title: Investigation of the "true" extraction recovery of analytes from multiple types of tissues and its impact on tissue bioanalysis using two model compounds.
abstract: LC-MS/MS has been widely applied to the quantitative analysis of tissue samples. However, one key remaining issue is that the extraction recovery of analyte from spiked tissue calibration standard and quality control samples (QCs) may not accurately represent the "true" recovery of analyte from incurred tissue samples. This may affect the accuracy of LC-MS/MS tissue bioanalysis. Here, we investigated whether the recovery determined using tissue QCs by LC-MS/MS can accurately represent the "true" recovery from incurred tissue samples using two model compounds: BMS-986104, a S1P1receptor modulator drug candidate, and its phosphate metabolite, BMS-986104-P. We first developed a novel acid and surfactant assisted protein precipitation method for the extraction of BMS-986104 and BMS-986104-P from rat tissues, and determined their recoveries using tissue QCs by LC-MS/MS. We then used radioactive incurred samples from rats dosed with3H-labeled BMS-986104 to determine the absolute total radioactivity recovery in six different tissues. The recoveries determined using tissue QCs and incurred samples matched with each other very well. The results demonstrated that, in this assay, tissue QCs accurately represented the incurred tissue samples to determine the "true" recovery, and LC-MS/MS assay was accurate for tissue bioanalysis. Another aspect we investigated is how the tissue QCs should be prepared to better represent the incurred tissue samples. We compared two different QC preparation methods (analyte spiked in tissue homogenates or in intact tissues) and demonstrated that the two methods had no significant difference when a good sample preparation was in place. The developed assay showed excellent accuracy and precision, and was successfully applied to the quantitative determination of BMS-986104 and BMS-986104-P in tissues in a rat toxicology study.
authors.affiliation: Bioanalytical Sciences, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA. Electronic address: long.yuan@bms.com.
authors.forename: Long
authors.initials: L
authors.lastname: Yuan
authors.affiliation: Biotransformation, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.affiliation: Discovery Toxicology, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Lisa
authors.initials: L
authors.lastname: Dillon
authors.affiliation: Metabolism and Pharmacokinetics, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: R Marcus
authors.initials: RM
authors.lastname: Fancher
authors.affiliation: Metabolism and Pharmacokinetics, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Huadong
authors.initials: H
authors.lastname: Sun
authors.affiliation: Biotransformation, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Mingshe
authors.initials: M
authors.lastname: Zhu
authors.affiliation: Discovery Toxicology, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Lois
authors.initials: L
authors.lastname: Lehman-McKeeman
authors.affiliation: Bioanalytical Sciences, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Aubry
authors.affiliation: Bioanalytical Sciences, Research & Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA. Electronic address: qin.ji@bms.com.
authors.forename: Qin C
authors.initials: QC
authors.lastname: Ji
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D008956:Models, Chemical; D051381:Rats; D012015:Reference Standards; D053719:Tandem Mass Spectrometry
keywords: Extraction recovery; Intact tissue QC; LC-MS/MS; QC in tissue homogenate; Radioactive incurred samples; Tissue bioanalysis