Figure no. 5.1.1.1.1.1.1: The transverse sectional …shodhganga.inflibnet.ac.in/bitstream/10603/72710/15/15...Formulation, characterization and evaluation of pharmaceutical dosage

Figure no. 5.1.1.1.1.1.1: The transverse sectional view of the skin.

Figure no. 5.1.1.1.1.2.1: Overview of the dreadful effects of ultraviolet radiations on

the skin.

Figure no. 5.1.1.1.1.3.1: Pictorial representation of the acne condition.

Figure no. 5.1.1.1.1.3.2: Psychological concern about the Acned looks.

Figure no. 5.1.1.2.1.9.1: Manilkara zapota plant at a glance.

M. zapota plantation in the Campus. M. zapota fruits

M. zapota leaves and bark

Figure no. 5.1.1.2.2.5.1: Photograph of Lemongrass herb.

Figure no. 5.1.4.2.1: Authentication certificate of Manilkara zapota plant.

Figure no. 5.1.4.4.1: Preliminary research on Ultraviolet Spectrums of the extracts.

Hydroalcoholic extract of M. zapota bark. Ethanol extract of M. zapota bark.

Ethyl acetate extract of M. zapota bark Petroleum ether extract of M. zapota bark

Ethanol extract of M. zapota seed Hydro-alcoholic extract of M. zapota leaves

Figure no. 5.1.4.4.2: Comparison of Spectrum of hydroalcoholic extract of M.

zapota leaves obtained by different methods of extraction.

Soxhlet extraction Maceration process

Figure no. 5.1.4.5.1.1: Manilkara zapota hydroalcoholic extracts of leaf and bark.

M. zapota leaf extract M. zapota bark extract

Figure no. 5.1.4.5.3.2.1: Photographs of the TLC studies.

Phenols Alkaloids

Bark Leaves Bark Leaves

Terpene Glocoside

Bark Leaves

Flavanoid Coumarin

Gallic acid

Leaves Bark

Quercetin Bark Leaves

Figure no. 5.1.4.6.2.1: FTIR spectra of hydroalcoholic extract of Manilkara zapota

leaves.

Figure no. 5.1.4.6.2.2: FTIR spectra of hydroalcoholic extract of Manilkara zapota

bark.

y = 0.013x + 0.448 R² = 0.966

Ab

sorb

ance

A

bso

rban

ce

y = 0.018x - 0.014 R² = 0.990

Figure no. 5.1.4.7.1.1: Standard curve of Gallic acid.

1.2

Standard curve of Gallic acid

1

0.8

0.6

0.4

0.2

0

0 10 20 30 40 50

Concentration (mcg/ml)

Figure no. 5.1.4.7.2.1: Standard curve of Quercetin.

0.7000

Standard curve of Quercetin

0.6000

0.5000

0.4000

0.3000

0.2000

0.1000

0.0000

0 5 10 15 20 25 30 35 40


% I

nhib

ition

%

Inh

ibiti

on

Figure no. 5.1.4.7.8.1: Comparison of graphs showing DPPH radical scavenging

potential.

DPPH radical scavenging potential of the extracts.

70

60

50

40

30

20

10

0

20 40 60 80 100


Hydroalcoholic bark

extract

Hydroalcoholic Leaves

extract

Figure no. 5.1.4.8.2.1: Comparison of graphs showing Hydroxyl radical scavenging

potential.

Hydroxyl radical scavenging activity of the extracts

90

80

70

60

50 Hydroalcholic M. zapota

40 leaves

30 Hydroalcoholioc M.

20 zapota bark

10

0

200 400 600 800 1000


% S

cave

ngin

g

Figure no. 5.1.4.8.3.1: Comparison of graphs showing Superoxide free radical

scavenging activity of extracts.

Superoxide free radical scavenging activity of the extracts

90

80

70

60 Gallic acid

50

40 Hydroalcoholic M.

30 zapota leaves extract

20 Hydroalcoholic M.

10 zapota bark extract 0

50 100 150 200 250

Concentrations (mcg/ml)

Figure no. 5.1.4.9.0: Authentication certificate of Cymbopogon citratus plant.

Figure no. 5.1.4.10.1.1: Effective absorption spectrums.

i. Spectra using hydroalcoholic ii. Spectra using hydroalcoholic M. zapota leaves extract M. zapota bark extract

iii. Spectra using lemongrass oil iv. Spectra using Combination of M. zapota leaves extract and Lemongrass oil

v. Spectra using Standard : vi. Spectra using Standard : Benzophenone Para amino benzoic acid.

vii. Spectra using EKRAN 30® lotion viii. Spectra using SUNKARE 50® Gel

Figure no. 5.1.4.11.1.1: Candida albicans yeast test.

i. Positive control: Melanocyl® Cream.

ii. M. zapota extract

iii. Negative control: Candida albicans seeded agar

Figure no. 5.1.4.11.2.1: Photodegradation curves

Formulation, characterization and evaluation of pharmaceutical dosage forms comprising Manilkara zapota extracts. 206



Abs

orba

nce

Figure no. 5.1.4.11.3.1: Standard curve of 1,1,3,3-tetraethoxypropane.

0.18

0.16

Standard Curve of 1,1,3,3-tetraethoxypropane

0.14

0.12

0.1

0.06

0.04

y = 0.0171x - 0.007 R² = 0.9965

0.02

0

0 2 4 6 8 10 12

Concentration of MDA (Micromoles/ml)

Figure no. 5.1.4.12.1.1: Experimental set up for antiacne studies of the M. zapota

extracts against S. epidermidis.


Figure no. 5.1.4.12.1.2: Antiacne studies of the M. zapota extracts against S.

epidermidis.

M. zapota leaves extracts

M. zapota bark extracts


Zon

e of

Inh

ibit

ion

(mm

)

Figure no. 5.1.4.12.1.3: Bar diagram showing comparison of Zone of Inhibitions

against S. epidermidis.

