同位素稀释-超高效液相色谱-串联质谱法测定人血浆中氟康唑的浓度

目的:建立测定人血浆中氟康唑浓度的方法。方法:血浆样品经乙腈沉淀蛋白后,以同位素氟康唑-d4为内标,采用超高效液相色谱-串联质谱法测定。色谱柱为ACQUITY UPLC BEH C₁₈,流动相为0.1%甲酸溶液-乙腈(梯度洗脱),流速为0.3 mL/min,柱温为40℃,进样量为3μL;采用电喷雾离子源,以多反应监测模式进行正离子扫描,用于定量分析的离子对分别为m/z 307.1→220.0(氟康唑)和m/z 311.1→223.0(内标)。结果:氟康唑血药浓度检测的线性范围为10～5 000 ng/mL(r=0.998 1),定量下限为10 ng/mL;日内、日间RSD均低于8%,准确度为95.8%～106.7%;提取回收率为97.3%～107.3%(RSD<5.0%,n=6),基质效应、稀释效应及残留效应均不影响待测物的定量分析。结论:该方法简便、快速、专属性强、准确度高,可用于氟康唑治疗药物监测及药动学研究。

Determination of Fluconazole in Human Plasma by UPLC-MS/MS with Isotopes Dilution

OBJECTIVE:To establish a method for determination of fluconazole concentration in human plasma.METHODS:UPLC-MS/MS method was adopted to determine plasma after precipitated with acetonitrile.Using isotope fluconazole-d4 as internal standard,the determination was performed on ACQUITY UPLC BEH C18 column with mobile phase consisted of 0.1%formic acid-acetonitrile(gradient elution)at the flow rate of 0.3 mL/min.The column temperature was 40℃,and the sample size was 3μL.ESI was used for positive ion scanning by multiple reaction monitoring mode.The ion pairs for quantitative analysis were m/z 307.1→220.0(fluconazole)and m/z 311.1→223.0(internal standard).RESULTS:The linear range of fluconazole was 10-5 000 ng/mL(r=0.998 1).The limits of quantitation was 10 ng/mL.RSDs of intra-day and inter-day were less than 8%;accuracy ranged 95.8%-106.7%.The extraction recovery ranged 97.3%-107.3%(RSD<5.0%,n=6),and matrix effect,dilution effect and residual effect didn’t influence quantitative analysis of the substance to be measured.CONCLUSIONS:The method is simple,rapid,specific and accurate,which can be used for therapeutic drug monitoring and pharmacokinetic study of fluconazole.