Formulation and Evaluation of Emulgel loaded with leaf extract of Artemisia princeps for its Anti-Inflammatory potential
Madhurima Thakur1, Sunny Dhiman2*
1School of Pharmacy, Abhilashi University, Mandi, HP, India.
2Chandigarh College of Pharmacy CGC Landran, Mohali, Punjab, India.
*Corresponding Author E-mail: sdsdhiman1@gmail.com
ABSTRACT:
Indian Systems of Medicine have their roots in ayurveda. The potential of naturally occurring bioactive components is of tremendous interest to researchers. Different plant extracts have remedial properties.North India, Japan, China, Korea and Nepal are among the countries where Artemisia princeps can be found.Apigenin, diosmetin, eriiodicytol, eupafolin, kaempferol, vitex, jaceosidine, and quercetins have all been discovered in plant extracts of Artemisia princeps.It is used to treat fever, apoptosis, pain, ulcers, inflammation, and other conditions as well as Diarrhoea and Asthma.Recent in-vivo and in-vitro research have shown that it has anti-inflammatory and analgesic properties. The objectives of proposed research work are to formulate, evaluate and optimize the Emulgel loaded with optimised extract of Artemisia princepsand assess its In-Vitro Anti-Inflammatory Activity. Collected leaves of Artemisia princeps was extracted using hydro-alcoholic solvent. On the basis of their Phytochemical evaluation, the extract was optimized. Additional 6 batches of Emulgel were created utilizing a chosen extract. The prepared Emulgel was assessed for pH, spreadability, viscosity, consistency, appearance, colour, infrared spectral assignment, and appearance. Using the HRBC Membrane stabilisation assay, extract and formulation were tested for in vitro anti-inflammatory efficacy. The suggested study project's findings support the potential for future topical application of the newly developed Emulgel, which contains Artemisia princeps leaf extract, to reduce inflammation.
KEYWORDS: Artemisia princeps, Anti-Inflammatory, Inflammation, Phytoconstituents, Herbs, Pharmacological action.
INTRODUCTION:
Since ancient times, people have used medicinal herbs to treat illness. They were aware of the value of medicinal herbs in the creation of drugs. Plants include a variety of phytoconstituents, each of which has a specific medicinal effect. India is frequently referred as the emporium of medicinal plants because there are over 50000 plant species in its different regions. Medicinal herbs are supplied to industry from wild sources. Traditional folk medicine can be considered while creating novel medications.
The majority of the public's primary healthcare needs are met by phytoconstituents.1,2 Plant-based ingredients are effective in treating a variety of ailments and illnesses and are also affordable. In order to treat a wide range of diseases in a big patient population, various plant parts are utilised. Wide range of advantages exist for herbal medications.3,4 Herbal medicines work well in the human body and have relatively less adverse effects. Additionally, a number of herbs are being examined for their effectiveness against inflammation. Traditional herbal remedies for inflammation include Artemisia princeps, Curcuma longa, Zingiber officinalis Roscoe, Borago officinalis Linn, Oenothera biennis Linn, and Harpagophytum procumbens.5-8 China, Japan, and Korea are the countries of origin of Artemisia princeps. It thrives in a range of environments, including riverbanks, hillsides, and roadside edges. Apigenin, Diosmetin, Eriodicytol, Eupafolin, Kaempferol, Vitex, Jaceosidine, and Querecetine are some of the phytoconstituents found in it.Asthma Treatment, Anti-Spasmodic, Anti-Diarrheal, Anti-Malarial, Antiseptic, Improves Digestion, and Neurological Disorder Treatment are some of the typical uses of Artemisia princeps in medicine. Recently in-vivo and in-vitro studies have indicated its anti-inflammatory and analgesic property.9 Emulgels are a class of formulations that are mostly utilized for topical application. Emulgel is the name given to the product created when an emulsion is combined with a gel and exhibits dual control release properties. Emulgel is used to relieve aches and pains, including arthritis, backaches, and muscle aches.10-12 The purpose of the proposed research is to develop and assess a herbal emulgel containing Artemisia princeps leaf extract for its in vitro anti-inflammatory activity.
