Rapid characterization and determination of isoflavones and triterpenoid saponins in Fu-Zhu-Jiang-Tang tablets using UHPLC-Q-TOF/MS and HPLC-UV†
Abstract
In this study, a rapid ultra-high performance liquid chromatograph coupled with quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) was used for qualitative analysis of chemical constituents in Fu-Zhu-Jiang-Tang tablets, a new six-herb preparation for treating type II diabetes. Qualitative analysis was performed on a Kromasil C18 column (4.6 mm × 150 mm, 5 μm), using gradient elution with 0.1% formic acid–water and acetonitrile mobile phase. As a result, 39 compounds were identified or tentatively characterized, including 13 flavonoids, 12 fatty acids, 8 triterpenoid saponins, 2 phenols and 4 unknown compounds. Among them, 17 compounds were confirmed by comparing with standards. In addition, an HPLC-UV method was developed for quantitative determination of ten major compounds in the Fu-Zhu-Jiang-Tang tablet, including 3′-hydroxypuerarin, puerarin, 3′-methoxypuerarin, daidzin, rutin, astragalin, daidzein, ginsenoside Rg1, astragaloside IV and 20(S)-ginsenoside Rg3. The HPLC chromatograms were monitored at 254 nm and 210 nm. The method was well validated with respect to intra-day and inter-day precision and stability, with RSD values less than 1.64%, 1.19% and 1.77%, respectively. All the ten major compounds showed good linearity (r2 > 0.9971), and their average recoveries varied between 97.6% and 104.3%. The HPLC-UV method was successfully applied to ten batches of Fu-Zhu-Jiang-Tang tablets.