LC-MS/MS法测定血浆中呋塞米浓度的研究

Study on Quantification of Furosemide in Patient Serum Using UFLC-MS/MS

目的建立UFLC-MS/MS测定血浆中呋塞米浓度的方法。方法通过电喷雾离子化,采用多反应检测方式进行负离子检测,用于定量分析的检测离子为m/z 328.8→284.6。采用Shim-pack XR-ODS(3.0 mm×75 mm,2.0μm)柱分离,以乙腈-水-甲酸(60∶40∶0.1)为流动相,流速为0.40 ml.min-1。结果在3 min内完成呋塞米定量分析。线性范围为2.5～1000 ng.ml-1,最低定量限为2.5 ng.ml-1;日内、日间测定的相对标准偏差分别小于2.13%、2.68%。结论本方法重复性好,灵敏度高、分析速度快、操作简单,可作为血浆样品中呋塞米浓度的测定方法,适用于人体呋塞米血药浓度监测。

Objective To establish an easy UFLC-MS/MS method for the quantification of furosemide in se- rum. Methods LC-MS/MS detection was performed using a triple-stage quadrupole mass spectrometer working in the multiple reaction monitoring mode with negative electrospray ionization using the transitions of m/z 328.8→284.6. Chromatographic separation was achieved using a Shim-pack XR-ODS column with acetonitrile-water-formic acid （60： 40：0.1 ） as the mobile phase at a flow-rate of 0.4 ml .rain-1. Resutls The analysis was finished in a sin- gle run within 3 min. Linearity was demonstrated over the expected concentration range 2.5-1000 ng ·ml^-1. The lower limit of quantification was 2.5 ng ·ml^-1. Intra-day precision expressed as RSD was ≤2.13% or less while in- ter-day result was ≤2.68% for furosemide. Conclusion The method has good repeatability, high sensitivity, and can be successfully applied to the determination of furosemide in patients.