HPLC测定贝母中尿嘧啶和3种核苷的含量及其在品种鉴定中的应用

HPLC Quantitative Analysis of Uracil and Three Nucleotides in Fritillariae Bulbus and the Application for Variety Discrimination

关键词:  贝母  核苷  高效液相色谱法  品种鉴定

Key words:  Fritillariae Bulbus  nucleotides  HPLC  variety discrimination

摘要:

目的 建立测定浙贝、川贝、平贝和新疆贝母中尿嘧啶、尿苷、鸟苷、腺苷含量和鉴定该4种药材品种的高效液相色谱法。方法 色谱柱为Alltima C₁₈(4.6 mm×250 mm，5 μm)；流动相为甲醇-水溶液；梯度洗脱，洗脱条件为0.0 min→ 5.0 min→10.0 min→20.0 min→30.0 min→40.0 min，甲醇：1%→1%→5%→12%→24%→31%；流速为0.8 mL·min-1；检测波长为260 nm；柱温为25 ℃。进样体积为20 μL。结果 尿嘧啶和3种核苷均达到基线分离，线性良好，4种贝母中指标成分含量差异明显。结论 该方法快速、准确、重复性好，可作为贝母类药材的核苷类成分的含量测定方法，同时亦可用于准确、可靠的鉴定该4种贝母药材。

Abstract:

OBJECTIVE To establish an HPLC method for simultaneously determination of uracil, uridine, granosine and adenosine in bulbus of F. thunbergii Miq., F. cirrhosa D.Don, F. ussuriensis Maxin, and F. walujewii Regel, and for identification of these varieties. METHODS The HPLC method was used with Alltima C₁₈ column(4.6 mm×250 mm, 5 μm). The gradient elution solvent system was MeOH/H2O with time program as follows: 0.0 min→5.0 min→10.0 min→20.0 min→30.0 min→ 40.0 min, 1%→1%→5%→12%→24%→31%(MeOH). The UV wavelength was 260 nm; flow rate was 0.8 mL·min-1; column temperature was 25 ℃. RESULTS Uracil, uridine, granosine and adenosine were baseline separated. The method had good linearity. CONCLUSION The method is rapid, precise and repeatable, and can be used for the determination of nucleosides and supplying evidence for variety discrimination of bulbus of F. thunbergii Miq., F. cirrhosa D.Don, F. ussuriensis Maxin, and F. walujewii Regel.