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Preparation and characterization of rodent intestinal microsomes: Comparative assessment of two methods

Author(s): Anagha Damre1, SR Mallurwar1, D Behera2
1 Drug Metabolism and Pharmacokinetics, Piramal Life Sciences Limited, Goregaon (E), Mumbai-400 063, India 2 School of Pharmacy and Technology Management, NMIMS University, Vile-Parle (W), Mumbai- 400 056, India

Correspondence Address:
Anagha Damre Drug Metabolism and Pharmacokinetics, Piramal Life Sciences Limited, Goregaon (E), Mumbai-400 063 India [email protected]

Small intestine plays an important role in the first-pass metabolism of orally ingested xenobiotics as a result of expression of both Phase I and Phase II metabolic enzymes, together with associated transporters. Intestinal microsomes thus can be used to study susceptibility of compounds to metabolism in vitro. The present study was undertaken to have a comparative assessment between different methods of preparation of rodent intestinal microsomes. Mouse and rat intestinal microsomes were prepared by two methods, in method A intestines were homogenized, while in method B mucosal cells were scrapped followed by homogenization. Further, microsomes were prepared by centrifugation (10000xg) followed by ultra centrifugation (100000xg) of the homogenates. The prepared microsomes were characterized for protein concentration using Bradford's method and CYP450 content using carbon monoxide bubbling method. The protein concentration and CYP450 content in microsomes prepared by method B was significantly higher than method A. In conclusion, superior quality intestinal microsomes can be obtained from rodents by using scrapped intestinal mucosal cells as compared to the intestinal homogenates.

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