title: Bioanalysis of propylparaben and p-hydroxybenzoic acid, and their sulfate conjugates in rat plasma by liquid chromatography-tandem mass spectrometry.
abstract: Two rugged liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods for the determination of propylparaben, its major metabolite, p-hydroxybenzoic acid (pHBA), and their sulfate conjugates have been developed and validated in citric acid-treated rat plasma. To prevent propylparaben being hydrolyzed to pHBA ex vivo, rat plasma was first treated with citric acid; then collected and processed at a reduced temperature (ice bath). Stable isotope labeled internal standards, d4-propylparaben, (13)C6-pHBA, and the d4-labeled internal standards of their sulfate conjugates were used in the methods. The analytes were extracted from the matrix using protein precipitation, followed by chromatographic separation on a Waters ACQUITY UPLC HSS T3 column. Quantification using negative ion electrospray was performed on a Sciex API 4000 mass spectrometer. The analytical ranges were established from 2.00 to 200 ng/mL for propylparaben, 50.0-5000 ng/mL for pHBA, 50.0-10,000 ng/mL for the sulfate conjugate of propylparaben (SPP) and 200-40,000 ng/mL for the sulfate conjugate of pHBA (SHBA). Inter- and intra-run precision for the quality control samples were less than 5.3% and 4.4% for all analytes; and the overall accuracy was within ±5.7% of the nominal values. The validated bioanalytical methods demonstrated excellent sensitivity, specificity, accuracy and precision and were successfully applied to a rat toxicology study under the regulations of Good Laboratory Practices (GLP). Strategies have been developed and applied toward overcoming the challenges related to analyte stability, and environmental and endogenous background.
authors.affiliation: Bioanalytical Sciences Department, Research & Development, Bristol-Myers Squibb Co, Route 206 & Province Line Road, Princeton, NJ, 08543, United States of America.
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Bioanalytical Sciences Department, Research & Development, Bristol-Myers Squibb Co, Route 206 & Province Line Road, Princeton, NJ, 08543, United States of America. Electronic address: guowen.liu@bms.com.
authors.forename: Guowen
authors.initials: G
authors.lastname: Liu
authors.affiliation: Drug Safety Evaluation, Research & Development, Bristol-Myers Squibb Co, One Squibb Drive, New Brunswick, NJ, 08903, United States of America.
authors.forename: Hongwu
authors.initials: H
authors.lastname: Shen
authors.affiliation: Bioanalytical Sciences Department, Research & Development, Bristol-Myers Squibb Co, Route 206 & Province Line Road, Princeton, NJ, 08543, United States of America.
authors.forename: Jim X
authors.initials: JX
authors.lastname: Shen
authors.affiliation: Bioanalytical Sciences Department, Research & Development, Bristol-Myers Squibb Co, Route 206 & Province Line Road, Princeton, NJ, 08543, United States of America.
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Aubry
authors.affiliation: Drug Safety Evaluation, Research & Development, Bristol-Myers Squibb Co, One Squibb Drive, New Brunswick, NJ, 08903, United States of America.
authors.forename: Lakshmi
authors.initials: L
authors.lastname: Sivaraman
authors.affiliation: Bioanalytical Sciences Department, Research & Development, Bristol-Myers Squibb Co, Route 206 & Province Line Road, Princeton, NJ, 08543, United States of America.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005520:Food Preservatives / Q000737:chemistry*; D062385:Hydroxybenzoates / Q000097:blood / Q000737:chemistry*; D015394:Molecular Structure; D010226:Parabens / Q000737:chemistry*; D010949:Plasma / Q000737:chemistry; D051381:Rats; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005520:Food Preservatives; D062385:Hydroxybenzoates; D010226:Parabens; C006068:propylparaben
keywords: LC–MS/MS; Propylparaben; Regulated bioanalysis; Validation