Simultaneous Quantification of 18 Bioactive Constituents in Tripterygium wilfordii Using Liquid Chromatography-Electrospray Ionization-Mass Spectrometry

Abstract

A liquid chromatography-electrospray ionization-mass spectrometry method with multiple reaction monitoring was established for simultaneous quantification of 18 bioactive constituents from the stem and root of Tripterygium wilfordii Hook. f. collected from different places in China and various commercial preparations. The chromatographic separations were achieved on an Agilent Poroshell SB-C₁₈ column (150 × 4.6 mm, 3.5 µm) with gradient elution using acetonitrile and 0.03 % formic acid aqueous solution in 45 min. Detection was performed in the positive ionization mode by monitoring the precursor-product combination. The validation of the method included tests of linearity, sensitivity, precision, repeatability, stability, and accuracy. All calibration curves showed good linearity (r > 0.9990) within the test range. The established method showed good precision and accuracy with intraday and interday variations of 2-5 % and 1-4 %, respectively, and recoveries of 95.5-104.5 %.

Integrative Approaches for the Identification and Localization of Specialized Metabolites in Tripterygium Roots

“…4B). This annotation also was consistent with the elution order and fragmentation patterns of sesquiterpene alkaloids reported previously for reverse-phase HPLC-MS and MS/MS (molecular ion as base peak, with abundant fragments at m/z 206 and 856; Zeng et al, 2013;Cai et al, 2015;Su et al, 2015;Luo et al, 2016). No NMR data were acquired in this study.…”

“…4A; retention time and exact mass [480.251 g mol 21 ] and MS/MS fragmentation patterns [base peak at m/z 231 and additional lower abundance fragments at m/z 245, 203, and 109]) were consistent with those reported for demethylzeylasteral (Table I; Fig. 4B; Zeng et al, 2013). The same metabolite record was retrieved with a high score in searches of the exact mass (480.251 g mol 21 ) and 1 H-NMR spectral data against Spektraris.…”

“…2A). These concentrations are in the same range as those reported by others (toward the lower end for triptolide and among the highest for celastrol; Zeng et al, 2013;Zhuo et al, 2013;Guo et al, 2014). Triptolide also was detected in extracts of stems (5.2 mg g…”

“…Previous studies that used reversephase chromatography with Tripterygium root extracts consistently listed wilfortrine as the earliest eluting abundant peak associated with this formula (Zeng et al, 2013;Cai et al, 2015;Su et al, 2015;Luo et al, 2016). The MS/MS fragmentation patterns we obtained at a collision energy of 30 eV were almost identical to those reported for wilfortrine when recorded at 25 eV (major product ion at m/z 846; Cai et al, 2015).…”

“…While the contents of bioactive specialized metabolites in commercial Tripterygium root extracts vary significantly between growing areas in China, triptolide is always present in very low concentrations (less than 0.008% of dry biomass), sesquiterpene pyridine alkaloids range between 0.0001% and 0.1% of dry biomass, and celastrol can accumulate up to about 1% of dry biomass (Zeng et al, 2013;Zhuo et al, 2013;Guo et al, 2014). To improve the yields of active principles extractable from Tripterygium roots, an understanding of the factors that control their accumulation is highly desirable.…”