Background
Methods
Drugs and chemicals
The plant material
Experimental animals
Extraction of the plant material
Preparation of 80ME
Preparation of solvent fractions
Acute oral toxicity test
Grouping and dosing
Determination of antidiarrheal activity
Castor oil (CO) induced diarrhea
CO induced gastrointestinal motility
CO induced enteropooling activity
The in vivo anti-diarrheal index
Preliminary phytochemical screening
Statistical analysis
Results
Acute oral toxicity test
Effects on castor oil induced diarrheal model
Dose (mg/kg) | Onset of diarrhea (Min) | No of wet feces | Total No of feces | Average weight of wet feces (gm) | Average weight of total feces (gm) | % inhibition of defecation | % WWFO | % WTFO |
---|---|---|---|---|---|---|---|---|
Control | 76.67 ± 7.99 | 6.67 ± 0.49 | 7.00 ± 0.52 | 0.36 ± 0.02 | 0.37 ± 0.02 | ------- | ----- | ---- |
80ME 100 | 109.67 ± 12.15 | 3.83 ± 0.70a2b1f1
| 4.50 ± 0.72b1f1
| 0.20 ± 0.02a2b2f2
| 0.21 ± 0.02a1b1f1
| 42.58 | 55.56 | 56.76 |
80ME 200 | 145.00 ± 21.77a1
| 2.50 ± 0.50a3
| 3.00 ± 0.68a2
| 0.14 ± 0.03a3
| 0.15 ± 0.03a2
| 62.52 | 38.89 | 40.54 |
80ME 400 | 173.83 ± 18.03a2
| 1.67 ± 0.49a3
| 2.67 ± 0.72a2
| 0.08 ± 0.02a3
| 0.10 ± 0.03a3
| 74.96 | 22.22 | 27.03 |
Loperamide 3 | 161.50 ± 16.93a2
| 1.83 ± 0.40a3
| 2.83 ± 0.60a2
| 0.09 ± 0.02a3
| 0.11 ± 0.02a3
| 72.56 | 25.00 | 29.73 |
Control | 80.17 ± 4.34 | 6.50 ± 0.43 | 6.83 ± 0.54 | 0.35 ± 0.03 | 0.36 ± 0.04 | ----- | ----- | ---- |
CF 200 | 123.33 ± 23.81 | 4.00 ± 1.00 | 4.33 ± 1.08 | 0.20 ± 0.05 | 0.21 ± 0.06 | 38.46 | 57.14 | 58.33 |
CF 300 | 140.50 ± 19.99 | 3.17 ± 0.65a1
| 3.67 ± 0.76 | 0.16 ± 0.03a1
| 0.17 ± 0.04a1
| 51.23 | 45.71 | 47.22 |
CF 400 | 152.00 ± 21.01a1
| 2.67 ± 0.76a1
| 3.17 ± 0.83a1
| 0.13 ± 0.04a1
| 0.15 ± 0.04a1
| 58.92 | 37.14 | 41.67 |
Loperamide 3 | 165.83 ± 33.17a1
| 1.67 ± 0.76a2
| 2.33 ± 1.05a1
| 0.08 ± 0.04a2
| 0.09 ± 0.04a2
| 74.31 | 22.86 | 25.00 |
Control | 69.33 ± 8.98 | 7.50 ± 1.34 | 8.17 ± 1.28 | 0.42 ± 0.05 | 0.45 ± 0.05 | ------ | ----- | --- |
MF 200 | 104.33 ± 6.14 | 5.17 ± 0.40b1
| 5.67 ± 0.49b1
| 0.29 ± 0.03b1
| 0.30 ± 0.04b1
| 31.07 | 69.05 | 66.67 |
MF 300 | 136.00 ± 29.02 | 3.83 ± 0.87a1
| 4.33 ± 1.05 | 0.22 ± 0.06a1
| 0.24 ± 0.06a1
| 48.93 | 52.38 | 53.33 |
MF 400 | 155.50 ± 26.89a1
| 2.83 ± 0.95a1
| 3.50 ± 1.23a1
| 0.14 ± 0.06a2
| 0.17 ± 0.06a2
| 62.67 | 33.33 | 37.78 |
Loperamide 3 | 166.83 ± 25.23a1
| 1.83 ± 0.70a2
| 2.33 ± 0.92a2
| 0.09 ± 0.03a3
| 0.11 ± 0.04a3
| 75.56 | 21.43 | 24.44 |
Control | 69.33 ± 8.98 | 7.50 ± 1.34 | 8.17 ± 1.28 | 0.42 ± 0.05 | 0.45 ± 0.05 | ------ | ---- | --- |
AF 200 | 70.83 ± 6.53b2j1n1
