Simultaneous determination of five compounds in Ganmaoning granules by UPLC-MS/MS

Abstract

Objective To establish a method for the simultaneous determination of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoning granules. Methods The UPLC-MS/MS method was adopted. The separation was performed on ZOBAX SB C18 (2.1 mm×150 mm, 5 μm) column with mobile phase consisted of methanol-2 mmol/L ammonium acetate aqueous solution (isocratic elution) at the flow rate of 0.2 ml/min. The column temperature was set at 35 ℃, and the injection volume was 2 μl. The electrospray ionization source (ESI) was used. Multiple reaction monitoring (MRM) mode was adopted, using positive and negative ions scanning. The ion source temperature was 300 ℃, the drying gas temperature was 400 ℃, the drying gas flow was 20 L/min, the atomizing gas pressure was 55 psi, and the capillary voltage was 4000 V. Results The linear ranges of adenosine, chlorogenic acid, caffeic acid, baicalin and wogonin in Ganmaoling granules were 0.20-12.80 ng (r=0.999 6), 1.00-64.00 ng (r=0.999 8), 0.40-25.60 ng (r=0.999 6), 4.00-256.00 ng (r=0.999 2), 0.20-12.8 ng (r=0.999 1), which showed the linear relationship was good. The limits of detection were 0.02, 0.10, 0.04, 0.40, 0.02 ng, respectively. The limits of quantitation were 0.04, 0.20, 0.08, 0.80, 0.04 ng, separately. The RSDs of precision, reproducibility and stability (24 h) tests were no more than 3.5%. The recovery rates were 99.3%-100.75% (RSD=2.09%-3.17%). Conclusions The method established in this experiment is simple, rapid, sensitive and accurate. It can be used to simultaneously determine the contents of five components in Ganmaoling granules, and can be used for quality control of Ganmaoning granules. Key words: Quality control; Ganmaoning granules; UPLC-MS/MS; Adenosine; Chlorogenic acid; Caffeic acid; Baicalin; Wogonin