植入宫内节育器(IUD)后Beagle犬血浆中吲哚美辛的LC-MS/MS测定法

Development of a liquid chromatography-tandem mass spectrometry method for measuring plasma levels of indomethacin in Beagle dog treated with intrauterine device (IUD)

目的建立快速、灵敏的液相色谱-大气压化学电离-串联质谱（LC-APCI-MS／MS）法测定植入含药宫内节育器（mo）后Beagle犬血浆中的吲哚美辛质量浓度。方法血浆样品经乙腈沉淀蛋白后，采用DiamonsilC18柱（150mm×4．6mm，5μ）分离，流动相为甲醇-乙腈-水-甲酸（体积比为45：45：10：0．5），流速为0．45mL·min^-1。采用大气压化学电离（APCI）源，以选择反应监测（SRM）方式进行检测，用于定量分析的离子反应分别为m／z358-m／z111（吲哚美辛）和m／z419-m／z343（内标，尼莫地平）。结果血浆中吲哚美辛的线性范围为1．0—200．0μg·L^-1，定量下限为1．0μg·L^-1，提取回收率为83．3％～91．4％，日内精密度不大于9．8％，日间精密度不大于5．9％，准确度（RE）在93．6％～100．0％内。结论该法简便、快速、灵敏，适用于植入含吲哚美辛的IUD的Beagle犬血浆中吲哚美辛的测定。

Objective To develop and validate a rapid and sensitive liquid chromatography-tandem mass spectrometric （LC-MS/MS） method for measuring plasma levels of indomethacin in Beagle dogs after in- sertion of intrauterine device （IUD）. Methods Indomethacin and internal standard were extracted from plas- ma with acetonitrile and separated on a Kromasil C18 column using a mobile phase consisting of methanol, acetonitrile,water and formic acid （V： V： V： V =45： 45： 10： 0. 5 ） at a flow rate of 0. 45 mL .m in-1. Mass spectrometric detection was achieved by a triple-quadrupole mass spectrometer equipped with an atmospheric pressure chemical ionization source （APCI） operating in positive ionization mode. Selected reaction monito- ring （SRM） using the precursor-product ion transitions of m/z 358→m/2 111 and m/z 419→m/z 343 were performed to quantify indomethacin and internal standard （nimodipine）, respectively. Results The assay was linear over the concentration range of 1.0- 200. 0 μg·L^-1 for indomethacin. The lower limit of quantifica- tion （LLOQ） was 1.0 μg·L^-1. The extraction recovery was more than 83.3%. The intra-day and inter-day precision were less than 9. 8% and 5.9% ,respectively. The accuracy was within 93.6% - 100. 0%. Conclu- sions The method is simple, rapid and sensitive, and can be successfully applied to determine indomethacin in Beagle dogs plasma after insertion of IUD.