| 6.83 ± 0.70b2i1j1m1n1
| 7.67 ± 0.67b2g1i1j1n1
| 0.39 ± 0.02b3g2i2j2n2
| 0.42 ± 0.02b3g2i1j2n2
| 8.93 | 92.86 | 93.33 |
AF 300 | 77.67 ± 4.86b1j1n1
| 6.33 ± 0.72b2j1n1
| 6.67 ± 0.76b1j1n1
| 0.35 ± 0.03b2g1i1j1n1
| 0.36 ± 0.04b2i1j1n1
| 15.60 | 83.33 | 80.00 |
AF 400 | 81.00 ± 3.53b1
| 5.67 ± 0.62b1j1n1
| 6.50 ± 0.62b1j1
| 0.32 ± 0.02b2j1n1
| 0.33 ± 0.02b1j1n1
| 24.40 | 76.19 | 73.33 |
AF 800 | 134.67 ± 32.67 | 3.50 ± 0.99a1
| 4.33 ± 1.22a1
| 0.18 ± 0.05a2
| 0.21 ± 0.05a2
| 53.33 | 42.86 | 46.67 |
Loperamide 3 | 166.83 ± 25.23a1
| 1.83 ± 0.70a2
| 2.33 ± 0.92a2
| 0.09 ± 0.03a3
| 0.11 ± 0.04a3
| 75.56 | 21.43 | 24.44 |
Effects on castor oil induced intestinal transit in mice
Dose (mg/kg) | Length of small intestine (cm) | Distance moved by the charcoal meal (cm) | Peristaltic index (%) | % inhibition |
---|---|---|---|---|
Control | 56.17 ± 1.42 | 36.67 ± 1.94 | 65.09 ± 2.25 | ------- |
80ME 100 | 58.33 ± 0.80 | 25.17 ± 2.43a3b2f2
| 43.26 ± 4.37a3b2f2
| 33.54 |
80ME 200 | 58.67 ± 1.12 | 20.05 ± 1.09a3
| 35.07 ± 2.27a3
| 46.12 |
80ME 400 | 58.17 ± 2.01 | 14.33 ± 1.65a3
| 24.53 ± 2.53a3
| 62.31 |
Loperamide 3 | 56.33 ± 1.28 | 14.83 ± 0.91a3
| 26.29 ± 1.34a3
| 59.61 |
Control | 57.67 ± 1.76 | 36.17 ± 4.18 | 62.33 ± 6.06 | ------ |
CF 200 | 56.33 ± 1.08 | 25.17 ± 3.46b1
| 44.88 ± 6.39b1
| 27.99 |
CF 300 | 59.50 ± 1.09 | 24.00 ± 1.39a1
| 40.44 ± 2.60a1
| 35.11 |
CF 400 | 60.17 ± 1.89 | 20.67 ± 2.43a2
| 34.37 ± 3.91a2
| 44.86 |
Loperamide 3 | 56.83 ± 1.11 | 14.00 ± 1.41a3
| 24.58 ± 2.32a3
| 60.56 |
Control | 58.50 ± 0.67 | 36.83 ± 3.21 | 62.81 ± 5.11 | ------- |
MF 200 | 59.17 ± 1.38 | 29.17 ± 1.96b2
| 49.47 ± 3.47b2
| 21.24 |
MF 300 | 59.50 ± 1.41 | 24.00 ± 3.33a1
| 40.29 ± 5.48a1
| 35.85 |
MF 400 | 59.00 ± 1.59 | 19.17 ± 3.27a3
| 32.95 ± 5.99a3
| 47.54 |
Loperamide 3 | 56.33 ± 1.28 | 13.67 ± 1.52a3
| 24.20 ± 2.55a3
| 61.47 |
Control | 58.50 ± 0.67 | 36.83 ± 3.21 | 62.81 ± 5.11 | ------ |
AF 200 | 56.83 ± 0.95 | 34.00 ± 1.83b3g1j1n1
| 59.79 ± 2.98b3g1j1n2
| 4.81 |
AF 300 | 56.83 ± 1.92 | 31.00 ± 2.05b2g1
| 54.91 ± 4.33b2g1n1
| 12.58 |
AF 400 | 59.17 ± 0.98 | 29.50 ± 3.09b2
| 49.79 ± 4.95b2
| 20.73 |
AF 800 | 56.83 ± 1.17 | 21.83 ± 3.55a2
| 38.56 ± 6.29a2
| 38.61 |
Loperamide 3 | 56.33 ± 1.28 | 13.67 ± 1.52a3
| 24.20 ± 2.55a3
| 61.47 |
Effects on castor oil induced enteropooling
Dose (mg/kg) | Volume of intestinal contents (ml) | % inhibition | Weight of intestinal contents (gm) | % inhibition |
---|---|---|---|---|
Control | 0.86 ± 0.07 | ------- | 1.12 ± 0.03 | ------- |
80ME 100 | 0.64 ± 0.04a2b1f1
| 25.58 | 0.91 ± 0.05a2b1f1
| 18.75 |
80ME 200 | 0.53 ± 0.03a3
