4 components from Salviae Miltiorrhizae extracted by HPLC-UV/ESI-MS

Objective To establish the HPLC fingerprint of 4 components from Radix Salviae Miltiorrhizae(RSM) extract,to qualitatively analyze the major marker peaks in the HPLC profile by MS,and to simultaneouly and quantitatively determine danshensu,salvianolic acid B,cryptotanshinone,and tanshinone ⅡA,and further study the relationship between their fingerprint and pharmacological effect.Methods HPLC analysis was carried out on an ODS-2 column with linear gradient elution.The mobile phases were composed of formic acid aqueous solution(0.5%)(A) and acetonitrile(B).The gradient was as follows: 0 min,80% A,20% B;50 min,15% A,85%,B,strarting with 20% B,and injection volume of 20 μL.The fingerprint profiles were recorded at an optimized wavelength of 280 nm.And 0.2 mL·min-1 flow phase was transferred into the MS system,and 150~750 total positive and negative irons were scanned to qualitatively analyze the marker peaks in HPLC profile on line.Molecular ions were monitored to quantitatively determine danshensu,salvianolic acid B,cryptotanshinone,and tanshinone,respectively.Results Most of the the characterized peaks in the HPLC fingerprint were well separated and provided much chemical information of the pharmacological compounds,and 14 marker peaks were qualitatively determined.The quantitative method was valid,and the linear range of danshensu,salvianolic acid B,cryptotanshinone,and tanshinone,ⅡA was 0.05~10.0,0.05~20.0,0.05~20,and 0.05~20 μg·mL-1 with good correlation coefficient(r≥0.999 8).The mean recovery rates from fortified samples(n=5) of dihydrotanshinone Ⅰ,cryptotanshinone,tanshinone Ⅰ,and tanshinone ⅡA were 98.1%,99.0%,98.9%,and 99.5%,respectively,with variation coefficient RSD≤1.5%.Conclusion The established fingerprints of 4 components from SM extract are suitable for the screening of pharmacological active constituents and effect-fingerprint study.The quantitative method is accurate,simple and suitable for the determination of the above 4 components in crude herbal of SM.