Simultaneous determination of bioactive constituents in Danggui Buxue Tang for quality control by HPLC coupled with a diode array detector, an evaporative light scattering detector and mass spectrometry

Abstract

Danggui Buxue Tang (DBT), a classical traditional Chinese formula comprising Radix Angelicae Sinensis (RAS) and Radix Astragali (RA), has been widely used to treat menopausal irregularity in Chinese women for nearly 800 years. In this study, a comprehensive analytical method of simultaneously determining the main types of bioactive constituents, eighteen in all from the formula, involving flavonoids, saponins, organic acid and some volatile compounds, was developed. This method was based on HPLC coupled to a diode array and evaporative light scattering detectors (HPLC-DAD-ELSD) on a common reverse-phase C(18) column. Liquid chromatography coupled with on-line electrospray ionization mass spectrometry (LC-ESI-MS) was also used to further validate and analyze the constituents. It was found that 0.3% aqueous formic acid and acetonitrile was the optimum mobile phase for gradient elution. This method, which showed good precision and accuracy, was successfully used to quantify the bioactive constituents in six products. As a result, the validated HPLC method, together with the LC-ESI-MS analysis, provided a new basis for assessing the quality of traditional Chinese medicinal compound preparations (TCMCPs) consisting of many bioactive components.

Simultaneous determination of calycosin-7-O-β-d-glucoside, ononin, astragaloside IV, astragaloside I and ferulic acid in rat plasma after oral administration of Danggui Buxue Tang extract for their pharmacokinetic studies by liquid chromatography–mass spectrometry

“…As the chemical constituents of the formula are complex, the pharmacokinetic study is usually focused on the main active constituents. In particular, calycosin-7-O-␤-d-glucoside, ononin, astragaloside IV, astragaloside I and ferulic acid were usually used in quality control studies of DBT [8,9]. Therefore, the current investigation was designed to administer DBT to animal subjects, and then to determine the plasma profiles and pharmacokinetic parameters of the five analytes in DBT.…”

“…Thirty grams of mixed powder of Radix Astragali and Radix Angelica Sinensis (5:1, w/w) was immersed in 500 mL 75% ethanol for 1 h, and then refluxed for 2 h at 85 • C. The solvent was removed with rotary evaporation under vacuum at 45 • C, and the residue was dissolved in 30 mL phosphate buffer and filtered through a 0.22 m membrane, and then 10 L of this solution was injected into the HPLC system for analysis. The HPLC analysis of the five constituents in DBT was previously method established by the authors [8]. The contents of calycosin-7-O-␤-d-glucoside, ononin, astragaloside IV, astragaloside I and ferulic acid were 0.408, 0.116, 0.120, 0.681 and 0.037 mg/mL in extract, respectively.…”

Rapid and sensitive quantitation of major constituents in Danggui Buxue Tang by ultra-fast HPLC–TOF/MS

“…To date, various analytical methods have been reported for analysis of active ingredients in DBT, including HPLC with diodearray detection (DAD) [7], evaporative light scattering detection (ELSD) [8], etc. Time-consuming procedure, no structure information and insensitivity are the major obstacles for these methods.…”

Quality evaluation ofYanghuo Sanqitablet through a simultaneous determination of five major active flavonoids and three main saponins by HPLC-DAD-ELSD

“…In recent years, an on-line coupled HPLC-DAD-ELSD method has attracted ever-increasing attention and has been successfully applied to simultaneous quantification of multi-components in traditional Chinese herbal medicines and their preparations [14][15][16][17][18][19][20][21][22][23]. This method is potentially ideal for routine analysis and quality evaluation of YST since UV and ELSD signals contain complementary information for each other.…”