LC-MS/MS分析生物检材中的呋喃酚葡萄糖醛酸结合物

Analyzing intact carbofuran-7-phenyl glucuronic acid in biological samples by LC-MS/MS

目的建立生物检材中呋喃酚葡萄糖醛酸结合物的LC-MS/MS检测方法。方法血和肝等检材经乙腈沉淀蛋白,采用ZORBAX HILIC Plus(4.6mm×100mm,3.5μm)亲水性色谱柱,以水和含0.1%甲酸的乙腈为流动相,梯度洗脱,以负离子多反应检测(MRM)模式检测,扫描时间8min。结果血和肝中呋喃酚葡萄糖醛酸结合物的线性范围分别是20ng/ml^5μg/ml和50ng/g^5μg/g,线性关系良好,r^2均大于0.999,血和肝中最低检出限分别为10ng/ml和9.93ng/g,最低定量限分别为15.63ng/ml和16.56ng/g;批内及批间精密度分别在1.53%~5.02%和5.06%~6.85%;相对回收率为86.13%~92.71%和87.01%~94.06%。对一例克百威中毒死亡人体检材进行检测,克百威及呋喃酚的血浓度分别为0.77ug/g和29.65ug/g,呋喃酚葡萄糖醛酸结合物的血浓度为1.059ug/g,肝组织浓度为4.056 ug/g。结论本研究所建方法特异性强、灵敏度高,可满足法医学中对于血中呋喃酚葡萄糖醛酸结合物的检测需求。

Objective To establish a detection method of intact carbofuran-7-phenyl glucuronic acid in biological samples by LC-MS/MS. Method Blood and liver samples were precipitated with acetonitrile. ZORBAX HILIC Plus(4.6×100 mm, 3.5 μm), a hydrophilic column, was used to analyze. The mobile phases were water and 0.1% formic acid in acetonitrile, gradient elution. Then the eluant was detected in multiple reaction monitoring(MRM) mode with negative ionization, scanning for 8 min. Results The linear range of carbofuran-7-phenyl glucuronic acid in the blood and liver is 20 ng/ml^5 ug/ml and 50 ng/g^5 ug/g, respectively and the linear relationship is good. Regression coefficient R is greater than 0.999, and the minimum limit of detection in blood and liver is 10 ng/ml and 9.93 ng/ml, respectively, the minimum limit of quantification is 15.63 ng/ml and 16.56 ng/g, respectively;the intra-assay and inter-assay precision are 1.53%~5.02% and 5.06%~6.85%;the relative recovery rate is 86.13%~92.71% and 87.01%~94.06%. A case of carbofuran poisoning is analyzed, the blood content of carbofuran and carbofuran-phenol is 0.77 ug/ml and 29. 65 ug/ml. The concentration of carbofuran-7-phenyl glucuronic acid in blood is 1.059 ug/ml, while in the liver is 4.056 ug/g. Conclusion The method can meet the requirements of analyzing carbofuran-7-phenyl glucuronic acid in biological samples, due to its high specificity and sensitivity.