HOD of B. Pharm. (Practice), Abhinav Education Society’s College of Pharmacy, Narhe, Pune- 411041, Maharashtra, India.
Syzygiumcumini (L.) Skeels, commonly known as Jamun or Indian blackberry, is a medicinal plant widely recognized for its diverse pharmacological properties, particularly its antioxidant potential. This project investigates the antioxidant activity of bark extract of Syzygiumcumini using standard in vitro method such as Superoxide Dismutase Assay (SOD). The phytochemical screening revealed the presence of phenolic compounds, flavonoids, and tannins, which contribute significantly to its antioxidant activity. The results demonstrated that the ethanolic seed extract showed the highest free radical scavenging activity, indicating its potential as a natural antioxidant source. The present study focuses on the formulation and evaluation of an antioxidant herbal gel using Syzygiumcumini (L.) Skeels, a medicinal plant known for its rich phytochemical profile and potent antioxidant properties. The antioxidant herbal gel demonstrated significant free radical scavenging activity, indicating its potential to protect the skin from oxidative stress and environmental damage. The findings suggest that Syzygiumcumini-based herbal gel could be a promising natural alternative for skincare applications and further studies may explore its therapeutic potential in dermatological conditions.
The project mainly aims in preparing pharmaceutical gel dosage form which is the “Formulation and evaluation of herbal antioxidant gel” it has antioxidant activity, which prevents free radicals from the skin and helps as moisturizing and hydrating medication.
Plant profile:
Binomial nomenclature:
Fig 1: Syzygiumcuminibark
Syzygiumcumini (syn. Eugenia Jambolana), a member of family Myrtaceae is commonly known as Jamun or Jambul in Hindi and Black Plum or Indian Blackberry in English. S.Cumini is an evergreen tree distributed in the Indian sub-continent, south-east Asian countries and eastern Africa. S. cumini, is widely used in different countries including India for the treatment of many disorders including diabetes. Various parts of this plant have been recognized to posses several medicinal properties in the traditional system of medicine. The bark of the plant is carminative, digestive, anti-oxidant, anti-helminthic and antibacterial.[2]
MATERIALS AND METHODS:
MATERIALS:
The bark of syzygiumcumini was collected from local source.
Method:
(1) Preparation of crude drug powder:
Bark of syzygiumcumini is collected from village and cleaned through washing. Then shade dried for 15 days under room temperature and grinded well. Fine powdered drug is obtained and used for extraction process.[4]
(2) Preparation of ethanolic extract:
The powdered bark of syzygiumcumini were used for extraction. The bark of syzygiumcumini were carefully cleaned/washed and then using a soxhlet apparatus with ethanol. The extraction process was carried out for 5 hours, 7 cycle of ethanolic extract were runnedrepeatively. The resulting filtrate was used for phytochemical test and preparation of formulation. The product is stored in desiccator for further studies. [4]
Fig 2: Soxhletapparatus Fig 3: Ethanolic extract of Syzygiumcumini
Phytochemical screening:
The preliminary phytochemical screening of Syzygiumcumini bark was done by the following tests;
Test for Carbohydrates:
a) Molisch’s test: To 2-3ml aq extract, add few drops of alpha-naphthol solution in alcohol, shake add conc. H?SO? from side test tube. violet ring is formed at junction of two liquid.
Test for Alkaloids
Test for Tannins:
a) Ferric chloride test: Add a few magnesium turnings and concentrated hydrochloric acid to the extract. Appearance of pink or red color indicates flavonoids.
b) Potassium Dichromate: To 1-2 ml extract, add pinch of Potassium Dichromate appearance red ppt.
Test for Steroids:
a) Salkowski reaction: To 2 ml of extract, add 2ml chloroform and 2ml conc. H?SO?. Shake well chloroform layer appears red and acid layer shows greenish yellow fluorescence.
b) Liebermann-Burchard Reaction: Mix 2 ml extract with chloroform, Add 1-2 ml aceteic anhydride and 2 drops conc. H?SO? from the side of test tube, First red, then blue and finally green colour appear.
Test for Flavonoids:
Test for Saponin:
Foam test: About 2 ml of the extract was shaken with 5ml of distilled water Foam shows the presence of Saponins [3]
Physico-chemical Test:
The method is designed to measure the total amount of material remaining after ignition. This includes both ‘physiological ash’, which is derived from the plant tissue itself, and ‘nonphysiological ash’ which is the residue of the extraneous matter (e. g. sand and soil) adhering to the plant surface.
