title: Simultaneous quantitation of delamanid (OPC-67683) and its eight metabolites in human plasma using UHPLC-MS/MS.
abstract: Delamanid (OPC-67683) is a novel nitro-dihydroimidazo-oxazole derivative that is being developed by Otsuka Pharmaceutical Co., Ltd., Japan (referred to as Otsuka hereafter) for the treatment of tuberculosis (TB). An ultra-high performance liquid chromatographic-tandem mass spectrometry (UHPLC-MS/MS) method has been developed and validated for the determination of OPC-67683 and its eight metabolites, DM-6704, DM-6705, DM-6706, DM-6717, DM-6718, DM-6720, DM-6721 and DM-6722 in human plasma to support regulated clinical development. During method development several technical challenges such as poor chromatography, separation of structural isomers, conversion of the analytes, instability in matrix and long cycle time were encountered and overcome. A protein precipitation extraction (PPE) was used to extract plasma samples (50μL) and the resulting extracts were analyzed using reversed phase UHPLC-MS/MS with a electrospray (ESI) and selected reaction monitoring (SRM). The method was fully validated over the calibration curve range of 1.00-500ng/mL for all nine analytes with linear regression and 1/x(2) weighting according to regulatory guidance for bioanalysis. Based on three inter-day precision and accuracy runs, the between-run % relative standard deviation (RSD) for all nine analytes varied from 0.0 to 11.9% and the accuracy ranged from 92.7% to 102.5% of nominal at all quality controls (QC) concentrations, including the lower limit of quantitation (LLOQ) QC at 1.00ng/mL. The extraction recovery of OPC-67683 and its eight metabolites were above 95%. Various short term and long term solution and matrix stability were established including the stability of OPC-67683 and its eight metabolites in human plasma for 708 days at -70°C. Although this method has been used to support regulated clinic studies during the last decade and over ten thousand samples have been analyzed, this is the first time that the method development process and validation data have been published.
authors: Covance, 1121 East 3900 South, Salt Lake City, UT 84124, USA. Electronic address: mengm@labcorp.com.
authors: Min
authors: M
authors: Meng
authors: Covance, 1121 East 3900 South, Salt Lake City, UT 84124, USA.
authors: Benjamin
authors: B
authors: Smith
authors: Covance, 1121 East 3900 South, Salt Lake City, UT 84124, USA.
authors: Brad
authors: B
authors: Johnston
authors: Covance, 1121 East 3900 South, Salt Lake City, UT 84124, USA.
authors: Spencer
authors: S
authors: Carter
authors: Otsuka Pharmaceutical Development & Commercialization (OPDC), 2440 Research Boulevard, Rockville, MD 20850, USA.
authors: Jerry
authors: J
authors: Brisson
authors: Otsuka Pharmaceutical Development & Commercialization (OPDC), 2440 Research Boulevard, Rockville, MD 20850, USA.
authors: Sharin E
authors: SE
authors: Roth
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004311:Double-Blind Method; D006801:Humans; D057230:Limit of Detection; D009593:Nitroimidazoles / Q000097:blood*; D010080:Oxazoles / Q000097:blood*; D010919:Placebos; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D009593:Nitroimidazoles; C516022:OPC-67683; D010080:Oxazoles; D010919:Placebos
keywords: Delamanid (code-named OPC-67683); Human plasma; Selected reaction monitoring (SRM); Tuberculosis; Ultra-high performance liquid chromatography and tandem mass spectrometry (UHPLC–MS/MS)