Abstract
Danggui Buxue Tang (DBT) is a Traditional Chinese Medicine (TCM) formula primarily used to treat symptoms associated with menopause in women. Usually, DBT is composed of one portion of Radix Angelicae Sinensis (RAS) and five portions of Radix Astragali (RA). Clinically, Radix Hedysari (RH) is sometimes used by TCM physicians to replace RA in DBT. In order to verity whether the chemical constituents of the DBT1 (RA:RAS = 5:1, w/w) and DBT2 (RH:RAS = 5:1, w/w) share similarities the chemical profiles of the two DBTs crude extracts and urine samples were analyzed and compared with the aid of HPLC-DAD-ESI-IT-TOF-MSn, which determines the total ion chromatogram (TIC) and multi-stage mass spectra (MSn). Then, the DBT1 and DBT2 were identified and compared on the basis of the TIC and the MSn. In the first experiment (with crude extracts), 69 compounds (C1–C69) were identified from the DBT1; 46 compounds (c1–c46) were identified from the DBT2. In the second experiment(with urine samples), 44 compounds (M1–M44) were identified from the urine samples of rats that had been administered DBT1, and 34 compounds (m1–m34) were identified from the urine samples of rats that had been administered DBT2. Identification and comparison of the chemical compositions were carried out between the DBT1 and DBT2 of the crude extracts and urine samples respectively. Our results showed that the two crude extracts of the DBTs have quite different chemical profiles. The reasons for their differences were that the special astragalosides in DBT1 and the isoflavonoid glycosides formed the malonic acid esters undergo single esterification and acetyl esters undergo acetylation in DBT1. In contrast, the urine from DBT1-treated rats strongly resembled that of DBT2-treated rats. These metabolites originate mainly from formononetin, calycosin and their related glycosides, and they were formed mainly by the metabolic process of reduction, deglycosylation, demethylation, hydrogenation and sulfation. The HPLC-DAD-ESI-IT-TOF-MSn method was successfully applied for the rapid chemical profiles evaluation of two DBTs and their related urine samples.
Highlights
Danggui Buxue Tang (DBT) is a Traditional Chinese Medicine (TCM) formula primarily used to treat symptoms associated with menopause in women
In order to obtain desirable HPLC and mass spectrometry chromatograms, the procedures for preparation of the urine samples and crude extracted samples of the two DBTs were optimized in terms of the extraction solvents and extraction times
Since MS and MSn fragmentations gave more information about the isoflavones in positive ion (PI) mode but about saponins in NI mode, the analysis was simultaneously conducted in both PI and NI mode
Summary
Danggui Buxue Tang (DBT) is a Traditional Chinese Medicine (TCM) formula primarily used to treat symptoms associated with menopause in women. It is believed to invigorate ‘Qi’ (vital energy) and nourish the ‘Blood’ (body circulation) [1] Nowadays, it is commonly used in China as an efficacious medicinal prescription and a healthy food supplement. The plants RA and RH belong to the same botanical family but different genus, and have long been widely used as the same crude herb in DBT [1]. This is always a question of whether RH can replace. RA-containing DBT showed higher amounts of calycosin-7-Oβ-D-glucoside, ferulic acid, ononin, calycosin, astragaloside IV, astragaloside III, and Z-ligustilide.
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