Qualitative Analysis and Determination of Four Components in Ganmao Qingre Granules by LC-MS/MS and HPLC

OBJECTIVE To establish a LC-MS/MS method and a HPLC method for qualitative analysis and determination of amygdalin, puerarin, prim-o-glucosylcimifugin and 5-o-methylvisammioside in Ganmao Qingre granules. METHODS The components were confirmed by LC-MS/MS. The analysis was performed on an Agilent Zorbax SB C₁₈ column(4.6 mm×250 mm, 5 μm) under the gradient elution with methanol-water. The detection of puerarin, prim-o-glucosylcimifugin and 5-o-methylvisammioside was set as 252 nm, the detection of amygdalin was set as 207 nm. The flow rate was 1.0 mL·min^-1 and the column temperature was 35℃. RESULTS The primary and secondary mass spectrograms of amygdalin, puerarin, prim-o-glucosylcimifugin and 5-o-methylvisammioside in Ganmao Qingre granules were consistent with the reference solution. Each component showed good linear relationship in the ranges of 5.278-105.558, 12.868-257.351, 0.938-18.761 and 0.493-9.861 μg·mL^-1respectively(r ≥ 0.999 9). The average recoveries were 100.86%, 100.09%, 100.06% and 99.58%. CONCLUSION The established method is simple and rapid with the good separation and reproducibility, and high sensitivity. It provides a fast, efficient and reliable method for the determination of the main components of Ganmao Qingre granules.