丹参制剂中丹参水溶性成分的指纹对照法质量分析

Quality assessment of hydrophilic components of Salvia miltiorrhiza Bge.in Danshen formulations by HPLC reference fingerprints

目的：建立指纹对照测定法以及定性和定量相似性评价体系，研究丹参及其复方制剂中的丹参水溶性成分，并用于评价中药制剂整体质量。方法：采用Lichrospher ODS-2（250mm×4．6mm，5μm）色谱柱，甲醇（A）-1．0％冰醋酸溶液（B）线性梯度程序洗脱：0min（25％A）～70min（75％A），流速为1．0mL／min，柱温30℃，检测波长254nm，建立丹参水溶性成分高效液相色谱指纹图谱。采用夹角余弦（Cosθ）法和相对欧氏距离（RED）法，分别从化学成分分布相似性和含量相似性两个方面评价不同丹参制剂与指纹对照品的相似程度。结果：丹参水溶性成分高效液相色谱指纹图谱中，以丹酚酸B为参照物峰，确定了16个指纹共有峰。不同厂家生产的丹参片剂定性相似度没有显著性差异，但定量相似度有较大差异。结论：高效液相色谱指纹图谱的同时定性和定量相似度分析可全面反映中药制剂的整体质量，可用于丹参制剂质量分析。

Aim： To establish a stable and repeatable HPLC reference fingerprint chromatogram standard and an evaluation system for qualitative and quantitative similarities of hydrophilic components of Salvia miltiorrhiza Bge. in different Danshen formulations, and to assess the quality of Chinese patent medicines. Methods： Hydrophilic components of Salvia mihiorrhiza Bge. were separated on a Lichrospher ODS-2 （ 250 mm × 4.6 mm, 5 μm） column with linear gradient elution by the mobile phase consisting of methanol （A） and 1.0% acetic acid solution （B） at a flow rate of 1.0 mL/min. Gradient elution began with A-B （25：75,0 min ） and ended with A-B （75：25, 70 min）. The detection wavelength was set at 254 nm and the column temperature was 30 ℃. The similarities of the corresponding HPLC fingerprint chromatograms were evaluated by cosine （ Cos θ） of vectors and relative Euclidean distance （RED） methods between Danshen formulations and the fingerprint reference substance, in terms of the chemical constituents and the contents, respectively. Results： The hydrophilic components of chromatographic fingerprints contained 16 common peaks while salvianolic acid B was chosen as a referential peak. The similarity analyses indicated that no significant difference was found in the qualitative similarity between different Danshen formulations. But there existed obvious difference in the quantitative similarity. Conclusion： The simultaneous qualitative and quantitative similarity analyses of the HPLC fingerprint chromatograms of herbal medicines are more practical for their quality control. It has been successfully applied in the quality evaluation of Danshen formulations.