Establishment of TLC identification and HPLC fingerprint of Inonotus obliquus and analysis of chemical pattern recognition / 中国药房

ABSTRACT

OBJECTIVE To establish thin-layer chromatography （TLC） identification method and high-performance liquid chromatography （HPLC） fingerprint of Inonotus obliquus， and to evaluate the quality of I. obliquus by chemical pattern recognition. METHODS TLC method was used to identify trametenolic acid and inotodiol in I. obliquus qualitatively. HPLC fingerprint of I. obliquus was established； Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition） was used to determine the common peaks and evaluate the similarity； chemical pattern recognition analysis [cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis （OPLS-DA）] of 22 batches of I. obliquus was performed with SPSS 23.0 software and SIMCA14.1 software. RESULTS In the TLC， the same color spots were found at the same position in the chromatograms of test sample and substance control. A total of 10 common peaks were marked in the HPLC fingerprints of 22 batches of I. obliquus， with similarities of 0.942-0.995. No. 3 peak was identified as trametenolic acid， No.4 peak as inotodiol， No. 9 peak as ergosterol and No. 10 peak as lanosterol. Results of cluster analysis showed that S1-S15， S19， S21 and S22 could be clustered into the first category， and S16-S18 and S20 were clustered into the second category. Results of principal component analysis showed that top 4 samples in the list of comprehensive score were S17， S18， S16 and S20. Results of OPLS-DA showed that three marking components that may affect the quality of I. obliquus were screened according to the standard of VIP＞1， i.e. No. 4 peak （inotodiol， VIP value of 1.86）， No. 3 peak （trametenolic acid， VIP value of 1.62） and No. 7 peak （VIP value of 1.27）. CONCLUSIONS This study establishes TLC method and HPLC fingerprint of I. obliquus successfully， which can provide reference for the quality control of I.obliquus by combining with chemical pattern recognition.