Zone of Inhibitions against S. epidermidis using Cup plate method

70

Hydroalcoholic M. zapota

60 leaves extract

Combination of M. zapota 50 leaves and Lemongrass oil

Hydroalcoholic M. zapota 40

bark extract

30 Combination of M.zapota

bark and Lemongrass oil

20 Clindamycin

10 Erythromycin

0

0.1 0.25 0.5 1 2 4

Concentration (%w/v)

FACECLIN Cream

Figure no. 5.1.4.12.1.4: Experimental set up for antiacne studies of the M. zapota

extracts against P. acnes.

Maintaining an Anaerobic Incubation of petriplates

Environment. in Anaerobic condition.


Zon

e of

Inh

ibiti

on (m

m)

Figure no. 5.1.4.12.1.5: Antiacne studies of the M. zapota extracts against P. acnes.

Figure no. 5.1.4.12.1.6: Bar diagram showing comparison of Zone of Inhibitions

against P. acnes.

Zone of Inhibitions against P. acnes by Cup plate method.

45

40

35

30

25

20

15

10

5

0

0.1 0.25 0.5 1 2 4

Concentration (%w/v)


leaves extract

Combination of M. zapota

leaves and lemongrass oil


bark extract

Combination of M. zapota

bark and lemongrass oil

Clindamycin

Erythromycin

FACECLIN Cream


Figure no. 5.1.4.12.2.1: Determination of Minimum Inhibitory Concentration of the

M. zapota extracts against S. epidermidis and P. acnes.

Against S. epidermidis

Against P. acnes

Figure no. 5.1.4.13.1: Appearance of the Formulation.

Cream top view Side view of the Cream


Figure no. 5.1.4.14.2.1: Observation of the skin staining test.

Before application of any product Post application of crude extract

Post application of 1%w/w cream


Figure no. 5.1.4.14.5.1: Photograph of the formulation observed under the

microscope.

Formulation F4 Formulation F5

Figure no. 5.1.4.14.7.1: Experimental set up for the In vitro Diffusion studies.


% C

on

stit

ue

nts

Pe

rme

ate

d a

cro

ss t

he

sk

in

She

ar R

ate

(RP

M)

Figure no. 5.1.4.14.7.2: Graph showing In vitro permeation extent of the actives in

the cream.

In vitro Diffusion study of the Formulated cream

70

60

50

40

30

20

10

0

0 0.25 0.5 1 2 4 6 8 10

Time (Hours)

diffusion of the

constituents at 310 nm

Diffusion of the

consituents at 279.5 nm

Diffusion of the

constituents at 255.5

nm

Figure no. 5.1.4.14.8.1: Rheogram of Cream.

Rheogram of Viscosity (cP) vs Shear rate (RPM)

60

50

40

30 Up Curve

20 Down Curve

10

0

0 50000 100000 150000 200000 250000 300000

Viscosity in Centipoise (cP)


Figure no. 5.1.4.14.9.1: Stability studies of the cream formulation at the sixth month.

Spectra obtained of the cream formulation at refrigeration temperature.

Spectra obtained of the cream formulation at 25ºC

Spectra obtained of the cream formulation at 40ºC


Figure no. 5.1.4.15.1.1: Sun Protection Factor Scan I.

Figure no. 5.1.4.15.1.2: Sun Protection Factor Scan II.

Figure no. 5.1.4.15.1.3: Sun Protection Factor Scan III.



Figure no. 5.1.4.17.1.1: Observations for skin reactions in Acute Dermal Irritation

test.

Individually housed animals on application of the formulation

Day 1: Skin Observation Day 14: Skin Observation


Figure no. 5.1.4.17.1.2: Histopathology of the skin sample in Dermal Irritation test.

(E: Epidermis, D: Dermis)

Test Animal No. 1

Test Animal No. 2


Figure no. 5.1.4.17.2.1: Observations for skin reactions in Acute Dermal Toxicity

Studies.

Day 1:Test dose 1 Day 14:Test Dose 1

Day 1:Test Dose 2 Day 14: Test Dose 2


Figure no. 5.1.4.17.2.2: Histopathology of skin for group 1 (test dose – 1000 mg/kg) in

Acute Dermal Toxicity study.

(E: Epidermis; D: Dermis)

Figure no. 5.1.4.17.2.3: Histopathology of skin for group 2 (test dose – 2000 mg/kg) in

Acute Dermal Toxicity study.

(E: Epidermis; D: Dermis)


Table No. 5.2.1.1.2.1: Fitzpatrick classification for Skin types.

Skin type Skin type Suggested SPF

I Always burns, never tans 15 >

II Burns easily, tans minimally 15

III Burns moderately (avg Caucasian) 10-15

IV Burns minimally, tans well (Olive skin) 6-10

V Rarely burns, tans profusely (Brown skin) 4-6

VI Rarely burns, tans easily,

deeply pigmented.(Black skin)

none

Table no. 5.2.1.1.3.1: Marketed Products for treating Acne.

Product Active

Erytop® Clindamycin

Deriva-CMS® Adapalene and

Clindamycin

Clearasil® Benzoyl peroxide

Retino-A® Tretinoin


Table no. 5.2.1.1.3.2: Sensitivity and Resistance reported against Antibiotics

(Bahmani et al., 2008).

Active Sensitivity (%) Resistance (%)

Clindamycin 50 50

Tetracycline 65 35

Erythromycin 48 52

Benzoyl peroxide 75 25

Clindamycin + Benzoyl peroxide 63 27

Erythromycin + Benzoyl peroxide 67 23

Cephalexin 40 60

Neomycin 20 80

Table no. 5.2.4.3.0: Observed proximate values.

Sr.

No.

Parameter Determined value

M. zapota leaves M. zapota bark

Extractive value

1. Alcohol soluble extractive value 5.6 % w/w 38.4 % w/w

2. Water soluble extractive value 16 % w/w 31.2 % w/w

Ash value

3. Total Ash Value 3.0 ±0.3 % 6.5 ±0.5 %

4. Acid insoluble ash value 2 ±0.1 % 2 ±0.2 %

5. Water soluble ash value 0.01 ±0.001 % 5.5 ±0.1 %

* Values expressed as mean ± SD of three replicate analyses.

