Simultaneous determination of six isoflavonoids in rat plasma after administration of total flavonoid from Gegen by ultra‐HPLC‐MS/MS

Abstract

A simple, specific, and sensitive ultra-performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous determination of 3'-hydroxypuerarin, 6''-O-xylosylpuerarin, mirificin, puerarin, 3'-methoxypuerarin and daidzin in rat plasma. After the addition of methanol containing 0.1% formic acid and 10% ascorbic acid, the analytes and rutoside were obtained by protein precipitation, then separated on a Thermo Syncronis C18 column (2.1 mm × 10 cm, 1.7 μm) by gradient elution and monitored using an electrospray ionization interface operating in positive ion and selective reaction monitoring acquisition mode. The calibration curves of these analytes showed good linearity (r > 0.99) within the test ranges. The lower limit of quantification was 0.0200 μg/mL for 3'-hydroxypuerarin, 0.0101 μg/mL for 6''-O-xylosylpuerarin, 0.0100 μg/mL for mirificin and puerarin, 0.0098 μg/mL for 3'-methoxypuerarin, and 0.0090 μg/mL for daidzin. The intraday and interday precision and accuracy were all within 15%. The extraction recoveries were from 74.0 to 95.8%. The validated method was successfully applied to pharmacokinetic studies of the six isoflavonoids in rat plasma after intravenous administration of total flavonoids from Gegen.

A UPLC–MS/MS approach for simultaneous determination of eight flavonoids in rat plasma, and its application to pharmacokinetic studies of Fu‐Zhu‐Jiang‐Tang tablet in rats

“…The four compounds achieved the maximum plasma concentration between 0.28 and 0.67 h. Meanwhile, the plasma concentrations of the four compounds were eliminated rapidly, within 4 h of intragastric administration. However, the pharmacokinetic parameters of 3′-methoxypuerarin were not entirely in accordance with the data reported in previous literature (Zhao, Zhao, Liu, Han, & Simultaneous, 2012). Double peaks emerged in curves of mean plasma concentration for 3′-methoxypuerarin (see Figure 3D), which is reported for the first time.…”

HPLC‐MS/MS analysis of anthocyanins in human plasma and urine using protein precipitation and dilute‐and‐shoot sample preparation methods, respectively

“…Our study demonstrated that protein precipitation can be used to analyze plasma samples from humans ingested low doses of anthocyanins. Protein precipitation as the only sample preparation procedure has been successfully used to analyze isoflavonoids (Zhao, Zhao, Liu, Han, & Yu, ), flavanone (Liu, Xu, Zhang, Song, & Tian, ) and puerarin (Prasain et al, ) in rat plasma or serum with HPLC‐MS/MS. For the analysis of small molecules, plasma protein precipitation was commonly facilitated by adding an acid.…”

Simultaneous determination of puerarin and its active metabolite in human plasma by UPLC-MS/MS: Application to a pharmacokinetic study

“…Among them, Wang et al [20] presented the LC-MS/MS method with a comparably higher sensitivity of relatively LLOQ 0.39 ng/mL than other studies [9,10,18,19] with a LLOQ of 2-18.6 ng/mL for puerarin. Also, further studies provided information about the simultaneous determination of puerarin and daidzein in rat plasma by using LC-MS [2], UPLC-MS [21] or UPLC-MS/MS [22], serum [23] and urine [24] in humans by using HPLC.…”