The Analysis Of Chemical Components And The Study Of Quality Control For Traditional Chinese Medicinal Materials Derived From Animal Bile

In traditional Chinese medicinal materials derived from animal bile,seven bile-based crude drugs,including pig bile,cattle bile,sheep bile,bear bile,Calculus bovis,in vitro cultivated Calculus bovis,artificial Calculus bovis,are widely used.Bile-based crude drugs have been used clinically for 2000 years and their therapeutic functions are acknowledged in traditional Chinese medicine theory.Although they contain similar chemical components,their therapeutic functions and target organs are different.Due to the limited supply and high demand,the prices of Calculus bovis and bear bile are high and some counterfeit drugs have appeared in market.Based on material base in bile-based crude drugs(bile acids,bilirubin,amino acids),some specific quality control studies including qualitative and quantitative were accomplished in the paper.The characteristic chromatograms of TLC for the seven frequently-used bile-based crude drugs(pig bile powder,cattle bile powder,sheep bile powder,bear bile powder,Calculus bovis,in vitro cultivated Calculus bovis,artificial Calculus bovis)were established.The result indicated seven crude drugs could be identificated accurately and fastly with the characteristic chromatograms of TLC.The characteristic chromatograms of HPLC-ELSD for the seven frequently-used bile-based crude drugs were established.Seventeen bile acids could be tested accurately under the separation and detection conditions.The conditions and parameters of the characteristic chromatograms of HPLC-ELSD not only were used in the identification of the seven bile-based crude drugs,but also were used in the studies on some other bile-based drugs.Due to the extremely high sensitivity,the method of UPLC-QTOF-MS applies to the test of compound preparations derived from animal bile.Based on these facts,the characteristic chromatograms of UPLC-QTOF-MS were studied.Some chemometrics methods were used in the study.On the other hand,some selective ion chromatograms were studied and some characteristic ions(m/z:407.28,391.29,464.30,448.31,498.29,375.29,432.29)were ascertained.In addition,the characteristic chromatograms were used in the tests of two compound preparations.The NIR spectra and IR spectra of the seven bile-based crude drugs were studied.Based on these spectra,some chemometrics methods were used in the characteristic identification and pattern discriminant of the seven crude drugs.Methods,including Hierarchical clustering analysis,principal component analysis,K-nearest neighbours,Partial least square-discriminant analysis,support vector machine-discriminant analysis,were used in the study.The results showed that support vector machine-discriminant analysis and K-nearest neighbours were the best mathematical models for the NIR spectra and the IR spectra respectively.Because bile-based crude drugs contained high amount of bile acids,a method for the determinations of the major bile acids in the seven crude drugs based on HPLC-ELSD were studied.After all samples tested,the contents of the major bile acids in the seven different bile-based crude drugs were obtained.The HPLC-UV methods for the quantification of bilirubin in the bile-based crude drugs were studied additionally.Because of its wide use,an accurate and sensitive HPLC-UV method coupled with pre-column derivatization was developed for the simultaneous quantification of the major bile acids in artificial Calculus bovis.The amounts of bile acids in samples were analyzed by the methods of hierarchical agglomerative clustering analysis and principal component analysis.The results of the chemometric analysis indicated that the contents of the major bile acids reflect the intrinsic quality of artificial Calculus bovis,and two compounds(HDCA,CDCA)were the most important markers for quality evaluating.In addition,a stable,sensitive HPLC-UV method coupled with pre-column derivatization for the quantification of taurine was established and validated and the result showed the method was more suitable than the national standards(GB/T 5009.169-2003)for the quantification of taurine.Finally,some quality standard drafts on the seven bile-based crude drugs were revised.