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

Karthik Arumugam, Mallikarjuna Rao Chamallamudi, Ravindranath Reddy Gilibili, Ramesh Mullangi, Subramanian Ganesan, Sidhartha S. Kar, Ranjithkumar Averineni, Gopal Shavi, Nayanabhirama Udupa

Research output: Contribution to journalArticle

15 Citations (Scopus)

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 tertiary methyl butyl ether. The chromatographic separation of RSM and IS was achieved with 20mm ammonium acetate buffer (pH 6.5) and acetonitrile (65:35, v/v) delivered at flow-rate of 1mL/min on a Kromasil KR-100. The method was in linear range from 50 to 5000ng/mL. The validation was done as per FDA guidelines and the results met the acceptance criteria. The method was successfully applied for the quantification of RSM in rat urine. Besides method validation, we have identified two metabolites 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.

Original languageEnglish
Pages (from-to)353-361
Number of pages9
JournalBiomedical Chromatography
Volume25
Issue number3
DOIs
Publication statusPublished - 03-2011

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Rivastigmine
Metabolites
Rats
High Pressure Liquid Chromatography
Urine
Pyridostigmine Bromide
Methyl Ethers
Liquid-Liquid Extraction
Personal digital assistants
Liquids
Ether
Buffers
Flow rate
Guidelines

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Cite this

Arumugam, Karthik ; Chamallamudi, Mallikarjuna Rao ; Gilibili, Ravindranath Reddy ; Mullangi, Ramesh ; Ganesan, Subramanian ; Kar, Sidhartha S. ; Averineni, Ranjithkumar ; Shavi, Gopal ; Udupa, Nayanabhirama. / 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. In: Biomedical Chromatography. 2011 ; Vol. 25, No. 3. pp. 353-361.
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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; Chamallamudi, Mallikarjuna Rao; Gilibili, Ravindranath Reddy; Mullangi, Ramesh; Ganesan, Subramanian; Kar, Sidhartha S.; Averineni, Ranjithkumar; Shavi, Gopal; Udupa, Nayanabhirama.

In: Biomedical Chromatography, Vol. 25, No. 3, 03.2011, p. 353-361.

Research output: Contribution to journalArticle

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