高效液相色谱-串联质谱法测定猪尿液中盐酸可乐定和盐酸赛庚啶

建立了高效液相色谱-串联质谱法检测猪尿中盐酸可乐定和盐酸赛庚啶的分析方法。样品用高氯酸溶液酸解及沉淀蛋白质,用乙酸乙酯-异丙醇提取,经MCX固相萃取小柱净化、富集后,采用Venusil MP C18色谱柱(100mm×2.1mm,3μm),以0.1%甲酸(体积分数,下同)5%乙腈水溶液和0.1%甲酸95%乙腈水溶液作为流动相进行梯度洗脱,采用多反应监测(MRM)模式进行定性和定量分析。盐酸可乐定和盐酸赛庚啶在1.0～100.0μg.L-1的范围内线性关系良好(r≥0.999 5)。在0.2、2.0、10.0μg.L-1添加水平下的回收率为83.5%～95.1%,相对标准偏差(RSD,n=6)为3.5%～8.7%,定量限(以信噪比>10计)为0.2μg.L-1。该方法精密度好,灵敏度高,能简便准确地测定猪尿中的盐酸可乐定和盐酸赛庚啶。

HPLC-MS/MS Determination of Clonidine Hydrochloride and Cyproheptadine Hydrochloride Contents in Pig Urine

A methodology applying HPLC-MS/MS was developed for the determination of clonidine hydrochloride and cyproheptadine hydrochloride contents in pig urine.Samples were hydrolyzed with perchloric acid,extracted with ethyl acetate-isopropanol,and purified through an MCX SPE cartridge before feeding to a Venusil MP C18 column(100 mm×2.1 mm,3 μm).On the column,the sample solution was flushed with a 0.1%(v/v) formic acid and 5%(v/v) acetonitrile aqueous solution,as well as a 0.1%(v/v) formic acid and 95%(v/v) acetonitrile aqueous solution as the mobile phase at the flow rate of 0.3 mL·min-1 and the column temperature of 30℃.The multiple reaction monitoring mode was used for the qualitative and quantitative analyses,and the external standard method for the quantitative determination.When the concentration of clonidine hydrochloride or cyproheptadine hydrochloride was 1.0-100.0 μg·L-1,a linear correlation(r≥0.9995) existed.At the addition of 0.2,2 and 10.0 μg·L-1 of the standard chemicals,the recovery rates were 83.5%-95.1% with an RSD(n=6) of 3.5%-8.7% and a quantity limit of 0.2 μg·L-1(the signal to noise ratio >10).The newly developed method appeared to be precise,sensitive,accurate and simple to operate for the determination of clonidine hydrochloride and cyproheptadine hydrochloride contents in pig urine.