论文部分内容阅读
建立了在线固相萃取(on-line SPE)结合液相色谱-线性离子阱多级质谱(LC-LIT/MS~n)同时测定全血、尿液和肝组织样品中7种乌头类生物碱的分析方法,并根据7种乌头类生物碱的多级质谱碎片对裂解规律进行了推测和总结。用乙腈沉淀样品中的蛋白质,于稀释和离心后直接进样。经Waters Oasis~ HLB在线SPE柱富集纯化,以0.1%(v/v)甲酸/乙酸铵溶液-0.1%(v/v)甲酸/甲醇溶液为流动相,以Accucore C18为分析柱进行梯度洗脱;在电喷雾电离(ESI)正离子模式下测定;扫描方式为连续反应监测(CRM)。在考察的质量浓度范围内,7种乌头类生物碱的标准曲线符合二阶方程(权重因子1/x),相关系数为0.999 1~0.999 9;在全血和尿液中的方法检出限为0.02~0.60 ng/mL,在肝组织中的方法检出限为0.02~0.40 ng/g;加标回收率为91.1%~104.7%,日内精密度和日间精密度分别为0.2%~10.7%、1.0%~13.7%(n=6)。该方法简单准确,灵敏度高,能够满足生物样品中7种乌头类生物碱的快速分析。
An online on-line SPE coupled with liquid chromatography-ion trap multi-stage mass spectrometry (LC-LIT / MS ~ n) was developed for the simultaneous determination of seven species of Aconitum species in whole blood, urine and liver tissue samples Alkali analysis method, and according to seven kinds of Aconitum alkaloids multi-stage mass spectrometry fragments were speculated on the rupture law and concluded. The protein in the sample is precipitated with acetonitrile and injected directly after dilution and centrifugation. The column was purified by Waters Oasis ~ HLB on-line SPE column. The mobile phase consisted of 0.1% (v / v) formic acid / ammonium acetate solution - 0.1% (v / v) formic acid / methanol and gradient of Accucore C18 Elution; electrospray ionization (ESI) positive ion mode; scanning mode is continuous reaction monitoring (CRM). The standard curve of the seven aconite alkaloids conformed to the second-order equation (weighting factor 1 / x) with a correlation coefficient of 0.999 1 to 0.999 9 in the investigated mass concentration range; the method was detected in whole blood and urine The limit of detection was 0.02-0.40 ng / g in the range of 0.02-0.60 ng / mL, and the recoveries were 91.1% -104.7%. The intra-day precision and intra-day precision were 0.2% 10.7%, 1.0% ~ 13.7% (n = 6). The method is simple, accurate and sensitive, and can meet the rapid analysis of seven aconite alkaloids in biological samples.