Rapid separation and identification of 54 major constituents in Buyang Huanwu decoction by ultra‐fast HPLC system coupled with DAD‐TOF/MS

Abstract Buyang Huanwu Decoction (BYHWD), is a well‐known traditional Chinese preparation consisting of Radix Astragali, Radix Angelicae Sinensis, Rhizoma Ligustici Chuanxiong, Radix paeoniae Rubra, Flos Carthami, Semen Persicae and Lumbricus. An ultra‐fast high‐performance liquid chromatography (HPLC) method coupled with diode array detection (DAD) and electrospray ionization time‐of‐flight mass spectrometry (ESI‐TOF/MS) has been developed for rapid separation and structural identification of constituents in BYHWD. Using an ultra‐fast HPLC system with short columns (4.6 × 50 mm, 1.8 µm), the total analysis time for this complex prescription is less than 30 min. With various fragmentor voltages in TOF/MS, accurate mass measurements (less than 5 ppm error) for molecular ions and characteristic fragment ions could represent reliable identification criteria for these compounds. Fifty‐four major constituents from BYHWD sample, including four C ‐glycosyl quinochalcones, four flavonoid O ‐glycosides, sixteen isoflavones, six monoterpene glycosides, eight saponins, four organic acids and five amino acids, were identified or tentatively characterized based on their retention times, DAD and TOF/MS data. All the compounds were further assigned in the seven individual crude drugs. In conclusion, the ultra‐fast HPLC with DAD‐TOF/MS is a highly useful and efficient technique to separate and identify constituents in complex matrices of herbal medicines or preparations. Copyright © 2009 John Wiley & Sons, Ltd.