血浆中 BZ类失能剂的高效液相二极管矩阵检测分析方法的研究

Study on simultaneous separation and determination of 3 quinuclidinyl benzilate group compounds in human plasma by HPLC DAD

目的：建立血浆中 Ｂ Ｚ类失能剂的高效液相二极管矩阵检测分析方法，为 Ｂ Ｚ类失能剂中毒病人的临床诊断提供分析手段。方法：采用 Ｓｅｐ Ｐａｋ Ｃ１８ 柱固相提取，乙酸乙酯洗脱， Ｒｅｓｏｌｖｅ Ｃ１８ 反相柱分离，０．１２％ 三氟乙酸（ Ｔ Ｆ Ａ）的 ５０％ 甲醇溶液梯度洗脱，二极管矩阵检测器（ Ｄ Ａ Ｄ） 三波长（２２０±４）ｎｍ ，（２５４±４）ｎｍ ，（３００±４）ｎｍ 检测。结果：血浆中 Ｈ Ｐ Ｌ Ｃ Ｄ Ａ Ｄ 法检测 ４ 种失能剂的线性范围为 ０．１～１．２ μｇ，回收率为６２．７６％ ～９２．２０％ ，标准偏差为 ３．４６％ ～１１．８４％ ，最低检出限为 １００～４００ ｎｇ。流动相中 Ｔ Ｆ Ａ 和甲醇浓度及梯度洗脱是影响分离效果和色谱峰形的主要因素。结论：本文为临床诊断提供简便、快速、灵敏的检测方法，血浆蛋白不干扰测定。４ 种失能剂在血浆中采用 Ｈ Ｐ Ｌ Ｃ Ｄ Ａ Ｄ 法同时分离测定目前未见报道。

Objective: To develope a reversed phase high performance liquid chromatography method for the simultaneous determination of four 3 quinuclidinyl benzilate compounds in human plasma. Methods: The human plasma samples were extracted with Sep Pak C18 cartridge and eluted with ethyl acetate. The elute was blown dry by nitrogen gas, and then dissolved with ethanol. Chromatographic separation was achieved with a Resolve C18 column (150 mm×4.6 mm ID),and the effluent was monitored at 220±4, 254±4, 300±4 nm. Results: The assay was linear over the range of 100-1 200 ng/ml of the four 3 quinuclidinyl benzilate compounds in plasma and the inter and intra day precision (RSD) did not exceed 14.04% in the range. The recoveriy rates were in the range of 62.76-92.20%. The minimum detectable limit was 100-400 ng. Conclusion: The method is simple, rapid and highly sensitive.