Quantification of fudosteine in human plasma by high-performance liquid chromatography-electrospray ionization mass spectrometry employing precolumn derivatization with 9-fluorenylmethyl chloroformate.
This paper describes a novel method for the sensitive and selective determination of fudosteine in human plasma. The method involves a derivatization step with 9-fluorenylmethyl chloroformate (FMOC-Cl) in borate buffer and detection based on high-performance liquid chromatography-electrospray ionization mass spectrometry (LC/ESI/MS). After acetonitrile-induced protein precipitation of plasma samples, fudosteine was derivatized with FMOC-Cl, then extracted by ethyl acetate, evaporated, reconstituted and injected using an LC/ESI/MS instrument. Separation was achieved using an ODS column and isocratic elution. Excellent linearity was obtained for the entire calibration range from 0.05 to 20 microg/ml. Validation assays of the lower limit of quantification (LLOQ) as well as for the intra- and inter-batch precision and accuracy met the international acceptance criteria for bioanalytical method validation. Using the developed analytical method, fudosteine could be detected for the first time in human plasma with a low limit of detection (LLOD) of 0.03 microg/ml. The proposed method has been successfully applied to study the pharmacokinetics of fudosteine in healthy Chinese volunteers after single and multiple oral administration.
Xu
Fengguo
F
Key Laboratory of Drug Quality Control and Pharmacovigilance, China Pharmaceutical University, Ministry of Education, Nanjing 210009, China.
Zhang
Zunjian
Z
Jiao
Haoyan
H
Tian
Yuan
Y
Zhang
Beibei
B
Chen
Yun
Y
D000284:Administration, Oral; D001881:Borates; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D003553:Cystine / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D005260:Female; D005449:Fluorenes / Q000737:chemistry*; D006801:Humans; D008297:Male; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods* / Q000592:standards
D001881:Borates; D002021:Buffers; D005449:Fluorenes; D003553:Cystine; C054007:1-(9-fluorenyl)methyl chloroformate; C113797:fudosteine