a) 5.3 x 10-3 8.5 x 10-3 -9.9 x 10-3 -4.6 x 10-3 -3.1 x 10-3

Correlation coefficient (r) 0.9998 0.9999 0.9998 0.9996 0.9999

RSD ** % 0.66 1.01 0.82 0.51 0.73

• With respect to A = a + b C where C is concentration of drug in µg ml-1 and A is absorbance.

** Relative standard deviation for six determinations

Table 2. Evaluation of the accuracy and precision of the Proposed procedure of amlodipine besylate.

Dye Taken µg ml-1 Recovery, % RSD a % RE b % Confidence limits C

MB

8.0 10 12

100.1 100.2 99.9

0.86 0.89 0.53

0.90 0.93 0.55

8.01 ± 0.0724 10.02 ± 0.0933 11.99 ± 0.0661

AB

8.0 10 12

99.8 100.5 99.7

0.74 0.46 0.38

0.78 0.48 0.39

7.98 ± 0.0619 10.05 ± 0.0483 11.96 ± 0.0472

AR

8.0 10 12

100.3 99.6 99.8

0.65 0.71 0.40

0.68 0.95 0.42

8.02 ± 0.0546 9.96 ± 0.0745 11.98 ± 0.0504

AM

8.0 10 12

99.5 99.9

100.3

0.70 0.88 0.67

0.74 0.92 0.71

7.96 ± 0.0588 9.99 ± 0.0923 12.04 ± 0.0850

AO 8.0 10 12

100.4 100.6 100.6

0.80 0.77 0.33

0.84 0.800.34

8.03 ± 0.0672 10.06 ± 0.0808 12.07 ± 0.0409

a: Relative standard deviation for six determinations , b Relative error , c: 95 % confidence limits and five degrees of freedom



- 99 -

Interference A systematic quantitative study was undertaken by

measuring the absorbance of solutions containing 10 µg ml-1

of ADB with varying concentration of the additives and

excipients such as calcium hydrogen phosphate, magnesium

stearate and starch. Under the experimental conditions, the

effect of excipients frequently found in formulations was

evaluated using the proposed method. The additives and

excipients in all tablets are not interfere.

Analytical applications The proposed method was successfully applied to determine

ADB in its dosage forms. The results obtained were

compared statistically by Student’s t-test (for accuracy), and

variance ratio F-test (for precision) [31], with the official

method [32] at 95 % confidence level as recorded in Table 3.

The results showed that the t- and F- values were lower than

the critical values indicating that there was no significant

difference between the proposed and official methods. The

proposed method was more accurate with high recoveries

compared to the official method (depended on liquid

chromatography using stationary phase, octadecylsilyl silica

gel 5.0 µm and mobile phase, mix 15 volumes of

acetonitrile, 35 volumes of methanol and 50 volumes of a

solution prepared as follows: dissolve 7.0 ml of

triethylamine in 1000 ml bidistilled water and adjust to pH

3.0 ± 0.1 with phosphoric acid), so the proposed method can

be recommended for routine analysis of ADB in pure and

dosage forms in the majority of drug quality control

laboratorie.

Table 3. Determination of ADB in pharmaceutical formulations using the proposed and official methods.

Proposed methods Pharmaceuticalformulations MB AB AR

Recovery

%

t-

value*

F- ratio* Recovery

%

t-

value*

F- ratio* Recovery

%

t-

value*

F- ratio*

Norvasc 5 mg1 99.2 0.27 1.36 99.8 0.58 2.11 100.1 0.82 2.85

Amilo 5 mg2 100.2 1.02 3.12 99.6 0.80 2.69 100.4 0.62 1.84

Alkapress 5 mg3 99.8 0.58 1.45 100.1 0.62 2.11 99.6 0.36 1.14

Amlodipin 5 mg4 100.5 0.19 1.21 99.6 0.37 1.27 99.7 0.34 1.64

Myodura 5 mg5 100.3 0.92 1.58 99.8 0.36 1.89 99.6 0.64 1.91

Official Pharmaceuticalformulations AM AO

Recovery

%

t-

value*

F- ratio* Recovery

%

t-

value*

F- ratio* Recovery

%

Recovery

%

Norvasc 5 mg1 99.5 0.90 2.58 99.7 0.57 1.89 100.1 98.9

Amilo 5 mg2 99.9 0.15 1.27 99.7 1.08 2.93 100.4 99.4

Alkapress 5 mg3 100.4 0.48 2.12 99.7 0.54 1.58 99.6 99.5

Amlodipin 5 mg4 99.8 0.39 1.77 100.2 0.95 1.89 99.7 99.4

Myodura 5 mg5 99.5 0.54 1.85 100.1 0.34 2.11 99.6 99.4 * Theoretical value for t- and F- values for five degrees of freedom and 95 % confidence limits are 2.57 and 5.05, espectively.

(1) Pfizer S.A.E. Cairo, Egypt under authority of Inc., USA.

(2) Alpha Chem Advanced of Pharmaceutical Industries Company (ACAPI), Bader Industrial City, Cairo, Egypt.



- 100 -

(3) Alkan Pharmaceutical Company, Cairo, Egypt.

(4) Pharaonia Pharmaceutical, Pharo-pharma Company, Cairo, Egypt.

(5) Global Napi Pharmaceuticals Company (GNP) under license from Merck & Co. Inc. USA, Egypt

4. Conclusion The proposed method was advantageous over other reported

visible spectrophotometric and colorimetric methods, related

to their high reproducibility, high sensitivity, less time

consuming and using simple and inexpensive reagents.

Moreover, this method allowed the determination of ADB up

to 0.9 µg ml-1, in addition to simplicity, rapidity, precision

and stability of colored species for more than 48 h. The

proposed method may be applied for routine analysis and in

quality control laboratories for the quantitative determination

of the ADB in raw materials and in pharmaceutical

formulations. The stability constant was determined and the

free energy change was calculated potentiometrically. The

positive value of ∆G reveals that the dissociation of this drug

is not spontaneous.

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Pharmacopoeia", 31st Edn., Royal Pharmaceutical

Society, London, 1996, 819.

[2] Baranda, A. B.; Jiménez R. M.; Alonso, R. M., J.

Chromatogr. 2004,1031, 913.

[3] Baranda, A.B.; Etxebarria, N.; Jiménez, R. M.; Alonso,

R. M., Talanta 2005, 67, 933.

[4] Sudhakar, P.; Nirmala, M.; Babu, J. M.; Vyas, K.;

Reddy, G. M.; Bhaskar, B. V.; Reddy P. P.; Mukkanti,

K., J. Pharm. & Biomed. Anal. 2006, 24, 605.

[5] Zarghia, A.; Foroutanb, S. M.; Shafaatia A.; Khoddam,

A., IL Farmaco 2005, 60, 789.

[6] Josefsson, M.; Zackrisson, A.-L.; Norlander, B., J.

Chromatogr., B: Biomed. Appl. 1995, 672, 310.

[7] Naidu, K. R.; Kale U. N.; Shingare, M. S., J. Pharm. &

Biomed, Anal.2005, 39, 147.

[8] Avadhanul, A. B.; Srinivas, J. S.; Anjaneyul, Y., Indian

[9] Sankar, S. R.; Nanjan, M. J.; Vasudevan, M.; Shaat, N.;

Suresh, B., Indian J. Pharm. Sci., 1997,59, 171.

[10] Yasuda, T.; Tanaka, M.; Iba, K., J. Mass Spectrom.

1996, 31, 879.

[11] Bersford, A. P.; V.Macrae, P.; Stopher, D. A.; Wood, B.

A., J. Chromatogr. 1987, 420, 178.

[12] Feng, Y.; Gou, X.; Yand, D.; Ne, Y., Guangdong

Yaoxueyuan Xuebao 1998, 14, 111.

[13] Chandrashekhar, T. G.; Rao, P. S. B.; Smrita, K.; Vyas,

S. K.; Dutt, C., J.

Planar Chromatogr. -Mod. TLC 1994, 7 458.

[14] Pandya, K. K.; Satia, M.; Gandhi, T. P.; Modi, I. A.;

Modi, R. I.; Chakravarthy, B. K., J. Chromatogr. B:

Biomed. Appl. 1995, 667, 315.

[15] Ilango, K.; Kumar, P. B.; Prasad, V. R. V., Indian J.

Pharm. Sci. 1997, 59, 336.

[16] Jiang, T.F.; liang, B.; Li, B.J.; Li C.; Ou, Q.Y., Fenxi

Kexue Xuebao 2003, 19, 33.

[17] Mohamed, Y. E.; Naglaa, M. E. K.; Bahia, A. M.;

Nashwa, G. M., Bull. Fac. Pharm. 1998, 36, 1.

[18] Sridhar, K.; Sastry, C. S. P.; Reddy, M. N.; Sankar, D.

G.; Srinivas, K. R., Anal. Lett. 1997, 30, 121.

[19] Rahman, N.; Singh M.; Hoda, M. N., IL Farmaco 2004,

59, 913.

[20] Gazy, A. A., Talanta 2004, 62, 575.

[21] Lokesh, B. V. S.; Reddy, M. N.; Sanker, D. G.,

Sreedhar, K., East. Pharm. 1996, 39, 125.

[22] Reddy, M. N.; Rani, G. T.; Rao, K. V. S. P.; Sankar, D.

G.; Sreedhar, K., Indian J. Pharm. Sci. 1997, 59, 188.

[23] Singhvi I.; Chaturvedi, S. C., Indian J. Pharm. Sci.

1998, 60, 309.



- 101 -

[24] Singhvi I.; Chaturvidi, S. C., Indian J. Pharm. Sci. 1999,

61, 190.

[25] Rahman N.; Azmi, S. N. H., Anal. Sci. 2000, 16, 1353.