MATERIAL AND METHOD:
Collection of plant materials:
The leaves of the plant Artemisia princeps were collected from the local area of Bagsiad district Mandi Himachal Pradesh and was authenticated from School of Ayurveda Abhilashi University, Mandi HP.
Processing of plant material:
The freshly collected Artemisia princeps leaves were washed for the removal of dirt under running tap water after that kept for shade drying for 3–4 weeks at room temperature. After authentication, Artemisia princepswas dried and powdered.13 This powder was further used to obtain the extract.
Extraction of plant material:
The powder of Artemisia princeps was extracted with two solvents including water and 70% ethanol using Soxhlet apparatus. Both of the resulting extracts were concentrated, and optimization was carried out using photochemical testing.
Phytochemical Screening:
Phytochemical screening is performed to identify presence of different Phytoconstituents for selecting optimized extract. The extract was dissolved in respective solvents, filtered and filtrate was used to carry out test for alkaloids, flavonoids, glycoside, saponins, tannins, steroids, phenols, reducing sugar and terpenoid.
Preparation of herbal Emulgel:
As shown in Table 1, multiple batches of emulgel were created by changing the concentration of the gelling agents and employing different gelling agents. The oil phase of the emulsion was prepared by dissolving Span 80 in glycerine and aqueous phase was prepared by dissolving Tween 80 in purified water. Coconut oil was added into oil phase. Both the oily and aqueous phases were separately heated up to 40–50°C, then the oily phase was mixed with the aqueous phase with continuous stirring until it cools at room temperature. In a beaker, a specified quantity of Carbopol 934 was taken into 25ml of purified water and beaker was kept at bay for 1˝ h to stir the Carbopol. In another beaker, 5 ml of purified water was taken and heated with water bath by adding the required quantity of methylparaben and propylparaben. The solution was cool, and then, propylene gl ycol 400 was added. An additional amount of extract required was added to the above mixture and mixed correctly with a continuous stirring. Finally, a volume of up to 50ml was added to formulas for adjustment of the required mouth pH (6.8–7.3) to obtain the required consistency of gel by adding remaining distilled water and triethanolamine.To obtain an Emulgel, combine the gel with emulsion in a ratio of 1:1.14,15 (Figure 1)
Figure:1 Preparation of herbal Emulgel
Table 1: Composition of Emulgel formulations containing ethanolic extract of Artemisia Princeps
|
S. No. |
Composition |
F1 (g) |
F2 (g) |
F3 (g) |
F4 (g) |
F5 (g) |
F6 (g) |
|
1. |
Plant Extract |
1 |
1 |
1 |
1 |
2 |
1 |
|
2. |
Coconut oil(ml) |
1 |
2 |
2 |
1 |
1 |
1 |
|
3. |
Carbopol 934 |
0.1 |
0.15 |
1.5 |
1 |
0.2 |
2.5 |
|
4. |
Span 80(ml) |
0.5 |
0.1 |
1 |
1 |
0.6 |
0.15 |
|
5. |
Tween 80(ml) |
1 |
0.05 |
0.5 |
1 |
0.9 |
1 |
|
6. |
Methyl Paraben |
0.03 |
0.05 |
1 |
0.06 |
0.04 |
0.1 |
|
7. |
Propyl Paraben |
0.01 |
0.03 |
0.05 |
0.03 |
0.04 |
0.03 |
|
8. |
Propylene Glycol(ml) |
4 |
3 |
5 |
5 |
2 |
5 |
|
9. |
Triethanolamine(ml) |
1.2 |
1.4 |
1.3 |
0.1 |
0.1 |
0.3 |
|
10. |
Glycerine(ml) |
7.4 |
6.3 |
7.1 |
6.5 |
6.7 |
6.5 |
|
11. |
HPMC (g) |
0.5 |
1.0 |
1.5 |
2.0 |
