Effect of Steaming Time and Pressure on Contents of Six Ginsenosides in Ginseng Radix et Rhizoma by UPLC-QqQ-MS/MS / 中国实验方剂学杂志

ABSTRACT

Objective:The processing method of red ginseng was determined by comparing the effects of different steaming time and pressure on the total content of six ginsenosides. Method:The contents of ginsenoside Rg1, Re, Rf, Rb1, Rc and Rb2 were determined by ultra high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-QqQ-MS/MS). The Waters ACQUITY UPLC BEH C8 column (2.1 mm×100 mm, 1.7 μm) was used. The mobile phase was 0.1% formic acid aqueous solution (A) and 0.1% formic acid acetonitrile solution (B) for gradient elution (0-4 min, 81%-79%A; 4-6.3 min, 79%-75%A; 6.3-6.5 min, 75%-71%A; 6.5-9.5 min, 71%A; 9.5-16.5 min, 71%-68.5%A; 16.5-16.6 min, 68.5%-60%A; 16.6-19 min, 60%-100%A). The flow rate was set at 0.4 mL·min-1 and the column temperature was set at 35 ℃. The mass spectrographic analysis employed electrospray ionization (ESI) and negative ion collection mode with capillary ionization voltage of 2.5 kV, desolvation temperature of 350 ℃, desolvation gas flow of 700 L·h-1 and cone gas flow of 50 L·h-1. Multiple reaction monitoring (MRM) mode was used to collect information, the collection range was m/z 100-1 500, detection was performed by MRM mode at m/z 799.59-637.49 for ginsenoside Rg1, m/z 945.54-475.79 for ginsenoside Re, m/z 799.59-475.49 for ginsenoside Rf, m/z 1 107.59-783.97 for ginsenoside Rb1, m/z 1 077.58-783.96 for ginsenoside Rc, m/z 1 077.75-191.19 for ginsenoside Rb2. Result:When the steaming time was 3 hours, the total mass fraction of six ginsenosides in each sample group was 7.099 8-16.768 5 mg·g-1, and the total amount of the six ginsenosides in atmospheric steaming was 2.5-12.6 times of that in pressurized steaming, which was obviously better than that in pressurized steaming. Conclusion:Under the conditions of this experiment, the best processing method of red ginseng is atmospheric steaming for 3 hours with fresh ginseng.