HPLC-DAD-ESI/MS^n-DPPH在线筛选与鉴别丹参和康定鼠尾草中抗氧化活性成分  

On-line Screening and Identification of Free Radical Scavenging Compounds in Salvia miltiorrhiza Bunge and Salvia prattii Hemsl. by HPLC-DAD-ESI / MS^n-DPPH

作　　者：耿丹丹[1,2] 董琦[1] 谭亮[1] 迟晓峰[1] 许旭[1,2] 胡风祖[1] 

机构地区：[1]中国科学院西北高原生物研究所,青海西宁810008 [2]中国科学院大学,北京100049

出　　处：《分析测试学报》2015年第3期314-320,共7页Journal of Instrumental Analysis

基　　金：中国科学院仪器功能开发项目(yg2012086)

摘　　要：本研究建立了在线高效液相色谱-质谱-二苯基三硝基苯肼(HPLC-DAD-ESI/MSn-DPPH)快速筛选和鉴别丹参和康定鼠尾草中抗氧化活性成分和含量的方法。经液相色谱、质谱和文献报道综合分析鉴定出丹参和康定鼠尾草中的3种抗氧化活性化合物,分别为咖啡酸、异迷迭香酸苷和迷迭香酸。比较了热回流、超声辅助提取和快速溶剂萃取的提取效果,并对色谱条件进行优化。在优化条件下,这3种化合物均可有效分离,并在1.7~35.3μg/m L范围内线性关系良好,相关系数为0.999 5~0.999 8,检出限为0.05~1.85μg/m L,定量下限为0.18~6.16μg/m L,平均回收率为96.6%~97.2%,相对标准偏差为1.1%~1.3%。运用该方法测定丹参和鼠尾草样品中这3个化合物的含量分别为:咖啡酸0.303,0.254 mg/g;异迷迭香酸苷1.019,1.401 mg/g;迷迭香酸17.279,8.104 mg/g。本方法简便、快速、准确、重现性好,适用于从复杂天然产物中快速筛选与鉴别抗氧化活性成分。A new method was used for the rapid screening and identification of free radical scavengers in the extracts of Salvia miltiorrhiza Bunge and Salvia prattiiHemsl. , and the determination of the con- tent of the analytes in the samples based on HPLC - DAD - ESI/MS - DPPH was developed. The three compounds were identified to be eaffeic acid, salviaflaside and rosmarinic through comprehen- sive analysis of HPLC spectra, MS spectra and literature data. The extraction effects of the hot re- flux, ultrasonic assisted extraction and accelerated solvent extraction （ASE） were compared and the chromatographic conditions were optimized. Under the optimized conditions, three compounds could be effectively separated. Good linear relationships were obtained for three compounds in the concen- tration range of 1.7 - 35.3 I^g/mL, with correlation coefficients of 0. 999 5 - 0. 999 8. The detec- tion limits were in the range of O. 05 - 1.85 Ixg/mL, and the quantitation limits were in the range of 0. 18 -6. 16 μg/mL. The average recoveries of three compounds were between 96.6% and 97.2% , with relative standard deviations of 1.1% - 1.3% . The method was applied in the determination of three compounds contents in Salvia miltiorrhiza Bunge and Salvia prattiiHemsl. , with caffeic acid： 0. 303, 0. 254 mg/g; salviaflaside： 1. 019, 1. 401 mg/g; rosmarinic： 17. 279, 8. 104 mg/g. The method is convenient, fast, accurate and reproducibile. Otherwise, it gives a powerful tool for the rapid screening and identification of free radical scavenging compounds in complex natural products.

关 键 词：丹参 康定鼠尾草 抗氧化活性成分 HPLC-DAD-ESI/MSn-DPPH在线筛选 SALVIA prattiiHemsl. 

分 类 号：O657.72[理学—分析化学] TQ460.72[理学—化学]