Quality Evaluation of Reineckia Carnea by Fingerprint Combined with Chemical Pattern Recognition and Determination of 4 Chemical Components in Reineckia Carnea by HPLC

Based on the HPLC fingerprint of Reineckia carnea, the quality of 20 batches of Reineckia carnea (S1-S20) was evaluated with chemical pattern recognition, and 4 chemical components, including rutin, ginsenoside Rb1, dioscin and diosgenin, were determined. The sample of Reineckia carnea was extracted twice by reflux with 75% (φ) ethanol solution and filtered, and the combined filtrate was evaporated by rotation to dryness. The residue obtained was dissolved and made its volume up to 10 mL with methanol. The solution was passed through 0.45 μm microporous membrane, and the filtrate was used for HPLC analysis. The HPLC fingerprints of 20 batches of Reineckia carnea were established and their similarity was evaluated with the similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2012A version). Cluster analysis, principal component analysis and partial least squares-discriminant analysis were performed combined with chemical pattern recognition, and the differential components were screened. The results showed that 13 common peaks were selected from the HPLC fingerprints of 20 batches of Reineckia carnea, and 4 components were identified from them, including rutin (peak 5), ginsenoside Rb1 (peak 11), dioscin (peak 12) and diosgenin (peak 13). The similarity between the fingerprints of 20 batches of Reineckia carnea and the control profile was in the range of 0.546-0.942. Cluster analysis and partial least squares-discriminant analysis showed that 20 batches of Reineckia carnea were divided into 3 classes, and S7, S10, S12, S16, S18, S17, S15, S3, S4, S1, S13, S2, S14, S19, S6, S8, S20 and S9 were class 1, S11 was class 2, and S5 was class 3. Principal component analysis showed that the cumulative variance contribution rate of principal components 1-4 was 85.374%, and S5 had the highest comprehensive score among 20 batches of Reineckia carnea, followed by S11. Variable importance projection method was used to screen out 7 differential components, which were peak 13 (diosgenin), peak 2, peak 7, peak 5 (rutin), peak 12 (dioscin), peak 1 and peak 6. Linear relationships between the masses of rutin, ginsenoside Rb1, dioscin and diosgenin and their corresponding peak areas were kept in definite ranges, with detection limits (3S/N) in the range of 0.005-0.020 mg·g^-1. Test for recovery was made by standard addition method on S2, giving results in the range of 97.4%-102%, and RSD (n=6) of the determined values were ranged from 1.1% to 2.6%. This method was applied to the analysis of 20 batches of Reineckia carnea, and the determined values of rutin, ginsenoside Rb1, dioscin and diosgenin were 0.249-0.984 mg·g^-1, 0.431-5.851 mg·g^-1, 0.007-0.261 mg·g^-1 and 0.003-0.095 mg·g^-1, respectively.