[26] Prabhakan A. H.; Giridhar, R., Indian Drugs 2002, 39,

204.

[27] Rahman N., Azmi, S. N. N., IL Farmaco 2001, 56, 731.

[28] Basset, J.; Denney, R. C.; Jeffery, G. H.; Mendham, J.,

“Vogels Text Book of Quantitative Inorganic Analysis”

4th Ed., 1986, 350.

[29] Basset, J.; Jeffery G. H.; Mendham, J., “Vogels Text

Book of Quantitative Inorganic Analysis” I, 1978,

308.

[30] IUPAC, Spectrochim. Acta, (B) 1978, 33, 242.

[31] Miller, J. C.; Miller J. N., "Statistics in Analytical

Chemical", 3rd Edn., Ellis Horwood, Chichester, UK,

1993.

[32] The British Pharmacopoeia (BP), Her Majesty's

Stationary Office, London, I, 2005, 125.



- 102 -

Arabian J. Chem. Vol.2, No. 1, 103-112(2009) - 103 -

Synthesis and Characterization of New Poly(ester-amide)s containing Diarylidenecyclohexanone in the

Main Chain. Part: II

Khalid S. Khairou, Mohamed A. Abdullah, Kamal I. Aly*, Nariman M. Nahas

and Ameena M. Al-Bonian.

Chemistry Department, Faculty of Applied Science, Umm Al-Qura University, Saudi Arabian.

*Polymer Lab 122, Chemistry Department, Faculty of Science, Assiut University, Assiut , 71516, Egypt

* E-mail: [email protected]

Abstract A new category from poly(ester-amide)s based on diarylidenecyclohexanone in the main chain, were

synthesized via interfacial polycondensation technique of two monomers namely: 2,6-bis (4-

hydroxybenzylidene) cyclohexanone I and 2,6- bis (4-hydroxy-3-methoxybenzylidene) cyclohexanone II

with diacid chlorides IIIa-c. The model compounds were synthesized by reacting one mole of compound

IVa-c with the two monomers I and II . The structure of the model compounds was confirmed by correct

elemental and spectral analyses. The various characteristics of the resulting polymers including: solubility,

viscosity, thermal analysis, X-ray diffraction analysis were determined and discussed. The majority of the

polymers were insoluble in most common organic solvents. The viscosity measurements in

dimethylsulphoxide showed the values 0.58-0.79 dL/g.. Thermal analysis shows that they are thermally stable

up to 500°C. X-Ray analysis showed that polymers having some degree of crystallinity in the region 2θ = 5 –

50o.

Keywords: Poly(ester-amide)s, diarylidenecycloalkanones, Synthesis, Characterization.

1. Introduction. Recently much attention on high-performance polymers

that have excellent thermal stability and solubility has

provided researchers with the impetus that has led to the

discovery of a variety of thermostable and processable

polymers. Poly(ester-amide)s

(PEAs) attracted scientific interest, since they may be

designed to couple the excellent mechanical properties of

polyamides and the biodegradability of polyesters [1].

PEAs have found a wide range of applications, such as

disposable bags, agricultural films, drug carriers or matrix

resins for biomedical materials [2]. (PEAs) can crystallise

rapidly if the amide segments have an ordered structure as

in alternating (PEAs) [3,4] and (PEAs) with uniform

diamide segments [5,6]. Several kinds of poly(ester-

amide)s copolymers based on lactic acid, 3-caprolactone

and amino acids have been previously studied [7,9].

Another linear poly(ester-amide)s derived from adipic acid,

1,4-butanediol, hexamethylene diamine and caprolactam,

emphasizing its thermal processing behavior and

composites with inorganic fillers [10]. The work reported

in this paper, outlines the synthesis and characterization of

some new poly(ester-amide)s based on

diarylidenecyclohexanone moiety in the main chain. The

major aim of this work has been to investigate the effect of

inclusion of cyclohexanone moiety on the polymer

Khalid S. Khairou, Mohamed A. Abdullah, Kamal I. Aly*, Nariman M. Nahas…….

- 104 -

properties. In addition other characteristic of these new

polymers such as thermal stability, solubility, and

crystallinity, were discussed.

2. Experimental 2.1. Instrumentation

Elemental analyses were carried out using an Elemental

Analyses system GmbH, VARIOEL, V2.3 July 1998 CHN.

Melting points were determined on a Perkin-Elmer 240C

electrothermal melting point apparatus and are uncorrected.

Infrared spectra were recorded on a Shimadzu 2110 PC

Spectrophotometer with KBr pellets. The 1H-NMR spectra

were recorded on a GNM-LA 400 MHz NMR

spectrophotometer at room temperature in DMSO or

CHCl3 using TMS as the internal reference. Viscosity

measurements were made with 0.5% (w/v) solution of

polymers in sulfuric acid (9 M) at 25oC using an

Ubbelohde suspended level viscometer. The X-ray

diffractograms of the polymers were obtained with a

Phillips X-ray unit (Phillips generator Pw-1710) and Ni-

Filtered CuKα radiations. TGA and DTG measurements

were performed on V 5.1 A Du Pont 2000 thermal analyzer

at a heating rate 10oC/min in air. The solubility of the

polymers was determined using 0.02 g of polymer in 3.5

ml of solvent. Electronic spectra were recorded for

solutions in DMSO in the region 200-600 nm with a

Shimadzu 2110 PC scanning spectrophotometer. The

morphology of the polymers was examined by scanning

electronic microscopy (SEM) using a Jeol JSM-5400 LV-

ESM.

2.2. Reagents and Solvents Cyclohexanone (Merck), p-hydroxy-benzaldehyde and

Vanilline were used without purification. Terephthaloyl

chloride (Aldrich) was recrystallized from n-hexane (m.p

83-84oC). Amino acids and all other solvents were of high

purity and were further purified by standard method [11].

2.3. Monomer Synthesis:

2.3.1. Synthesis of Monomers I and II: 2,6-Bis (4-hydroxybenzylidene) cyclohexanone I and 2,6-bis (4-

hydroxy-3-methoxybenzylidene) cyclohexanone II were

prepared as described in previous works [ 12,13].

2.3.2. Synthesis of Diacid chlorides IIa-c: These monomeric compounds were prepared by similar

methods that used in Literature [14]. In a conical flask 250

ml, a mixture of 0.2 mol of ℓ -alanine was dissolved in 25

ml of sodium hydroxide 10% and then a 0.1 mol of

isophthaloyl was added in one hour in five portions with

vigorously shaking after each addition. At the end of

reaction time 50 grams of crushed ice was added and

acidified with dilute HCl acid to Congo red paper. Whereas

a white precipitate was isolated, washed well with water,

dried and recrystallised from a mixture of 1:3 H2O/ethanol

and gave white needle crystals

2.4. Synthesis of Model Compounds Va,b:

General method: In a round bottomed flask 250 ml a 0.02 mol of compounds

III- V, was dissolved in a mixture of ratio 1:1 thionyl

chloride-benzene 60 ml and refluxed for one hour on water

bath. After this time few drops of pyridine was added and

refluxed more for another hour. At the end of the reaction

time, the mixture was evaporated under reduced pressure

whereas a hemi-solid and solid product was obtained and

used as it without purification.

2.5. Polymer synthesis

General procedure: A three-necked flask, equipped with a mechanical stirrer

(200 rpm/min) and dropper, was charged with a mixture of

0.02 mol monomer I or monomer II, 50 ml methylene

chloride and a suitable quantity of sodium hydroxide. A

stoichiometric quantity of (0.04 mol) of the latter dissolved

in 100 ml of water was also introduced. After mixing, 0.01

Synthesis and Characterization of New Poly(ester-amide)s containing Diarylidenecyclohexanone in ……… - 105 -

mol of acid chlorides IIIa-c dissolved in 25 ml methylene

chloride was added over 2-min at 25°C and vigorously

stirred. After complete addition of the acid chloride, stirred

was continued for 60 min whereby a highly-yellowish solid

separated out. The solid was filtered off, washed with

water, hot ethanol and dried under reduced pressure 1

mmHg at 90°C for one day.

By using the above general procedure the following

poly(ester-amide)s VIa-c and VIIa-c were obtained:

2.5.1. Poly(ester-amide) VIa : Obtained by the polymerization of 2,6-bis (4-

hydroxybenzylidene) cyclohexanone I (0.002 mole) with

acid chloride IIIa (0.002 mole) for 4hrs as yellow powder,

yield 98% Anal. Calcd. for C34

H30

N2O

7: C,70.59; H,5.19;

N,4.84. Found: C,69.61; H,5.02; N,4.21

2.5.2. Poly(ester-amide) VIb : Obtained by the polymerization of 2,6-bis (4-


acid chloride IIIb (0.002 mole) for 4hrs as yellow powder,

yield 92% . Anal. Calcd. for C38

H38

N2O

7: C,71.92; H,5.99;

N,4.41. Found: C,69,90; H,5.58; N,4.18

2.5.3. Poly(ester-amide) VIC : Obtained by the polymerization of 2,6-bis (4-


acid chloride IIIc (0.002 mole) for 4hrs as yellow powder,

yield 88% .Anal. Calcd. for C46

H38

O7N

2: C,75.62; H,5.21;

N,3.84. Found :C,75.01;H,5.11; N,3.32

2.5.4. Poly(ester-amide) VIIa : Obtained by the polymerization of 2,6- bis (4-hydroxy-3-

methoxybenzylidene) cyclohexanone II (0.002 mole) with

acid chloride IIIa (0.002 mole) for 4hrs as yellow powder,

yield 95% .

