摘要
目的：建立液相色谱-大气压化学离子化-串联质谱联用（LC-APCI-MS／MS）法测定大鼠尿液和胆汁中的BAPoTA-AM浓度。方法：尿液和胆汁样品经乙腈沉淀去蛋白，离心后取上清液进行LC-APCI-MS／MS分析。色谱柱：Zorbax C18柱（150mm×4．6mm，5μm）；流动相：含0．5％乙酸和75％甲醇的水溶液；流速：1mL／min。大气压化学离子化（APCI）离子源，以选择离子反应检测（SRM）方式进行检测。用于定量分析的检测离子对分别为：m／z 765．15→647．25（BAPTA-AM）和m／z 419．10→343．05（内标，尼莫地平）。结果：尿液和胆汁中BAPTA-AM在0．25-50n异／mL范围内线性关系良好，回收率大于90％，定量下限为0．25ng／mL．批内与批间RSD均小于10％。结论：建立的LC-APCI-MS／MS法符合生物样本测定要求，可用于大鼠静脉给予BAPTA-AM脂质体后，大鼠尿液和胆汁中BAPTA-AM浓度的测定。
Aim-To develop a sensitive and specific LC-APCI-MS/MS method for the determination of BAPTA-AM in rat urine and bile. Methods： The urine and bile collected from the samples were deproteinized by acetonitrile and centrifuged for the clean-up. The supematant was then injected onto the LC-MS/MS system. Chromatographic separation of BAPTA-AM was carried out on a Zorbax C18 column using mobile phase consisting of 0.5% acetic acid and methanol （25： 75） at a flow-rate of 1 mL/min. Elution was monitored by Finnigan TSQ tandem mass spectrometer equipped with APCI source, which was operated in positive ion mode. Selected reaction monitoring （SRM） using the precursor product ion combination ot m/z 765.15→647. 25 aria m/z 419.10→343.05 was used to quantify BAPTA-AM and the internal standard （nimodipine）, respectively. Results： Calibration curves of BAPTA-AM in rat urine and bile showed linearity in the range of 0.25 - 50 ng/mL. The lower limit of quantification was 0.25 ng/mL, and the recovery of BAPTA-AM was above 90%. Inter- and intra-batch RSD was less than 10%, which indicated good precision. Conclusion： The method is proved to be suitable for the determination of BAPTA-AM in rat urine and bile after intravenous administration of BAPTA-AM liposomes.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
2008年第3期243-247,共5页
Journal of China Pharmaceutical University
