Preparation, characterization and evaluation of hydrogels from different fractions of diverse medicinal plants for management of pain and inflammation

Figures & data

Table 1. Formulation of Hydrogels with and without Polyherbal extract.

Ingredients	G	MG 1%	MG 3%	M 5%	NG 1%	NG 3%	NG 5%
Carbopol 940 (g)	1	1	1	1	1	1	1
Propylene glycol (g)	5	5	5	5	5	5	5
Poly herbal n-hexane extract (g)	-	-	-	-	1	3	5
Poly herbal methanolic extract (g)	-	1	3	5	-	-	-
Triethanolamine (drops)	3–4	3–4	3–4	3–4	3–4	3–4	3–4
Distilled water (g)	94	93	91	89	93	91	89
Total weight	100 g	100 g	100 g	100 g	100g	100g	100g

Table 2. Percentage yield of n-hexane and methanol extract.

Sr. No	Plant name	Quantity in (grams)	Solvent	Extract (grams)	Extract percentage values (%)
1	Black cumin	400	n-hexane	23.4	5.85
		373	Methanol	15	4.02
2	Fenugreek seeds	400	n-hexane	24	6
		370	Methanol	19.4	5.24
3	Cinnamon	400	n-hexane	10	2.5
		380	Methanol	26.4	6.94
4	Moringa	400	n-hexane	30	7.5
		360	Methanol	18	5

Table 3. pH of hydrogel formulations with and without Polyherbal extract.

Sr. No.	Formulations	pH
1	G	7.00
2	MG 1%	7.1
3	MG 3%	7.0
4	MG 5%	6.9
5	NG 1%	6.8
6	NG 3%	7.0
7	NG 5%	7.1

Table 4. Appearance, homogeneity, spreadability, and extrudability of hydrogel formulations.

Sr. No.	Formulation	Appearance	Homogeneity	Spreadability	Extrudability
1	G	White	Clear	6.9	Good
2	MG 1%	Dark greenish	Clear	6.9	Good
3	MG 3%	Dark greenish	Clear	6.2	Good
4	MG 5%	Dark greenish	Clear	6.2	Better
5	NG 1%	Light greenish	Clear	6.6	Good
6	NG 3%	Light greenish	Clear	6.2	Good
7	NG 5%	Dark greenish	Clear	6.7	Better

Figure 1. Rheological studies of hydrogel formulations with and without polyherbal extract (PNE, PME). Sample G is hydro-gel formulation without polyherbal extract, Sample MG 1% is hydrogel formulation containing 1.0 g of polyherbal methanolic extract, Sample MG 3% is hydrogel formulation containing 3.0 g of polyherbal methanolic extract and sample MG 5% is hydrogel formulation containing 5.0 g of polyherbal methanolic extract, Sample NG 1% is hydrogel formulation containing 1.0 g of polyherbal n-hexane extract, Sample NG 3% is hydrogel formulation containing 3.0 g of polyherbal n-hexane extract and sample NG 5% is hydrogel formulation containing 5.0 g of polyherbal n-hexane extract

Table 5. Stability study of hydrogel formulations with and without different extracts.

	Storage condition
	2–8°C/ 60–70%RH ± 5%			32°C/ 60–70%RH ± 5%			40°C/ 60–70%RH ± 5%
Name of Formulation	days
	15	30	60	15	30	60	15	30	60
G	No change	No change	No change	No change	No change	No change	No change	No change	No change
MG 1%	No change	No change	No change	No change	No change	No change	No change	No change	No change
MG 3%	No change	No change	No change	No change	No change	No change	No change	No change	No change
MG 5%	No change	No change	No change	No change	No change	No change	No change	No change	No change
NG 1%	No change	No change	No change	No change	No change	No change	No change	No change	No change
NG 3%	No change	No change	No change	No change	No change	No change	No change	No change	No change
NG 5%	No change	No change	No change	No change	No change	No change	No change	No change	No change

Table 6. Skin irritation test of hydrogel formulations with and without polyherbal extract.

Sr. No.	Formulations	Observation for 7 days
		Day 1	Day 2	Day 3	Day 4	Day 5	Day 6	Day 7
1	G	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
2	MG 1%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
3	MG 3%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
4	MG 5%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
5	NG 1%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
6	NG 3%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction
7	NG 5%	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction	No reaction

Figure 2. Analgesic activity of polyherbal n-hexane extracts (PNE) by tail flick method.

