LC-MS/MS法测定人血浆中的辅酶Q10

LC-MS/MS determination of coenzyme Q10 in human plasma

目的:建立一种快速、灵敏的LC-MS/MS法测定人血浆中的辅酶Q10。方法:血浆样品经正己烷液-液萃取2次,以甲醇为流动相,CapcellPakC18柱(35mm×2.0mm,5μm)进行分离,采用大气压化学电离源,以多反应监测(MRM)方式进行正离子检测。用于定量分析的离子反应分别为m/z864→197(辅酶Q10)和m/z796→197(内标辅酶Q9)。结果:测定血浆中辅酶Q10的线性范围为10.0～1000ng·mL-1,定量下限可达10.0ng·mL-1,日内、日间精密度(RSD)均小于8.9%,准确度(RE)在-0.9%～3.8%之间,单个样品分析时间为4.5min。结论:本方法分析测试时间短,灵敏度较高,血浆用量少,适用于人血浆样品中辅酶Q10的测定和药物动力学研究。

Objective：To develop a rapid,sensitive and convenient liquid chromatographic-tandem mass spectrometric（LC-MS/MS）method for quantification of coenzyme Q10（CoQ10）in human plasma.Methods：The plasma sample was extracted twice with n-hexane,then separated on a Capcell Pak C18（35 mm×2.0 mm,5 μm）column using methanol as the mobile phase.A triple quadrupole mass spectrometry equipped with atmospheric pressure chemical ionization（APCI）source was used as the detector and operated in the positive ion mode.Quantitation was performed using multiple reaction monitoring（MRM）of the transitions of m/z 864→197 and m/z 796→197 for CoQ10 and the internal standard CoQ9,respectively.Results：The linear concentration range of the calibration curve for CoQ10 was 10.0~1000 ng·mL-1.The lower limit of quantification was 10.0 ng·mL-1.The intra-day and inter-day relative standard deviation over the entire concentration range were less than 8.9%.The accuracy was in the range of-0.9% to 3.8% in terms of relative error.Each sample was chromatographed within 4.5 min.Conclusion：The method is sensitive,rapid and suitable for the determination of CoQ10 in human plasma and investigation of coenzyme Q10 pharmacokinetics.