Liquid-liquid Extraction of Everolimus an Immunosuppressant from Human Whole Blood and its Sensitive Determination by UHPLC-MS/MS
Vivek Upadhyay, Vikas Trivedi, Priyanka A. Shah, Jaivik V. Shah, Pranav S. Shrivastav
Corresponding Author : Pranav S. Shrivastav,
Chemistry Department, Kadi Sarva Vishwavidyalaya, Sarva Vidyalaya Campus, Gandhinagar, India.
Email ID : email@example.com
Received : 2014-05-29 Accepted : 2014-06-25 Published : 2014-06-25
Abstract : A sensitive and precise ultra high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method has been developed and fully validated for therapeutic drug monitoring of everolimus in human whole blood. Sample preparation involved liquid-liquid extraction of everolimus and its deuterated internal standard (IS, everolimus-d4) from 100 ÂµL blood sample using diethyl ether: ethyl acetate (30:70, v/v) solvent system under alkaline conditions. The chromatography was conducted on a COSMOSIL 2.5C18-MS-II (50 mm Ã— 2.0 mm, 2.5 Âµm) analytical column. The analyte and IS were eluted within 2.5 min under isocratic conditions using 10mM ammonium acetate, pH 6.00 adjusted with formic acid and acetonitrile (20:80, v/v). Multiple reaction monitoring was used for the quantitation of everolimus (m/z 975.5 â†’ 908.5) and everolimus-d4 (m/z 979.6 â†’ 912.6) in the positive ionization mode. The method was shown to be linear over the entire concentration range from 0.10-50.0 ng/mL. The recovery ranged from 90.9-94.8 % for everolimus and 91.4-95.6 % for everolimus-d4. The selectivity of the method is demonstrated in six different sources of blank human blood. The method is free from matrix effect as apparent from the post-column analyte infusion experiment, absolute and relative matrix effect results. The stability of everolimus in whole blood was thoroughly established under different storage conditions.
Keywords : Everolimus; everolimus-d4; UHPLC-MS/MS; liquid-liquid extraction; sensitivity; matrix effect
Citation : Pranav S. Shrivastav, et al (2014). Liquid-liquid Extraction of Everolimus an Immunosuppressant from Human Whole Blood and its Sensitive Determination by UHPLC-MS/MS. J. of Advancement in Medical and Life Sciences. DOI : 10.5281/zenodo.999754
Copyright : © 2014 Pranav S. Shrivastav. 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 credit
Volume / Issue
ScienceQ Publishing GroupDownload Article