高效液相色谱法测定灵芝提取物中尿苷和腺苷含量

Determination of Uridine and Adenosine Content of Ganoderma Lucidum Extract by HPLC

目的：建立一种测定灵芝提取物中尿苷和腺苷含量的高效液相色谱法，测定不同产地灵芝提取物中的尿苷和腺苷的含量。方法：色谱柱：Diamonsil C18（2）（5μm，250 mm ×4.6 mm）；流动相：乙腈-0.04 MKH2PO4＝9∶91；流速：1 mL＆#183;min-1，检测波长：260 nm，柱温：35℃。结果：尿苷和腺苷分别在0.2～50μg·mL^-1和0.1～20μg·mL^-1范围内，峰面积与浓度的线性关系良好；尿苷和腺苷的平均准确度均在99.91％～102.12％之间；日内、日间精密度 RSD 均小于1.76％，样品回收率均在94.42％～100.50％之间。6个不同产地的灵芝提取物中，以福建灵芝中尿苷含量和吉林灵芝中腺苷含量最高，其他5个产地灵芝中尿苷和腺苷含量差异无统计学意义。结论：该方法特异性强、准确度高、重复性好，可用于测定灵芝提取物中的尿苷和腺苷含量，为灵芝提取物的评价提供参考和依据。

To establish a HPLC method to determine uridine and adenosine content of Ganoderma lucidum extract,and determine content of uridine and adenosine in Ganoderma lucidum extract in different habitats.Methods：The chromatographic column was Diamonsil C18（2）（5 μm,250 ×4.6 mm）;the mobile phase acetonitrile was -0.04 MKH2PO4=9：91;the flow rate was 1 mL, min-1;the detection wavelength was 260 nm;the column temperature was 35 ℃.Results：The range of uridine and adenosine was respectively in 0.2-50 μg·mL^-1 and 0.1 ~20 μg·mL^-1 ,and the linear relationship between concentration and peak area were good;the average accuracy of uridine and adenosine were both between 99.91 to 1 02.1 2%;day and inter day RSD were less than 1.76%,and sample recovery rate were within 94.42%to1 00.50%.Among the extract of Ganoderma lucidum in the 6 different areas, the highest uridine content contained in Ganoderma lucidum in Fujian and highest adenosine content appeared in Jilin,and there were no statistical differences of uridine and adenosine content of Ganoderma lucidum in the other 5 places.Conclusion：The method is specific, accurate,reproducible,and can be used for determination of uridine and adenosine in ganoderma extract,which provides the reference and basis for evaluation of Ganoderma lucidum extract.