title: Validation of an LC-MS/MS method for simultaneous detection of four HDAC inhibitors - belinostat, panobinostat, rocilinostat and vorinostat in mouse plasma and its application to a mouse pharmacokinetic study.
abstract: A sensitive and rapid LC-MS/MS method was developed and validated for the simultaneous quantitation of four HDAC inhibitors, namely belinostat (BST), panobinostat (PST), rocilinostat (RST) and vorinostat (VST), in mouse plasma as per regulatory guidelines. The analytes and internal standard were extracted from 50 μL mouse plasma by protein precipitation, followed by chromatographic separation using an Atlantis C18column with an isocratic mobile phase comprising 0.1% formic acid-acetonitrile (25:75, v/v) at a flow rate of 0.5 mL/min within 2.5 min. Detection and quantitation were done by multiple reaction monitoring on a triple quadrupole mass spectrometer following the transitions: m/z 319 → 93, 350 → 158, 434 → 274 and 265 → 232 for BST, PST, RST and VST, respectively, in the positive ionization mode. The calibration curves were linear from 2.92 to 2921 ng/mL for BST and PST and from 1.01 to 1008 ng/mL for RST and VST with r2≥ 0.99 for all of the analytes. The intra- and inter-batch accuracy and precision (CV) across quality controls varied from 85.5 to 112% and from 2.30 to 12.5, respectively, for all of the analytes. Analytes were found to be stable under different stability conditions. The method was applied to an i.v. pharmacokinetic study in mice.
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Kalpesh Kumar
authors.initials: KK
authors.lastname: Giri
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: P S
authors.initials: PS
authors.lastname: Suresh
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Syed Mohd
authors.initials: SM
authors.lastname: Saim
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Mohd
authors.initials: M
authors.lastname: Zainuddin
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Ravi Kanth
authors.initials: RK
authors.lastname: Bhamidipati
authors.affiliation: Department of Medicinal Chemistry, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Purushottam
authors.initials: P
authors.lastname: Dewang
authors.affiliation: Department of Medicinal Chemistry, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Mahanandeesha S
authors.initials: MS
authors.lastname: Hallur
authors.affiliation: Department of Medicinal Chemistry, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Sridharan
authors.initials: S
authors.lastname: Rajagopal
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Sriram
authors.initials: S
authors.lastname: Rajagopal
authors.affiliation: Drug Metabolism and Pharmacokinetics, Jubilant Biosys Ltd, Industrial Suburb, Bangalore, India.
authors.forename: Ramesh
authors.initials: R
authors.lastname: Mullangi
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D056572:Histone Deacetylase Inhibitors / Q000097:blood* / Q000493:pharmacokinetics; D057230:Limit of Detection; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D056572:Histone Deacetylase Inhibitors
keywords: LC-MS/MS; belinostat; method validation; mouse plasma; panobinostat; pharmacokinetics; rocilinostat; vorinostat