Anal. Calcd. for C36

H34

N2O

9: C,67.71; H,5.33;

N,4.39.Found : C,68.69; H,5.25; N,4.16

2.5.5. Poly(ester-amide) VIIb : Obtained by the polymerization of 2,6- bis (4-hydroxy-3-


acid chloride IIIb (0.002 mole) for 4hrs as yellow powder,

yield 96% .


H42

N2O

9: C,69.16; H,6.05; N,4.03.

Found : C,69.14; H,5.95; N,4.21 2.5.6. Poly(ester-amide) VIIc : Obtained by the polymerization of 2,6- bis (4-hydroxy-3-


acid chloride IIIc (0.002 mole) for 4hrs as yellow powder,

yield 90% .


H42

O9N

2: C,75.49; H,5.51; N,3.67.

Found: C,74.48; H,5.21; N,3.04


3.1 Synthesis of Monomers I,II. The preparation of these poly(ester-amide)s VIa-c and VII

a-c were based on 2,6-bis (4-hydroxy benzylidene)-

cyclohexanone I and 2,6 bis (4-hydroxy-3-

methoxybenzylidene) cyclohexanone II. These monomeric

units were synthesized by condensation of two moles of 4-

hydroxybenzaldehyde or 4-hydroxy-3-methoxy-

benzaldehyde with one mole of cyclohexanone in presence

of ethanol and catalytic amount of conc. HCl as shown in

Scheme 1. O

+OHC

R

OH HO

R

HC CH

O R

OH

( I: R = H ; II, R = OCH3 )

Scheme 1. Synthesis of Monomers I and II.


- 106 -

These monomeric units were purified by recrystallization

twice before using in the polymerization. The structures of

these compounds were elucidated by elemental and spectral

analysis (IR and 1H-NMR).

3.2. Synthesis of Diacid chlorides IIIa-c: These monomeric units were prepared according to the

literature [15] by interaction of one mole terephthaloyl

chloride with two mole of amino acid namely ℓ- alanine, ℓ-

valine or ℓ-phenylalanine in presence of 10% NaOH with

vigorously shaking and 1/2 hr stirring, as shown in

Scheme 2.

Cl C

O

C

O

NaOH

HOOC CH HN C

O

R

O

C NH CH COOH

R

h stirringCl + NH2 CH COOH

R 21

2

Scheme 2. Synthesis of Diacid chlorides IIIa-c.

3.3. Synthesis of Model Compounds compounds

Va,b. Before attempting polymerization, the model compounds

for the desired polymers were prepared. This was

performed by interaction of one mole of any amino acid

and ℓ-phenylalanine ( as example) with one mole of

benzoyl chloride to producing benzoyl ℓ-phenylanine, The

former was converted to acid chloride by dissolved the acid

compound in a mixture of thionyl chloride – benzene and

few drops of pyridene as catalyst was added. At the end of

the reaction time, the mixture was evaporated under

reduced pressure, to give the acid chloride IV ( Scheme 3).

Scheme 3. Synthesis of precursor compound IV.

COCl + Ph - CH2 - CH - COOH

NH2

NaOH 10%

CO - NH - CH - COOH

CH2 - Ph

HCl 10%CO - NH - CH - COCl

CH2 - PhIV

V


The model compounds Va,b was synthesized by

interaction of the monomers I or II with the previous acid

chloride IV On the basis of good agreement between

calculated and found elemental analyses, IR, IH-NMR

spectra, the possible reaction is depicted in Scheme 4.

C NH-CH-COCl + HO

CH2Ph

R

CH

O

CH

R

OH

O

CH2CH2 / NaOH

PhC-NH-CH-C-O

R

HC

O O

CH2Ph

O

CH

R

O-C-CH-NHC-Ph

O O

CH2Ph

Va: R=H; Vb: R=OCH3

Scheme 4. Synthesis of the Model Compounds Va,b.

3.4. Synthesis of Poly(ester-amide)s VI a-c and VII

a-c. One of the aims of studies presented in this work is to

synthesize a new series of poly(ester-amide)s VIa-c and

VII a-c by using interfacial polycondensation technique

which proved to be useful for the synthesis of polyesters

and their analogues [16-18]. These new polymers were

synthesized by condensation of 2,6-bis (4-

hydroxybenzylidene) cychohexanone I or 2,6-bis (4-

hydroxy-3-methoxy-benzylidene) cyclohexanone II with

diacid chlorides IIIa-c as represented in Scheme 5.

HO

R

CH CH

R

OH

O

ClC-HC-HN-C

O OC-NH-CH-C-Cl

R

OO

C-N-C-C

R

OO

O

R

CH CH

R

O-C-CH-NH-C

O

R

O O

n

H H

+

VI a-c: (R=H): a: R'=CH3; b: R'=CH (CH3)2; c: R'= -CH2Ph.

VII a-c: (R=OCH3): a: R'=CH3; b: R'=CH (CH3)2; c: R'= -CH2Ph.

Scheme 5. Synthesis of Poly(ester-amide)s VI a-c and VII a-c.


- 108 -

The structure of these polymers was also established from

elemental and spectral analyses. The elemental analyses of

all the polymers coincided with the characteristic repeating

units of each polymer; the data are included in the

experimental part. It should be noted that the elemental

analyses of these polymers deviated up to 1.12% from the

theoretical values. However, it is not uncommon for

polymers to trap solvent molecules within the polymer

matrix [19].

Spectral data support the structural assignment of the

poly(ester-amide). IR spectra recorded from pellets of KBr

mixed with respective polymer showed characteristic

absorption bands due to -NH groups at 3210-3250 cm-1 ;

C=O of esters at 1745-1735

cm-1; C=O of cyclohexanone at 1690-1700 cm-1; C=C

stretching at 1590-1600 cm-1; phenylene rings at 1590-

1510cm-1 and C-O-C bonds (ether linkage) at 1250-1260

cm-1 and other characteristic bands were appeared in the IR

spectra.

3.5. Characterization of Poly(ester-amide)s VI a-c

and VII a-c. The various characteristics of the resulting poly(ester-

amide)s VI a-c and VII a-c including: solubility, X-ray

diffraction analysis, TGA and DTA were also determined

and all the data are discussed as described below.

3.5.1. Solubility

Room temperature solubility characterizations of

poly(ester-amide)s VI a-c and VII a-c were tested using

various solvents including : THF, DMF, DMSO, NMP

Tetrachloroacetylene, chloroform – acetone (1:1; v/v),

formic acid + phenol (1:1; v/v), and conc. H2SO4. A 5%

(w/v) solution was taken as a criterion for solubility. All

the poly(ester-amide)s VI a-c were insoluble in most

simple organic solvents such as: alcohols, benzene, and

acetone. It can be clarified from Table 1 that, the majority

of the polymers were completely soluble in polar aprotic

solvents like DMSO, DMF or NMP except polymers VIa-c

are partially soluble. In strong protic solvent like H2SO4, all

the synthesized poly(ester-amide)s are freely soluble and

gave reddish color. From these data it reveals that the

incorporation of R'= CH2ph in poly(ester-amide)s VIc and

VIIc backbone induce some extent toward higher solubility.

Moreover, it was found that all the polymers VIa-c were

completely insoluble in chloroform – acetone mixture and

TCE except polymers VIIb,c which are partially soluble.

On comparison between the solubility of the polymers

based on divanilyidenecyclohexanone with those based on

diarylidenecyclohexanone, it was found that, the latter

series are s more soluble in most solvents than the former

polymers. This may be attributed to the higher flexibility of

the cyclohexanone moiety as described in our previous

works [20, 21].

Table 1. Solubility characteristics of poly(ester-amide)s VI a-c and VII a-c.

Polymer THF DMF DMSO NMP TCE* CHCl3 +

acetone

(1:1)

HCOOH

+Phenol

(1:1)

Conc.

H2SO4

VIa

VIb

VIc

VIIa

VIIb

VIIc

±

±

-

±

±

±

±

±

±

+

+

+

±

+

+

+

±

+

+

+

+

+

+

+

-

-

-

-

-

-

-

-

-

-

±

±

-

-

±

+

+

±

+

+

+

+

+

+

+ Soluble at room temperature (RT), ±, partially soluble; - , insoluble. , * Tetrachloroethane


3.5.2. Determination of Viscosity The reduced viscosity of poly(ester-amide)s VI a-c and

VII a-c were determined by ubbelohde suspended level

viscometer using dimethylsulfoxide ( DMSO ) at 25°C ±

0.5°C and gave the value, 0.58 dL/g, 0.65dL/g , 0.75 dL/g

and 0.79 dL/g respectively.

3.5.3. X- ray analysis The X-ray diffractograms of poly(ester-amide)s VI a-c and

VII a-c. are shown in figure 1. It can be clarified from this

figure, that the majority of the polymers showed few

reflection peaks in the region 2θ =5-60o , this indicate that

these polymers are semicrystalline except the polymer VIc

showed a halo-pattern in the same region. Also

diffractographs indicated that the polymers VIa,b,c have

high degree of crystallinity in comparison with those

polymers VIIa,b,c. Moreover, the presence of C=O, C=C,

polar groups, induces some order between two adjacent

chains of the polymers, leading to some extended of

crystallinity [22]. On comparison between all the

poly(ester-amide)s VI a-c and VII a-c, it was found that the

presence of methoxy group as a substituent in the polymers

backbone caused some hindering between the repeating

units and inforced it to unsymmetrical orientation in the

polymers chain and reduced the crystallinity [23].

