Background
Methods
Plant material
Preparation of the extract and fractions
Animals
Acute toxicity
Evaluation of antioxidant study
Estimation of total phenolic content
Estimation of total flavonoid content
Phosphomolybdate assay (total antioxidant capacity)
DPPH radical scavenging activity assay
Anti-inflammatory activity
Analgesic activity
Antipyretic activity
Anti-diabetic activity
Statistical analysis
Results
Acute toxicity
Evaluation of antioxidant study
Estimation of total phenolic content
Extract or fraction | Total extract | Petroleum ether fraction | Dichloromethane fraction | Ethyl acetate fraction | n-Butanol fraction |
---|---|---|---|---|---|
Total phenolic content (mg/g gallic acid equivallent) | 5.76 ± 0.23 | 4.75 ± 0.22 | 8.81 ± 0.04 | 31.03 ± 0.15 | 4.54 ± 0.04 |
Estimation of total flavonoid content
Extract or fraction | Total extract | Petroleum ether fraction | Dichloromethane fraction | Ethyl acetate fraction | n-Butanol fraction |
---|---|---|---|---|---|
Total flavonoidal content (mg/g rutin equivallent) | 192.24 ± 0.33 | 13.09 ± 0.11 | 295.45 ± 0.13 | 811.91 ± 0.07 | 84.29 ± 0.017 |
Phosphomolybdate assay (total antioxidant capacity)
Extract or fraction | Total extract | Petroleum ether fraction | Dichloromethane fraction | Ethyl acetate fraction | Butanol fraction |
---|---|---|---|---|---|
Free radical scavenging activity | 24.50 ± 0.37 | 26.35 ± 0.31 | 27.05 ± 0.28 | 103..40 ± 0.05 | 12.58 ± 0.60 |
DPPH radical scavenging activity assay
Extract or fraction | Total extract | Petroleum ether fraction | Dichloromethane fraction | Ethyl acetate fraction | Butanol fraction | BHT |
---|---|---|---|---|---|---|
IC50 | 1.33 ± 0.03 mg/ml | 1.21 ± 0.21 mg/ml | 1.03 ± 0.06 mg/ml | 0.03 ± 0.01 mg/ml | 1.28 ± 0.10 mg/ml | 0.96 ± 0.03 μg/ml |
Anti-inflammatory activity
Group | Thickness of the paw (mm) | ||||||
---|---|---|---|---|---|---|---|
0 h | 30 min | 1 h | 2 h | 3 h | 4 h | 5 h | |
Control (−ve) | 7.38 ± 0.25 | 7.25 ± 0.29 | 7.25 ± 0.29 | 7.63 ± 0.25 | 7.75 ± 0.29 | 7.75 ± 0.29 | 7.75 ± 0.29 |
Total extract (350 mg/kg) | 7.25 ± 0.29 | 7.00 ± 0.00 | 6.50 ± 0.41 * | 6.25 ± 0.29 *** | 6.00 ± 0.41 *** | 6.25 ± 0.29 *** | 6.75 ± 0.29 ** |
Petroleum ether fraction (350 mg/kg) | 7.25 ± 0.29 | 7.13 ± 0.25 | 6.88 ± 0.25 | 6.75 ± 0.29 *** | 6.63 ± 0.48** | 6.38 ± 0.25 *** | 7.13 ± 0.25 * |
Dichloromethane fraction (350 mg/kg) | 7.00 ± 0.00 * | 7.00 ± 0.00 | 6.38 ± 0.48 * | 5.75 ± 0.29 *** | 4.63 ± 0.75 *** | 4.25 ± 0.29 *** | 5.63 ± 0.48 *** |
Ethyl acetate fraction (350 mg/kg) | 7.38 ± 0.25 | 7.13 ± 0.25 | 6.63 ± 0.48 | 6.13 ± 0.48 ** | 5.50 ± 0.41 *** | 4.63 ± 0.48 *** | 6.00 ± 0.41 *** |
Butanol fraction (350 mg/kg) | 7.25 ± 0.29 | 7.13 ± 0.25 | 6.75 ± 0.29 * | 6.50 ± 0.41 ** | 6.38 ± 0.25 *** | 5.755 ± 0.29 *** | 6.50 ± 0.41 ** |
