Dose calculation

1) Preparation of 1% v/v Tween 80 in saline: (for 150 mL) 100 mL saline contains 1 mL Tween 80 150 mL saline contains Tween 80 = 1.5 mL Tween 80 So, 1.5 mL Tween 80 takes in a volumetric flask and volume adjust to 150 mL with saline.

2) Preparation of positive control:

Dose: Diazepam 1mg/kg average body weight Let average body weight = 25 g = (25 1000) mg =25000 mg So, for (1000 1000) mg body weight requires 1 mg 25000 mg body mg body weight requires = 0.025 mg mg

0.5 mL contains 0.025 mg Diazepam 10 mL contains mg Diazepam

= 0.5 mg Diazepam = 0.0005 g Diazepam So, 0.0005 g Diazepam takes in a test tube and volume adjust to 10 mL with saline.

3) Preparation of plant extract preparation:

A) Dose: 400 mg/ kg average body weight Let average body weight = 25 g = (25 1000) mg =25000 mg So, for (1000 1000) mg body weight requires 400 mg

25000 mg body mg body weight requires = 10 mg

mg

0.5 mL contains 10 mg plant extract 5 mL contains mg plant extract

= 100 mg plant extract = 0.1 g plant extract So, 0.1 g plant extract takes in a test tube and volume adjust to 5 mL with saline.

B) Dose: 200 mg/ kg average body weight Let average body weight = 25 g = (25 1000) mg =25000 mg So, for (1000 1000) mg body weight requires 200 mg 25000 mg body mg body weight requires = 5 mg mg

0.5 mL contains 5 mg plant extract 5 mL contains mg plant extract = 50 mg plant extract = 0.05 g plant extract So, 0.05 g plant extract takes in a test tube and volume adjust to 5 mL with saline.

4) Calculation for acetic acid: (0.1 mL/ 10g) Let average body weight = 25 g So, for 10 g body weight requires 0.1 mL acetic acid 25 g body mg body weight requires mL acetic acid = 0.25 mL acetic acid

Serial dilution
0.004 g plant extract dissolves in 200 l of DMSO

Volume adjust to 5 ml by saline (Concentration 800 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 400 g/ml)

Volume adjust to 5 ml by saline (Concentration 400 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 200 g/ml)

Volume adjust to 5 ml by saline (Concentration 200 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 100 g/ml)

Volume adjust to 5 ml by saline (Concentration 100 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 50 g/ml)

Volume adjust to 5 ml by saline (Concentration 50 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 25 g/ml)

Volume adjust to 5 ml by saline (Concentration 25 g/ml)

2.5 ml solution + 2.5 ml saline + shrimp (Concentration 12.5 g/ml)

(Up to 0.7813 5 g/ml)