柱前衍生化结合UHPLC-MS/MS法同时测定Beagle犬血浆中的红景天苷和酪醇  

Simultaneous determination of salidroside and tyrosol in Beagle dog plasma using UHPLC-MS/MS after pre-column dansyl chloride derivatization

作　　者：陈帅[1,2] 夏媛媛[2] 魏广力[2] 李全胜[2] 刘梦杰[1,2] 陈勇[1] 司端运[2] 

机构地区：[1]湖北大学中药生物技术省重点实验室,湖北大学绿色资源生物转化协同创新中心,湖北武汉430062 [2]天津药物研究院释药技术与药代动力学国家重点实验室,天津300193

出　　处：《药学学报》2017年第2期296-301,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金青年科学基金资助项目(81503154)

摘　　要：本文建立了同时测定Beagle犬血浆中红景天苷及酪醇浓度的柱前衍生化结合UHPLC-MS/MS的分析方法。血浆样品用乙腈沉淀蛋白后,上清液与丹磺酰氯进行衍生化反应,终止反应后采用甲基叔丁基醚进行提取,提取物复溶后经反相色谱分离,色谱柱为ACQUITY UPLC~?BEH C_(18),(100 mm×2.1 mm,1.7μm),流动相为乙腈(含0.1%甲酸)-水(含10%乙腈,0.1%甲酸),梯度洗脱,分析时间为9 min。在正离子条件下采用多反应监测模式对两种待测物进行同时测定。用于定量分析的离子对分别为m/z 534.2→372.0(红景天苷衍生物),m/z372.0→171.0(酪醇衍生物)和m/z 506.0→171.0(内标熊果苷衍生物)。在上述条件下,红景天苷和酪醇分别在0.02~20和0.1~10μmol·L^(-1)内线性良好,定量下限分别为0.02和0.1μmol·L^(-1);批内、批间精密度均小于8.68%;准确度在±11.4%范围之内。本方法灵敏度高,专属性好,适合用于Beagle犬灌胃给药后血液中红景天苷、酪醇的药动学研究。A pre-column derivatization method combined with UHPLC-MS/MS was developed for the simultaneous determination of salidroside and tyrosol in Beagle dog plasma. After protein precipitation by acetonitrile,the liquid supernatant was treated with dansyl chloride under dark conditions at 60 ℃ for 30 min,and then,the sample solution was extracted using methyl tertiary butyl ether. The multiple reaction monitoring in positive ion mode was used for MS detection of the tested analytes with the specific ion transitions of m/z 534.2→372.0 for salidroside derivative,m/z 372.0→171.0 for tyrosol derivative and m/z 506.0→171.0 for arbutin derivative. The chromatograph separation was achieved on an ACQUITY UPLC BEH C18 column（100 mm × 2.1 mm,1.7 μm） with a gradient mobile phase consisting of acetonitrile（0.1% formic acid）-water（10% acetonitrile,0.1% formic acid） for 9 min. The assay showed a good linearity over the range of 0.02/0.1- 20/10 μmol·L^-1 with a lower limit of quantitation of 0.02 and 0.1 μmol·L^-1 for salidroside and tyrosol in dog plasma,respectively. The intra- and inter-day precisions were all less than 8.68%,and the accuracy was within ±11.4%. The established method with a high sensitivity,good specificity and reliability was appropriate for simultaneous determination of salidroside and tyrosol in dog plasma and successfully applied to a pharmacokinetic study after intragastric administration of salidroside to Beagle dogs.

关 键 词：红景天苷 酪醇 丹磺酰氯 衍生化 UHPLC-MS/MS 药代动力学 

分 类 号：R917[医药卫生—药物分析学]