Indomethacin (8 mg/kg) | 7.25 ± 0.29 | 6.88 ± 0.25 | 5.25 ± 0.65 *** | 3.88 ± 0.25 *** | 3.75 ± 0.29 *** | 3.75 ± 0.29 *** | 5.75 ± 0.29 *** |
Group | Dose mg/kg | Percentages of inhibition (%) | |||||
---|---|---|---|---|---|---|---|
30 min | 1 h | 2 h | 3 h | 4 h | 5 h | ||
Total extract | 350 | 3.45 | 10.34 | 18.09 | 22.58 | 19.35 | 12.90 |
Petroleum ether fraction | 350 | 1.66 | 5.10 | 11.53 | 14.45 | 17.68 | 8 |
Dichloromethane fraction | 350 | 3.45 | 12 | 24.64 | 40.26 | 45.16 | 27.35 |
Ethyl acetate fraction | 350 | 1.66 | 8.55 | 19.66 | 29.03 | 40.26 | 22.58 |
Butanol fraction | 350 | 1.66 | 6.90 | 14.81 | 17.68 | 25.81 | 16.13 |
Indomethacin | 8 | 5.10 | 27.59 | 49.15 | 51.61 | 51.61 | 25.81 |
Analgesic activity
Group | Time (sec) | ||||||
---|---|---|---|---|---|---|---|
0 h | 30 min | 1 h | 2 h | 3 h | 4 h | 5 h | |
Control (−ve) | 87.00 ± 8.32 | 83.67 ± 7.57 | 80.33 ± 4.16 | 69.67 ± 4.16 | 70.00 ± 3.61 | 80.67 ± 4.73 | 81.67 ± 3.06 |
Total extract (350 mg/kg) | 75.67 ± 11.68 | 94.33 ± 7.23 | 106.33 ± 3.06*** | 130.67 ± 18.50** | 140.67 ± 10.60*** | 128.33 ± 21.20* | 101.33 ± 4.93** |
Petroleum ether fraction (350 mg/kg) | 72.00 ± 6.56 * | 100.33 ± 4.16* | 191.00 ± 31.08** | 270.67 ± 8.50*** | 289.00 ± 3.00*** | 227.00 ± 28.84* | 99.00 ± 10.00* |
Dichloromethane fraction (350 mg/kg) | 78.67 ± 10.60 | 86.67 ± 8.08 | 170.33 ± 21.39** | 206.33 ± 16.29*** | 288.33 ± 20.82*** | 199.67 ± 9.87*** | 89.00 ± 13.86 |
Ethyl acetate fraction (350 mg/kg) | 71.00 ± 10.58 | 91.67 ± 3.06 | 107.33 ± 11.93* | 129.67 ± 2.08*** | 139.33 ± 5.69*** | 103.33 ± 1.53** | 91.33 ± 12.42 |
Butanol fraction (350 mg/kg) | 65.00 ± 8.00 ** | 83.33 ± 7.37 | 85.67 ± 13.01 | 74.33 ± 5.03 | 74.33 ± 5.03 | 79.67 ± 2.03 | 80.33 ± 4.62 |
Paracetamol (100 mg/kg) | 70.67 ± 5.51** | 107.67 ± 5.77* | 311.00 ± 19.28*** | 321.67 ± 25.01*** | 386.33 ± 8.08*** | 258.33 ± 41.59*** | 122.33 ± 15.01** |
Group | Dose mg/kg | Percentage of protection against external stimulus (%) | |||||
---|---|---|---|---|---|---|---|
30 min | 1 h | 2 h | 3 h | 4 h | 5 h | ||
Total extract | 350 | 12.74 | 32.37 | 87.56 | 100.96 | 59.08 | 24.07 |
Petroleum ether fraction | 350 | 19.91 | 137.77 | 288.50 | 312.86 | 181.39 | 21.22 |
Dichloromethane fraction | 350 | 3.59 | 112.04 | 196.15 | 311.90 | 147.51 | 8.98 |
Ethyl acetate fraction | 350 | 9.56 | 33.61 | 86.12 | 99.04 | 28.09 | 11.83 |
Butanol fraction | 350 | −0.4 | 6.65 | 6.69 | 6.19 | −1.2 | −1.6 |
Paracetamol | 100 | 28.68 | 287.15 | 361.71 | 451.43 | 220.23 | 49.79 |
Antipyretic activity
Group | Rectal temperature (°C) | ||||||
---|---|---|---|---|---|---|---|
0 h | 30 min | 1 h | 2 h | 3 h | 4 h | 5 h | |
Control (−ve) | 39.83 ± 0.10 | 39.88 ± 0.05 | 39.88 ± 0.05 | 39.88 ± 0.05 | 39.83 ± 0.10 | 39.88 ± 0.05 | 39.28 ± 0.09 |
