基于HPLC指纹图谱研究清炒和盐炙对泽泻中化学成分的影响

Effect of simple stir-frying and salted stir-frying on the chemical constituents in Alisma Rhizoma by HPLC fingerprints

目的建立生泽泻、清炒泽泻和盐泽泻的HPLC指纹图谱,通过比较三者的差异考察清炒和盐炙对泽泻中化学成分的影响。方法样品用甲醇5 m L超声提取30分钟,采用HPLC建立指纹图谱,色谱条件为Hypersil BDS C18(250 mm×4.6 mm,5μm)色谱柱,乙腈-水梯度洗脱,检测波长208 nm,柱温25℃,流速1.0 m L/min,分析时间120分钟。分别采用"中药色谱指纹图谱相似度评价系统(2004A版)"和SPSS软件对谱图进行相似度分析和聚类分析。结果生泽泻、清炒泽泻和盐泽泻的HPLC指纹图谱的相似度分别为0.905~0.948、0.902~0.955、0.904~0.960,三者分别有14、15和15个共有峰,其峰面积存在不同程度的差异。其中,生泽泻的2号峰在炮制品中未见,而炮制品中出现的8和16号峰为未知成分,尚待进一步研究。在聚类分析中可聚为两大类,生泽泻S1~S10号样品聚为一类,清炒泽泻C1~C10和盐泽泻Y1~Y10号样品聚为一类。结论清炒和盐炙对泽泻化学成分的组成和含量均可产生影响,其中盐炙的影响明显大于清炒。

Objective To establish HPLC fingerprints of crude Alisma Rhizoma, stir-fried Alisma Rhizoma and salted Alisma Rhizoma and to investigate the effect of simple stir-frying and salted stir-frying on the chemical constituents in Alisma Rhizoma by comparing the difference among them. Methods Samples were extracted with methanol 5 mL by ultrasonic wave and then separated on Hypersil BDS C18 （250 min×4.6 mm, 5μm）column. Gradient elution was ean-ied out with a mobile phase of methyl eyanides-water. The detection wavelength was 208 nm, the column temperature was set at 25 ℃, the flow rate was 1.0 mL·min-1 and the analytic time was 120 rain. The software ＂Similarity Evaluation System tor Chromatographie fingerprint of TCMs＂ （ Version 2004A ） was employed to generate the mean chromatogram and carry out the similarity analysis of the samples. SPSS was employed to carry out the cluster analysis. Results Similarity of crude Alisma Rhizoma was 0.905 - 0.948. Similarity of stir-fried Alisma Rhizoma was 0.902-0.955. Similarity of salted Alisma Rhizoma was 0. 904- 0. 960. There were 14 common peaks in HPLC fingerprints of crude Alisma Rhizoma. There were respectively 15 common peaks in HPLC Finger-prints of stir-fried Alisma Rhizoma and salted Alisma Rhizoma. Their peak areas had distinctions in different degrees. The peak 2 which was in HPLC fingerprints of crude Alisma Rhizoma could not be tound in HPLC Fingerprints of processed produces.The peak 8 and peak 16 were unknown constituents in HPLC fingerprints of processed products.These unknown constituents should be studied further.They belonged to two main categories in cluster analysis.The sample S1-S10 which were crude Alisma Rhizoma could be clustered together.The sample C1-C10 which were stir-fried Alisma Rhizoma and the sample Y1-Y10 which were salted Alisma Rhizoma could be clustered together.Conclusion Simple stir-frying and salted stir-frying could affect the compositions and contents of chemical constituents in Alisma Rhizoma.The effect of salted stir-frying was obviously larger than that of simple stir-frying.