Table no. 5.2.4.5.3.1.1: Results of the Chemical test.

S.No.

Constituents

Extracts M. zapota leaves M. zapota bark

1. Triterpenoids:Hirshonn reaction + + 2. Saponins:Foam test + +

3.

Steroids Lieberman-Burchard test + + Lieberman test - - Salkowski reaction + +

4. Phenols + +

5.

Flavones Shinoda test - + NaOH + -

6. Flavanones & coumarins + - 7. Anthocyanins - -

8.

Carbohydrates Molish‟s test + + Reducing sugars

Benedict‟s test

+

+

9.

Alkaloids Dragendroff‟s test + + Wagner‟s test + + Mayer‟s test + + Hager‟s test + +

10. Quinones - +

11.

Tannins Ferric chloride test + + Basic lead acetate + +

12.

Proteins Biuret - - Millon‟s test + +

13.

Amino acids Ninhydrin + - Tyrosin - -

14.

Gycosides i)Anthraquinones glycosides Borntrager‟s test - + Modified Borntrager‟s test - - ii)Cardiac glycosides Legal‟s test (Cardenoloids) - + Keller-Killiani test(deoxy sugars) + + Liebermann‟ test (Bufadenoloids) - - iii)Saponin Glycosides

Foam test

+

+



Table no. 5.2.4.6.1.1: Physicochemical characteristics of the hydroalcoholic extracts of M. zapota.

Sr.

No.

Tests

Hydroalcoholic extracts


1. Color Dark green Dark brown

2. Odor Characteristic Characteristic

3. Consistency Free powder Free powder

4. Solubility Freely soluble in 70% ethanol, sparingly soluble in absolute ethanol and water.

Freely soluble in 70% ethanol, sparingly soluble in absolute ethanol and water.

Table no. 5.2.4.6.2.1: Observations of FT-IR spectra of the hydroalcoholic extracts of M. zapota.

Sr. No.

Hydroalcoholic extracts


Peak (cm-1) Interpretation Peak (cm-1) Interpretation

1. 3385.6 O-H stretch 3293.7 O-H stretch

2. 2927.7 C-H stretch 2933.5 C-H stretch

3. 1617.9 C=O stretch 1619.2 C=O stretch

4. 1448.6 -CH2-CH bend 1444.5 C-H bend and C-C stretch

5. 1376.3 Carboxylate anion stretch

-- --

6. 1238.5 C-N Aromatic 1248.3 C-O/O-H bend of phenol

7. 1045.5 C-O/O-H bend of phenol

1067.8 O-H bend , primary alcohol

8. 899.8 One H atom Aromatic ring

826.1 C-H bend, two adjacent H


Table no. 5.2.4.7.1.1: Total phenol contents of hydroalcoholic M. zapota extracts.

Sr. No. Hydroalcoholic Extract (mg/g) of Gallic acid equivalent

per gm of crude extracts.

1. M. zapota leaves 330.77 ± 0.71

2. M. zapota bark 333.08 ± 0.96

* Values are expressed as mean ± SD of three replicate analyses.

Table no. 5.2.4.7.2.1: Total flavonoid contents of hydroalcoholic M. zapota extracts.

Sr. No. Hydroalcoholic Extract (mg/g) of Quercetin equivalent

per gm of crude extracts.

1. M. zapota leaves 183.89± 2.3

2. M. zapota bark 47.39± 1.2


Table no. 5.2.4.8.1.1: DPPH radical scavenging potential of hydroalcoholic M.

zapota extracts.

Standard Ascorbic acid M. zapota leaves extract M. zapota bark extract

Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition

4 39.90±0.004 20 42.49±0.007 20 14.72±0.008

8 43.79±0.005 40 47.49±0.009 40 22.54±0.004

12 47.87±0.004 60 49.01±0.002 60 34.49±0.003

16 56.36±0.007 80 54.95±0.001 80 46.84±0.005

20 60.92±0.008 100 57.71±0.006 100 56.01±0.007

IC50 value:

12.5 ± 0.004 µg/ml

IC50 value:

61.2 ± 0.002 µg/ml

IC50 value:

85.4 ± 0.005µg/ml



Table no. 5.2.4.8.2.1: Hydroxyl radical scavenging activity of hydroalcoholic M.

zapota extracts.

Standard Gallic acid M. zapota leaves extract M. zapota bark extract

Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition

20 9.95±0.524 200 47.55±0.005 200 19.25±0.004

40 26.79±0.096 400 53.15±0.009 400 27.35±0.005

60 34.54±0.865 600 67.01±0.007 600 39.87±0.002

100 55.74±0.752 800 72.54±0.001 800 50.74±0.006

120 64.29±0.652 1000 85.18±0.008 1000 63.45±0.005

IC50 value:

86.85 ± 0.046 µg/ml

IC50 value:

210.5 ± 0.005 µg/ml

IC50 value:

752.44 ± 0.003µg/ml


Table no. 5.2.4.8.3.1: Superoxide anion free radical scavenging activity of

hydroalcoholic M. zapota extracts.

Standard Gallic acid M. zapota leaves extract M. zapota bark extract

Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition Conc.

(µg/ml)

% Inhibition

50 20.45±0.034 50 16.53±0.009 50 8.72±0.057

100 37.24±0.025 100 22.34±0.001 100 17.34±0.003

150 48.40±0.008 150 32.27±0.008 150 24.37±0.009

200 59.37±0.004 200 48.76±0.004 200 40.54±0.004

250 85.42±0.001 250 52.74±0.003 250 47.51±0.002

IC50 value:

154.95 ± 0.008 µg/ml

IC50 value:

205.08 ± 0.004 µg/ml

IC50 value:

265.39 ± 0.002µg/ml



Table no. 5.2.4.11.3.1: Concentration of Malondialdehyde (MDA) formed.

Sr.

No

Test Substance Absorbance Concentration of MDA

formed (µmoles/ml)

1 M. zapota leaves extract 0.171 10.47

2 Quercetin 0.123 7.65

3 Gallic acid 0.110 6.88

4 Faceguard® 0.086 5.47

Table no. 5.2.4.11.3.2: A detail note on the marketed formulations comprising

combination of multi extracts.