2.5 |
3.0 |
|
12. |
Purified water(ml) |
qs |
qs |
qs |
qs |
qs |
qs |
(a) Spreadability (b) Viscosity
(c) pH
Figure:2 Preparation of herbal Emulgel
Characterization and Optimization of Emulgel:
The pH, spreadability, viscosity, consistency, appearance, colour, washability, and appearance of the prepared herbal Emulgel were assessed.16-18 (Figure 2)
Infrared Spectral Assignment:
The infra-red spectral assignment of optimized Extract and prepared emulgel was carried out for estimating any interaction using FTIR/ATIR spectrophotometer
Physical appearance:
Studying colour, homogeneity, uniformity, look, etc., is included in the category of physical appearance. By using ocular observation, colour was recorded. By rubbing Emulgel between fingertips, homogeneity was verified. Emulgel's appearance was evaluated through visual inspection.19
Spreadability:
To measure the spreadability, the one-gram Emulgel was placed between two petri plates and its circle's diameter was measured. Weight of 50grams was placed on the top of petri plate for about 60 seconds and the diameter of circles formed were measured in triplicate. The average of the reading was calculated. The reading was put into the following formula.20,21 S = M x L/Twhere S: Spreadability; M: Mass; L: Diameter; T: Times
Viscosity:
Using a Brookfield viscometer, the viscosity of Emulgel was calculated. The selected spindle speed was 50 rpm. Readings were taken in triplicate and the average of the readings was determine.22
pH:
By dissolving 0.5grams in 10 ml of distilled water and dipping an electrode into it, a digital pH meter was used to measure the pH of the batches of Emulgel that had been created.23,24
Stability studies:
The stability studies of Emulgel were carried out by keeping the samples at 5°C, 25°C/60% RH, 30°C/ 65% RH, and 40°C/75% RH for a period of 3 months. The samples were tested at interval of 15 days. They were evaluated for its appearance, pH, viscosity and spreadability.23
In vitro anti-inflammatory (Human Red Blood Cell (HRBC) Membrane Stabilization Method):
The anti-inflammatory activity was assessed using the HRBC membrane stabilization technique. 2-3ml of blood was used and equal quantity of Alsever’s solution was added to blood. iso-saline was added to the above mixture. This mixture was centrifuge for 5 minute to get Human Red Blood Cell (HRBC) suspension. Equal amount of sample was added to HRBC suspension. 100, 200, 300µg/ml of concentration of sample were prepared. It was incubated at 370°C for 30 minutes. The mixtures were centrifuged for 5 minutes. Alsever’s solution and blood were taken as negative control. Aspirin was taken as a standard. The supernatant solution obtained from centrifugation was used to carry out estimation using UV Spectroscopy at wavelength 560nm.25,26
RESULT AND DISCUSSION:
The Artemisia princeps Leaf was gathered and verified. The leaves were dried, and ground into a coarse powder. The powder was subjected to Soxhlet extraction. The extract obtained was used to study the phytochemical screening.
The presence of alkaloids, tannins, carbohydrate, flavonoids and terpenoids were determined by performing phytochemical test. Aqueous Extract of plant showed high occurrence of alkaloids, tannins, flavonoids, and terpenoids as shown in Table 2, hence it is selected for formulation of emulgel.
