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Development and validation of a HPLC method for quantification of rivastigmine in rat urine and identification of a novel metabolite in urine by LC-MS/MS†

Arumugam, Karthik and Rao, Mallikarjuna C and Reddy, Ravindranath Gilibili and Mullangi, Ramesh and Ganesan, Subramanian and Kar, Sidhartha S and Averineni, Ranjith Kumar and Shavi, Gopal and Udupa, Nayanabhirama (2011) Development and validation of a HPLC method for quantification of rivastigmine in rat urine and identification of a novel metabolite in urine by LC-MS/MS†. Biomedical chromatography, 25. pp. 353-361.

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Abstract

A sensitive, specific and accurate HPLC method for the quantification of rivastigmine (RSM) in rat urine was developed and validated. The method involves the simple liquid–liquid extraction of RSM and pyridostigmine as an internal standard (IS) from rat urine with tertiarymethyl butyl ether. The chromatographic separation of RSM and IS was achieved with 20 mM ammonium acetate buffer (pH 6.5) and acetonitrile (65:35, v/v) delivered at flow-rate of 1 mL/min on a Kromasil KR-100. Themethod was in linear range from 50 to 5000 ng/mL. The validation was done as per FDA guidelines and the results met the acceptance criteria. Themethod was successfully applied for the quantification of RSM in rat urine. Besides method validation, we have identified twometabolites of RSM in urine. Both the metabolites were characterized by HPLC-PDA and LC-MS/MS and it was found that one metabolite is novel. Copyright © 2010 JohnWiley & Sons, Ltd.

Item Type: Article
Uncontrolled Keywords: rivastigmine; rat urine; validation; HPLC; metabolites; characterization; LC-MS/MS
Subjects: Pharmacy > MCOPS Manipal > Pharmacology
Depositing User: KMC Manipal
Date Deposited: 23 Jan 2012 04:15
Last Modified: 02 Nov 2016 09:26
URI: http://eprints.manipal.edu/id/eprint/2661

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