Product name Active SPF

Protective sunscreen lotion (Himalaya®) Spiked ginger li ly, greater galangal extract, aloe vera

15

Ayur® herbal sunscreen Wheat germ oil, vit E 15

Bio carrot sun protective face and body lotion. (Biotique®)

Carrot oil, carrot seed extract, lodhra bark, quince seed, aloe vera

25

Khadi® herbal sunscreen lotion Aloe Vera, carrot seed oil 30

Badgers Aloe Vera mineral sunscreen Aloe Vera & zinc oxide 16

Korres® watermelon sunscreen Watermelon rind extract 30

Shahnaz® forever sun protective Cream* Sunflower oil, Indian madder, cucumber seed, wild cherry

40

Natural sun SPF 30 Green tea antioxidant 4oz (Aubrey®)

Green tea extract, zinc oxide, titanium di-oxide

30

Bio sandal-red sandalwood lotion (Biotique®)

Sandalwood oil, ashwagandha extract, wheatgerm extract, lodhra

extract, Mesua ferrea extract

50

*Also contains synthetic ingredients: benzotriazolyl, tetramethylbutylphenol


Table no. 5.2.4.12.1.1: Zone of Inhibitions against Staphylococcus epidermidis.

Concentrations Zone of

Inhibition

using

hydroalcoholic

M. Zapota

leaves extract

Zone of

Inhibition using

combination of

hydroalcoholic

M. zapota leaves

extracts and

lemongrass oil

Zone of

Inhibition

using

hydroalcoholic

M. zapota bark

extract

Zone of

Inhibition using

combination of

hydroalcoholic

M. zapota bark

extracts and

lemongrass oil

Zone of

Inhibition

using

Clindamycin

Zone of

Inhibition

using

Erythromycin

Zone of

Inhibition

using

FACECLIN

Cream®

(%w/v) (mm) (mm) (mm) (mm) (mm) (mm) (mm) 0.1 3 ±0.7 9.7 ± 0.3 2.8 ±0.1 5.7 ± 0.2 15.0 ± 0.6 8.2 ± 0.4 9.9 ± 0.4

0.25 8 ± 0.4 17.4 ± 0.5 5.9 ± 0.5 9.3 ± 1.8 22.3 ± 1.8 14.5 ± 0.6 19.4 ± 0.5 0.5 14 ± 0.8 25.3 ± 0.2 11.4 ± 1.2 19.2 ± 0.6 35.8 ± 1.5 26.8 ± 0.6 28.3 ± 0.4

1 21.5 ± 1.1 44.3 ± 1.2 17.3 ± 0.1 28.4 ± 1.4 50.7 ± 0.4 34.5 ± 0.9 49.6 ± 0.1

2 30.2 ± 0.3 55.3 ± 0.2 22.2 ± 0.4 40.2 ± 0.3 62.7 ± 0.5 48.3 ± 1.6 58.5 ± 0.3

4 36.5 ± 0.6 -- 25.2 ± 1.5 -- -- -- --

*Values are expressed as mean ± SD (n = 3)

Table no. 5.2.4.12.1.2: Zone of Inhibitions against Propionibacterium acnes.

Concentrations Zone of

Inhibition

using

hydroalcoholic

M. Zapota

leaves extract

Zone of

Inhibition using

combination of

hydroalcoholic

M. zapota leaves

extracts and

lemongrass oil

Zone of

Inhibition

using

hydroalcoholic

M. zapota bark

extract

Zone of

Inhibition using

combination of

hydroalcoholic

M. zapota bark

extracts and

lemongrass oil

Zone of

Inhibition

using

Clindamycin

Zone of

Inhibition

using

Erythromycin

Zone of

Inhibition

using

FACECLIN

Cream®

(%w/v) (mm) (mm) (mm) (mm) (mm) (mm) (mm) 0.1 1 ±0.2 4.7 ± 0.3 0.8 ±0.5 1.5 ± 0.2 8.0 ± 0.6 5.2 ± 0.4 5.9 ± 0.3 0.25 5 ± 0.5 13.2 ± 0.2 4.3 ± 0.4 9.1 ± 0.5 15.3 ± 0.4 14.5 ± 0.6 14.6 ± 0.2

0.5 11 ± 0.2 22.3 ± 0.4 8 ± 1.0 15.2 ± 0.2 24.8 ± 1.6 26.8 ± 0.6 24.2 ± 0.5

1 18.4 ± 0.6 34.3 ± 0.5 14.3 ± 0.5 26.2± 0.8 36.4 ± 0.2 34.5 ± 0.9 36.3 ± 0.4 2 22.2 ± 0.3 38.3 ± 0.5 19.2 ± 0.6 31.4 ± 0.2 39.4 ± 0.4 38.3 ± 1.1 39.8 ± 0.5

4 30.5 ± 0.4 -- 22.2 ± 1.1 -- -- -- --

*Values are expressed as mean ± SD (n = 3)


Table no. 5.2.4.13.1: Ingredients of Cream Formulation.

Ingredient F 1 F2 F3 F4 F5 Role

M. zapota Leaf extract

1%w/v 1%w/v 1%w/v 1%w/v 1%w/v Active

Lemongrass oil 0.5 % v/v

0.5 % v/v

0.5 % v/v

0.5 % v/v

0.5 % v/v

Active

Stearic acid 12 g 15 g 18 g 20 g 25 g In situ emulsifiers Triethanolamine 0.8 g 1 g 1 g 1.2 g 1.6 g

Liquid Paraffin - - 5 g - 2 g Emollient

Lanolin - - 1 g - 0.5 g Emollient

Isopropyl myristate

1.5 g 1.8g 1 g 2 g 1.5 g Speadability enhancer

Glyceryl Monostearate

- - 4 g - 3 g Emollient

Glycerine 4 g 4 g 4 g 5 g 5 g Humectant

EDTA 0.1 g 0.1 g 0.1 g 0.1 g 0.1 g Chelating agent

Methyl Paraben 0.18 g 0.18 g 0.18 g 0.18 g 0.18 g Preservative

Propyl Paraben 0.02 g 0.02 g 0.02 g 0.02 g 0.02 g Preservative

Purified water Qs. to 100 g

Qs. to 100 g

Qs. to 100 g

Qs. to 100 g

Qs. to 100 g

Vehicle

Table no. 5.2.4.14.4.1: pH of the formulations.