Total extract (350 mg/kg) | 39.75 ± 0.13 | 38.63 ± 0.39 *** | 38.53 ± 0.50 ** | 37.73 ± 0.17 *** | 37.65 ± 0.13 *** | 37.83 ± 0.10 *** | 38.00 ± 0.27 *** |
Petroleum ether fraction (350 mg/kg) | 39.83 ± 0.10 | 39.20 ± 0.60 | 38.80 ± 0.34 *** | 37.68 ± 0.21*** | 37.60 ± 0.08 *** | 37.43 ± 0.10 *** | 38.08 ± 0.41 *** |
Dichloromethane fraction (350 mg/kg) | 39.83 ± 0.10 | 39.25 ± 0.17 *** | 38.53 ± 0.10 *** | 37.78 ± 0.15 *** | 37.63 ± 0.13 *** | 37.60 ± 0.08 *** | 38.88 ± 0.47 ** |
Ethyl acetate fraction (350 mg/kg) | 39.80 ± 0.14 | 38.73 ± 0.17 *** | 38.73 ± 0.17 *** | 37.73 ± 0.40 *** | 37.45 ± 0.31 *** | 37.70 ± 0.18 *** | 38.25 ± 0.41 *** |
Butanol fraction (350 mg/kg) | 39.85 ± 0.10 | 39.73 ± 0.17 | 39.13 ± 0.17 *** | 38.33 ± 0.35 *** | 37.73 ± 0.17 *** | 37.78 ± 0.15 *** | 38.75 ± 0.19 *** |
Acetyl salicylic acid(330 mg/kg) | 39.83 ± 0.10 | 38.28 ± 0.49 *** | 37.58 ± 0.10 *** | 37.48 ± 0.05 *** | 37.43 ± 0.17 *** | 37.13 ± 0.15 *** | 37.08 ± 0.10 *** |
Anti-diabetic activity
Group | Blood glucose level (mg/dl) | ||||||
---|---|---|---|---|---|---|---|
0 h | 30 min | 1 h | 2 h | 3 h | 4 h | 5 h | |
Control (−ve) | 197 ± 7.21 | 214.33 ± 5.03 | 214 ± 7.81 | 212.67 ± 19.66 | 300.33 ± 2.31 | 309 ± 4 | 322.33 ± 13.01 |
Total extract (350 mg/kg) | 209 ± 14.42 | 136 ± 13.45*** | 108.33 ± 3.06*** | 99.67 ± 1.15*** | 95.67 ± 5.77*** | 120 ± 1.73*** | 224.33 ± 15.01*** |
Petroleum ether fraction (350 mg/kg) | 211 ± 18.33 | 117 ± 7.21*** | 107.67 ± 4.16*** | 93.67 ± 7.57*** | 87.67 ± 10.26*** | 106 ± 6.08*** | 237 ± 13.86** |
Dichloromethane fraction (350 mg/kg) | 213 ± 16.37 | 213 ± 11.13 | 163.67 ± 15.01** | 121.67 ± 3.06*** | 190.33 ± 1.15*** | 271 ± 3.46*** | 323.67 ± 2.31 |
Ethyl acetate fraction (350 mg/kg) | 201.67 ± 3.06 | 205.67 ± 5.03 | 215 ± 4 | 216.33 ± 1.15 | 310.33 ± 1.15** | 311.67 ± 8.08 | 312.33 ± 4.16 |
Butanol fraction (350 mg/kg) | 204.33 ± 5.03 | 215 ± 5.29 | 221.67 ± 9.02 | 268 ± 29.44*** | 293.33 ± 7.51 | 305.33 ± 3.51 | 327 ± 15.20 |
Vildagliptin (50 mg/kg) | 208.67 ± 7.77 | 125 ± 12.12*** | 102.33 ± 2.31*** | 94.67 ± 6.66*** | 64 ± 11.36*** | 85 ± 15.20*** | 178 ± 20.66*** |
Group | Dose mg/kg | Percentage of decrease in blood glucose level (%) | ||||||
---|---|---|---|---|---|---|---|---|
0 h | 30 min | 1 h | 2 h | 3 h | 4 h | 5 h | ||
Total extract | 350 | −6.09 | 36.54 | 49.38 | 53.13 | 68.22 | 61.17 | 30.40 |
Petroleum ether fraction | 350 | −7.11 | 45.41 | 49.69 | 55.96 | 70.78 | 65.70 | 26.47 |
Dichloromethane fraction | 350 | −16 | 0.62 | 23.52 | 42.79 | 36.56 | 12.30 | −0.42 |
Ethyl acetate fraction | 350 | −2.37 | 4.04 | −0.47 | −1.72 | −3.44 | − 0.64 | 3.10 |
Butanol fraction | 350 | −3.72 | −0.31 | − 3.58 | −26.02 | 2.22 | 1.19 | −1.45 |
Vildagliptin | 50 | −5.90 | 41.68 | 52.18 | 55.49 | 78.69 | 72.49 | 44.78 |