Procedure:
Total ash percentage = Pw-Fw x 100 /W
Where,
Pw = Pre weight of the crucible
Fw= Final weight of the crucible
W= Total weight of powdered plant material. [3]
(b) Determination of alcohol-soluble extractive value:
Alcohol- soluble extractive value is applied for the drug which contain alcohol soluble constituents such as tannins, resin, and alkaloids.
Procedure:
(c) Determination of water-soluble extractive value:
Water soluble extractive value is applied for the drug which contain water soluble constituents such as tannins, sugars, plant acid.
Procedure:
1. About 4 gm of coarsely powdered drug in a weighing bottle and transfer it to a dry 250 ml conical flask.
2. Fill a 100ml graduated flask to a delivery mark with the water.
3. Wash out the weighing bottle and pour the washing, together with the remainder of the solvent into the conical flask.
4. Cork the flask and set aside for 24 hours, shaking frequently.
5. Filter into a 50 ml cylinder.
6. When sufficient filtrate has collected, transfer 25 ml of the filtrate to a weighed, thin porcelain dish, as used for the ash values determination.
7. Evaporate to dryness on a water bath and complete the drying in an oven at 105degree Celsius for 6 hours.
8. Cool in a desiccator for 30 min and weight immediately.
9. Calculate the percentage w/w of extractive with reference to the air dried drug. [3]
Formulation:
(a) Preparation of Gel formulation:
1 g of carbapol-940 was dispersed in 50 ml of distilled water kept the beaker a side to swell the carbapol-940 to form gel. Take 5ml of distilled water and required quantity of propyl paraben were dissolved by heating on water bath solution was cooled and propylene glycol 400 and CMC added. Further required quantity of extract was mixed to the above mixture and add this solution into the carbapol 940 gel with continuous stirring and add triethanolamine was added to the formulation to adjust pH required to skin.[4]
Chemicals: syzygiumcumini extract, Carbopol940, Propylene Glycol, Tri-ethanolamine, Methyl paraben, Glycerin, Carboxy methyl cellulose
Apparatus: Beaker, stirrer, tripod stand, burner
Instruments: Brookfield viscometer, pH meter.
Excipient functions:
(a)Carbopol 940: Gelling Agent
(b) Propylene Glycol: Penetration Enhancer & Humectant
(c)Triethanolamine: pH Adjuster
(d) Methylparaben: Preservative
(e) Glycerin: Moisturizing agent
(f)Carboxy methyl cellulose: Thickening agent
Formulation table:
Table No.1: Formulation Table
Ingredient |
|
Quantity |
|
|
Roles of Ingredient |
||
F1 |
F2 |
F3 |
F4 |
F5 |
F6 |
||
Drug Extract |
0.5g |
1g |
1.5g |
0.5g |
1g |
1.5g |
Antioxidant Activity |
Carbopol940 |
1g |
1g |
1g |
1.5 g |
1.5 g |
1.5 g |
Gelling Agent |
Propylene Glycol |
2.5ml |
2.5 ml |
2.5 ml |
2.5 ml |
2.5 ml |
2.5 ml |
Moisturizer |
Methylparaben |
0.1g |
0.1 g |
0.1 g |
0.1 g |
0.1 g |
0.1 g |
Preservative |
Triethanolamine |
0.1ml |
0.1ml |
0.1ml |
0.1ml |
0.1ml |
0.1ml |
Surfactant, Neutralizer |
Glycerin |
1ml |
1ml |
1ml |
1ml |
1ml |
1ml |
Moisturizing agent |
Carboxy methyl cellulose |
0.5g |
0.5g |
0.5g |
0.5g |
0.5g |
0.5g |
Thickening agent |
Distilled water |
Q. S |
Q. S |
Q. S |
Q. S |
Q. S |
Q. S |
vehicle |
(3) Evaluation parameters of gel formulation:
Visual inspection: The gel were examined for their physical properties by visual inspection of colour, clarity, homogenesity, odour etc. [4]
Appearance: All the formulation of antioxidant gel was pale yellow/yellow in colour. [4]
Consistency: The consistency was checked by applying on the skin. [4]
Greasiness: The greasiness was assisted by the application on the skin and the slide [4]
Irritancy test (patch test): Skin irritation test was performed for the selected gels on human volunteers to find out any irritation problems which could make it unsuitable for topical use. Skin irritation test was performed, for each gel on three volunteers. Approximately 1gm of gel was topically applied to the hand near the wrist over 2 square inch area and observed for any lesions, irritation, allergy or edema etc. [4]
Washability: Gel Formulation was applied on hand and was observed continuously under running water [4]
Determination of pH: The pH of gel was determined using digital pH meter by dipping the glass electrode completely into the gel and by using pH paper [4]
Determination of Spreadability: Spreadability was measured on the basis of “slip” and “drag” characteristics of the gels ground glass slide was fixed on wooden block.an weight is provided by pulley ground slide was fixed on this block. 2gm of gel placed on the slide make sandwich weight of 100 gm. placed on the top of the two slides for 5 minutes excess of gel (about 2 gm.) was scrapped out from edges top plate subjected to pull of 20 gm. weight with help of stirring attached hook. Time in seconds required by the top slide cover a distance 7.5was noted. Spreadability was determined using following formula, S=M.L/T where,
S is the Spreadability in grams.cm/sec, M is the mass in grams, T is time in seconds [4]
Determination of viscosity: Viscosities of the formulated gels was determined using viscometer (speed 60 rpm. At 250 c. was used for gels, corresponding dial reading on the meter was noted. [4]
g) Clarity: The prepared gel formulation was evaluated in glass container and observed under the glass. [4]
RESULT AND DISCUSSION:
As we performed all evaluation parameters tests, the results are also found for every single tests of the gel formulation. Evaluation of herbal gel has been performed with parameters given for the evaluation purpose of the gel. It is evaluated by all factors like physicochemical as well as phytochemical factors.
