title: Development and validation of a liquid chromatography/tandem mass spectrometry assay for the simultaneous determination of nateglinide, cilostazol and its active metabolite 3,4-dehydro-cilostazol in Wistar rat plasma and its application to pharmacokinetic study.
abstract: Nateglinide (NTG), an insulin secretogogue, has been studied in rats for drug-drug interaction with cilostazol (CLZ), an antiplatelet agent commonly used in diabetics. We developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) based method that is capable of simultaneous monitoring plasma levels of nateglinide, cilostazol, and its active metabolite 3,4-dehydro-cilostazol (DCLZ). All analytes including the internal standard (Repaglinide) were chromatographed on reverse phase C(18) column (50 mm x 4.6mm i.d., 5 microm) using acetonitrile: 2mM ammonium acetate buffer, pH 3.4 (90:10, v/v) as mobile phase at a flow rate 0.4 ml/min in an isocratic mode. The detection of analyte was performed on LC-MS/MS system in the multiple reaction monitoring (MRM) mode. The quantitations for analytes were based on relative concentration. The method was validated over the concentration range of 20-2000 ng/ml and the lower limit of quantitation was 20 ng/ml. The recoveries from spiked control samples were >79% for all analytes and internal standard. Intra- and inter-day accuracy and precision of validated method were with in the acceptable limits of <15% at all concentration. The quantitation method was successfully applied for simultaneous estimation of NTG, CLZ and DCLZ in a pharmacokinetic drug-drug interaction study in Wistar rats.
authors.affiliation: Department of Pharmacokinetics and Drug Metabolism, Glenmark Research Centre, Glenmark Pharmaceuticals Ltd., A-607, TTC Industrial Area, MIDC, Mahape, Navi Mumbai 400709, Maharashtra, India. kanthikiranv@gmail.com
authors.forename: Kanthi Kiran V S
authors.initials: KK
authors.lastname: Varanasi
authors.forename: V
authors.initials: V
authors.lastname: Sridhar
authors.forename: Suresh
authors.initials: S
authors.lastname: Potharaju
authors.forename: R
authors.initials: R
authors.lastname: Shraddha
authors.forename: S P N
authors.initials: SP
authors.lastname: Sivakumar
authors.forename: S Kanaga
authors.initials: SK
authors.lastname: Sabapathi
authors.forename: T R S
authors.initials: TR
authors.lastname: Satheeshmanikandan
authors.forename: V V S Swaroop
authors.initials: VV
authors.lastname: Kumar
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D000077407:Cilostazol; D003510:Cyclohexanes / Q000097:blood* / Q000493:pharmacokinetics; D000077715:Nateglinide; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013777:Tetrazoles / Q000097:blood* / Q000493:pharmacokinetics
chemical_list: D003510:Cyclohexanes; D013777:Tetrazoles; D000077715:Nateglinide; D010649:Phenylalanine; C519131:3,4-dehydrocilostazol; D000077407:Cilostazol