Determination of nine kinds of nucleosides in Spirodelae Herba from different habitats by UPLC-MS/MS / 中草药

ABSTRACT

Objective: To establish a UPLC-MS/MS method for the simultaneous determination of nine kinds of nucleosides in Spirodelae Herba, and analyze and investigate the difference of nucleosides ingredients in samples from 13 habitats. Methods: The determination was performed on the Waters XbridgeTM Amide-C18 (150 mm × 4.6 mm, 3.5 μm), the 0.1% formic acid-water (A) and 0.1% acetonitrile-water (B) in gradient elution as mobile phase. The flow rate was 0.5 mL/min. The column temperature was 30 ℃. MS instrument was equipped with ESI+ ion source. Gaining the extracted ion chromatograms, then the peak area for quantitative, principal component analysis (PCA), and hierarchical clustering analysis (HCA) were used to comprehensive evaluation. Results: All nine kinds of nucleoside showed good linearity (r ≥ 0.998 9). The RSD of the precision, repeatability, and stability tests were less than 3.5%. The average recovery rates were in the range of 94.4%-101.9%, RSD were in the range of 1.73% - 3.58%. There are differences in the composition and content from different habitats. The content of 2'-deoxyuridine and 2'-deoxyinosine in the lower levels, cytosine was the lowest, while the contents of uridine, inosine, and xanthine were in higher levels. The largest content was in Spirodelae Herba from Huangshan, Anhui province. Conclusion: The method for the simultaneous determination of the nine kinds of nucleosides is simple, accurate, and reproducible, that will provide the basis for the formulation of herbs Spirodelae Herba for quality control standards.