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Identification of Multiple Ingredients for a Traditional Chinese Medicine Preparation (Bu-yang-huan-wu-tang) by Liquid Chromatography Coupled with Tandem Mass Spectrometry

by 1,†, 2,3,† and 1,4,5,*
Institute of Traditional Medicine, National Yang-Ming University, Taipei 112, Taiwan
Division of Gastroenterology and Hepatology, Department of Internal Medicine, Chang Gung Memorial Hospital, Chiayi 61363, Taiwan
Graduate Institute of Clinical Medical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
Graduate Institute of Acupuncture Science, China Medical University, Taichung 40402, Taiwan
Department of Education and Research, Taipei City Hospital, Taipei 10629, Taiwan
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Molecules 2013, 18(9), 11281-11298;
Received: 19 July 2013 / Revised: 5 September 2013 / Accepted: 10 September 2013 / Published: 12 September 2013
(This article belongs to the Section Natural Products Chemistry)
Bu-yang-huan-wu-tang (BYHWT) is a popular Traditional Chinese Medicine formula consisting of seven herbal medicines (Astragalus membranaceus, Angelica sinensis, Paeonia lactiflora, Ligusticum chuanxiong, Carthamus tinctorius, Amygdalus persica and Pheretima aspergillum), that has been used in China for centuries to overcome stroke-induced disability. To ensure the consistency of quality, a reliable analytical method is required, therefore, we developed a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for quantitative analysis of the major constituents in BYHWT. The herbal ingredients consisting of the cycloartane-type triterpene glycosides of astragaloside I, astragaloside II and astragaloside IV; isoflavones of formononetin, ononin calycosin, calycosin-7-O-β-d-glucoside; ligustilide and paeoniflorin were separated on a C18 column with gradient elution of methanol/10 mM ammonium acetate buffer–formic acid (100:0.1, v/v). This study was performed by a mass spectrometer using electrospray ionization (ESI) with positive ionization ions monitored in the multiple reaction-monitoring (MRM) mode. The linearity, accuracy, precision, limit of detection (LOD) and lower limit of quantification (LLOQ) were validated for this quantification method, and the sensitivity, reliability and reproducibility were all confirmed. The experiments provided a good method for analyzing BYHWT extracts. This study also quantitated the active components in various brands of commercially available products. The results indicated that the pharmaceutical industrial products of BYHWT exhibited considerable variation in their contents of the herbal compounds. View Full-Text
Keywords: bu-yang-huan-wu-tang; herbal analysis; LC-MS/MS; traditional Chinese medicine bu-yang-huan-wu-tang; herbal analysis; LC-MS/MS; traditional Chinese medicine
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Shaw, L.-H.; Chen, W.-M.; Tsai, T.-H. Identification of Multiple Ingredients for a Traditional Chinese Medicine Preparation (Bu-yang-huan-wu-tang) by Liquid Chromatography Coupled with Tandem Mass Spectrometry. Molecules 2013, 18, 11281-11298.

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