LC-MS/MS测定紫草中醇溶性成分和水溶性成分的含量

Determination Content of Alcohol-soluble Components and Water-soluble Components in Arnebiae Radix by LC-MS/MS

目的:建立液相色谱串联质谱法测定紫草中醇溶性成分和水溶性成分(左旋紫草素,β,β’-二甲基丙烯酰阿卡宁,β-乙酰氧基异戊酰阿卡宁,紫草酸,咖啡酸,迷迭香酸)的含量,用于紫草的质量控制。方法:采用Agilent ZORBAX SB C18色谱柱(4.6 mm×50 mm,1.8μm),以0.1%甲酸乙腈(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱(0~3 min,5%~95%A;3~7 min,95%A;7~7.01 min,95%~5%A;7.01~8.50 min,5%A);流速0.2 mL·min^-1,柱温35℃,进样量5μL;采用负离子电离模式,MRM扫描模式进行定量。结果:左旋紫草素,β,β’-二甲基丙烯酰阿卡宁,β-乙酰氧基异戊酰阿卡宁,紫草酸,咖啡酸,迷迭香酸分别在10.12~1012μg·L^-1(r=0.9981),10.88~1088μg·L^-1(r=0.9912),10.08~806.4μg·L^-1(r=0.9976),20.32~1016μg·L^-1(r=0.9966),10.37~1037μg·L^-1(r=0.9996),10.26~1026μg·L^-1(r=0.9978)均有良好线性关系(r≥0.9912);平均加样回收率分别为95.8%(RSD 3.2%),103.5%(RSD 2.3%),105.3%(RSD 2.1%),96.1%(RSD 3.3%),98.9%(RSD 2.7%),100.8%(RSD 3.4%)。13批次紫草样品中6个种成分含量存在较大差异。结论:所建立的方法合理可行,为综合评价紫草质量提供一定依据。

Objective:To establish an LC-MS/MS method for the determination of alcohol-soluble components and water-soluble components in Arnebiae Radix(L-shikonin,β,β’-dimethylacrylshikonin,β-acetoxy-isovalerylshikonin,lithospermic acid,caffeic acid,rosmarinic acid),for the purpose of quality control of the herb.Method:Chromatographic separation was carried out at 35℃on Agilent ZORBAX SB C18(4.6 mm×50 mm,1.8μm),with 0.1%formic acid acetonitrile(A)-0.1%formic acid solution(B)as the mobile phase for gradient elution(0-3 min,5%-95%A;3-7 min,95%A;7-7.01 min,95%-5%A;7.01-8.5 min,5%A).The flow rate was 0.2 mL·min^-1,and the injection volume was 5μL.In a negative ionization mode,MRM scanning mode was adopted for quantification.Result:The calibration curves of L-shikonin,β,β’-dimethylacrylshikonin,β-acetoxy-isovalerylshikonin,lithospermic acid,caffeic acid,rosmarinic acid showed a good linearity,and the linear ranges of the above six compounds were 10.12-1012μg·L^-1(r=0.9981),10.88-1088μg·L^-1(r=0.9912),10.08-806.4μg·L^-1(r=0.9976),20.32-1016μg·L^-1(r=0.9966),10.37-1037μg·L^-1(r=0.9996),10.26-1026μg·L^-1(r=0.9978),respectively.The average recoveries of the analysts were 95.8%(RSD 3.2%),103.5%(RSD 2.3%),105.3%(RSD 2.1%),96.1%(RSD 3.3%),98.9%(RSD 2.7%),100.8%(RSD 3.4%).The contents of six components in 13 batches of samples showed significant differences.Conclusion:The established method is feasible and simple,and provides a basis for comprehensive quality evaluation of Arnebiae Radix.