Figure 1: X-ray Diffraction patterns of poly(ester-amide)s Via-c and VIIa-c


- 110 -

3.5.4. Thermal Analyses The thermal stabilities of poly(ester-amide)s VI a-c and VII

a-c were evaluated by TGA and DSC. Figure 2 presents

typical TGA traces of poly(ester-amide)s in N2. The initial

decomposition temperature (IDT), the polymer

decomposition temperature (PDT) and the maximum

polymer decomposition temperature (PDTmax ) are listed in

Table (2). The PDT corresponds to the temperature at

which a weight loss of 10% was recoded. The PDTmax

corresponds to the temperature at which the maximum rate

of weight loss occurred. In Figure 2 the TGA curves

showed a small weight loss in the range 1-2% starting at

125°C up to 180°C, which may be attributed to loss of

absorbed moisture and entrapped solvent, respectively. All

the poly(ester-amide)s showed similar decomposition

pattern. The expected nature of decomposition of these

polymers are the scission of many bonds of

olefinic groups and ester groups and a pyrolytic

oxidation of amidic bonds. The PDT for all poly(ester-

amide)s ranged from 276-465°C. Therefore the data in

Table ( 2) indicate that the thermal stabilities of these

poly(ester-amide)s are in the order:VI a > VIIa >VI c ≈VII c

> VIIb > VIb .

Figure 3 shows typical DSC traces of poly(ester-amide)s

VI a-c and VII a-c . Poly(ester-amide)s VIIa and VIIb

showed large ascending exothermic curves without definite

Tg , Tc and Tm and may be attributed to curing reactions

involving the olefenic bonds 71. Polymer VII c shows a

brood exotherm with Tc from 125-148 °C and Tm at 160 °C

and this reflected the existence of some degree of

crystallinity inside the polymer bulk and this confirmed by

the data of X-ray as shown in Figure 1.

The DSC curves of VI a , VIc showed different brood

exotherms with Tc from 215-225 °C and followed with Tm

at 230 °C and Tc from 117-150°C accompanied by Tm at

175 °C respectively and this indicated the presence of some

degree crystallinity order in polymer backbone see X-ray

Figure 1.

Table2: decomposition temperature of poly(ester-amide)s VI a-c and VII a-c.

Sample IDTa (°C ) PDT b (°C ) PDT c max (°C )

VIa 196 289 455

VIb 142 250 440

VIc 203 301 493

VIIa 208 304 495

VIIb 250 384 570

VIIc 283 410 575 a Initial decomposition temperature.

b Polymer decomposition temperature. c Maximum polymer decomposition temperature.


Figure 2: TGA races of poly(ester-amide)s Via-c and VIIa-c in air at a heating rate of 10 oC/min

Figure 3: DSC curves of polymers (Via-c – VIIa-c)


- 112 -

4. Conclusions A new series of poly(ester-amide)s VI a-c and VII a-c was

synthesized using interfacial polycondensation technique.

The various characteristics of the resulting polymers were

tested. The presence of methoxy group as substituent in the

polymer backbone reduced the crystallinity and caused

some hindering between the repeating units and inforced it

to unsymmetrical orientation in the polymers chain. All the

poly(ester-amide)s were yellowish to pale-yellow , and

had inherent viscosity in the range 0.56-0.79 dL/g. They

are soluble in polar aprotic solvents like DMSO or NMP.

X-ray difractograms of poly(ester-amide)s showed some

degree of crystallinity in the region 2θ = 5-50°.

References [1] Okada Masahiko. Chemical syntheses of biodegradable

polymers.Progress in Polymer Science 2002;27:87–

133.

[2] Grigat E, KoCh R, Timmermann R. BAK 1095 and

BAK 2195:1998;59:223–6.

[3] Qian ZY, Li S, He Y, Liu XB. Biomaterials 2004;25:

1975–81.

[4] Qian ZY, Li S, He Y, Liu XB. 2004;84:41–7.

[5] Qian ZY, He Y, Liu XB. 2004;83:127–32.

[6] Qian ZY, Li S, Liu XB. Polymer Degradation and

Stability 2003;81:279–86.

[7] Qian ZY, Li S, Liu XB. Colloid and Polymer Science

2003;281:869–75.

[8] He Y, Qian ZY, Liu XB. Colloid and Polymer Science

2004;282:972–8.

[9] Qian ZY, Li S, Liu XB.. Colloid and Polymer Science

2004;282: 1083–8.

[10] Zou YB, Wang L, Liu Xiaobo.. Polymer Degradation

and Stability 2004;83:87.

[11] Perrin, D.D.; Armarego, W.L.F.; Perrin, D.R.;

Purification of Laboratory Chemicals 2nd edn New

York: Pergamon, 1980.

[12] Aly K. I. Polymer International 47,773- , (1999).

[13] Aly K. I. Polymer International 45, 483, (1998).

[14] Aly K. I. High Performance Polymers 10, 353, 1998.

[15] I.V.Vogel " Textbook of practical organic Chemistry

" 4th ed.,.885, 1978.

[16] Wittbecker, E.L. and Morgan, P.W.:. 40, 289 (1959).

[17] Imai, Y.; Ueda, M. and Li, M.: Die Makromolekulare

Chemie 179, 2085, (2003).

[18] Imai, Y.; Sato, N. and Ueda, M..Die Macromolekulare

Chemie, Rapid Communications. 1, 419,

(1980);Andrew S. M., Bass R. G. Journal of Polymer

Science Part A: Polymer Chemistry, 27, 1225, (1989).

(R)

[19] Aly, K.I.; Khalaf, A.A. J. of Applied Polymer Science,

2000, 77, 1218.

[20] Aly, K.I. Ph.D. Thesis; Assiut University, Egypt,

1989.

[21] Aly, K. I.; Khalaf, A. A. and Mohamed, I. A.

European Polymer Journal, 2002, 39, 1273.

[22] Mandelkern, L. Crystallization of Polymers,

McGraw-Hill, New York, 1964.

Arabian J. Chem. Vol. 2, No. 1,113-126(2009)


- 113 -

دراسة تأثري بعض أنواع مشروبات الطاقة على مؤشرات حيوية ونسيجية يف اجلرذان Study of Effect of Energy Drinks on Biochemical and Histological Markers in Rats

الرشيدي. أماين ع العزيز جبدة لك عبدجامعة امل– كلية التربية لالقتصاد املرتيل والتربية الفنية-قسم التغذية وعلوم األطعمة

Amani A. Al-Rasheedi Nutrition Department, Faculty of Science, Girls college of Education, King Abdul-Aziz University,

P. O. Box 18886 Jeddah- 21425, Saudi Arabia

E-mail: [email protected] وذلك نتيجة للميزانيات الضخمة املرصـودة بني األشخاص العاديني ، مشروبات الطاقة من أكثر املشروبات استهالكا تعترب

من قبل الشركات املنتجة على اإلعالنات اجلذابة واليت تصور منتجاهتا كعالج لتحسني مستوى األداء وعدم الشعور بالتعب . ية وعقلية أفضل صول على قدرات بدنوللح

احليويـة املؤشـرات مشروبات الطاقة على بعض لثالثة من املختلفة التأثرياتمقارنة وهتدف هذه الدراسة إىل عن جرعة وقد مت تقسيم ستة وثالثون من ذكور اجلرذان البيضاء إىل أربع جمموعات ، مت إعطائها يوميا . للجرذانوالنسيجية

، ٢٠ ، أو ١٠ أو املاء للمجموعة الضابطة ملدة الريد بول أو البور هورس ، أو الطاقة البايسون ، طريق الفم من أحد مشروبات Bilirubin البيلروبني و ، Hemoglobinاهليموجلوبني دم لتقدير مستويات عينات أخذوقد مت ذبح احليوانات و . يوما ٣٠أو

إلجـراء حتليـل القلبكما مت أخذ جزء من أنسجة . Triglycerideاجلليسريدات الثالثيةو ، Cholesterol، والكوليسترول الالكتيـت و ،Xanthine Oxidaseالزانثني أوكسيديز و ، Nitric Oxide أكسيد النتريك نسيجي وكذلك لتقدير مستوى

. Lactate dehydrogenaseدي هيدروجينيز أيضا جرذان و والنسيجية لل احليوية على املؤشرات وقد أظهرت النتائج تأثريات واضحة ملشروبات الطاقة املختلفة

. باجملموعة الضابطة ةأنسجة القلب مقارنعلى Energy drinks are one of the most common beverages consumed by average citizens, probably as a result of

the huge budgets allocated by the manufacturing companies to produce enticing commercials to promote

their products as the cure for feeling good, never get tired, and/or superior physical and mental abilities. This study aimed at comparing the effect of three kinds of energy drinks on certain biochemical and

histological markers in rats. Thirty six male Wister Albino rats were divided into four groups, each group

received a daily drinking dose of either Bison®, Power Horse®, Red bull®, or water (control group) for 10, 20,

or 30 days. The animals were then sacrificed, and samples of their blood were evaluated for hemoglobin,

bilirubin, cholesterol, and triglyceride levels. Heart tissues were also excised and used for histological

examination and/or determination of nitric oxide, xanthine oxidase, and lactate dehydrogenase levels. The

results showed clear effects of the different energy drinks on the biochemical and histological parameters in

rats as well as on heart tissue of the rats in comparison with the control group.

