title: Development and validation of an UFLC-MS/MS method for enantioselectivity determination of d,l-thero-methylphenidate, d,l-thero-ethylphenidate and d,l-thero-ritalinic acid in rat plasma and its application to pharmacokinetic study.
abstract: A chiral UFLC-MS/MS method was established and validated for quantifying d-threo-methylphenidate (d-threo-MPH), l-threo-methylphenidate (l-threo-MPH), d-threo-ethylphenidate (d-threo-EPH), l-threo-ethylphenidate (l-threo-EPH) and d,l-threo-ritalinic acid (d,l-threo-RA) in rat plasma over the linearity range of 1-500ng/mL. Chiral separation was performed on an Astec Chirobiotic V2 column (5μm, 250×2.1mm) with isocratic elution using methanol containing 0.003% ammonium acetate (w/v) and 0.003% trifluoroacetic acid (v/v) at a flow of 0.3mL/min. All analytes and IS were extracted from rat plasma by a one-step liquid-liquid extraction (LLE) method. The intra- and inter-run accuracies were within 85-115%, and the intra- and inter-run precision were <10% for all analytes. Extraction recoveries were 55-62% for d-threo-MPH, 54-60% for l-threo-MPH, 55-60% for d-threo-EPH, 53-57% for l-threo-EPH and 25-30% for d,l-threo-RA. The validated UFLC-MS/MS method successfully applied to the pharmacokinetic interaction study of oral d-threo-MPH and l-threo-MPH (alone or in combination) in female Sprague Dawley rats. The EPH was not detected in rat plasma following oral administrated MPH without EtOH. As far as it is known to the authors, this study is the first one step liquid-liquid extraction method to extract and UFLC-MS/MS method to quantify d-threo-MPH, l-threo-MPH, d-threo-EPH, l-threo-EPH and d,l-threo-RA simultaneously.
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Chenghao
authors.initials: C
authors.lastname: Zhang
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Huafei
authors.initials: H
authors.lastname: Luo
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Yubo
authors.initials: Y
authors.lastname: Wu
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Junyun
authors.initials: J
authors.lastname: Zhang
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Furong
authors.initials: F
authors.lastname: Zhang
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China.
authors.forename: Guobei
authors.initials: G
authors.lastname: Lin
authors.affiliation: National Pharmaceutical Engineering Research Center, China State Institute of Pharmaceutical Industry, 1111 Ha Lei Road, Shanghai 201203, China. Electronic address: 13162596759@163.com.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D008774:Methylphenidate / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D008774:Methylphenidate
keywords: Chiral separation; Drug interaction; Ethylphenidate; Methyphenidate; Pharmacokinetics; UFLC-MS/MS