HERMIN HARDYANTI UTAMI (111 304 0202) CHEMISTRY DEPARTMENT MATHEMATIC AND SCIENCE FACULTY STATE UNIVERSITY OF MAKASSAR 2012
Apr 16, 2015
HERMIN HARDYANTI UTAMI (111 304 0202)
CHEMISTRY DEPARTMENT
MATHEMATIC AND SCIENCE FACULTY
STATE UNIVERSITY OF MAKASSAR
2012
APPROVAL SHEET
Complete report of organic chemistry II with the tittle “Piperine Isolation
from Pepper and Chemical Properties of Piperine” was created by :
Name : Hermin Hardyanti Utami
Reg. Number : 1113040202
Group : VII (seventh)
Class : ICP Chemistry “B”
after corrected and consulted by assistant and assistant coordinator, so it was
accepted.
Makassar, December 2012
Assistant Coordinator Assistant
Fandi Ahmad, S.Pd Yanti Madong, S.Si.
Known by
Responsibility Lecturer
Drs. Hj. Ramdani, M.Si.
NIP : 19630221 198803 2 001
A. Title of experiment
Piperine Isolation from Pepper and Chemical Properties of Piperine
B. Purposes of experiment
At the last experiment, student can be understand about :
1. Isolation principle of organic compound from nature material, especially
alkaloid group.
2. Isolation technique in nature material, continue extraction, separating, and
it purification.
3. Knowing chemicals properties of piperine based on the structure and it
degradation result.
C. Background
A natural product is a compound synthesized by a plant or an animal.
Alkalords are natural products that contain one or more nitrogen hetero atoms and
are found in leaves, bark, roots or seeds of plants. Examples include caffeine
(found in tea leaves, coffee beans, and cola nuts) and nicotine (found in tobacco
leaves). Morphine is an alkaloid obtained from opium, the juice derived from a
species of poppy.
Morfphine is so times stronger than aspirin as an analgesic but it is addictive
and suppresses respiration. Heroin is a syntetic compound that is made by a
cetylating morphine. (Bruice, P.Y. 2003:884).
With identities and amounts of amino acids known, the peptide is sequenced
to find out in what order the amount amino acid are linked together. Much peptide
sequencing is now done by mass spectrometry, using either electropspay
ionization (ESI) or matrix-assisted laser desoroption ionization (MALDI) linked
to a time of flight (TOF) mass analyzer as described. Also in common use is a
chemical method of peptide seqencing by called the Edman degradation.
Edman degradation involves treatment of a peptide with phenil isothiocyanale
(PLTC), C6H5-N = C = S, followed by treatmen with trifluoroastic acid. The first
step splits tne N-terminal residue from the peptide chain, yielding an anilinotr
zlinone (ATZ) derivative with shortened peptide. Futher acid catalyzed
rearrangement of the ATZ derivative with aqueous acid converts it into a
phenylthrohydantion (PTH), which is derivative with aquous acid converts it into
by chromatographicaly by comparasion if its solution time with the known elution
times of PTH derivaties of the 20 common amino acids. The chain-shortened
peptide is then automatically resubmitted to anther round of Edman degradation
(Mc Murry, John. 2008 : 1031).
Protein metabolism. Catabolism of proteins: transamination
1. The pathway to amino acid catabolism
a. The amino group is removed by transamination
b. What remains is converted a compound that enters the citric acid cycle.
2. Transamination
a. The NH2 group of an amino acid adds to the aldehyde group of pyridoxal
phospate to form an imine.
b. The imine tautomerizes to different imine.
c. The second imine is hydrolyzed to give an keto acid and an amino
derivative of pyridoxal phosphate.
d. The pyridoxal derivate transfers its amino group to -ketoglutarate, to
regenerate pyridoval phasphate and form glutamate.
3. Deamination
The glutamate from transamination undergoes oxidative deamination to yield
ammonium ion and -ketoglutarate (McMurry, Susan. 2008:796).
Spectrophotometric metods, involving UU absorbance measurements at the
pipenrine absorption maximum near 343 nm measure predominantly piperine but
include some contribution from piperyline, piperettine and any other components
having substantial absorption at this wave lenght. Piperine is extracted from
ground pepper by refluxing 3 hour with ethanol (96% v/v) whilst mogenized
oleoresins are simply dissolve solvent (Wood, A.B. 988:55).
