Simultaneous Determination Of Multiple Components Of Cistanches Herba Based On Reversed Phase-Hydrophilic Interaction Chromatography-Tailored Multiple Reaction Monitoring

Reversed phase-hydrophilic interaction chromatography-tailored multiple reaction monitoring (RPLC-HILIC-tailored MRM) was developed for simultaneous determination of 27 components with wide polarity and wide range of content in Cistanches herba. RPLC was directly coupled with HILIC to achieve comprehensive retention of numerous compounds regardless of the polarity. Acquity UPLC HSS T3 column (100 mmx2.1 mm, 1.8 mu m) and Xbridge Amide column (150 mmx4.6 mm, 3.5 mu m) were employed for reversed phase separation and HILIC mode, respectively. To overcome the obstacle of incompatible mobile phases, dilution pumps were involved between two columns via a mixer. Pump A and pump B were responsible for delivering 0. 1 % formic acid and acetonitrile , respectively , into the RPLC column with a flow rate of 0. 15 mL/min by gradient elution. Pump C and pump D, served as dilution pumps, introduced the same mobile phase as pump A and pump B into the mixer at a flow rate of 1. 0 mL/min, which contributed to retention of polar compounds on HILIC column. For mass detection, tailored MRM with optimal collision energy was carried out by online energy-resolved MS to suppress mass response of abundant constituents, and to advance mass response of trace compounds. A total of 27 components in thirty-six batches of Cistanches Herba, such as amino acids , organic acids, phenylethanoid glycosides , lignans, and iridoids, were determined by RPLC-HILIC-tailored MRM. Satisfactory correlation coefficients were observed for all calibration equations with greater than 0. 9929 over their corresponding concentration ranges. The limits of detection (LODs) were 0. 0032-160 mu g/g and the limits of quantification (LOQs) were 0. 032-320 mu g/g. The average recoveries at three spiked levels were in the range of 74. 7% -125. 1% with relative standard deviations (RSDs) of 1. 6% -13. 7% . The developed method showed wide linear range, good accuracy and wide applicability , and had the potential for large-scale quantitative analysis of traditional Chinese medicine.