Quality difference analysis of Tribuli Fructus from different producing areas based on HPLC multi-component quantitative combined with chemical pattern recognition
Objective To establish a method for the simultaneous determination of quercetin,kaempferol,isorhamnetin,chrysin,tribuloside,terrestrosin K,terrestrosin D,terrestroside B,(25R)-spirost-4-ene-3,12-dione,hecogenin,diosgenin and tigogenin in Tribuli Fructus,and to explore the application of principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA)and grey relational analysis(GRA)in the quality difference evaluation of Tribuli Fructus.Method HPLC method was used with acetonitrile as mobile phase A and 0.05％phosphate acid solution as mobile phase B in gradient elution mode;the detection wavelengths were 360 and 203 nm.Through the chemometric analysis of the results of multi-index component content,the GRA model was established to comprehensively analysis the quality differences of Tribuli Fructus from different producing areas.Results The results of methodological verification were in line with the requirements of Chinese Pharmacopoeia;There were good linear relationships(r＞0.999)when the 12 components were in their respective ranges,and the average recovery rate(n=9)were 96.92％-100.18％(RSD＜2.0％).Chemometric analysis showed that(25R)-spirost-4-ene-3,12-dione,isorhamnetin,kaempferol and quercetin were the main potential markers affecting the quality of Tribuli Fructus.The relative correlation degree of GRA were 0.3397-0.5971,and there was a certain difference between producing areas of Tribuli Fructus.Conclusion The HPLC method can simultaneously determine the 12 components in Tribuli Fructus,which is simple and practical,PCA,OPLS-DA and GRA can be used to analyze the quality differences of Tribuli Fructus from different producing areas.
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