Study On Developing Fingerprint Of Salvia Miltiorrhiza Bunge And Determining Water-soluble Components In It

With the development of Chinese traditional medicine (CTM) and the associated subjects, research on quality control of CTM is turning from content determination of single component to that of multi-component and multi-data. The technology of Fingerprint Spectrum based on modern analytical technology has been the main direction of quality control of Chinese herbs and Chinese medicinal preparations. As a most used method in developing fingerprint of CTM, high-performance liquid chromatography with ultraviolet detector cannot separate co-eluting peaks. Therefore, the sensitivity of it declined. However, high-performance liquid chromatography with Coulometric electrode array system (HPLC-CEAD) can solve these problems and it has never been applied in fingerprinting Salvia miltiorrhiza Bunge until now.In this study, HPLC-CEAD was developed to detect the contents of four water-soluble compounds in Salvia miltiorrhiza Bunge from Hebei province (Danshensu, protocatechuic acid, protocatechuic aldehyde and salvianolic acid) in the first section. This analysis was carried on a column of Hypersil C18 (250mm×4.6mm, 5μm) with a mobile phase of sodium acetate (pH2.5, 50mM) and acetonitrile in gradient mode as follows: 0~15min, 6%B; 15~45min, 6~22%B; 0.8mL/min. Potentials of four electrodes in series were set at 100, 150, 200 and 250mV, respectively. The results revealed that calibration curves showed good linearity with correlation coefficients (r) more than 0.9937, average recoveries of the four compounds were more than 97% and relative standard deviations were less than 6.6%.In the second section, the fingerprints of Salvia miltiorrhiza Bunge from Shandong and other places were studied by HPLC-CEAD system. The analysis column is the same as above with a mobile phase of sodium acetate (pH2.7, 50mM) and acetonitrile in gradient mode as follows: 0~15min, 6%B; 15~45min, 6~20%B; 45~60min, 20%~53%B; 60~120min, 53%B; 0.8mL/min. Potentials of four electrodes in series were set at–600mV, 100mV, 250mV and 500mV, respectively. The results of method validation showed that relative standard deviations of each compound in precision, stability and repeatability test were all less than 3.0% which met the requirement of fingerprint. 25 components in total were separated from extracts of Salvia miltiorrhiza Bunge using this method. From the results, it was concluded that HPLC-CEAD system provide super sensitivity and stability in development of...