In vivo drug metabolism assay

Summary

In vivo drug metabolism experiments are important experiments used to assess drug toxicity, etc. during the development of new drugs. Currently, the most common method used for in vivo drug metabolism experiments is the oral route of administration.

Principle

The basic principle of in vivo drug metabolism assay is to measure various indicators of drug in animals using oral administration.

Appliance

In vivo drug metabolism assays can be used for drug development and toxicity assessment.

Operation method

In vivo drug metabolism assay

Principle

The basic principle of in vivo drug metabolism assay is to measure various indicators of drug in animals using oral administration.

Materials and Instruments

Laboratory animals: rats
Equipment: test tubes, centrifuge tubes, hard glass blood collection tubes
Reagents:
① Heparin solution
② Ethyl acetate
③ Methanol

Move

The method of drug metabolism in vivo is as follows:
A. Choose 30 rats of 200-250 g, 6-10 rats in a group, half of male and half of female.
B. No food or water is allowed 12 hours before drug administration, and it is better for the animals not to be anesthetized in the course of the experiment.
C. Blood is taken before drug administration: Insert the glass cannula into the inner corner of the eye and between the eyeball and the fundus of the eye, which is treated and dried by 1 % heparin solution. Insert the glass cannula between the inner corner of the eye and the eyeball after being treated with 1% heparin solution, gently pierce it toward the fundus of the rat's eye, stop poking when you feel resistance, rotate the blood tube to cut the venous plexus, and the blood will flow into the blood tube.
D. After blood collection, pull out the blood tube and relax the left hand, and the bleeding will stop. 0.51.0 ml of blood can be collected at one time.
E. Drug administration: administer by gavage (<4 ml), timing.
F. Blood collection after drug administration: blood is collected in the same way as in step C at 1, 5, 10, 15, 30, 45, 60, 90 and 120 minutes after drug administration, and put into centrifuge tubes, mixing.
G. Centrifuge the tube for 5 minutes at 3000×g and then remove the tube. G. Centrifuge for 5 minutes at 3000×g and then remove the centrifuge tube. The supernatant (plasma) is transferred from the centrifuge tube to a 5 ml tube.
H. The plasma is vortexed with four times the volume of ethyl acetate and centrifuged for 10 minutes at 2000 × g. The plasma is then evaporated under nitrogen and reconstituted in methanol and then centrifuged for 10 minutes at 2000 × g. The plasma is then analyzed by HPLC.

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