Improved liquid chromatography-tandem mass spectrometry method for the analysis of eptifibatide in human plasma.
A rapid, selective and highly sensitive high performance liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed and validated for the determination and pharmacokinetic investigation of eptifibatide in human plasma. Eptifibatide and the internal standard (IS), EPM-05, were extracted from plasma samples using solid phase extraction. Chromatographic separation was performed on a C(18) column at a flow rate of 0.5 mL/min. Detection of eptifibatide and the IS was achieved by tandem mass spectrometry with an electrospray ionization (ESI) interface in positive ion mode. Traditional multiple reaction monitoring (MRM) using the transition of m/z 832.6-->m/z 646.4 and m/z 931.6-->m/z 159.4 was performed to quantify eptifibatide and the IS, respectively. The calibration curves were linear over the range of 1-1000 ng/mL with the lower limit of quantitation validated at 1 ng/mL. The intra- and inter-day precisions were within 13.3%, while the accuracy was within +/-7.6% of nominal values. The validated LC-MS/MS method was successfully applied for the evaluation of pharmacokinetic parameters of eptifibatide after intravenous (i.v.) administration of a 45 microg/kg bolus of eptifibatide to 8 healthy volunteers.
Zhou
Zhi-Ling
ZL
Medical Research Center, Guangdong General Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
Yu
Xi-Yong
XY
Yang
Min
M
Mai
Li-Ping
LP
Lin
Qiu-Xiong
QX
Deng
Chun-Yu
CY
Shan
Zhi-Xin
ZX
Kuang
Su-Juan
SJ
Zhu
Ping
P
Huang
Xiao-Zhong
XZ
Li
Xiao-Hong
XH
Chen
Tie-Feng
TF
Lin
Shu-Guang
SG
D044466:Asian People; D002853:Chromatography, Liquid; D004355:Drug Stability; D000077542:Eptifibatide; D005260:Female; D006262:Health; D006801:Humans; D008297:Male; D010455:Peptides / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D010975:Platelet Aggregation Inhibitors / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors
D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D000077542:Eptifibatide