Amani A. Al-Rasheedi


- 114 -

المقدمة - ١تعترب مشروبات الطاقة يف الوقت احلاضر من أكثر املنتجات الغذائيـة

إىل اإلعالنـات شخاص العاديني ، وقد يرجع ذلـك انتشارا بني األ التجارية املكثفة اليت حتظى هبا وامليزانيات الضخمة اليت تنفق عليها من

ـ قبل الشركات املنتجة هبدف الترويج هلا وإبراز دورهـا د زويف التبالطاقة وتأخري الشعور بالتعب البدين والذهين وبالتايل حتسني مستوى

جملـال إىل وقد أدى التنافس بني الشركات العاملة يف هـذا ا .األداء املشروبات الرياضية ومـشروبات أنواع بالعديد من امتالء األسواق

الطاقة مع تعمد بعض الشركات املنتجة إىل عدم كتابـة املكونـات ة على العبوات أو إغفال ذكر بعض املكونات الضارة صـحيا ياحلقيقهنا تؤثر ويدعي منتجوها أ .[1]ذلك نسبة املواد املكونة للمشروب وك

تأثريا جيدا على وظائف حمدودة يف اجلسم وتنـشط اجلـسم واملـخ وحتسن األداء النفسي وترفع املعنويات وهي إدعاءات حمـل جـدال

.[2]اآلنعلمي كبري مل يتم حسمه حىت Energyوكثريا ما يتم اخللط ما بني مشروبات الطاقة

drinks واملشروبات الرياضيةSport drinks ويف احلقيقـة فـإن هـذين النـوعني مـن االعديد من اإلدعاءات اليت يقدمها مـصنعو

املشروبات متشاهبة مما جيعل نسبة كـبرية مـن املـراهقني تتنـاول وختتلف مـشروبات . شروبات الرياضية مشروبات الطاقة بديال للم

األوىل على نسب عالية مـن باحتواءالطاقة عن املشروبات الرياضية الكربوهيدرات والكافيني يف حني تعمل الثانية على تزويـد اجلـسم

.[3]بااللكتروليتات واملاء واملغذياتوتعرف مشروبات الطاقة بأهنا املشروبات اليت هتـدف إىل

والطاقة الذهنية "الكربوهيدرات" الناجتة عن أيض لطاقة تزويد اجلسم با أمحاض أمينيـة ، وبعض العناصر األخرى "الكافيني"الناجتة عن أيض

وهتدف مشروبات الطاقة إىل تزويد .خل ا ... ، وأعشاب وفيتاميناتالعضالت العاملة بنسبة كبرية من الكربوهيدرات لتعـويض الطاقـة

ت اليت تستمر لفترات طويلة وبالتايل اإلسـراع املستنفذة أثناء التدريبا اجلهـاز العـصيب املركـزي تنبيه باإلضافة إىل االستشفاءمن عملية

وحتسني وظائف املخ ، وكذلك تعويض النقص يف بعض الفيتامينات ومشروبات الطاقة من املـواد الغذائيـة .[1]واملواد الغذائية األخرى

ثافة جزيئاهتا أكرب من كثافـة أي أن ك Hypertonicفائقة التناضح . [4]سوائل اجلسم ، كما تتميز باخنفاض معدل تفريغها من املعـدة

وحتتوي مشروبات الطاقة على تركيزات عالية مـن الكربوهيـدرات Carbohydrates ــوز ــل اجللوك ــوز و ، Glucose مث الفركت

Fructose،اجلاالكتوز و Galactose ،الـسكروز وSucrose [5] . Ephedrineى نسبة عالية من املنبهات مثل اإلفيدرين كما حتتوي عل

على تنبيه اجلهاز العصيب املركزي الن يعم نللذاا Caffeineوالكافينياليقظة والشعور بالنشاط واحليوية ، كما أنه يزيد من التركيـز مسببا

Taurineالتيـورين و. [6]الة املزاجيـة حلوسرعة االنفعال وحيسن اا يف التوازن االمسـوزي وانقبـاض العـضالت يلعب دورا هام الذي

وأمحــاض أمينــة أخــرى مثــل ،[7]نتــاج الطاقــة وزيــادة إ Leucineوالليوسني Arginine واألرجنني Glutamineاجللوتامني

على تقليل واليت تعمل Valine. والفالني Isoleucine وأيزوليوسني واملـرتبط اقل العصيب يف الـدماغ الن Serotoninإنتاج السريوتونني

. [8]بتقليل الشعور باإلجهاد والتعب herbsتوي مشروبات الطاقة على بعض األعشاب كما حت

yerbaيوربا ماتو، cola الكوال و ،Gurana مثل حبوب اجلورانا

mate كمــصادر طبيعيــة للكــافيني ، وكــذلك اســتراجالس Astragalus ،ســــــــــيزاندراك وSchizandrac ،

كمواد داعمة جلهاز املناعة وأيضا جينكـو Echinaceaإيشيناكياوكمـواد مقويـة Ginseng ، جينسينج Ginkgo biloba بيالوبا

، هيدروكـسي سـيترات Ciwujiaللذاكرة ، وكذلك سـيوجيا Hydroxycitrate واإلفيدراEphedra اصـية زيـادة واليت هلا خ

إىل Pyruvateويـضاف البريوفـات . [9]معدل احتراق الـدهون كمـا . ]10[طاقة حملاربة اإلجهاد والشعور بالتعـب مشروبات ال

زالة الـشعور إل Creatineتضاف كميات صغرية جدا من الكرياتني وتضاف أيضا كمية قليلة من اجلليسريدات الثالثيـة . ]11[بالتعب

ملـشروبات Medium Chain Triglyceridesمتوسطة السلسلة ا اجلسم كمصدر للطاقة الطاقة إلزالة الشعور بالتعب حيث يستخدمه

وكذلك نسبة حمددة مـن . Glycogen [12]بدال من اجلليكوجني ، واإلينوسيتول Vitamin C مثل فيتامني ج Vitaminsالفيتامينات

Inositolــامني ب ــامني ب Vitamin B2 ٢ ، وفيت ١٢، وفيت

Vitamin B12 والنياسنيNiacin [13] القوة والنشاطإلعطاء . ثالثة أنواع من مـشروبات اختيارة احلالية مت ويف الدراس

: يف اململكة العربية السعودية وهيانتشاراالطاقة األكثر

Study of Effect of Energy Drinks on Biochemical and Histological Markers in Rats


- 115 -

على ) مل١٠٠ ( حيتوي يف عبوتهو Bisonالبايسون • جـم ١٣ جم كربوهيدرات ، ١٣ سعر حراري ، ٥١

جم تيـورين ، ل م ٣٠٠جم كافيني ، ل م ٢٤سكريات ، جـم ل م ٢,٥ نياسني ، جمل م ٦جم فيتامني ج ، ل م ٢٥

، ٦جـم فيتـامني ب ل م ٠,٦حامض البانتوثينيـك ، .[14]جم حامض الفوليك لم٠,٠٥٣

وحيتـوي يف عبوتـه Power Horseالباور هورس • جـم ٢٨ سـعر حـراري ، ١١٠علـى ) مل ٢٥٠(

ملجـم ٦٠٠ ملجم تيـورين ، ١٠٠كربوهيدرات ، ملجـم ٥٠ ملجم كـافيني ، ٨٠جلوكونوالكتون ،

ملجـم ٥ ، ٢ ملجـم فيتـامني ب ٠,١٥ل ، إينوسيتو ملجـم ٢٠ ، ١٢ ملجم فيتامني ب ٠,٠٥ ، ٦فيتامني ب

.[15] ملجم كالسيوم بانتوسنات ٢,٢ ، نينياس ٢٥٠( عبوتـه وحيتوي يف Red bull ولالريد ب •

جم كربوهيدرات ، ٢٧ سعر حراري ، ١٠٩على ) مل ملجــم كــافيني ، إضــافة إىل التيــورين ٨٠

ــض واجللوكون ــيتول وبعـ ــون واإلينوسـ والكتـ .[16]الفيتامينات

:المواد والطرق المستخدمة- ٢ Male Albino) التجـارب من جرذان٣٦تناول البحث عدد

Rats) ــراوح ــا تتـــ ــنيأوزاهنـــ ــا بـــ مـــ جم ، وكانت سليمة وخالية من األمراض ، وأمكـن ١٠٠ – ٧٠

أربعة وقسمت إىل . من مركز امللك فهد لألحباث جبدة احلصول عليها : كالتايل جرذان٩كل جمموعة تتكون من جمموعات ان سليمة وتركت حىت هنايـة جرذ :(A)اجملموعة األوىل •

Control group )ابطةاجملموعة الض(التجربة تعطى يوميا عن طريق الفم جرذان :(B)اجملموعة الثانية •

. يوما ٣٠وملدة ) بايسون(مشروب الطاقة تعطى يوميا عن طريق الفم جرذان :(C)ة موعة الثالث اجمل •

يوما٣٠وملدة ) بور هورس( طاقة مشروب التعطى يوميا عن طريق الفم جرذان :(D)ة اجملموعة الرابع •

. يوما ٣٠وملدة ) بولريد (مشروب الطاقة ث ين اجلرعات عن طريق الفم حب وقد أعطيت مجيع اجلرذا

. جم من وزن اجلسم ١٠٠/ مل ١,٥ حسب وزنه ذ منها اجلرذ يأخان ذرومت ذبح ثالثة ج . [17]ومت ذلك بشكل يومي طوال مدة التجربة

ثالثـون يومـا إلجـراء وعشرون ، ومن كل جمموعة بعد عشرة ، : التالية احليويةالتقديرات : في الدم

عـشرون ، ومت ذبح ثالثة فئران من كل جمموعة بعد عشرة ، ة كما مت مجع عينـات الـدم ، ثالثون يوما ، من بداية التجرب و

: لتقدير (Plane tube)ووضعت يف أنابيب فارغة مت القياس تبعا لطريقةوقد : Hemoglobin اهليموجلوبني

(Koch & Akingbe, 1981) واليت تعتمد على التفاعل التايل:

Methaemoglobin ) Hemoglobin + K3 (Fe(CN)6 Cyanometahaemoglobin + Hg(CN)2 Methaemoglobin

حيث نانوميتر ٥٦٧ طول موجي يقدر الناتج من التفاعل طيفيا عند و . [18] ثانية١٢٠ل يستغرق التفاع :يف مصل الدم لفة ٤٠٠٠ من الدم بالطرد املركزي عند (Serum)مت فصل املصل

