Stadardized Processing Conditions Of Salvia Militorriza Pieces And Simultaneous Determination Of Six Major Active Components In Its Related Species By HPLC

The dried root and rhizome of Salvia miltiorrhiza (Lamiaceae, Salvia), one of the popular traditional Chinese medicine (TCM), is officially listed in Chinese Pharmacopoeia under the name Danshen. It was widely used for its good treatments to many diseases and Because of its red skin, thick flesh and high content of active components, DanShen cultivated in Sichuan was better than others. Accordingly, Sichuan province was considered as the genuine producing area of DanShen, which was mainly cultivated at Zhongjiang County. This study researched the processing procedure and quatitative standard of Salvia miltiorrhiza cultivated in Sichuan.In additional, the RP-HPLC methods were established to simultaneous analysis six active components in S. miltiorrhiza and its related species. The main studies as follows:1. We employed the L9(34) orthogonal design to optimize the processing procedure about fired S. miltiorrhiza, wine fired S. miltiorrhiza and vinegar fired S. miltiorrhiza by determine the content of water-soluble as Protocatechuic aldehyde, Salvianolic acid A, Salvianolic acid B and liposoluble constituents as Tanshinone I, Cryptotanshinone, Tanshinone II A. The optimum condition for processing of Stir-frying S. miltiorrhiza is:the prepared pieces of S. miltiorrhiza are roast at 70-80℃for 5 min; The optimum condition for processing of Wine Fired S. miltiorrhiza is:Add 20% wine per 100g S. miltiorrhiza, moisten for 60min, roast at 100-110℃for 5 min; The optimum condition for processing of Vinegar Fired S. miltiorrhiza is:Add 20% vinegar per 100g S. miltiorrhiza, moisten for 60min, roast at 70-80℃for 20 min.2. Based on the quality standard of S. miltiorrhiza in Chinese Pharmacopoeia(2010), the micro-identification, water, total ash, acid-insoluble ash, extractives and heavy metal the in different processed produces were assayed, the results showed that the content of water was 8.0%-8.9%; the contents of total ash, acid-insoluble ash were 5.971%-6.047% and 0.4239%-0.8634%, the contents of extractives were 55.81-57.94% and 41.77%-43.12.% At the same time, the TLC chromatography showed that there has the corresponding dot with Salvianolic acid B and Tanshinone II A, respectively. Moreover, the contents of liposoluble constituents and water-soluble constituents were determined. The average recoveries of Protocatechuic aldehyde, Salvianolic acid A, Salvianolic acid B, Tanshinone I, Cryptotanshinoneand, Tanshinone II A are 95.17%,94.06%,91.43%, 96.50%,97.88% and 98.02%; the RSD are 1.7%,1.9%,2.0%,1.8%,1.4%,1.0%, respectively. The contents tests showed that Salvianolic acid A and Salvianolic acid B are raised 294.9%-316.8% and 17.01%-18.12% than S. miltiorrhiza, but all others reduced. Therefore, the quality standards of processing S. miltiorrhiza are as follows:the contents of water, total ash, and acid-insoluble ash should be not more than 13.0%,7.2%,0.82%, respectively. Water extract and ethanol ectract shouldn't be less than 45% and 34%, respectively. The contents of Protocatechuic aldehyde, Salvianolic acid A, Salvianolic acid B, Tanshinone I, Cryptotanshinone and Tanshinone IIA in stir-Fired S. miltiorrhiza, Wine Fired S. miltiorrhiza, Vinegar Fired S. miltiorrhiza shouldn't less than 0.04%,0.04%,0.03%,1.8%,1.8%,1.8%,0.19%,0.18%,0.18%,0.007%,0.008%,0.008%,0.03%,0.04%,0.04%,0.08%,0.09%,0.09%, respectively.3. Study on fingerprint by HPLC of S. miltiorrhiza pieces for total quality control. In the HPLC fingerprint of S. miltiorrhiza by stir-firing,28 characteristic peaks were detected, 33 characteristic peaks were detected in Wine Fired S. miltiorrhiza and 34 characteristic peaks were detected in Vinegar Fired S. miltiorrhiza. The method has repeatability and the relative retention time of chromatography peak is consistent, providing a scientific basis for quality control of S. miltiorrhiza and its pieces by different processing.4. This paper we developed a high performance liquid chromatograhy method for simultaneous determination of Protocatechuic aldehyde, Salvianolic acid A, Salvianolic acid B, Cryptotanshinone, Tanshinone I and Tanshinone IIA in S. miltiorrhiza and its related species; moreover, the simultaneous extraction method of liposoluble constituents and water-soluble constituents were optimized. The average recovery and the RSD of the method in the range of 91.4%-98.0% and 1.0%-2.0%. It was concluded that the developed HPLC assay has been applied successfully to determination of the six biological ingredients in root of S. miltiorrhiza and it related species. The optimal extraction procedure is that:0.500g power of medical was extracted with water in Soxhlet's apparatus extracting for 4 hour, firstly; then the residue extracted by 80% ethanol with ultrasonic extraction for 20 minutes, merged and concentrated these two extract solution to 50.00 mL with deionized water. Six active major components in 20 samples of Rhizoma et Radix S. miltiorrhiza and it related species were quantitatively determined by the developed HPLC method. The results indicated that the contents of Protocatechuic aldehyde in S. miltiorrhiza from SiChuan Province(NO.4); Salvianolic acid B in S. prezwalskii; Salvianolic acid A in S. miltiorrhiza from SiChuan Province (NO.2); Cryptotanshinone in S. miltiorrhiza from ShanDong Province; Tanshinone I and Tanshinone IIA in S. trijuga are the highest in all samples.