Quantitative determination of thirteen nucleosides and nucleobases in pseudostellariae radix in different harvest periods by QTRAP LC-MS/MS / 中国药学杂志

ABSTRACT

OBJECTIVE: To establish a QTRAP UPLC-MS/MS method for simultaneous determination of thirteen nucleosides and nucleobases in Pseudostellariae Radix, and analyze their dynamic changes at different periods. METHODS: The separation was carried out on a Waters Atlantis T3 column (2.1 mm × 150 mm, 3 μm)eluted by a mobile phase of 5 mmol · mL-1 of ammonium acetate (B)-methanol(A) at a flow rate of 0.4 mL · min-1. The elution program is as follows. 0 -4.5 min, 3% -4% A; 4.5-8 min, 4% -18% A; 8 -10 min, 18%A; 10 -10.1 min, 18% -3% A; 10.1 -13 min, 3%A. The target compounds were analyzed by the positive ion multiple reaction monitoring (MRM) mode. RESULTS: The calibration curves of the 13 nucleosides and nucleobases showed good linearity (r >0.991 0) in the ranges of the tested concentrations, and the average recoveries were between 96.46% - 105.07%. The contents of cytidine, uridine, inosine, guanosine, and thymidine in Pseudostellariae Radix harvested at different periods showed some differences, and the contents were higher in mid-June. CONCLUSION: The method is simple, sensitive, accurate and credible, and provides the basis for exploring the quality forming mechanism of Pseudostellariae Radix medicinal materials and the suitable harvest time.