Validation of a liquid chromatography-electrospray ionization-tandem mass spectrometry method for determination of all-trans retinoic acid in human plasma and its application to a bioequivalence study.
A sensitive, reliable and specific LC-MS-MS method was developed and validated for the identification and quantitation of all-trans retinoic acid (ATRA) in human plasma. Acitretin was used as the internal standard (IS). After liquid-liquid extraction of 500 μL plasma with methyl tert-butyl ether (MTBE), ATRA and the IS were chromatographed on a HyPURITY C18 column (150 mm×2.1 mm, 5 μm) with the column temperature set at 40 °C. The mobile phase was consisted of 40% phase A (MTBE-methanol-acetic acid, 50:50:0.5, v/v) and 60% phase B (water-methanol-acetic acid, 50:50:0.5, v/v) with a flow rate of 0.3 mL/min. The API 4000 triple quadrupole mass spectrometer was operated in multiple reaction monitoring (MRM) mode via the positive electrospray ionization interface using the transition m/z 301.4→123.1 for ATRA and m/z 326.9→177.1 for IS, respectively. The calibration curve was linear over the range of 0.45-217.00 ng/mL (r≥0.999) with a lower limit of quantitation (LLOQ) of 0.45 ng/mL. The intra- and inter-day precisions values were below 8% relative standard deviation and the accuracy was from 98.98% to 106.19% in terms of relative error. The validated method was successfully applied in a bioequivalence study of ATRA in Chinese healthy volunteers.
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. jingbopeng@csu.edu.cn.
Jing-Bo
JB
Peng
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. lchhr0107@163.com.
Chen-Hui
CH
Luo
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. leewanaimuting@sohu.com.
Yi-Cheng
YC
Wang
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. endeavor34852@aliyun.com.
Wei-Hua
WH
Huang
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. cbohua@163.com.
Yao
Y
Chen
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. hhzhou2003@163.com.
Hong-Hao
HH
Zhou
Institute of Clinical Pharmacology, Central South University, Changsha, Hunan 410078, China. tanzr@163.com.
Zhi-Rong
ZR
Tan
D000284:Administration, Oral; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D001774:Blood Chemical Analysis; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction; D008297:Male; D008738:Methyl Ethers / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013810:Therapeutic Equivalency; D014212:Tretinoin / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics
D000970:Antineoplastic Agents; D008738:Methyl Ethers; D012997:Solvents; C043243:methyl tert-butyl ether; D014212:Tretinoin