Piperine [(1-5-(1,3) – benzodioxol -5 – yl.) . 1 -0 x 0 – 2, 4 – pentadieny]
piperidine) an alkalord responsible for the pungency of black that pepper and long
pepper. Systemic pharmacological studies on the piperine have revealed that this
compound elicited diverse was pharmacological activities, analgesic, anty-pyretic,
antrinflammatory, anti convulsant and CNS depressant activities. Piperine was
isolated from piper nigium Linn. (Piperaceae) and identified by TLC. Piperine
was fabricated into alginate beads by using sodium alignate. The main aim of this
study was to demonstrate that the sutained release of piperie from alginate beads
by invitro evalution. The drug release studies were showed that the alginate beads
sustained the release of drug with % drug released in hours (Madhavi, B.B.
2009:156).
D. Chemical and Equipment
1. Chemical
a. Pepper sample
b. Aquadest (H2O)
c. Ethanol (C2H5OH) solution
d. Alcoholist potassium hydroxide (KOH alcoholist) solution
e. Potassium Permanganate (KMnO4) solution
f. Concentrated hydrochloric acid (HCl) solution
g. Aqueous hydrochloric acid (HCl) soution
h. Aqueous sodium hydroxy (NaOH) solution
i. Molisch reagent
j. Benedict reagent
k. Aluminium foil
l. Matches
m. Litmus paper
n. Universal indicator
o. Filter paper
p. Whatmann filter paper.
2. Equipment
a. Soxclet equipment (1 set)
b. Analytical scale (1 piece)
c. Buchner funnel (1 piece)
d. Test tube (4 pieces)
e. Test tube rack (1 piece )
f. Stir bar (2 pieces)
g. Dropping pipette (5 pieces)
h. Beaker glass 50 ml and 250 ml (2 pieces)
i. Funnel (2 pieces)
j. Graduated cylinder 10 ml, 25 ml and 100 ml (1 piece)
k. Bunsen burner (1 piece)
l. Triangle (1 piece)
m. Asbestoz Gauze (1 piece)
n. Spray flask (1 piece)
o. Water bath (2 pieces)
p. Round flask 250 ml (2 pieces)
q. Soft cloth and rough cloth (1 piece)
E. Work Procedure
1. Piperine Isolation
a. 20 grams of pepper sample was balanced on analytical scale and was filled
into soxchlet that has covered by filter paper.
b. It was added by ethanol with 3 times of circulation.
c. It was soxcheletd by 15 times circulation.
d. It was let stand and added by 60 ml alcoholist KOH.
e. It was heated on water bath until concentrated and stirred.
f. It was filtered by funnel and divided into 3 part (for piperine isolation,
degradation, and it reaction).
g. It was decreased the volume and let stand on the temperature until 1 day.
h. It was filtered on Buchner funnel and the residue was took.
2. Piperine degradation
a. 1/3 part of isolation result was decreased the volume by reflux it.
b. It was added by HCl 10 M and stirred until dissolve and forming yellow
precipitate.
c. It was let stand on the temperature until 1 day.
d. It was filtered on Buchner funnel and the residue was took.
3. Piperine reaction
a. 3 pieces of test tube was filled by 2 ml of piprine isolation result.
b. It was added by HCl and reacted with :
1)Test tube 1st was reacted by KMnO4
2)Test tube 2nd
was reacted by benedict reagent
3)Test tube 3rd
was reacted by molisch reagent.
c. Each was neutralized the pH and was checked by litmus paper.
F.Observation result
No Activity Result
1.
2.
3.
4.
5.
6.
Isolation of Piperine
Balanced dust paper on the
analitycal scale
Covered with filter paper, filled
to soxlet and added ethanol
The mixture soxlet
1st circulation
2nd
circulation
3rd
circulation
4th
circulation
5th
circulation
6th
circulation
7th
circulation
8th
circulation
9th
circulation
10th circulation
11th circulation
12th circulation
13th circulation
14th circulation
15th circulation
Soxlet mixture heat 10 minutes
Added 60 ml KOH alkoholis
Stir and filtered
25.0124 g
3 times circulation of soxlet
11 minutes
7 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
6 minutes
Yellow solution
Yellow solution
Yellow solution
7.
8.
9.
10.
Devided the solution
Vaporated until saturated (b)
Let it stand 1 day
Filtered it with ethanol
a. 6 ml reaction
b. 59 ml for vapor (isolation)
c. 59 ml for reflux (degradation)
Brown solution
Brown solution
Not crystall
1.
2.
3.
4.
Degradation (c)
59 ml solution reflux
Vapored it until 10 minutes
Added 10 M 10 ml HCl and 7
ml H2O
Filtered with ethanol
Dark yellow solution
Yellownish brown solution
Yellownish brown
Not crystall
1.
