HPLC Fingerprint Analysis of Substance Benchmark of Xiao Chengqitang / 中国实验方剂学杂志

ABSTRACT

Objective:To establish a high performance liquid chromatography （HPLC） fingerprint of the substance benchmark of Xiao Chengqitang and evaluate its quality with chemical pattern recognition method. Method:Diamonsil C₁₈ column （4.6 mm×150 mm， 5 μm） was used， mobile phase was consisted of methanol （A）-0.1% phosphoric acid solution （B） for gradient elution （0-60 min， 20%-90%A； 60-70 min， 90%-100%A）， the flow rate was 1 mL·min^-1， the column temperature was 25 ℃， and the detection wavelength was 254 nm. The similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine （2012 edition） was used to evaluate the similarity of HPLC fingerprint of 15 batches of substance benchmark of Xiao Chengqitang， and the chromatographic data were analyzed by cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis， in order to evaluate the quality difference between different batches of substance benchmarks of Xiao Chengqitang and find out the main chemical components that caused the quality difference. Result:The HPLC fingerprint of Xiao Chengqitang substance benchmarks was established， 31 common peaks were identified， and 18 components were identified by comparing with the reference substances. The similarities of 15 batches of HPLC fingerprint of Xiao Chengqitang substance benchmarks were >0.92. The samples could be divided into two categories by three chemical pattern recognition methods. Nine main components leading to the quality discrepancy of samples between batches were screened out， including rhein， chrysophanol-8-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， aloe-emodin-8-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， sennoside A， chrysophanol-1-<italic>O</italic>-<italic>β</italic>-<italic>D</italic>-glucoside， rhein-8-<italic>O</italic>-glucoside and others. Conclusion:The established fingerprint analysis method is accurate， stable and reproducible， which basically reflects the overall chemical composition characteristics of Xiao Chengqitang， and can be used for the quality control of Xiao Chengqitang preparations.