Formulations F 1 F2 F3 F4 F5

Mean pH 6.7 ± 0.2 6.8 ± 0.2 6.8 ± 0.2 6.8 ± 0.2 6.7 ± 0.2

Table no. 5.2.4.14.7.1: In vitro Diffusion studies of the formulated cream.


Table no. 5.2.4.14.8.1: Rheology of the formulation.

Shear Increasing shear

(Up curve)

Dial reading (D1)

Dial reading,

(D1)* Factor

Decreasing shear

(Down curve)

Dial reading (D2)

Dial reading,

(D2)* Factor

0.5 13 260000 4.25 85000

2.5 21.75 87000 12 48000

5 25.75 51500 14.75 29500

10 31 31000 19 19000

20 37.25 18625 24.75 12375

50 41.5 8300 41.5 8300



Table no. 5.2.4.14.9: Stability studies of the formulations.

Table no. 5.2.4.14.9.1: Stability studies of the formulations at refrigeration condition.

Storage Parameters Formulae Day 0 1 month 3 month 6 month

Refrigeration

Color F2 Arylide yellow Arylide yellow Arylide yellow Arylide yellow

F3 Arylide yellow Arylide yellow Arylide yellow Arylide yellow


Odor F2 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil

Faint lemongrass oil

F3 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil


F4 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil


Visual Appearance

F2 Smooth,Uniform Smooth,Uniform Smooth,Uniform Smooth,Uniform



Drug content

F2 324 nm

36.35% 34.36% 33.25% 33.02%

287.5 nm

26.24% 26.25% 24.56% 24.32%

275.5 nm

51.65% 50.57% 48.72% 48.24%

F3 324 nm

36.35% 34.36% 33.25% 33.02%

287.5 nm

26.24% 26.25% 24.56% 24.32%

275.5 nm

52.25% 50.57% 48.72% 45.24%

F4 324 nm

36.35% 34.36% 33.25% 33.02%

287.5 nm

26.24% 26.25% 24.56% 24.32%

275.5 nm

53.78% 51.57% 50.72% 49.46%

pH F2 6.8 ± 0.2 6.8 ± 0.2 6.7± 0.2 6.8 ± 0.2


F3 6.7 ± 0.2 6.7 ± 0.2 6.8 ± 0.2 6.9 ± 0.2

F4 6.7 ± 0.2 6.8 ± 0.2 6.8 ± 0.2 6.8 ± 0.2

Table no. 5.2.4.14.9.2: Stability studies of the formulations at 25ºC.



25ºC F3 Arylide yellow Arylide yellow Arylide yellow Arylide yellow


Odor F2 Strong

Lemongrass oil

F3 Strong

Lemongrass oil

F4 Strong

Lemongrass oil

Strong

Lemongrass oil

Strong

Lemongrass oil

Strong

Lemongrass oil

Medium lemon grass oil Medium lemon grass oil Medium lemon grass oil

Faint lemongrass oil Faint lemongrass oil Faint lemongrass oil

Visual Appearance

F2 Smooth,Uniform Smooth,Uniform Smooth,Uniform Smooth,Unifor m

F3 Smooth,Uniform Smooth,Uniform Smooth,Uniform Smooth,Unifor

m


m

Drug content

F2 324 nm

287.5 nm

36.35% 34.36% 33.25% 33.02% 26.24% 26.25% 24.56% 24.32%

275.5 nm

F3 324

nm

51.65% 49.65% 46.72% 43.24% 36.35% 33.46% 31.89% 30.03%

287.5 nm

26.24% 24.89% 23.73% 21.64%

275.5 nm

52.25% 48.95% 46.78% 44.32%


F4 324 nm

36.35% 33.34% 32.39% 30.68%

287.5 nm

26.24% 24.26% 22.96% 21.75%

275.5 nm

53.78% 51.54% 49.35% 48.65%

pH F2 6.8 ± 0.2 6.7 ± 0.2 6.8 ± 0.2 6.7 ± 0.2

F3 6.7 ± 0.2 6.8 ± 0.2 6.9 ± 0.2 6.9 ± 0.2

F4 6.8 ± 0.2 6.9 ± 0.2 6.8 ± 0.2 6.8 ± 0.2

Table no. 5.2.4.14.9.3: Stability studies of the formulations at 40 ºC.



40 ºC F3 Arylide yellow Arylide yellow Arylide yellow Arylide yellow


Odor F2 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil

Faint

lemongrass oil

F3 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil

Faint

lemongrass oil

F4 Strong

Lemongrass oil

Strong

Lemongrass oil

Medium

lemon grass oil

Faint

lemongrass oil

Visual

Appearance



m


Drug

content

F2 324

nm 36.35% 33.25% 31.68% 29.34%

287.5

nm 26.24% 25.87% 22.64% 20.34%

275.5

nm 51.65% 47.71% 46.27% 40.74%

F3 324

nm 36.35% 34.79% 31.69% 28.63%

287.5

nm 26.24% 24.26% 23.72% 20.58%

275.5

nm 52.25% 48.56% 45.76% 40.87%

F4 324

nm 36.35% 35.57% 31.35% 28.95%

287.5

nm 26.24% 25.22% 19.35% 16.58%

275.5

nm 53.78% 50.23% 47.56% 40.28%

pH F2 6.7 ± 0.2 6.7 ± 0.2 6.7 ± 0.2 6.8 ± 0.2

F3 6.8 ± 0.2 6.8 ± 0.2 6.9 ± 0.2 6.8 ± 0.2

F4 6.8 ± 0.2 6.9 ± 0.2 6.9 ± 0.2 6.9 ± 0.2



Table no. 5.2.4.15.1: Details of Sun Protection Factor Determination.

Sr. No

Test sample

Parameters

Average values

1.

