Preparation of salvianolic acid A by the degradation reaction of salvianolic acid B in subcritical water integrated with pH-zone-refining counter-current chromatography.
Salvianolic acid A is the major bioactive compound in Danshen, however, due to the chemical instability and low content in Danshen, it is difficult to extract amount of salvianolic acid A. Therefore, this study was to establish an effective strategy for obtaining adequate amount of salvianolic acid A, subcritical water extraction was used to degrade salvianolic acid B and prepare salvianolic acid A. Different reaction conditions including temperature, time, concentration and pH value in subcritical water were investigated. Under 40mg/mL of reactant concentration, 180°C of temperature, 4.0 of pH value and 60min of reaction time, the highest yield rate of salvianolic acid A reached 34.86%. Then, the degradation products were successfully separated by pH-zone-refining counter-current chromatography with the solvent system Pet-EtAc-n-BuOH-H2O (2:3:1:9, v/v), where 10mM TFA was added in stationary phase and 10mM NH3·H2O in mobile phase. As a result, a total of 227.3mg of salvianolic acid A at 98.2% purity, 38.9mg of danshensu at 99.3% purity, 9.5mg of salvianolic acid D at 92.7% purity, and 32.8mg of protocatechuic aldehyde at 93.1% purity were obtained from 1.2g degradation products of salvianolic acid B by one-step purification. The results demonstrated that the combinative application of subcritical water and pH-zone-refining counter-current chromatography is a potential technique for the preparative separation of salvianolic acid A from salvianolic acid B.
Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan, Shandong 250014, China; College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan, Shandong 250355, China.
Huaizhi
H
Li
Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan, Shandong 250014, China.
Yan
Y
Cheng
Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan, Shandong 250014, China.
Hongjing
H
Dong
Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan, Shandong 250014, China. Electronic address: wangx@sdas.org.
Xiao
X
Wang
College of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Street, Jinan, Shandong 250355, China.
Jia
J
Li
Key Laboratory of TCM Quality Control Technology, Shandong Analysis and Test Center, Shandong Academy of Sciences, 19 Keyuan Street, Jinan, Shandong 250014, China.
Qianshan
Q
Gao
D001572:Benzofurans / Q000138:chemical synthesis*; D002109:Caffeic Acids / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003377:Countercurrent Distribution; D006863:Hydrogen-Ion Concentration; D007773:Lactates / Q000378:metabolism*; D014867:Water / Q000737:chemistry*; D053630:Depsides
D001572:Benzofurans; D002109:Caffeic Acids; D007773:Lactates; D014867:Water; C066201:salvianolic acid A; C076944:salvianolic acid B; D053630:Depsides
Degradation; Salvianolic acid A; Salvianolic acid B; Subcritical water; pH-zone-refining counter-current chromatography