Table no.2: Preliminary Characteristics of Gel
Sr.no. |
Parameter |
Observation |
1 |
Colour |
Light Green |
2 |
Odour |
Slight |
3 |
Acid value |
2.45 |
4 |
Ester value |
36.75 |
5 |
Solubility in ethanol |
Freely soluble |
6 |
Solubility in chloroform |
Soluble |
Table No.3: Test For Carbohydrates, Alkaloids, Flavonoids, Tannins, Steroids and Saponin
Sr. No |
Test |
Observation |
Inference |
1 |
Test of Carbohydrate |
|
|
|
|
Violet ring observed |
Carbohydrate Present |
2 |
Test of Alkaloids |
|
|
|
|
Orange brown precipitate Yellow precipitate |
Alkaloids present |
3 |
Test of Tannins |
|
|
|
|
Dark green solution Red precipitate |
Tannins present |
4 |
Test of steroids
|
Chloroform layer red and acid layer green precipitate Green colour appeared |
Steroid Present |
5 |
Test of Flavonoids
|
Yellow precipitate, colourless liquid |
Flavonoids present |
6 |
Test of Saponin
|
Foam is observed |
Saponin present |
Table No.4: Physico-Chemical Test Results
Sr.no. |
Parameter |
Observation |
1 |
Ash value |
8 % w/w |
2 |
Alcohol-soluble extractive value |
20 g |
3 |
Water-soluble extractive value |
|
Calculation:
(a) Ash value:
Weight of empty dish (x) = 22.50g
Weight of powder (y) = 2 g
Weight of dish with powder = 24.50 g
After incineration weight of dish (z) = 22.34 g
Weight of ash (z-x) = 22.34- 22.50
= 0.16
Total Ash = (100 ×Weight of Ash) / Weight of Powder
= (100 ×0.16) / 2
= 8 % w/w
Fig 9: Ash Value
Weight of porcelain with dried extract = 66.62 g
Weight of dried extract = Weight of porcelain with dried extract - Weight of porcelain = 66.62 - 66.42 = 0.2g 25 ml of alcoholic extract gives = 0.2 g of dried extract 100 ml of alcoholic extract gives = 0.2/25 x 100
= 0.8g 4 g of air-dried drug gives = 0.8g of alcoholic soluble extractives 100g of air-dried drug gives = 0.8/4 x 100 = 20 g of alcoholic soluble extractivesFig 10: Alcohol-soluble extractive value
Weight of porcelain = 65.14g
Weight of porcelain with dried extract = 65.25g
Weight of dried extract = Weight of porcelain with dried extract - Weight of porcelain = 65.25 – 65.14 = 0.09 g 25 ml of aqueous extract gives = 0.09 g of dried extract 100 ml of aqueous extract gives = 0.09/25 x 100 = 0.36 g 4 g of air -dried drug gives = 0.36 g of water -soluble extractives 100 g of air -dried drug gives = 0.36/ 4x 100 = 9 g of water- soluble extractives.
