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 1×150

100 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 1×25000

1000×1000 mg

= 0.025 mg

0.5 mL contains 0.025 mg Diazepam

10 mL contains 0.025×10

0.5 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 400×250001000×1000

mg

= 10 mg

0.5 mL contains 10 mg plant extract

5 mL contains 5×10

0.5 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 200×250001000×1000

mg

= 5 mg

0.5 mL contains 5 mg plant extract

5 mL contains 5×50.5

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 25× .01

10 mL acetic acid

= 0.25 mL acetic acid

Serial dilution

0.004 g plant extract dissolves in 200 µl of DMSO

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 800 µg/ml) (Concentration 400 µg/ml)

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 400 µg/ml) (Concentration 200 µg/ml)

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 200 µg/ml) (Concentration 100 µg/ml)

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 100 µg/ml) (Concentration 50 µg/ml)

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 50 µg/ml) (Concentration 25 µg/ml)

Volume adjust to 5 ml by saline 2.5 ml solution + 2.5 ml saline + shrimp

(Concentration 25 µg/ml) (Concentration 12.5 µg/ml)

(Up to 0.7813 5 µg/ml)