: ملدة عشر دقائق لتقدير املؤشرات التالية

:Bilirubinالبيلروبني & Koch) مت تقدير البيلروبني يف البالزمـا باسـتخدام طريقـة

Akingbe, 1981) ويتم التفاعل على النحو التايل :

Bilirubin + 2-methoxy -4 nitrophenyl diazonium tetrafluoroborate Azobilirubin للوسـط باسـتخدام bilirubin بطريقـة غـري مباشـرة جروخي

Dyphilline ويقدر Azobilirubin الناتج من التفاعـل طيفيا ١٨٠وتـستغرق فتـرة التفاعــل نـانوميتر ٦٤٢ طول موجي عند

.[19]ثـانية



- 116 -

عتمد واليت ت (Van der Honing, et al, 1968) ةلطريقمت القياس تبعا :olesterol Chالكوليسترول : على التفاعل التايل

Cholesterol esters Cholesterol Cholesterol + RCOOH Esterase

Cholesterol + O2 Cholesterol Cholestenone + H2 O2

Oxidase

CH3

NH2

CH3

CH3

NH2

CH3

H2O2

CH3

CH3 CH3

NH2

CH3

H2N+ + H2OPOD +

((TTMMBB))

.[20] نانوميتر٦٤٢ طول موجيويتم تقدير تركيز الناتج طيفيا عند

:عتمد على التفاعل التايلواليت ت )Trinder,1969( مت القياس تبعا لطريقة :Triglyceride اجلليسريدات الثالثية Triacylglycerols + 3H2O esterase glycerol + 3RCOOH

Glycerol + ATP GK L - α - Glycerolphosphate + ADP

LL -- αα -- GGllyycceerroollpphhoosspphhaattee ++ OO22 GGPPOO hhyyddrrooxxyyaacceettoonnee pphhoosspphhaattee ++ HH22 OO22

IInnddiiccaattoorr ((ccoolloorrlleessss)) ++ HH22 OO22 PPOODD IInnddiiccaattoorr ((bblluuee))

.[21] نانوميتر٦٤٢ طول موجيميكن تقديره طيفيا عندوالـذي

Reflotron Type II وقد مت قياسهم مجيعا باستخدام جهاز الرفلترون

- Manual من شركةBoehringer-Mannheim GmbH, West Germany.

بعد احلصول على عينات الدم ، مت تـشريح : القلب يف •زئني ، حيـث اجلرذان واستئصال القلب وتقسيمه إىل ج

املؤشـرات استخدم جزء من خاليا النسيج القليب لتقدير : التالية احليوية

أكسيد النيتريـك مت تقدير : NO( Nitric Oxide(تريك يالنأكسيد واسـتخدام جهـاز Nitrateيف نسيج القلب بواسطة قياس تركيز

التحليل الطيفي لقراءة العينات ن حيث يـستخدم تفاعـل جـريس Griess reaction املتبع يف طريقة (Willams, 1984) [22].

ـ )XO (Xanthine Oxidase .EC( زانثني أوكـسيديز إنـزمي ال

إىل Hypoxanthineعملية أكـسدة حيفز XOأن حيث (1.1.3.22Xanthine مث إىل ، Uric Acid كميـا يف اإلنـزمي مت قياس نشاط و

. [23] (Bergemyer, 1974) أنسجة القلب تبعا لطريقة بروتني HLD (Lactate dehyrogenase( إنزمي الالكتيت دي هيدروجينيز

(EC.1.1.1.27) أن حيث LDH عامل حمفـز لعمليـة األكـسدة :واالختزال كما يف التفاعل التايل

CH3CHOH.COO¯ + Lactate + NAD CH3CO.COO¯ + NADH+H Pyruvate

[24] نسيج القلب تتبع طريقة بروتني كميا يفLDHولتقدير إنزمي

(Caband & Wroblewski ,1958). ومت قياسهم باسـتخدام قيـاس / Spectrophotometerالطيف يف جمال األشعة فـوق البنفـسجية

Visible موديــل UV minin-1240 مــن شــركةShimadzu Corporation, Kyoto-Japan



- 117 -

فحصها نـسيجيا تبعـا لطريقـة مت القلب واجلزء اآلخر من عينات (Bancroft & Stevens, 1996) [25]. ومجيع البيانات مت مجعهـا

عـن طريـق احلاسـب اآليل ANOVAوحساهبا بتحليل البيانات Excel كما مت استخدام برنامج ١٣ النسخة SPSSدام برنامج باستخ

. للرسوم البيانية النــتــائــج-٢

واألشكال من ) 1 ( رقم أظهرت النتائج املتمثلة يف اجلدول زيادة يف مستويات اهليموجلوبني بشكل عام وخاصة اجملموعة ) 1-6(

B ت وكذلك حدث . يوما ٢٠ و ١٠ حيث كانت الزيادة معنوية عند

وبني جلميع اجملموعات مقارنـة باجملموعـة ريزيادة يف مستويات البيل عنـد D يوما واجملموعة ٣٠ و ١٠ عند Cالضابطة وخاصة اجملموعة

أظهرت النتـائج اضـطراب يف مـستويات كما . يوما ٢٠و ١٠ ٢٠ معنويا عند والذي كان B,C,Dالكوليسترول جلميع اجملموعات

اجلليسريدات الثالثية لتوضح حـدوث وجاءت نتائج . يوما ٣٠و ، Aزيادة يف مستوياهتا جلميع اجملموعات مقارنة باجملموعة الـضابطة

، عنـد ثالثـني يومـا Bحيث كانت الزيادة معنوية للمجموعـة . يوما ٣٠ و ١٠ عند Dوللمجموعة

وريد بل (C) وبور هورس (B)واجلرذان اليت مت إعطائها مشروب الطاقة بايسون ) A( واالحنراف املعياري للجرذان السليمة املؤشرات احليوية يوضح قيم ) ١(جدول

(D) كال على حده ، واليت مشلت مؤشرات اهليموجلوبني والكوليسترول واجلليسريدات الثالثية والبيلريوبني يف مصل الدم . D C B A

Mean ±SD Mean ±SD Mean ±SD Mean ±SD Days Groups

Parameters 12.47±0.95 11.80±0.87 11.40±0.36** 9.05±2.81 10 days 14.13±1.33 13.27±0.78 14.67±0.76** 8.65±0.73 20 days 13.27±0.47 12.93±0.95 13.70±0.70 9.41±0.64 30 days

Hemoglobin

(g/dl) 0.31±0.01** 0.23±0.01* 0.26±0.03 0.17±0.03 10 days 0.23±0.01** 0.25±0.01 0.34±0.15 0.18±0.04 20 days

0.23±0.02 0.23±0.01** 0.22±0.01 0.18±0.02 30 days

Bilirubin (mg/dl)

104.33±4.01 97.67±4.51 83.33±7.64 110.67±13.61 10 days 95.55±10.70** 92.67±5.90** 103.33±2.08* 101.67±4.72 20 days 68.00±5.57** 75.00±6.08** 73.67±10.50** 104.33±8.39 30 days

Cholesterol

(mg/dl) 213.67±57.73** 121.70±33.79 99.00±10.00 98.93±37.39 10 days

77.73±1.70 111.40±19.59 114.23±38.48 72.13±3.00 20 days 167.67±47.90* 127.07±59.36 197.67±73.04* 126.30±36.95 30 days

Triglyceride

(mg/dl)

0

5

10

15

Hae

mog

lobi

ne

g/dl

A B C D

Groups

شكل (١) التأثيرات المختلفة لمشروبات الطاقة على مستوى الهيموجلوبين في دم الجرذان

10days

20days

30days

00.050.1

0.150.2

0.250.3

0.35

Bilir

ubin

mg/

dl

A B C D

Groups

شكل (٢): التأثيرات المختلفة لمشروبات الطاقة على مستوى البيلروبين في مصل دم الجرذان .

10 days

20 days

30 days



- 118 -

020406080

100120

Cho

lest

erol

m

g/dl

A B C D

Groups

شكل (٣) التأثيرات المختلفة لمشروبات الطاقة على مستوى الكوليسترول في مصل دم الجرذان .

10 days

20 days

30 days

0102030405060708090

NO

(nm

ol/m

in/m

gpr

otei

n)

A B C D

Groups

شكل (٥) التأثيرات المختلفة لمشروبات الطاقة على مستوى أآسيد النيتریك في قلب الجرذان .

10 days

20 days

30 days

0

50

100

150

200

250

Trig

lyce

ride

mg/

dl

A B C D

Groups

شكل (٤) التأثيرات المختلفة لمشروبات الطاقة على مستوى الجليسریدات الثالثية في مصل دم الجرذان .

10 days

20 days

30 days

0

5

10

15

20

25

XO (n

mol

/min

/mg

prot

ein)

A B C D

Groups

شكل (٦): التأثيرات المختلفة لمشروبات الطاقة على نشاط إنزیم الزانثين أوآسيدیز في قلب الجرذان

10 days

20 days

30 days

01234567

LDH

(nm

ol/m

in/m

gpr

otei

n)

A B C D

Groups

شكل (٧): التأثيرات المختلفة لمشروبات الطاقة على مستوى نشاط إنزیم الالآتيت دي هيدروجينيز في قلب الجرذان .

