Study on HPLC Fingerprint and Multiple Index Components Determination of Changtongshu Granules

OBJECTIVE  To establish the HPLC fingerprint of Changtongshu granules, and components determination methods of multi-index component.

METHODS  HPLC was used to create the fingerprint of Changtongshu granules, and to evaluate the similarity of 22 batches of samples from 3 companies by the similarity evaluation system software for traditional Chinese medicine fingerprint. With Agilent ZORBAX SB-C₁₈ chromatographic column(250 mm×4.6 mm, 5 μm), acetonitrile-0.2% phosphoric acid as mobile phase of gradient elution, column temperature of 35 °C, flow rate of 1 mL·min⁻¹. To determine the contents of gallic acid, protocatechin acid, paeoniflorin, 2,3,5,4’-tetrahydroxy stilbene-2-O-β-D-glucoside, benzoic acid and emodin. The quality of sample was evaluated using cluster analysis, principal component analysis and orthogonal partial least squares method-discriminant analysis.

RESULTS  A total of 12 common peaks were identified, and 7 components were identified. Comparing the 22 batches of samples from 3 different companies with the reference fingerprint, only 3 batches of samples had a similarity above 0.96, 1 sample greater than 0.87, while the similarity of the remaining samples ranged from 0.627 to 0.859. Though cluster analysis, the sample from manufacturer A were clustered into group Ⅰ, the sample from manufacturer B were clustered into group Ⅱ, and the sample from manufacturer C was clustered into group Ⅲ. Principal component analysis showed that the cumulative variance contribution rate of the six principal components was 94.447%, and the classification results of the three-dimensional model were consistent with cluster analysis. Based on orthogonal partial least squares method-discriminant analysis, 6 components with large differences were identified, including gallic acid, protocatechin acid, paeoniflorin, 2,3,5,4’-tetrahydroxy stilbene-2-O-β-D-glucoside, benzoic acid, and emodin.

CONCLUSION  The established method is accurate, simple and good repeatability, and it can be used for Chuantongshu granules quality control and evaluation.