离心式固相微萃取与液相色谱-质谱联用测定尿样中莨菪烷生物碱

Centrifugal solid-phase microextraction combined with LC-MS for the simultaneous determination of tropane alkaloids in urine sample

建立了离心式固相微萃取与液相色谱-质谱联用(LC-MS)同步测定尿样中4种莨菪烷烃类生物碱(山莨菪碱、氢溴酸樟柳碱、甲溴东莨菪碱和丁溴东莨菪碱)的方法。样品经C18微柱富集、分离解吸后进行LC-MS测定,前处理过程用时5 min。结果表明,山莨菪碱、氢溴酸樟柳碱和丁溴东莨菪碱在0.1~50μg/L浓度范围内线性关系良好,甲溴东莨菪碱线性范围为0.5~50μg/L,相关系数(R^(2))均大于0.99。4种莨菪烷碱的检出限(LODs)和定量限(LOQs)分别为0.02~0.15和0.08~0.50μg/L。同一柱、不同柱之间萃取10μg/L莨菪烷碱的精密度良好,相对标准偏差(RSDs)均低于10%。在尿样中分别添加4,20,80μg/L 3个浓度水平的莨菪烷碱,回收率为80.2%~99.6%,无明显基质效应。该方法适用于生物流体中莨菪碱类物质的同步测定。

A method based on centrifugal solid-phase microextraction combined with LC-MS was established for the simultaneous determination of tropane alkaloids(racanisodamine,anisodine hydrobromide,methscopolamine bromide,and N-butylscopolamine bromide)in urine.The sample solution was extracted by the C18 spin column,and the eluate was collected for determination by LC-MS.The pretreatment process took about 5 min.The linearity ranges of racanisodamine,anisodine hydrobromide,and N-butylscopolamine bromide were 0.1-50μg/L,and that of methscopolamine bromide was 0.5-50μg/L.The relative coefficients(R^(2))of the four tropane alkaloids were higher than 0.99.The limits of detection(LODs)and limits of quantification(LOQs)were 0.02-0.15 and 0.08-0.50μg/L,respectively.The precisions for extraction of tropane alkaloids(10μg/L)using one spin column and three different spin columns were lower than 10%.The extraction recovery rates of urine samples at three spiked concentration levels of 4,20 and 80μg/L were 80.2%-99.6%.No significant matrix effect was observed.This method can meet the requirement of simultaneous determination of various tropane alkaloids in biological samples.