Sample 1

Scans 1 2 3

SPF 2.73 2.72 2.65 2.70

Standard deviation 0.15 0.18 0.15 0.16

UVA/UVB ratio 0.667 0.664 0.655 0.662

Critical wavelength 385.17 385.17 385.00 385.11

Boots Star Rating *** *** *** ***

Table no. 5.2.4.17.1.1: Changes in body weight of Group I animals in Acute Dermal Irritation study.

Days Body weight of Animals (in Gm)

I II III IV V Day 1 213 191 207 204 204 Day 7 226 205 220 222 219 Day 14 230 216 231 236 228

Table no. 5.2.4.17.1.2: Scoring of skin reactions in Acute Dermal Irritation study.

Grading of skin reactions

Animal I II III IV V

Reaction Erythema Edema Erythema Edema Erythema Edema Erythema Edema Erythema Edema

C F C F C F C F C F C F C F C F C F C F

Observation Day

0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0

Day 1 – 3 hour

0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0

Day 1 – 6 hour

0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0

Day 2 0 0 0 0 0 0 0 0 0 1 0 0 1 1 0 0 1 0 0 0

Day 3 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0

Day 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0


Day 11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Total score 0 0 0 0 0 0 0 0 0 4 0 0 5 5 0 0 3 0 0 0

Final Score 0 0 0 0 4 0 0 0 0 0

Table no. 5.2.4.17.2.1: Changes in body weight of Group I Animals in Acute Dermal

Toxicity.


I II III IV V

Day 1 189 171 189 208 176

Day 4 190 175 194 207 180

Day 7 194 182 211 212 186

Day 11 202 189 221 220 190

Day 15 210 191 231 223 196

Table no. 5.2.4.17.2.2: Changes in body weight of Group II Animals in Acute

Dermal Toxicity.


I II III IV V

Day 1 214 169 171 195 166

Day 4 212 169 172 195 169

Day 7 219 178 180 213 173

Day 11 224 185 184 216 180

Day 15 228 190 189 220 185


Table no. 5.2.4.17.2.3: Scoring of skin reactions in Acute Dermal Toxicity study, Test dose 1.





Observatio n Day

0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 1 – 3 hour

0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 1 – 6 hour

0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0

Day 2 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0

Day 3 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0

Day 4 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0

Day 7 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 1 0 0

Day 11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Total score 0 0 0 0 0 4 0 0 0 0 0 0 2 4 0 0 2 4 0 0

Final Score

0 0 4 0 0 0 2 0 2 0

*C: Control applied; F: Formulation applied.

Table no. 5.2.4.17.2.4: Organ weights of Group I Animals in Acute Dermal Toxicity study.

Organs Organ weight of Animals (in Gm) I II III IV V

Brain 1.415 1.962 1.724 1.6 1.8 Heart 0.4 0.69 0.865 0.726 0.59 Lungs 1.215 1.428 1.355 1.667 1.224 Liver 5.78 7.1 8.54 7.9 7.956

Kidneys 1.373 1.44 1.66 1.512 1.515 Uterus-ovar ies 1.94 0.82 0.705 1.04 0.584

Spleen 0.36 0.66 0.832 0.64 0.914


Table no. 5.2.4.17.2.5: Organ to body weight ratio of Group I Animals in Acute

Dermal Toxicity study.

Organ: Body weight

I

II

III

IV

V

Mean

SD

Brain 0.006738 0.010272 0.007463 0.007175 0.009184 0.008166 0.001499

Heart 0.001905 0.003613 0.003745 0.003256 0.00301 0.003106 0.000731

Lungs 0.005786 0.007476 0.005866 0.007475 0.006245 0.00657 0.000845

Liver 0.027524 0.037173 0.03697 0.035426 0.040592 0.035537 0.004861

Kidneys 0.006538 0.007539 0.007186 0.00678 0.00773 0.007155 0.0005

Uterus-Ovaries 0.009238 0.004293 0.003052 0.004664 0.00298 0.004845 0.002566

Spleen 0.001714 0.003455 0.003602 0.00287 0.004663 0.003261 0.00108

Table no. 5.2.4.17.2.6: Scoring of skin reactions in Acute Dermal Toxicity study,

Test dose 2.





Observation Day

0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 1 – 3 hour

0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 1 – 6 hour

0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0

Day 2 0 2 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0

Day 3 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0

Day 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 11 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Day 14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Total score 0 6 0 0 0 0 0 0 0 0 0 0 0 3 0 0 0 0 0 0

Final Score 6 0 0 0 0 0 3 0 0 0

*C: Control applied; F: Formulation applied.


Table no. 5.2.4.17.2.7: Organ weights of Group II Animals in Acute Dermal Toxicity

study.

Organs Organ weight of Animals (in Gm)

I II III IV V

Brain 2.2 1.35 1.7 1.45 1.73

Heart 0.661 0.68 0.59 0.71 0.56

Lungs 1.69 1.3 1.154 1.6 1.12

Liver 6.414 5.95 6.22 5.85 6.36

Kidneys 2.212 1.546 1.3 1.68 1.33

Uterus-Ovaries 1.694 0.57 0.62 0.71 0.9

Spleen 0.814 0.48 0.68 0.661 0.53

Table no. 5.2.4.17.2.8: Organ to body weight ratio of Group II Animals in Acute

Dermal Toxicity study.

Organ:body

weight

I

II

III

IV

V

Mean

SD

Brain 0.0096 0.0071 0.0089 0.0065 0.0093 0.0083 0.0012

Heart 0.0028 0.0035 0.0031 0.0032 0.0030 0.0031 0.0002

Lungs 0.0074 0.0068 0.0061 0.0072 0.0060 0.0067 0.0005

Liver 0.0281 0.0313 0.0329 0.0265 0.0343 0.0306 0.0029

Kidneys 0.0097 0.0081 0.0068 0.0076 0.0071 0.0079 0.0009

Uterus-Ovaries 0.0074 0.003 0.0032 0.0032 0.0048 0.0043 0.0016

Spleen 0.0035 0.0025 0.0035 0.0030 0.0028 0.0031 0.0004


Table no. 5.2.4.17.2.9: Comparison of Organ to body weight ratios for Group I and

Group II in Acute Dermal Toxicity study.