| 38.37 | 0.77 ± 0.06a3
| 31.25 |
80ME 400 | 0.46 ± 0.02a3
| 46.51 | 0.70 ± 0.02a3
| 37.50 |
Loperamide 3 | 0.47 ± 0.04a3
| 45.35 | 0.71 ± 0.03a3
| 36.61 |
Control | 0.78 ± 0.08 | -------- | 1.02 ± 0.07 | -------- |
CF 200 | 0.61 ± 0.07b1
| 21.79 | 0.85 ± 0.07b1
| 16.67 |
CF 300 | 0.53 ± 0.04a1
| 32.05 | 0.75 ± 0.04a1
| 26.47 |
CF 400 | 0.48 ± 0.04a2
| 38.46 | 0.69 ± 0.03a2
| 32.35 |
Loperamide 3 | 0.43 ± 0.06a2
| 44.87 | 0.66 ± 0.05a3
| 35.29 |
Control | 0.83 ± 0.06 | ----- | 1.10 ± 0.03 | ------- |
MF 200 | 0.67 ± 0.04b1
| 19.28 | 0.93 ± 0.07b1
| 15.45 |
MF 300 | 0.58 ± 0.03a1
| 30.12 | 0.81 ± 0.04a1
| 26.36 |
MF 400 | 0.49 ± 0.08a2
| 40.96 | 0.73 ± 0.08a2
| 33.64 |
Loperamide 3 | 0.47 ± 0.04a3
| 43.47 | 0.68 ± 0.02a3
| 38.18 |
Control | 0.83 ± 0.06 | ------ | 1.10 ± 0.03 | ------- |
AF 200 | 0.76 ± 0.04b2g1j1n1
| 8.43 | 1.02 ± 0.05b3g2j2n1
| 8.18 |
AF 300 | 0.73 ± 0.07b1j1n1
| 12.05 | 0.98 ± 0.07b2g1j1
| 10.91 |
AF 400 | 0.68 ± 0.07 | 18.07 | 0.93 ± 0.05b1
| 15.45 |
AF 800 | 0.55 ± 0.02a2
| 33.74 | 0.76 ± 0.03a2
| 30.90 |
Loperamide | 0.47 ± 0.04a3
| 43.47 | 0.68 ± 0.02a3
| 38.18 |
The in vivo antidiarrheal index
Dose (mg/kg) | Delay in defecation (time of onset in min, Dfreq) (%) | Gut meal travel distance (Gmeq) (%) | Purging frequency in number of wet stools (%) | In vivo antidiarrheal index (ADI) |
---|---|---|---|---|
Control | ------- | ------- | ------- | ------- |
80ME 100 | 43.04 | 33.54 | 42.58 | 39.47 |
80ME 200 | 89.12 | 46.12 | 62.52 | 63.58 |
80ME 400 | 126.72 | 62.31 | 74.96 | 83.96 |
Loperamide 3 | 110.64 | 59.61 | 72.56 | 78.22 |
Control | ------- | ------ | ------ | ----- |
CF 200 | 53.84 | 27.99 | 38.46 | 38.69 |
CF 300 | 75.25 | 35.11 | 51.23 | 51.34 |
CF 400 | 89.59 | 44.86 | 58.92 | 61.87 |
Loperamide 3 | 106.85 | 60.56 | 74.31 | 78.34 |
Control | ------ | ------ | ------ | ------ |
MF 200 | 50.48 | 21.24 | 31.07 | 32.18 |
MF 300 | 96.16 | 35.85 | 48.93 | 55.25 |
MF 400 | 124.29 | 47.54 | 62.67 | 71.81 |
Loperamide 3 | 140.63 | 61.47 | 75.56 | 86.76 |
Control | ------- | ------- | -------- | ------- |
AF 200 | 2.16 | 4.81 | 8.93 | 4.53 |
AF 300 | 11.54 | 12.58 | 15.60 | 13.13 |
AF 400 | 16.83 | 20.73 | 24.40 | 20.42 |
AF 800 | 94.24 | 38.61 | 53.33 | 57.89 |
loperamide | 140.63 | 61.47 | 75.56 | 86.76 |
Preliminary phytochemical screening
Constituents | Crude extract | Solvent fraction | ||
---|---|---|---|---|
80ME | Chloroform | Methanol | Aqueous | |
Cardiac glycosides | + | _ | + | + |
Flavonoids | + | + | + | - |
Alkaloids | _ | _ | _ | _ |
Saponins | + | _ | + | + |
Steroids | _ | + | _ | _ |
Tannins | + | + | + | + |
Terpenoids | + | + | + | _ |