中药炮制辅料米醋的指纹图谱研究

Study on Fingerprints of Traditional Chinese Medicine Processing Accessories of Rice Vinegar

目的通过对不同产地和品牌的米醋进行指纹图谱研究,为建立中药炮制辅料米醋的药用质量标准提供基础。方法收集33批米醋,采用HPLC方法,通过对流动相种类、色谱柱、洗脱程序、流动相pH的考察,优化米醋HPLC指纹图谱的分析方法,并对其中20批黑醋指纹图谱进行相似度、聚类分析和主成分分析。结果米醋浓度以1∶3稀释进样,色谱条件为色谱柱:Thermo Hypersil GOLD(5μm,250mm×4.6mm),流动相:0.07%磷酸水(pH=2.28~2.38)-甲醇梯度洗脱,流速:1mL/min,柱温:30℃,进样量:10μL。此时特征峰分离效果最佳。米醋分白醋和黑醋,其指纹图谱差异较大,白醋几乎只有一个醋酸峰,单从HPLC方面无法判断配制白醋和酿造白醋的区别;20批黑醋指纹图谱相似度差异较大,聚类分析和主成分将其聚为几类。将同一品牌恒顺不同类型的米醋进行相似度比较,发现相似度较高,可见同一品牌内的米醋由于酿造工序、原料相似,品质比较均一稳定。结论市场上的米醋种类繁多,质量也参差不齐,实验选择20批常用黑醋进行指纹图谱分析,比较不同种类米醋的差异,为中药炮制辅料米醋的药用质量标准提供参考。

OBJECTIVE To provide the basis for the medicinal quality standard of Chinese medicine processing accessories of rice vinegar through analysis with fingerprints of different kinds of Origin and brand of rice vinegar. METHODS 33 batches of vinegar were collected. The mobile phase, column, elution, the mobile phase pH were investigated and the analysis method of HPLC fingerprint was optimazated, Among them, 20 batch of rice vinegar fingerprint were chosen to perform the similari- ty, cluster analysis and principal component analysis. RESULTS The concentration of rice vinegar was diluted at 1： 3 when it was injected. Chromatographic column was Thermo Hypersil GOLD （5 μm, 250 mm× 4.6 mm） ; the mobile phase was com- posed of methanol and 0.07% phosphoric acid aqueous solution and a gradient elution program at flow rate of 1 mL/min was applied. The column temperature maintained at 30 ℃. The injection volume was 10μL. Then, the separation of characteristic peaks was optimum. Rice vinegar was divided into black vinegar and white vinegar. Furthermore, there was a big difference between black vinegar and white vinegar. The white vinegar almost showed only one peak of acetic acid, so, it＇s impossible to distinguish between brewing white vinegar and formulated white vinegar by HPLC. Similarity evaluation showed that 20 batch of white vinegar＇s fingerprints had palpable difference. Besides, clustering analysis and principal component analysis splited them into several categories. Different types of Hengshun vinegar indicated high similarity. Hence it could be seen that the quality of rice vinegar from the same brand was uniform and stable because of similar brewing processes and similar materials . CONCLUSION There are varieties of rice vinegar on the market with extensive source and uneven quality. 20 batches of black rice vinegar were chosen to compare the differences by HPLC fingerprints analysis, which provides reference for the medicinal quality control standard of traditional Chinese medicine processing accessories of rice vinegar.