Identification and determination of the saikosaponins in Radix bupleuri by accelerated solvent extraction combined with rapid-resolution LC-MS.
A method based on accelerated solvent extraction combined with rapid-resolution LC-MS for efficient extraction, rapid separation, online identification and accurate determination of the saikosaponins (SSs) in Radix bupleuri (RB) was developed. The RB samples were extracted by accelerated solvent extraction using 70% aqueous ethanol v/v as solvent, at a temperature of 120 degrees C and pressure of 100 bar, with 10 min of static extraction time and three extraction cycles. Rapid-resolution LC separation was performed by using a C(18) column at gradient elution of water (containing 0.5% formic acid) and acetonitrile, and the major constituents were well separated within 20 min. A TOF-MS and an IT-MS were used for online identification of the major constituents, and 27 SSs were identified or tentatively identified. Five major bioactive SSs (SSa, SSc, SSd, 6''-O-acetyl-SSa and 6''-O-acetyl-SSd) with obvious peak areas and good resolution were chosen as benchmark substances, and a triple quadrupole MS operating in multiple-reaction monitoring mode was used for their quantitative analysis. A total of 16 RB samples from different regions of China were analyzed. The results indicated that the method was rapid, efficient, accurate and suitable for use in the quality control of RB.
Guangdong Provincial Public Laboratory of Analysis and Testing Technology, China National Analytical Center Guangzhou, Guangzhou, P. R. China.
Yun-Yun
YY
Yang
You-Zhi
YZ
Tang
Chun-Lin
CL
Fan
Hui-Tai
HT
Luo
Peng-Ran
PR
Guo
Jian-Xin
JX
Chen
D029972:Bupleurum / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004365:Drugs, Chinese Herbal / Q000032:analysis* / Q000737:chemistry; D000431:Ethanol / Q000737:chemistry*; D013058:Mass Spectrometry; D008516:Medicine, Chinese Traditional; D008968:Molecular Conformation; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D010936:Plant Extracts / Q000032:analysis* / Q000737:chemistry; D012503:Saponins / Q000032:analysis* / Q000737:chemistry; D013237:Stereoisomerism
C051274:Bupleurum root extract; D004365:Drugs, Chinese Herbal; D010936:Plant Extracts; D012503:Saponins; D000431:Ethanol; D009828:Oleanolic Acid; C025759:saikosaponin D