Study on Quality Evaluation of Famous Classical Formula Kaixin Powder Based on the Characteristic Chromatography， Content Determination Combined with Chemometrics Analysis

OBJECTIVE To establish the characteristic chromatogram and multi-component content determination method of Kaixin Powder, and to evaluate the quality of multiple batches of Kaixin Powder using chemometric methods.

METHODS The HPLC-UV-ELSD method was used to establish the characteristic chromatograms and multi-component content determination of Kaixin Powder, principal component analysis (PCA) was used for dimensionality reduction analysis of multiple batches of Kaixin Powder, and Hotelling’s T² and DModX methods were used to set the quality control range of different batches of Kaixin Powder.

RESULTS Fifteen batches of Kaixin Powder HPLC-UV characteristic chromatograms identified 26 characteristic peaks and generated a reference characteristic chromatogram, with a similarity greater than 0.90 compared to the reference characteristic chromatogram. HPLC-ELSD characteristic chromatograms identified 18 characteristic peaks, and the relative retention times of each characteristic peak were calculated based on ginsenoside Rb₁ as a reference peak. The RSD was 0.01% to 0.39%, indicating good inter-batch consistency. PCA analysis identified common peaks that contribute significantly to the overall quality of Kaixin Powder, among which Polygalasaponin Ⅲ 3,6′-disinapoyl sucrose, Ginsenoside Re, and Rb₁ were the indicators measured in this study. Hotelling’s T² and DModX control limits were 28.857, 14.433 and 1.605, 1.603, respectively. The contents of polygalaxanthone Ⅲ, 3,6′-disinapoyl sucrose, β-asarone, ginsenoside Rg₁, ginsenoside Re and ginsenoside Rb₁ in 15 batches of Kaixin Powder were 0.43 to 0.49 mg·g^-1, 1.57 to 2.15 mg·g^-1, 1.77 to 3.12 mg·g^-1, 0.45 to 0.65 mg·g^-1, 0.42 to 0.98 mg·g^-1, 0.59 to 1.04 mg·g^-1, respectively, with good linear relationship (r≥0.998 1). The results of cluster heat map analysis showed that 15 batches of Kaixin Powder could be clustered into two categories.

CONCLUSION The established HPLC-UV-ELSD characteristic chromatogram method and multi-component content determination method are convenient and accurate. Combined with chemometrics, they can be systematically and comprehensively used for the quality evaluation of Kaixin Powder. This study proposes that the HPLC-UV characteristic chromatogram similarity of Kaixin Powder should be no less than 0.90. Under the HPLC-ELSD characteristic chromatogram, ginsenoside Rb₁ should be used as the reference peak, and the limit of the average relative retention time of each characteristic peak of 15 batches of Kaixin powder should be ±10%. The content determination limits should be as follows: Polygalasaponin III (0.33-0.60) mg·g^-1, 3,6′-disinapoylsucrose (1.36-2.52) mg·g^-1, β-asarone (1.70-3.15) mg·g^-1, ginsenoside Rg₁ + ginsenoside Re (0.80-1.48) mg·g^-1, and ginsenoside Rb₁ (0.56-1.04) mg·g^-1.