Table No. 2: Phytochemical Screening of Artemisia princeps
|
S. No. |
Phytochemical |
Aqueous Extract |
Ethanol Extract |
|
1. |
Alkaloids |
++ |
+ |
|
2. |
Flavonoid |
+++ |
++ |
|
3. |
Glycosides |
++ |
+ |
|
4. |
Reducing Sugar |
- |
- |
|
5. |
Saponin |
+ |
+ |
|
6. |
Steroids |
- |
- |
|
7. |
Phenols |
+ |
+ |
|
8. |
Terpenoid |
++ |
+ |
|
9. |
Tannin |
++ |
+ |
Emulgel was prepared using different gelling agents shown in Table 1. Based on a survey of the literature, the gelling agent's concentration was selected. The formulated batches had green colour. The formulations F1, F3, and F5 had good consistency F2 had poor consistency whereas F4 and F6 had very good consistency. The nature of all batches was non-greasy. The washability of all the formulations was good. Viscosity of formulations ranged from1750-2959 The pH of formulation ranged from 6.22-6.39. Spreadability of formulations were found in range of 6.9-8.7. Stability studies of emulgel were carried per ICH guidelines. Formulation F4 was found to be optimized as demonstrated in Table 3. Additional Infrared Spectral testing demonstrates that there is no interaction between the plant extract and the excipients as shown in Figure 3.
Table No. 3: Evaluation of different formulation of Emulgel
|
Parameters |
F1 |
F2 |
F3 |
F4 |
F5 |
F6 |
|
Color |
Green |
Green |
Green |
Green |
Green |
Green |
|
Consistency |
Good |
Poor |
Good |
Very good |
Good |
Very Good |
|
Homogeneity |
Poor |
Good |
Good |
Good |
Poor |
Good |
|
Appearance |
Semi-solid |
Semi-solid |
Semi-solid |
Semi-solid |
Semi-solid |
Semi-solid |
|
Greasiness |
Non-Greasy |
Non-Greasy |
Non-Greasy |
Non-Greasy |
Non-Greasy |
Non-Greasy |
|
Washability |
Washable |
Washable |
Washable |
Washable |
Washable |
Washable |
|
pH |
6.22 ±0.10 |
6.18 ±0.09 |
6.15 ±0.08 |
6.35 ±0.13 |
6.39 ±0.10 |
6.37 ±0.11 |
|
Viscosity (Pa. S) |
2810 |
1750 |
2381 |
2959 |
2593 |
2330 |
|
Spreadability |
6.9 |
7.06 |
8.7 |
7.25 |
8.10 |
7.47 |
Figure:3 Infrared Spectral Assignment (A) Extract (B) F4 Emulgel
Table 4: Prevention of lysis by Artemisia princeps extractTable 5: Prevention of lysis by Emulgel
|
Concentration |
Absorbance |
Prevention of lysis |
|
300 |
0.154 |
37.65 |
|
200 |
0.157 |
36.43 |
|
100 |
0.167 |
32.38 |
|
Aspirin |
0.156 |
36.84 |
|
Negative control |
0.247 |
---------- |
|
Concentration |
Absorbance |
Prevention of lysis |
|
300 |
0.160 |
35.23 |
|
200 |
0.158 |
36.03 |
|
100 |
0.173 |
29.95 |
|
Aspirin |
0.156 |
36.84 |
|
Negative control |
0.247 |
------- |
Figure:4 HRBC membrane stabilization method
(A)
(B)
Figure 5 Result of HRBC membrane stabilization (A) HRBC of plant extract (B) HRBC of F4 Emulgel
In vitro Anti-inflammatory Activity:
In-vitro anti-inflammatory activity of Optimized extract and formulation F4 was accessed. HRBC membrane stabilization method was used to access anti-inflammatory activity (Figure 4). The extract and formulated emulgel F4 Both showed good anti-inflammatory activity (Table 4 and 5, Figure 5).
CONCLUSION:
Both developing and developed nations have used herbal remedies for the treatment of numerous diseases for thousands of years. Our research demonstrates that Artemisia princeps' aqueous extract has strong anti-inflammatory properties. We created emulgel utilising several gelling agents. The improved formulation was discovered to be reliable and exhibit strong anti-inflammatory properties. So anti-inflammatory properties of Artemisia Princeps emulgel can be utilized topically.
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Received on 12.08.2023 Modified on 24.08.2023
Accepted on 03.09.2023 ©Asian Pharma Press All Right Reserved
Asian J. Pharm. Tech. 2023; 13(4):257-262.
DOI: 10.52711/2231-5713.2023.00046