Simultaneous Quantification of Five Constituents in Asarum insigne Using Ultra-Fast Liquid Chromatography with Tandem Mass Spectrometry
LI Yao-li1, YAN Zi-shan2, GUO Fang1, HU Chuan-qin2*, SHANG Ming-ying1, XU Feng1, CAI Shao-qing1*
1. Pharmacognosy Research Laboratory, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China; 2. School of Light Industry, Beijing Technology and Business University, Beijing 100048, China
Abstract：OBJECTIVE To evaluate the effectiveness and safety of Asarum insigne Diels and develop a sensitive, accurate, and efficient ultra-fast liquid chromatography coupled with triple quadrupole mass spectrometer (UFLC-MS/MS) method for simultaneous quantification of five compounds. METHODS Chromatographic separation was performed on a Shimadzu shim-pack XR-ODS Ⅲ column(2.0 mm×75 mm,1.6 μm). The mobile phase was composed of 0.1% aqueous formic acid and methanol and the flow rate was 0.2 mL·min-1. Using the established method, all components could be easily separated within 14 min. All components were detected in multiple reaction monitor mode after positive atmospheric pressure chemical ionization. RESULTS The method was validated, and had good precision, linearity, lower limit of detection, lower limit of quantification, recovery, and stability. This method was used to determine and compare the contents of five components in different medicinal parts of 11 samples, and the contents of 3,4,5-trimethoxytoluene, 3,5-dimethoxytoluene, methyleugenol in different medicinal parts of Asarum insigne were reported for the first time, which (in ng·mg-1) were as follows: 3,4,5-trimethoxytoluene: 1.5-16.71; 3,5-dimethoxytoluene: 28.59-177.20; methyleugenol: 2.46-22.48; AL-I: 46.39-324.04; 7-OCH3-AL-IV: 12.95-251.04. The content of 7-OCH3-AL-IV and total contents of two aristololactams in roots were lower than those in leaves or flower parts, and the contents of AL-I in flowers (171.9-324.0 ng·mg-1) were higher than those in roots (78.44-124.56 ng·mg-1). For methyleugenol, the contents in roots were lower than those in the other parts. CONCLUSION It is recommended to remove the leaves, flowers, and rhizomes when using Asarum insigne and use it with extreme caution. The UFLC-MS/MS method established in this study can provide guidance for the quality evaluation and safe clinical use of traditional Chinese medicine derived from Asarum insigne.
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LI Yao-li, YAN Zi-shan, GUO Fang, HU Chuan-qin, SHANG Ming-ying, XU Feng, CAI Shao-qing. Simultaneous Quantification of Five Constituents in Asarum insigne Using Ultra-Fast Liquid Chromatography with Tandem Mass Spectrometry. Chinese Pharmaceutical Journal, 2020, 55(12): 1027-1033.
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