Organs Organ: Body weight

(Expressed as mean ± SD)

Group I Group II

Brain 0.0081 ± 0.0014 0.0083 ± 0.0012

Heart 0.0031 ± 0.0007 0.0031 ± 0.0002

Lungs 0.0065 ± 0.0008 0.0067 ± 0.0005

Liver 0.0355 ± 0.0048 0.0306 ± 0.0029

Kidneys 0.0071 ± 0.0005 0.0079 ± 0.0009

Uterus-Ovaries 0.0048 ± 0.0025 0.0043 ± 0.0016

Spleen 0.0032 ± 0.001 0.0031± 0.0004

*P<0.05 is considered as statistical significant analyzed by student t- test. Table no. 5.2.4.19.0: Nutritional value of M. zapota fruit and Recommended Dietary Intake.

Constituent Quanti ty provided per gm

of the M. zapota fruit. Dietary Reference

Intake/Day (U.S. DA)

Proteins 1 g 60 g

Fats 1.1 g 35 g

Carbohydrates 16 g 300 g

Total dietary Fibres 8 g 25 g

Calcium 210 mg 600 mg

Iron 0.8 mg 17 mg

Magnesium 12 mg 420 mg

Phosphorus 12 mg 1250 mg

Potassium 193 mg 4700 mg

Sodium 12 mg 1500 mg

Vitamin C 25 % of daily intake 90 mg


Vitamin A 2 % of daily intake 90 mcg

Total calories 73 kcal 2720 kcal

Table no. 5.2.4.21.1.1: Elemental analysis of the Freeze dried powder and comparison with

marketed products.

Sr. No.

Product and Company name

Element

Cu Zn Fe Ca K Pb As

1 Freeze Dr ied pulp, per 0.05 gm powder.

Per 100 gm powder.

0.064 ppm 128 mcg

2.983 ppm 5.9 mg

0.345 ppm 690 mcg

47.814 ppm 95.6 mg

325.961 ppm 651 mg

ND

ND

2 PEDIASURE,

per 100 gm powder. (Abbott)

400 mcg

3.5 mg

5.5mg 386 mg

512 mg - -

3 ENSURE,

per 100 gm powder. (Abbott)

410 mcg

3.5mg 35 mg 224 mg

536 mg - -

4 B-PROTIN, per 100 gm powder.

(British Biologicals)

- 10 mg 5 mg 800 mg

700 mg - -

5 CARDIOPRO, per 100 gm powder. *for Heart.


2 mg 12 mg - 750 mg

720 mg - -

6 PULMOCARE,

per 100 gm powder.


0.5 mg

5.63 mg

4.5 mg

250 mg

410 mg - -

7 KIDS-PRO,

per 100 gm powder.


0.12 mg

3 mg 1.25 mg

800 mg

900 mg - -

8 MACPROT,

per 100 gm powder.

(Macleods)

1.6 mg

10 mg 30 mg 1333.3 mg

- - -

9 XPLODE,

per 100 gm powder.

(Emcure Pharmaceuticals)

0.5 mg

4 mg - - - - -

10 GRD SMART, per 100 gm powder.

(Zydus Nutriva)

*for Pregnancy and Lactating mothers.

0.5 mg

4 mg 25 mg 900 mg

800 mg - -

11 NURTURE,

per 100 gm powder.

(Ranbaxy LTD.)

- - 20 mg 500 mg

- - -

12 ZEST, per 100 gm powder.

(Coral labs.)

84.6 mcg

- 22 mg 673 mg

- - -

13 MEGAMOM, per 100 gm powder.

(Alembic)

*for Pregnancy and Lactating mothers.

3.33 mg

5 mg 33.3 mg

666.66 mg

166.7 mg

- -

14 HANPRO-XP,

per 100 gm powder.

(Pax Healthcare)

2.5 mg

10 mg 20 mg 800 mg

150 mg - -

15 HIGH PRO, 2 scoops powder. (Nouveau Dietetique)

0.5 mcg

- 77 mcg

- 377 mg - -

16 PROTINEX, - - 6.5 449 - - -



per 100 gm powder. (Nutrica International Pvt. Ltd.)

mg mg

17 SOYGROWTH, per 100 gm powder. (Modi-Mundipharma) Pvt. Ltd.

1 mg 4 mg 20 mg 200 mg

100 mg - -

18 MAGNIFI, per 100 gm powder. (INTAS)

500 mcg

2 mg 10 mg 300 mg

100 mg - -

19 MAAPREG,

per 100 gm powder.

(BV Biocorp Pvt. Ltd.)

25 mcg

1 mg 2.5 mg

500 mg

- - -

20 AL BUMIN CARE, per 100 gm powder.

(BV Biocorp Pvt. Ltd.)

400 mcg

1 mg 2.5 mg

200 mg

- - -

21 ALPROVIT, per 100 gm powder. (Alkem Labs)

- 5 mg 5 mg 650 mg

- - -

22 PROGEM, per 100 gm powder.

(PLUS Pharma)

- 15 mg 10 mg 720 mg

- - -

*ND means less than 0.01 ppm, hence not detected.

Table no. 5.2.4.21.2.1.1: DPPH radical scavenging potential of freeze dried powder of

M. zapota fruit pulp.

Standard Ascorbic acid M. zapota fruit pulp

Conc. % Inhibition Conc. % Inhibition


(µg/ml) (µg/ml)

4 39.90±0.004 10 12.42±0.008

8 43.79±0.005 20 18.28±0.004

12 47.87±0.004 30 29.01±0.005

16 56.36±0.007 40 43.53±0.002

20 60.92±0.008 60 74.71±0.006

IC50 value: 12.5±0.004 µg/ml IC50 value: 65.54±0.004 µg/ml


Table no. 5.2.4.21.2.2.1: Total reducing power of freeze dried powder of M. zapota

fruit pulp.

Standard Ascorbic acid M. zapota fruit pulp

Conc.