Normal saline was given to control group, while diclofenac sodium 100mg/kg was given to standard group. Sample N 1 is 200 mg/kg of polyherbal n-hexane extract, sample N 2 is 400 mg/kg of polyherbal n-hexane extract, and sample N 3 is 800 mg/kg of polyherbal n-hexane extract. Each group was analyzed by using two-way ANOVA followed by Tukey’s test. When control and standard group are compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with standard and extracts and 2nd bar denoted by (#) indicates the comparison of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 3. Analgesic activity of polyherbal methanolic extracts (PME) by tail flick method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample M 1 is 200 mg/kg of polyherbal methanolic extract, sample M 2 is 400 mg/kg of polyherbal methanolic extract, and sample M 3 is 800 mg/kg of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by Tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 4. Analgesic activity of hydrogel formulations (NG) with polyherbal n-hexane extracts by tail flick method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample NG 1% is hydrogel formulation containing 1.0 g of polyherbal n-hexane extract, Sample NG 3% is hydrogel formulation containing 3.0 g of polyherbal n-hexane extract and sample NG 5% is hydrogel formulation containing 5.0 g of polyherbal n-hexane extract. Each group is analyzed by using ANOVA followed by Tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 5. Analgesic activity of hydrogel formulations (MG) with polyherbal methanolic extracts by tail flick method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample MG 1% is hydrogel formulation containing 1.0 g of polyherbal methanolic extract, Sample MG 3% is hydrogel formulation containing 3.0 g of polyherbal methanolic extract and sample MG 5% is hydrogel formulation containing 5.0 g of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 6. Analgesic activity of polyherbal n-hexane extracts (PNE) by hot plate method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample N 1 is 200 mg/kg of polyherbal n-hexane extract, sample N 2 is 400 mg/kg of polyherbal n-hexane extract and sample N 3 is 800 mg/kg of polyherbal n-hexane extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 7. Analgesic activity of polyherbal methanolic extracts (PME) by hot plate method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample M 1 is 200 mg/kg of polyherbal methanolic extract, sample M 2 is 400 mg/kg of polyherbal methanolic extract and sample M 3 is 800 mg/kg of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 8. Analgesic activity of hydrogel formulations (NG) with polyherbal n-hexane extracts by hot plate method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample NG 1% is hydrogel formulation containing 1.0 g of polyherbal n-hexane extract, Sample NG 3% is hydrogel formulation containing 3.0 g of polyherbal n-hexane extract and sample NG 5% is hydrogel formulation containing 5.0 g of polyherbal n-hexane extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 9. Analgesic activity of hydrogel formulations (MG) with polyherbal methanolic extracts by hot plate method.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample MG 1% is hydrogel formulation containing 1.0 g of polyherbal methanolic extract, Sample MG 3% is hydrogel formulation containing 3.0 g of polyherbal methanolic extract and sample MG 5% is hydrogel formulation containing 5.0 g of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 10. Anti-inflammatory activity of polyherbal n-hexane extracts (PNE).

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample N 1 is 200 mg/kg of polyherbal n-hexane extract, sample N 2 is 400 mg/kg of polyherbal n-hexane extract and sample N 3 is 800 mg/kg of polyherbal n-hexane extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 11. %inhibition of anti-inflammatory activity of polyherbaln-hexane extracts (PNE).

(Sample N 1 is 200 mg/kg of polyherbal n-hexane extract, sample N 2 is 400 mg/kg of polyherbal n-hexane extract and sample N 3 is 800 mg/kg of polyherbal n-hexane extract).

Figure 12. Anti-inflammatory activity of polyherbal methanolic extracts (PME).

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample M 1 is 200 mg/kg of polyherbal methanolic extract, sample M 2 is 400 mg/kg of polyherbal methanolic extract and sample M 3 is 800 mg/kg of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 13. %inhibition of anti-inflammatory activity of polyherbal methanolic extracts (PME).

(Sample M 1 is 200 mg/kg of polyherbal methanolic extract, sample M 2 is 400 mg/kg of polyherbal methanolic extract and sample M 3 is 800 mg/kg of polyherbal methanolic extract).

Figure 14. Anti-inflammatory activity of hydrogel formulations (NG) with polyherbal n-hexane extracts.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample NG 1% is hydrogel formulation containing 1.0 g of polyherbal n-hexane extract, Sample NG 3% is hydrogel formulation containing 3.0 g of polyherbal n-hexane extract and sample NG 5% is hydrogel formulation containing 5.0 g of polyherbal n-hexane extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 15. % inhibition of anti-inflammatory activity of hydrogel formulations (NG) with polyherbal n-hexane extracts.

(Sample NG 1% is hydrogel formulation containing 1.0 g of polyherbal n-hexane extract, Sample NG 3% is hydrogel formulation containing 3.0 g of polyherbal n-hexane extract and sample NG 5% is hydrogel formulation containing 5.0 g of polyherbal n-hexane extract)

Figure 16. Anti-inflammatory activity of hydrogel formulations (MG) with polyherbal methanolic extracts.

Normal saline was given to control group while diclofenac sodium 100mg/kg was given to standard group. Sample MG 1% is hydrogel formulation containing 1.0 g of polyherbal methanolic extract, Sample MG 3% is hydrogel formulation containing 3.0 g of polyherbal methanolic extract and sample MG 5% is hydrogel formulation containing 5.0 g of polyherbal methanolic extract. Each group is analyzed by using ANOVA followed by tukey’s test. When control group and standard group compared with sample groups, results are considered nonsignificant (ns) if p > 0.05; first bar denoted by (*) indicates the compared significance of control with sample groups and 2nd bar denoted by (#) indicates the compared significance of standard group (diclofenac sodium) with sample group. Results are significant (*, #) if p < 0.05, more significant (**, ##) if p < 0.01, very significant (***, ###) if p < 0.001, and highly significant (****, ####) if p < 0.0001.

Figure 17. %inhibition of anti-inflammatory activity of hydrogel formulations(MG) with polyherbal methanolic extracts.

(Sample MG 1% is hydrogel formulation containing 1.0 g of polyherbal methanolic extract, Sample MG 3% is hydrogel formulation containing 3.0 g of polyherbal methanolic extract and sample MG 5% is hydrogel formulation containing 5.0 g of polyherbal methanolic extract)