Table No.5: Some Basic Parameters Like; Ph, Spreadability, Viscosity, Homogeneity and Clarity Is Well Measured as Per IP
Formulation |
pH |
Spreadability |
Viscosity |
Homogeneity |
Clarity |
F1 |
6.52 |
5.32 |
2000 |
Average |
Not clear |
F2 |
6.51 |
6.08 |
3124 |
Good |
Not clear |
F3 |
6.54 |
6.04 |
3327 |
Very Good |
Clear |
F4 |
6.60 |
7.24 |
3749 |
Good |
Clear |
F5 |
6.60 |
7.60 |
6249 |
Good |
Partially Clear |
F6 |
6.64 |
7.60 |
7998 |
Good |
Not clear |
Result of activity determination:
In-vitro Antioxidant activity test:
Superoxide Dismutase (SOD):
Procedure:
(a) Critical Step: Add BSA first to prevent BCS precipitation
Table no.6: Solutions prepared for Superoxide Dismutase System
Solutions |
Final Concentrations |
Volume required for SOD System |
PB with DETAPAC and BSA |
0.05 M PB, 1 mM DETAPAC, 0.13 mg BSA |
12.9 ml
|
Catalase 40 U ml-1 |
1 U |
0.5 ml
|
Xanthine 1.18 mm |
100 μm |
1.7 ml
|
NBT 2.24 |
56 μm |
0.5 ml
|
PB |
0.05 m |
0.3 ml
|
BCS 10 mm |
50 μm |
0.1 ml
|
Total volume |
|
16.0 ml
|
(b) Preparation of three dilutions
Table no.6: Dilution Preparation
Dilution |
Volume of SOD |
Volume of PB |
Final Concentration (ng/tube) |
Dilution #1 (1:1000) |
1 μl |
999 μl |
2-10 |
Dilution #2 (1:100) |
3 μl |
297 μl |
25-50 |
Dilution #3 (1:10) |
5 μl |
45 μl |
500 |
Calculation:
Table no.7: Results observed for F3 batch
Sr.no |
Sample Type |
Dilutin Fator |
Protein Concentration (µg/ml) |
Control XO Rate (Abs/min ) |
Sample Rate (Abs/min ) |
Percent (100) |
% Inhibition |
|
Control XO |
- |
- |
|
|
|
|
1 |
Sample Dilution 1 |
1.10 |
20 |
0.52 |
0.25 |
100 |
51.92 |
2 |
Sample Dilutio n 2 |
1.08 |
50 |
0.36 |
0.11 |
100 |
69.44 |
3 |
Sample Dilutio n 3 |
1.06 |
100 |
0.41 |
0.22 |
100 |
46.34 |
4 |
Sample Dilutio n 4 |
1.04 |
150 |
0.96 |
0.36 |
100 |
62.50 |
5 |
Sample Dilutio n 5 |
1.03 |
250 |
0.56 |
0.32 |
100 |
42.86 |
6 |
Sample Dilutio n 6 |
1.02.05 |
500 |
0.37 |
0.21 |
100 |
43.24 |
7 |
Sample Dilutio n 7 |
1.02 |
1000 |
0.45 |
0.11 |
100 |
75.56 |
8 |
Sample Dilutio n 8 |
1.01.05 |
2000 |
0.51 |
0.24 |
100 |
52.94 |
9 |
Sample Dilutio n 9 |
1.01.02 |
3000 |
0.78 |
0.54 |
100 |
30.77 |
10 |
Sample Dilutio n 9 |
1.01.02 |
3000 |
0.82 |
0.26 |
100 |
68.29 |
11 |
Sample Dilutio n 10 |
1.01 |
5000 |
0.47 |
0.11 |
100 |
76.60 |
CONCLUSION:
This explains about overall information of herbal antioxidant gel formulation, its physicochemical, phytochemical parameters and evaluation of gel formulation.
Antioxidant gel formulation is an effective medication for restoring moisture and hydration to the skin. Antioxidant activity gel inhibits damage to the skin cells by reducing free radicals.
Future perspective:
Herbal gel formulation possessing antioxidant activity is helpful and effective to deliver moisturizing and hydrating effect on topical application onto the skin. Syzygiumcumini bark having more flavonoid and phenolic content essential to show antioxidant activity. This herbal gel formulation can reduce damage of skin cell form environmental or other free radicals.
REFERENCES
Dr. Hemalata S. Wadkar*, Krishna B. Kendre, Aditi A. Mardhekar, Tejashree P. Patil, Formulation and Evaluation of Herbal Gel of Syzygiumcumini of Antioxidant Activity, Int. J. of Pharm. Sci., 2025, Vol 3, Issue 6, 5332-5345. https://doi.org/10.5281/zenodo.15756483