alexa Development and Validation of a UPLC-MS/MS Assay for Si
ISSN: 1948-593X

Journal of Bioanalysis & Biomedicine
Open Access

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Research Article

Development and Validation of a UPLC-MS/MS Assay for Simultaneous Estimation of Raloxifene and its Metabolites in Human Plasma

D. H. Jadhav and C. S. Ramaa*

Department of Pharmaceutical Chemistry, Bharati Vidyapeeth’s College of Pharmacy, Navi Mumbai-400614, India

*Corresponding Author:
C. S. Ramaa
Department of Pharmaceutical Chemistry
Bharati Vidyapeeth’s College of Pharmacy
Navi Mumbai-400614, India
Fax: +91-2227574515
E-mail: [email protected]

Received Date: July 29, 2012; Accepted Date: August 20, 2012; Published Date: August 23, 2012

Citation: Jadhav DH, Ramaa CS (2012) Development and Validation of a UPLCMS/ MS Assay for Simultaneous Estimation of Raloxifene and its Metabolites in Human Plasma. J Bioanal Biomed 4:061-067. doi: 10.4172/1948-593X.1000064

Copyright: © 2012 Jadhav DH, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.



An aim of the study is development and validation of a method for the simultaneous estimation of raloxifene (RAL) and its two active metabolites, raloxifene-4-glucuronide (R4G) and raloxifene-6-glucuronide (R6G) in human plasma samples using raloxifene-D4 as an Internal Standard. Sample preparation was performed by solid phase extraction (SPE) and was followed by separation of the analytes on a UPLC system with a linear gradient and a mobile phase consisting of acetonitrile and ammonium formate. Detection was achieved by tandem mass spectrometry operated in the electrospray positive ion mode. The method had a short sample preparation time, as well as a chromatographic run time of just 4.2 min, the shortest so far reported for RAL, R4G and R6G determination. It was validated and fulfilled all preset criteria for sensitivity, specificity, linearity, within inter- and intra-accuracy and precision, stability studies for all molecules. The method was linear in the concentration range of 0.040 to 1.5 ng/mL, 0.6 to 50.0 ng/mL and 0.6 to 50.0 ng/mL for RAL, R4G and R6G, respectively. The proposed method could be applied to the rapid and reliable simultaneous determination of RAL, R4G and R6G in a bioequivalence study.


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