中药常山高效液相色谱检测方法的建立

Establishment of HPLC detection method for Radix Dichroae

建立常山药材的HPLC指纹图谱测定方法,为建立常山药材的质量标准提供依据。方法:采用Kromasil C18色谱柱(4.6 mm×250 mm,5μm);流动相:乙腈-0.03%H3PO4(20∶80);p H值:5.0~6.5;流速:1.0m L/min;检测波长:265 nm;柱温:25℃;进样量:10μL;采用以上条件进行线性关系考察、精密度试验、重复性试验、稳定性试验和加样回收率试验,并测定了3批样品中常山乙素的含量。结果:通过对常山药材HPLC指纹图谱结果进行分析,得出常山乙素标准品在0.5~50μg/m L范围内线性关系良好,相关系数为0.999 8,精密度试验、重复性试验和稳定性试验的RSD值均小于2.0%,符合《中国兽药典》的相关规定。结论:中药常山的高效液相色谱检测方法所得图谱清晰,特征直观,所建立的HPLC指纹图谱可作为评价常山药材质量的参考。

A test method of HPLC fingerprint on Radix Dichroae was established in order to provide basis for the quality standard of herbs Dichroa medicament. The optimal conditions of separation and detection were achieved on Kromasil C18 chromatographic column（4.6 mm × 250 mm,5 μm）with a mobile phase consisting of acetonitrile-0.03% phosphoric acid（20∶80）at the flow-rate of 1.0 m L/min. The p H value was 5.0~6.5. The detection wavelength was 265 nm. The column temperature was 25℃. Using the above conditions,we conduct experiments on linear relationship test,accuracy test,repeatability test,stability test as well as sample recovery rate test. Meanwhile the febrifugine content from three batches of samples was determined. The analysis of HPLC fingerprint on Radix Dichroae showed that the correlation coefficients of febrifugine reference materials in the range of0.5~50 μg/m L was 0.999 8,and the liner relationship was good. The RSD value of accuracy test,repeatability test and stability test were less than 2.0%. The above results were accord with the relevant provisions of Chinese Veterinary Pharmacopoeia. It can obtain a clear,obvious and intuitive fingerprint using HPLC detection methods of Radix Dichroae. HPLC fingerprints could be a reference to evaluate the quality of Radix Dichroa.