10 days

20 days

30 days

ة كـبرية يف معـدالت حدوث زياد ) 2(رقم دول اجليتضح من كما وإنزمي الزانثني أوكسيديز وذلك جلميع اجملموعـات أكسيد النيتريك

B,C,D يوما وكانت مجيعهـا ٣٠ و ٢٠ و ١٠ وعند مجيع الفترات بينما حدث اخنفاض كبري ومعنـوي يف معـدالت إنـزمي . معنوية



- 119 -

موعات وعند مجيـع الفتـرات الالكتيت دي هيدروجينيز جلميع اجمل .وذلك مقارنة باجملموعة الضابطة

املقارنات الثنائيـة ومـستوى املعنويـة بـني ) ٣(ويوضح اجلدول لبعض املؤشرات يف مصل الدم حيـث تظهـر A,B,C,Dاجملموعات

يف حالـة A(D), A(C), A(D)فروقات معنويـة واضـحة بـني ٣٠و١٠البيلروبني عند ويف حالة . يوما ٣٠و٢٠اهليموجلوبني عند

وتظهر . أيام ١٠ عند C(D), B(D)يوما ، وكذلك بني اجملموعات يومـا ٣٠ و ١٠ عند A(B)نتائج الكوليسترول فروقات معنوية بني

كما تظهر نتائج اجلليـسريدات . يوما ٣٠ عند A(D), A(C)وبني

C(D) يوما وبني ٢٠ عند A(B)الثالثية فروقات معنوية بني اجملموعة

B(D),A(D) املقارنات الثنائية ) ٤(ويوضح اجلدول . أيام ١٠ عند لنشاط النيتريك أوكسيد A,B,C,Dومستوى املعنوية بني اجملموعات

وإنزمي الزانثني أوكسيديز وإنيزم الالكتيت دي هيدروجينيز يف نسيج القلب حيث يظهر وجود فروقات معنوية واضحة بـني اجملموعـات

وعنـد ,B(C), A(D), A(C), A(B) C(D), B(D)وبعضها البعض ومل تكـن الفروقـات . يوما ٣٠ و ٢٠ و ١٠مجيع الفترات الزمنية عنـد B(D), B(C) يوما ، وبني ٢٠ عند C(D)معنوية بني اجملموعة

. يوما ٣٠ و٢٠ و ١٠ عند B(D) يوما ، وبني ٣٠

بول وريد (C) وبور هورس (B)واجلرذان اليت مت إعطائها مشروب الطاقة بايسون ) A(ري للجرذان السليمة واالحنراف املعيااملؤشرات احليويةيوضح قيم ) ٢(جدول (D) كال على حده ، واليت مشلت مؤشرات النتريك أوكسيد وإنزمي الزانثني أوكسيديز وإنزمي الالكتيت دي هيدروجينيز يف نسيج القلب .

D C B A Mean ±SD Mean ±SD Mean ±SD Mean ±SD

Days Groups Parameters

46.04±5.60** 52.38±2.02** 38.36±0.92** 10.72±1.31 10 days 60.31±3.16*** 68.30±8.43*** 46.69±1.59*** 10.98±1.48 20 days

69.26±2.88* 87.73±2.21** 54.25±3.77** 11.85±1.37 30 days

Nitric Oxide (µmol/gm tissue)

9.62±1.22** 11.95±1.61** 7.41±1.12** 2.01±0.18 10 days 14.13±1.62*** 18.48±2.19*** 10.81±1.02*** 1.93±0.16 20 days 20.28±1.31** 24.02±1.21** 22.06±1.67*** 1.95±0.19 30 days

Xanthine Oxidase (µmol/min/mg

protein) 2.82±0.32*** 3.77±0.19*** 2.23±0.23** 5.32±0.91 10 days 1.29±0.16*** 2.73±0.20*** 1.53±0.04*** 5.88±0.60 20 days 0.45±0.04** 1.81±0.16** 0.74±0.08** 6.15±0.32 30 days

Lactate dehyrogenase (µmol/min/mg

protein) Data are presented as mean ±SD. SD= Standard deviation, * Significant P<0.05 ** High significant P<0.01 *** Very highly significant P<0.000

وبور هورس (B)والجرذان التي تم إعطائها مشروب الطاقة بايسون ) A(يوضح المقارنات الثنائية ومستوى المعنوية بين الجرذان السليمة ) ٣(جدول (C) بول وريد (D) شملت نشاط بعض المؤشرات في مصل الدم وذلك باستخدام كال على حده، والتيANOVA .

Parameters Groups

Hemoglobin Bilirubin

A(B) 10days 20 days 30 days 10 days 20 days 30 days A(C) 23 NS 6.0 *** 4.3 *** 0.08 ** 0.15 N.S 0.04 * A(D) 27 NS 4.6 *** 3.5 *** 0.06 * 0.06 N.S 0.04 * B(C) 3.3 * 5.5 *** 3.9 *** 0.14 *** 0.05 N.S 0.05 * B(D) 0.4 NS 1.4 NS 0.77 NS 0.02 N.S 0.09 N.S 0.00 N.S C(D) 1.1 NS 0.53 NS 0.43 NS 0.05 * 0.10 N.S 0.01 N.S A(B) 0.67 NS 0.87 NS 0.33 NS 0.08 ** 0.01 N.S 0.00 N.S

Parameters Groups

Triglyceride Cholesterol

A(B) 10days 20 days 30 days 10 days 20 days 30 days A(C) 27.3 ** 1.7 N.S 30.7 ** 0.07 N.S 42.1 * 71.4 N.S A(D) 13.0 N.S 9.0 N.S 29.3 ** 22.8 N.S 39.3 N.S 0.77 N.S B(C) 6.3 N.S 6.3 N.S 36.3 *** 114.7 ** 5.6 N.S 41.4 N.S B(D) 14.3 N.S 10.7 N.S 1.3 N.S 22.7 N.S 2.8 N.S 70.6 N.S C(D) 21.0 N.S 8.0 N.S 5.7 N.S 114.6 ** 36.5 N.S 30.0 N.S A(B) 6.7 N.S 2.7 N.S 7.0 N.S 91.9 * 33.7 N.S 40.6 N.S



- 120 -

(B)يـسون والجرذان التي تم إعطائها مشروب الطاقة با ) A(يوضح المقارنات الثنائية ومستوى المعنوية بين الجرذان السليمة ) 4(جدول

كال على حده ، والتي شملت نشاط النيتريك أوكسيد وإنـزيم الـزانثين أوكـسيديز وإنـزيم الالكتيـت دي (D) وريد بل (C)وبور هورس . ANOVAهيدروجينيز في نسيج القلب وذلك باستخدام

Parameters Groups

Nitric Oxide (µmol/gm tissue)

Xanthine Oxidase (µmol/min/mg protein)

A(B) 10days 20 days 30 days 10 days 20 days 30 days A(C) 27.6 *** 35.7 *** 42.4 *** 5.4 *** 8.9 *** 20.1 *** A(D) 41.7 *** 57.3 *** 75.9 *** 9.9 *** 16.5 *** 22.1 *** B(C) 35.3 *** 49.3 *** 57.4 *** 7.6 *** 12.2 *** 18.3 *** B(D) 14.0 ** 21.6 *** 33.5 *** 4.5 ** 7.7 *** 1.9 N.S C(D) 7.7 * 13.6 ** 15.0 *** 2.2 * 3.3 * 1.8 N.S A(B) 6.3 * 8.0 N.S 18.5 *** 2.3 * 4.3 ** 3.7 **

Parameters Groups

Lactate dehyrogenase (µmol/min/mg protein)

A(B) 10days 20 days 30 days A(C) 3.1 *** 4.3 *** 5.4 *** A(D) 1.5 ** 3.1 *** 4.3 *** B(C) 2.5 *** 4.6 *** 5.7 *** B(D) 1.5 ** 1.2 ** 1.1 *** C(D) 0.58 N.S 0.24 N.S 0.29 N.S A(B) 0.96 * 1.4 ** 1.4 ***

. يوما ٣٠،٢٠،١٠الفحص النسيجي لقطاع يف قلب اجملموعة الضابطة بعد ) ٨(شكل



- 121 -

. يوما ٣٠،٢٠،١٠الفحص النسيجي لقطاع يف قلب اجملموعة اليت اعطيت مشروب الطاقة بايسون بعد ) : ٩(شكل

. يوما ٣٠،٢٠،١٠الفحص النسيجي لقطاع يف قلب اجملموعة اليت اعطيت مشروب الطاقة بور هورس بعد ) : ١٠(شكل

. يوما ٣٠،٢٠،١٠ بعد بوللنسيجي لقطاع يف قلب اجملموعة اليت اعطيت مشروب الطاقة ريد الفحص ا ) : ١١(شكل

وتظهر نتائج الفحص النسيجي لقطاعات يف قلب اجملموعات األربعة

ازدادتالتطورات احلادة اليت أحدثتها مشروبات الطاقة الثالثة والـيت حيث يظهـر .طةحدة مع طول فترة التجربة ومقارنة باجملموعة الضاب

واليتالفحص النسيجي لقطاع يف قلب اجملموعة الضابطة ) ٨(الشكل تظهر ألياف العضلة القلبية ذات السيتوبالزم األمحر واألنوية البيضاوية

). X 400( صبغة اهليماتوكسيلني واأليوسني ،املركزية



- 122 -

الفحص النسيجي لقطاعات يف قلب اجملموعـة ) ٩(ويوضح الشكل يومـا ، ٣٠ ، و ٢٠ ،و ١٠يت مشروب الطاقة بايسون بعد اليت أعط

التلف بطول مدة التجربة ، وخروج مكونات الدم ازديادحيث يتضح بني األلياف القلبية العضلية ، ورشح للخاليا االلتهابية حول األوعيـة

).X 400(صبغة اهليماتوكسيلني واأليوسني . الدموية قطاعـات يف الفحص النسيجي ل ) ١٠(ويوضح الشكل

مــشروب الطاقــة أعطيــتقلــب اجملموعــة الــيت يث يظهر رشح ملكونات الدم يوما ، ح ٣٠،٢٠،١٠باور هورس بعد