2
3
Piperin reaaction
2 ml piperin+HCl dilute
Added NaOH
5 drops KMnO4
2 ml piperin+HCl dilute
Added NaOH
5 drops Benedict
2 ml piperin+HCl dilute
Added NaOH
5 drops Molish
Acid solution
Unknown pH
2 layers :
1st :yellow
2nd: turbid (negative)
Acid solution
Unknown pH
Greenish brown (negative)
Acid solution
Unknown pH
Yellow solution (negative)
G. Data Analyzed
1. Isolation of piperine
Known. Weight of sample : 25.0124 g
Mr of Pyperine (C17H19O3N) : 285 g/mol
Mr of pyperidine (C5H10NH) : 85 g/mol
Question=….?
Mass of theory =
=
= 0.2982
2. Degradation of Piperin
Dik : Weight of sample : 25.0124 g
Weight molecule of pyperine (C17H19O3N) : 285 g/mol
Weight molecule of piperidine hydrokloride (C5H10NHCl): 120,5
gram/mol
Weight of theory =
=
= 0,4228
H. Discussion
1. Isolation of piperine
Isolation is seperating process organic compound from nature. In this
experiment would be isolated is pepper that have done smoothly and technique
that we used is continue extraction(soxletation), separation and purity from
another compound that contain in black pepper. The principle soxletation is
pulling chemical compound that done by the dust put on the filtering paper, the
liquid heated in round flask until vapor and condetation by condensor ball become
liquid molecule that drops until in the syfon surface, all of liquid will be down and
back to theflask occured capiler pipes until became circulation. In this experiment
the sample is black pepper that want to get the piperine. Piperine is the alkaloid
responsible for the pungency of black pepper and long pepper, along with
chavicine (an isomer of piperine). Alkaloids are a group of naturally occurring
chemical compounds that contain mostly basic nitrogen atoms.
At this experiment, solvent that used is etanol. Etanol as solvent to solute
all of compound that want in black pepper. Etanol used because has same polarity
is polar until ethanol can solute as the prinsiple like dissolve like. Ethanol used
because volatil properties where in hot condition easy soluble piperin and in cold
condition easy to formed crystall. Then we do extraction continue until 15 times
circulation. At circulation process used boiling stone in flask solvent that purpose
is to keep the pressure and temperature of solution for stable and not occured
explosion.
Process that happen belong circulation is ethanol solvent heated in round
flask until vapor and colded used condensor, until fall become liquid to sample
(black pepper) to soluble active element in sample black pepper and if liquid
solvent have done reached the bottom of syfon then all of liquid from ethanol will
be solute that have in syfon surface will be out occured small pipes to round flask
in this process will occured continue and occured soxletation process, that process
we call the principle of continue extraction. From soxletation process will be
produced 124 ml extraction and in soxletation process occurs 15 circulations
produced transparant yellow extract. After that, extract will dried belong 10
minutes until the solution concentration. It is because to vapor ethanol that contain
in solution. Then added KOH alkoholis 10%. The function is to bond piperine
from solution because occured hydrolysis and to avoid the compotition in
compound other it the function of KOH for get piperine from concentration
extract where in the extract exist another compound when added KOH alkoholis
and caused piperine in extract react become salt acid piperate and to eliminated
other compound because in the extract still contain purity compound. Purity
compound cause piperine compound is alkaloid compound from amida that
sustain hydrolisis reaction. So, alkoholis of KOH for isolated piperine in salt is
piperate acid. Then solution will stir and filter. Filtering process to filtrate can be
seperated from purity.
After that, the filtrate divede 3 part, first part 6ml for piperine reaction,
59ml for piperine isolation and another part for degradation. In isolation of
piperine, the extract will concentrated until saturated and easy formed crystall.
Then, keep it 1 days and not formed crystall. Then crystalitation with ethanol
based on the principle of recriztalitation based the different principle in solubilty
in the solvent and temperature. In the room temperature, piperine compound in the
crystall and polar properties and soluble in etanol as solvent, then piperine in the
filtrate will soluble but purity compound will be left in the residue. In this
process, we didnt get crystall it is because when heated apperantice not attend to
see the extract and the extract exhaused vaporated.
Reaction
N
O
O
O
CH 3CH 2OH+KOH
piperine
ethanol
NH
+O
OH
+ CH 3CH 2OK
. Equation reaction:
1.
N
O O
O
+C 2H 5OH
N+
O O
O
H
+ CH 3CH 2O-
2.
N+
O O
O
H
+ CH 3CH 2O-
NH
+
C+ O + CH 3CH 2O
-
3.
NH
+
C+ O + CH 3CH 2O
-
NH
+
CO
OH
+ CH 3CH 2OK
Piperidine piperate acid
2. Piperine degradation
Degradation is decompotition of compound become a simple compound.