(µg/ml)

Optical

density*

Conc.

(µg/ml)

Optical

density*

10 0.1893±0.003 10 0.1367±0.001

20 0.3581±0.001 20 0.3421±0.002

30 0.5244±0.002 30 0.4758±0.002

40 0.6421±0.004 40 0.5962±0.005

60 0.7521±0.004 60 0.7362±0.003


Table no. 5.2.4.22.1.1: Observations of Animals in Acute Oral Toxicity study.

Observations Group I - Vehicle control Group II - Treated

4h 24h 4h 24h

Skin and fur Normal Normal Normal Normal


Eyes Normal Normal Normal Normal

Mucus membrane Normal Normal Normal Normal

Behavioural patterns Normal Normal Normal Normal

Salivation Normal Normal Normal Normal

Lethargy Normal Normal Normal Normal

Sleep Normal Normal Normal Normal

Diarrhoea NO NO NO NO

Coma NO NO NO NO

Tremors NO NO NO NO

*NO indicates “Not Observed”

Table no. 5.2.4.22.1.2: Changes in body weight of Group I Animals in Acute Oral

Toxicity study, Vehicle Control.

Days Body weight (in Gm)

I II III IV V

Day 1 278 256 289 276 267

Day 7 286 269 295 285 278

Day 14 297 280 301 295 290

Table no. 5.2.4.22.1.3:Changes in body weight of Group II Animals in Acute Oral Toxicity.

Days Body weight (in Gm)

I II III IV V

Day 1 252 253 228 236 235

Day 7 266 271 244 252 260

Day 14 275 278 250 262 272

Table no. 5.2.4.22.1.4: Organ weights of Group I Animals in Acute Oral Toxicity

study, Vehicle Control.

Organs Organ weight of Animals (in Gm)


I II III IV V

Brain 1.86 1.6 1.9 2.1 1.73

Lungs 1.9 1.7 2 1.95 1.6

Liver 8.3 8.5 9.3 8.4 8

Kidney 1.74 1.8 1.95 1.85 1.62

Spleen 0.75 0.82 0.85 0.69 0.6

Heart 0.95 1.1 1.3 0.98 0.9

Ovaries-uterus 0.87 1.2 1.5 1.1 0.7

Table no. 5.2.4.22.1.5: Organ to body weight ratio of Group I Animals in Acute Oral

Toxicity study, Vehicle Control.

Organ:Body

weight

I

II

III

IV

V

Mean

SD

Brain 0.006263 0.005714 0.006312 0.007119 0.005966 0.006275 0.000474

Lungs 0.006397 0.006071 0.006645 0.00661 0.005517 0.006248 0.000419

Liver 0.027946 0.030357 0.030897 0.028475 0.027586 0.029052 0.001328

Kidney 0.005859 0.006429 0.006478 0.006271 0.005586 0.006125 0.000346

Spleen 0.002525 0.002929 0.002824 0.002339 0.002069 0.002537 0.000314

Heart 0.003199 0.003929 0.004319 0.003322 0.003103 0.003574 0.00047

Ovaries-Uterus 0.002929 0.004286 0.004983 0.003729 0.002414 0.003668 0.00092

*P<0.05 is considered as statistical significant analyzed by student t- test.

Table no. 5.2.4.22.1.6: Organ weights of Group II Animals in Acute Oral Toxicity

study.

Organs

Organ weights of Animals (gm)

I II III IV V


Brain 1.8 1.8 1.62 2 1.91

Lungs 1.8 2.17 2.2 1.93 3.69

Liver 8.6 9.7 7.94 8.6 8.1

Kidney 1.9 2.06 1.67 1.92 1.7

Spleen 0.68 0.76 0.59 0.77 0.8

Heart 1.02 1.04 1.05 0.8 1.28

Ovaries -Uterus 1.91 2 0.8 1.3 1.2

Table no. 5.2.4.22.1.7: Organ to body weight ratio of Group II Animals in Acute

Oral Toxicity study.

Organ:Body

weight

I

II

III

IV

V

Mean

SD

Brain 0.006545 0.006475 0.00648 0.007634 0.007022 0.006831 0.00045

Lungs 0.006545 0.007806 0.0088 0.007366 0.013566 0.008817 0.002484

Liver 0.031273 0.034892 0.03176 0.032824 0.029779 0.032106 0.001703

Kidney 0.006909 0.00741 0.00668 0.007328 0.00625 0.006915 0.000427

Spleen 0.002473 0.002734 0.00236 0.002939 0.002941 0.002689 0.000238

Heart 0.003709 0.003741 0.0042 0.003053 0.004706 0.003882 0.000551

Ovaries-Uterus 0.006945 0.007194 0.0032 0.004962 0.004412 0.005343 0.001523


Table no. 5.2.4.22.1.8: Comparison of Organ to body weight ratios for Group I and

Group II in Acute Oral Toxicity study.

Organs Organ: Body weight

Group 1 - Vehicle Group 2 - Treated

Brain 0.0062 ± 0.0004 0.0068 ± 0.0004


Lungs 0.0004 ± 0.0004 0.0088 ± 0.0024

Liver 0.029 ± 0.0013 0.0321 ± 0.0017

Kidney 0.0061 ± 0.0003 0.0069 ± 0.0004

Spleen 0.0025 ± 0.0003 0.0026 ± 0.0002

Heart 0.0035 ± 0.0004 0.0038 ± 0.0005

Ovaries-Uterus 0.0036 ± 0.0009 0.0053 ± 0.0015

Values are expressed as mean ± SD.


Table no. 5.2.4.23.1: Stability study of the freeze dried powder.

Test “0 Day” One Month Three month

Moisture content 2%w/w 2.1%w/w 2.1%w/w

pH 3.9 4.1 4

Ascorbic acid content 26.68±4.5

mg/100 gm

25.82±1.3

mg/100 gm

25.25±2.7

mg/100 gm


Figure no. 5.1.1.1.1.1.1: The transverse sectional …shodhganga.inflibnet.ac.in/bitstream/10603/72710/15/15...Formulation, characterization and evaluation of pharmaceutical dosage

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