不同产地和商品等级延胡索中尿苷、鸟苷、腺苷的含量分析

Determination of Uridine,Guanosine and Adenosine Contents in Corydalis Rhizoma from Different Regions and Commercial Grades

目的：建立高效液相色谱法同时测定延胡索药材中尿苷、鸟苷和腺苷3种核苷成分含量的方法,并比较不同产地、不同商品等级中3种核苷类成分含量的差异和各成分间的结构比。方法：采用Venusil MP C18（2）色谱柱（4.6 mm×250mm,5μm）,以甲醇（A）-水（B）为流动相梯度洗脱（0-10 min,1%-5%A;10-15 min,5%-15%A;15-22 min,15%-17%A;22-26 min,17%-20%A;26-32 min,20%-24%A）,流速1.0 m L·min^-1,柱温35℃,检测波长260 nm。运用上述方法对38批延胡索药材进行测定,并对测定结果进行聚类分析。结果：测定了延胡索药材中3种核苷类成分的含量,在线性范围内线性关系良好（r〉0.999 8）,平均回收率为99.63%-104.24%,RSD为0.7%-2.3%。38批延胡索样品中尿苷、鸟苷和腺苷均可检测到,不同产地延胡索药材中尿苷、鸟苷、腺苷量及组成结构比差异显著;同一产地不同种植户的延胡索药材其3种成分量及组成结构比也存在明显差异。采用聚类分析能将不同产地的38批延胡索药材分为3类。结论：总体上来说,不同产地延胡索药材成分的组成存在显著性差异,不同商品等级延胡索中核苷类含量无明显差异,延胡索药材的质量受自然环境、栽培技术因素的影响。该方法适用于延胡索中尿苷、鸟苷和腺苷的含量测定,为其质量控制提供保证。

Objective： To establish a HPLC method for simultaneously determining the contents of three nucleosides,namely uridine, guanosine and adenosine, and compare the content differences of the three nucleosides and the composition ratios in Corydalis Rhizoma from different regions and commercial grades.Method： A Venusil MP C18（ 2） chromatographic column（ 4. 6 mm × 250 mm,5 μm） was used,with methanol（ A）-water（ B） as the mobile phase for gradient elution（ 0-10 min,1%-5% A; 10-15 min,5%-15% A; 15-22 min,15%-17% A; 22-26 min,17%-20% A; 26-32 min,20%-24% A）,at a flow rate of 1. 0 m L·min- 1. UV detection wavelength was set at 260 nm,and the temperature of the column was maintained at 35 ℃. The above HPLC method was applied to determine 38 batches of Corydalis Rhizoma and a cluster analysis was carried out for the results. Result： The contents of three nucleosides in Corydalis Rhizoma were determined,and showed a good linear relationship within the linear range（ r 〉0. 999 8）. The average recovery rate was in the range of 99. 63%-104. 24%,with the RSD raging from 0. 7% to 2. 3%. The three nucleosides could be detected in all 38 batches of Corydalis Rhizoma. There were significant differences in contents of uridine,guanosine and adenosine and the composition ratios for Corydalis Rhizoma from different regions; there were significant differences in contents of the three nucleosides and composition ratios for Corydalis Rhizoma from different farmer households of a same region.38 batches of Corydalis Rhizoma were divided into three categories by cluster analysis. Conclusion： Overall,the contents of three nucleosides in Corydalis Rhizoma showed significant difference while there was no significant difference in nucleosides content for the Corydalis Rhizoma with different commercial grades.