In this thing, piperine degradation to get salt of piperate acid. In this degradation
process, extract will be reacted with KOH 10% (same with piperine isolation) then
concentrated belong 5minutes, produced dark yellow solution. Then, it mixture
reflux belong a hour. This reflux to occurred hydrolysis reaction perfectly. In this
process happen is solvent is KOH filled with piperin in round flask, then heat it.
Vapor of solvent condentation by condenser become solvent molecule until down
to round flask and will solute back piperine that in the flask until surface and all of
by continue. Then suspentition by water to dilute and hydrolysis piperine. Then
add HCl 10M to neutralization because hydrolysis occur in base condition and
must neutralization. Reaction:
O
OHOOC
O
OO-OC
+ H+
Salt piperate acid piperate acid
HCl 10M addition to get proton until formed piperate acid occurred
hydrolysis reaction. Next, solution keep silent belong one day then filtered and
crystallization with ethanol. This process not crystal formed because apperantice
mistaken when reflux and added HCl or water.
3. Reaction of piperine
a. KMnO4 reaction
This experiment we show that piperine easy oxidate because have double
bond. This experiment, alkoholis solution add with HCl dilute to decreasse pH
because solution in base condition to get netral solution. Then reacted with
KMnO4 to oxidate piperine and produce two layers, yellow in uplayer and turbity
in the down layer. It is not same with theory must be purple. It is caused while
netralization not universal indicator and only litmus paper and don’t know the
solution netral or not if only used litmus.
Reaction is
b. Benedict reaction
At this experiment to show that piperine have aldehide group that is
reducing group. Solution that get from isolation added with HCl diluted to
decreasse pH because solution in base condition to get netral solution. Solution
reacted with benedict reagent produce green brown solution. It is not same with
theory must result brick red, green or yellow. It is not same with theory because
piperine still contain purity compound and while netralization not universal
indicator and only litmus paper and don’t know the solution netral or not if only
used litmus.
Reaction :
c.Molish reaction
The purpose of this experiment to identify existence of epoxy group that
sign with furfural ring in piperine. Alkoholis solution from extraction added with
HCl dilute to decreasse pH because solution in base condition to get netral
solution. Then reacted with Molish reaction and formed yellow solution. It is not
same with theory because piperine still contain purity compound and while
netralization not universal indicator and only litmus paper and don’t know the
solution netral or not if only used litmus.
Reaction is
N
O
O
ON
H
HCl + CH3CH3
O
+O
O
piperidine piperanol epoksi
hydrochloride
I.Closing
1. Concusion
a. The principle of organic compound from chemical nature specially alkaloids
extraction alkaloid in acid solution in water and netral or base compound from
mixture to separate with solvent. After this solution base so alkaloid set with
extraction in solvent.
b. Technique isolation in chemical nature is extraction continue, purifying with
method crystalization and purifying step with determine melting point.
c. Piperine reaction with KMnO4 to show that piperine easy oxidezed. Piperine
reaction with benedict to test for carbonyl group in piperine can be reduced by
Cu2+
. Piperine reaction with molish reagent to find a ring that piperine furfural
in epoxy groups.
2.Suggestion
a. Apperantice must be attend when vapor the extraction,
b. Laboran must prepare our reagent before experiment.
BIBLIOGRAPHY
Bruice, P.Y. 2003.Orgnic Chemistry 4th
Edition.New York. Longman Hill.
Madhavi, B.B. 2009. Extraction, Identification, Formulation and Evaluation of
Piperine in Alginate Beads. India. International Journal of Pharmacy
and Pharmaceutical Sciences.
McMurry, John. 2008. Organic Chemistry 7th
edition.USA. Thomson Learning,
Inc.
McMurry, Susan. 2008. Study Guide and Solutions Manual Fundamental of
Organic Chemistry. USA. Thomson Learning, Inc.
Tim Dosen. 2012. Penuntun Organik 2. Makassar.Universitas Negeri Makassar.
Wood, A.B. 1988. Piperine Determination in Pepper (Pipper ningrum L.) and its
oleorensis A reversed phase High-performance Liquid
chromatographic Method. England. Flavour and Fragrance Journal.
Answer Question
1.Using potassium hydroxide to remove resin(said to contain chavicine an isomer
of piperine.
2. Degradation of piperine
O
OHOOC
O
OO-OC
+ H+
Salt piperate acid piperate acid
3. Reaction of Piperine
a. Reaction with KMnO4
b. Reaction with benedict
c.Reaction with Molish
NH
HCl + CH 3CH 3
O
+O
O
piperidine piperanol epoksi
hydrochloride
N
O
O
O