Establishment of high-performance liquid phase(HPLC)fingerprint of Centaurea behen.L root and analysis of chemical pattern recognition
Objective Toestablish a high-performance liquid phase(HPLC)fingerprint of Centaurea behen L.root medicinal material,toevaluate it using chemical pattern recognition method,and toestablish a multi index component content determination method.Methods WondaSil C18(250 mm ×4.6 mm,5 μm)chro-matography column was used,with a gradient elution of acetonitrile(A)-0.1％formic acid(B)as the mobile phase.The flow rate was 1.0 mL/min,the detection wavelength was 290 nm,the column temper-ature was 30℃,and the injection amount was 10 μL.Using the similarity evaluation system for traditional Chinese medicine chromatography fingerprint(2012 version),HPLC fingerprint overlay spectra were es-tablished for 15 batches of C.behen roots and similarity was analyzed.By comparing and identifying the control samples,chemical components were confirmed,and content was determined.Hierarchical cluster analysis(HCA),principal component analysis(PCA),and orthogonal partial least squares discriminant a-nalysis(OPLS-DA)were performed using SPSS 26.0 and SIMCA-P 14.1 softwares to evaluate the quality of different batches of thesamples.Results HPLC fingerprint spectra of Fifteen batches of C.behen L.root was established,and a total of 20 common peaks were matched.Chlorogenic acid(peak11),caffeic acid(peak13)and isochlorogenic acid A(peak19)were identified by comparison of reference substance.A method for determining the content of chlorogenic acid and isochlorogenic acid A has been established;The similarity range of the fingerprint spectrum was 0.601～0.983.HCA divided 15 batches of C.behen into 2 categories;The cumulative variance contribution of the first 4 principal components derived from PCA was exactly 85.260％;The OPLS-DA showed that 2 components,including chlorogenic acid and isochlorogenic acid A,were the symbolic components of the quality difference of C.behen L.root.Conclusion The com-bination of HPLC fingerprinting with chemical pattern recognition techniques allows for the rapid differen-tiation of characteristic components and effective evaluation of C.behen root.This integrated approach provides a robust basis for standardizing and ensuring the quality control of C.behen root.
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