لياف القلبية العضلية مع زيادة كبرية يف مسك جدار األوعيـة بني األ التاجية وتكاثر يف اخلاليا الطالئية املبطنة لألوعية من الداخل وخاصة

.X 400)(صبغة اهليماتوكسيلني واأليوسني . يوما٣٠يف القطاع بعد الفحص النسيجي لقطاعـات يف ) ١١(ويوضح الشكل

، ٢٠ ،و ١٠قلب اجملموعة اليت أعطيت مشروب الطاقة ريد بول بعد يوما ، ويظهر رشح للخاليا وحيدة النواة يف الشعريات الدموية ٣٠و

يـا التهابيـة املتالصقة مع احتقان يف األوعية التاجية مع ظهور خال ٣٠وحيدة النواة حميطة باألوعية احملتقنة وخاصة يف القطاعات بعـد

).X 400(صبغة اهليماتوكسيلني واأليوسني . يوما

:املناقشة -٤واضـحة يف من النتائج السابقة الذكر يتضح حدوث اضـطرابات

حدوث خلـل يف كما تظهر النتائج التقديرات البيوكيميائية املختارة وقـد . ت عديـدة ، وهذا ما أكدته دراسا جة ووظائف القلب أنس

أن التركيزات العالية للكربوهيدرات فيها تعمـل أظهرت الدراسات كما أن تناوهلا بكميات . [26]على إذابة طبقة املينا على سطح األسنان

كبرية يؤدي إىل ارتفاع أكسدة الكربوهيدرات واخنفاض يف أكـسدة وحتتوي . [27]ت األخرى مثل شراب الليمون الدهون مقارنة باملشروبا

مشروبات الطاقة على مادة الكافيني اليت تعترب منبهة للجهاز العـصيب وهو أحد مثريات اجملموعـة . [29] املركزي مما يعطي شعورا بالنشاط

العصبية السيمبثاوية الذي جيعل للقهوة والشاي والكوال والشوكوالته تمي إىل فئة العقاقري املثرية ، ويعتقـد وين. ومشروبات الطاقة آثارها

العديد من العلماء أن مضادات االكتئاب من هذه الفئة أيضا حيث أن آثارها السلوكية متاثل أو تشابه آثار األمفيتامينات واملثريات األخرى

واالستخدام املعتدل للمشروبات احملتوية على . للمجموعة السيمبثاوية

احلالة املزاجية ويزيد من االنتبـاه العقلـي الكافيني يؤدي إىل إنعاش والطاقة ، أما االستخدام بشكل مبالغ فيه فإنه قد يؤدي إىل النـشاط الزائد وعدم الراحة واألرق والقلق وحىت الغضب واملـشاجرة كمـا يؤدي تكرار استخدامها إىل اإلدمان حيث تظهر تغريات فـسيولوجية

ضغط الـدم والوظـائف تتضمن تغريات يف معدل ضربات القلب و املعدية املعوية واضطراب التنفس والتشتت ويف حاالت شدة تركيـز

.[29]الكافيني يف الدم يؤدي إىل اإلغماء كما أن استهالك كميات كبرية من مـشروبات الطاقـة

كما [30]جالغنية بالكافيني يسبب ارتفاع معدالت البيلريوبني يف الدم وبات الطاقة الغنية بالكافيني يؤدي إىل ثبت أن االنتظام يف تناول مشر وأظهرت معظـم الدراسـات . [31]اإلصابة بأمراض القلب املزمنة

[32,33]احتواء مشروبات الطاقة على نسب مرتفعة من مادة التيورينومن املعروف أن هذه املادة هلا خصائص مضادة لألكـسدة ومزيلـة

ـ سترول للشقوق احلرة كما تعمل على خفـض معـدالت الكولي بينما أظهـرت دراسـات [34].واجلليسريدات الثالثية يف مصل الدم

أخرى أن ماديت التيورين والكافيني تعمالن على رفع ضـغط الـدم وزيادة نبضات القلب واجلفاف واألرق وتناوهلا بكميات كبرية يسبب

، وال تؤثر على التركيز كما هو شائع عن مـشروبات [35]اإلدمان . [36]الطاقة

ما أن اإلفراط يف تناول ماديت الكافيني والتيورين يعمل كعلى رفع مستوى الكوليسترول يف بالزما الدم مما يزيد مـن خطـر اإلصابة بأمراض القلب الوعائية املزمنة لدى الرجال والنساء على حد

وحتتوي مشروبات الطاقة على نـسب متفاوتـة مـن . [37]سواءكافيني والتيورين تأثريا إجيابيا على اجللوكونوالكتون حيث تؤثر مع ال إضافة إىل أهنا تزيد وبدرجة [38].العمليات العقلية واملزاج لدى الناس

وتؤثر على التفكري والوظائف . [39]معنوية من قوة التركيز والذاكرة وأوضح أن استهالك كميات كـبرية مـن . [40]اخللوية يف الدماغ

حلمض االمـيين اهليموسيـستني مشروبات الطاقة تعمل على تركيز ا Hemocystine يف البالزما وهو من العوامل القوية واخلطرية حلدوث

. [41]أمراض القلب الوعائيةوأكدت الدراسات أن مشروبات الطاقة احملتويـة علـى الكافيني تسبب األرق وعدم القدرة على النوم ، ومعظم األعـشاب

يتم التعـرف علـى اليت تدخل ضمن مكونات مشروبات الطاقة مل



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كما وجد أن مـشروبات [42].آثارها اجلانبية وخاصة عند األطفالالطاقة رمبا تقلل من أضرار تناول الكحول باملقارنة بتناول الكحـول فقط ، حيث تقلل من أعراض التسمم الـيت تـسببها املـشروبات

ا وعند اختبار تأثري مادة الكافيني املوجودة هب. [43] الكحولية منفردةعلى منع النعاس لدى سائقي السيارات وجد أهنا قللـت وبدرجـة معنوية الرغبة يف النوم يف التسعني دقيقـة األوىل مقارنـة باجملموعـة

. [44] الضابطة أن العديـد مـن [45] وأوضحت دراسات عديدة منها

األعشاب واملضافات ملشروبات الطاقة تسبب زيادة تكـوين إنـزمي هو من أهم اإلنزميات املسئولة عن تكوين فـوق و .الزانثني أوكسيديز

األكاسيد وزيادة حدوث الضغط األكسيدي وبالتايل زيادة تكـوين النتريك أوكسيد وخاصة عند تناول تلك املشروبات بكميات كبرية

وحتتوي مشروبات الطاقة على مواد داعمة جلهاز املناعـة . ومتكررة ــتراجالس ــل اس ــيزاندارك Astragalusمث ، Schizandrac ، س

تعمل عن طريق ارتباطهـا باخلاليـا البلعميـة Echianceaإيشيناكيا النتريـك وارتفـاع أوكسيد ومن مث يؤدي ذلك إىل خروج .املناعية

كما أن العديد من األعشاب املـضافة [46]معدالته يف نسيج القلب

، هيدروكـسي سـيترات Ciwujiaملشروبات الطاقة مثل سيوجيا Hydroxyl Citrate واإلفيدرا ، Ephedra تعمـل علـى خفـض

مستويات الكوليسترول واجلليسريدات الثالثية والربوتينات الدهنية يف كمـا تـسبب اخنفـاض نـشاط إنـزمي الالكتيـت دي . الـدم

. [47]هيدروجينيزبات الطاقة مبا أن مشرو املتحصل عليها من النتائج يتضح

تامينـات والعديـد مـن ومواد سكرية وفي حتتويه من أمحاض أمينيه . إذا أخذت بكميات معتدلة بالغة أضرارااملضافات الغذائية ال تسبب

للحصول على أما اإلفراط يف تناوهلا كما هو شائع بني فئات الشباب اضطرابات شديدة يف معدالت الكوليسترول القوة واليقظة فإنه يسبب

وارتفـاع . الدم واجلليسريدات الثالثية والبيلروبني واهليموجلوبني يف مـن هوما يقابل النتريك وإنزمي الزانثني أوكسيديز أوكسيد معدالت

ز مع حدوث تلـف شـديد اخنفاض إلنزمي الالكتيت دي هيدروجيني مما يؤكد على خطورة تناوهلا . ووظائفهوأضرار بالغة ألنسجة القلب

بكميات كبرية وضرورة كتابة بعض التحذيرات على العبوات قبـل .ها تسويق



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المجلة العربية للكيمياء

א :א ƭلة Ɯوǫ علمية Ʈكمة أسسها اƠاد الكيميائيني العرب

بالتعاون مـȜ اجلمعيـة – قسم الكيمياǒ – كلية العلوم – جامعة امللك سعود بالرياȏ –اململكة العربية السعودية الكيميائية السعودية

:رئيȄ التحرير ȸ بȸ عبداǃ الورثان عبدالرƥ. د.أ

ǒجامعة امللك سعود – كلية العلوم –قسم الكيميا – ȏ١١٤ الرياĐ١ ȋ ، . ٢٤: بĐĐ اململكة العربية السعودية

E-mail: [email protected] , [email protected]

: هيǞة التحرير سلطان توȥيȨ أبوعراŸ . د.أ

، األردن اربد ، الريموȫرئيȄ جامعة E-mail: [email protected]

يسري عيسى . د.أ

ǒهورية مصر العربية – القاهرة – جامعة القاهرة –قسم الكيمياƤ E-mail : [email protected]

ǯراǹتنفيذ وإ: )سكرتري اجمللة (عبد الرȸƥ بȸ سعد الطليحي. أ

ǒجامعة امللك سعود – كلية العلوم –قسم الكيميا – ȏ١١٤ الرياĐ١ – ȋ . ٢٤بĐĐ E-mail : [email protected]

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