基于冬瓜皮HPLC特征图谱中核苷类活性成分研究

Study on the active components of nucleosides in Benincasae Exocarpium based on HPLC

目的建立不同产地冬瓜皮药材HPLC特征图谱, 并同时测定尿苷、鸟苷等核苷类成分含量, 并对不同产地冬瓜皮进行质量评价。方法采用Waters Atlantis?T3色谱柱(250 mm×4.6 mm, 5 μm), 流动相为甲醇-水溶液, 梯度洗脱, 检测波长254 nm, 流速1.0 ml/min, 柱温25 ℃。以含量、特征图谱共有峰相似度为主要评价指标, 分析不同产地冬瓜皮药材质量差异。结果 15批冬瓜皮药材特征图谱均呈现6个共有特征峰, 与各自对照特征图谱相似度均大于0.90, 指认出鸟嘌呤、黄嘌呤、尿苷、腺嘌呤、鸟苷5个成分。尿苷、鸟苷含量测定结果表明, 15批冬瓜皮药材尿苷含量范围为0.017%~0.036%、鸟苷含量范围为0.018%~0.031%。河南省焦作市冬瓜皮药材尿苷含量最高, 为0.036%, 湖南省常德市冬瓜皮药材鸟苷含量最高, 为0.031%。结论不同产地冬瓜皮药材核苷类功效成分含量差异较小, 该HPLC特征图谱方法精密度、重复性、稳定性良好, 可客观全面评价不同产地冬瓜皮药材质量。

Objective:To establish HPLC characteristics of Benincasae Exocarpium from different regions;To simultaneously determine the contents of uridine,guanosine and other nucleosides;To conduct overall evaluation on Benincasae Exocarpium from different regions.Methods:The chromatography was performed on Waters Atlantis?T3(250 mm×4.6 mm,5μm)column with methanol(A)and aqueous solution(B)as mobile phase gradient elution.The detection wavelength was 254 nm,the flow rate was 1.0 ml/min,and the column temperature was 25℃.The similarity of common peak of content and characteristic map was used as the main evaluation index to analyze the difference of the Benincasae Exocarpium in different producing areas.Results:15 batches of Benincasae Exocarpium showed 6 common characteristic peaks,and the similarity with the control characteristic maps was greater than 0.90.5 components of guanine,xanthine,uridine,adenine and guanosine were identified,and uridine and guanosine were quantitatively determined by HPLC.The results showed that,the contents of uridine and guanyl in 15 batches of Benincasae Exocarpium ranged from 0.017%to 0.036%and 0.018%to 0.031%.The highest content of uridine was 0.036%in Jiaozuo City of Henan Province,and 0.031%in Changde City of Hunan Province.Conclusions:This HPLC method has good precision,repeatability and stability,and the two components of uridine and guanosine have good linear relationship in the determination range,which can objectively and comprehensively evaluate the overall appearance of Benincasae Exocarpium from different regions.The difference of nucleoside efficacy components of Benincasae Exocarpium from different regions is small.