HPLC测定红曲中7种核苷类成分的含量

Determination of 7 nucleosides in red yeast rice by HPLC

目的建立同时测定红曲中尿嘧啶、胞苷、次黄嘌呤、腺嘌呤、肌苷、胸苷和腺苷7种核苷类成分的高效液相色谱法。方法采用Shiseido C18(4.6 mm×250 mm,5μm)色谱柱,乙腈(A)-0.05 mol/L磷酸二氢钾(B)二元线性梯度洗脱:0～9 min,0%A;9～40 min,0%～8%A;流速1 mL/min;柱温30℃;检测波长254 nm;进样量20μL。结果尿嘧啶、胞苷、次黄嘌呤、腺嘌呤、肌苷、胸苷和腺苷分别在1.00～5.00μg、3.08～15.35μg、2.78～13.95μg、9.37～46.87μg、4.58～22.86μg、1.06～5.29μg、4.41～22.28μg与其峰面积呈良好的线性关系(r≥0.9995),平均加样回收率分别为99.34%、99.56%、100.18%、97.95%、98.34%、98.02%、99.28%,RSD <2.00%。结论该方法重现性好、准确精密,可用于红曲中核苷类成分含量测定及质量评价。

Objective To establish a high performance liquid chromatography method for the simultaneous determination of seven nucleosides of uracil,cytidine,hypoxanthine,adenine,inosine,thymidine and adenosine in red yeast rice.Methods Shiseido C18(4.6 mm×250 mm,5μm)was adopted to separate the sample at the column temperature of 25°C.Acetonitrile(A)-0.05 mol/L KH2 PO4(B)was used as the mobile phase.Gradient elution method:0-9 min,0%A;9-40 min,0%-8%A.The injection volume was 20μL,and flow rate was 1 mL/min.The detection wavelength was 254 nm.Results Uracil,cytidine,hypoxanthine,adenine,inosine,thymidine and adenosine showed a good linear relationship with the peak area(r<0.9995)in the range of 1.00-5.00μg,3.08-15.35μg,2.78-13.95μg,9.37-46.87μg,4.58-22.86μg,1.06-5.29μg,4.41-22.28μg,respectively.The average recovery rates were 99.34%,99.56%,100.18%,97.95%,98.02%,99.28%,RSD<2.00%.Conclusion The method is reproducible and accurate,and can be used for the determination and quality evaluation of nucleotide in red yeast rice.