| Objective:In order to provide scientific evidence for studying the quality control and evaluation,pharmacodynamic material basis,compatibility regularity and preparation technology of traditional Chinese herbal compounds,the UPLC-MS/MS fingerprint chromatograms of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine were established and correlation between constituents which were represented by main common peaks from fingerprint chromatograms and pharmacodynamic informations which were acquired by the research on effective mechanism of the damp type ulcerative colitis model of rats were treated by Gegen Qinlian Decoction was confirmed with the help of study on spectrum-effect relationship.Mothods:1.The chromatograms of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine were acquired by the method of UPLC-MS/MS which was performed on an ACQUITY UPLC BEH C18 column(1.7μm,100 mm×2.1 mm)by gradient elution in 80min with the mobile phase of 0.2%formic acid water-acetonitrile at the speed of 0.3 mL·min-1 and the column temperature was 40℃.Besides,using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(version 2004A)which was issued by Chinese Pharmacopoeia Commission,the similarity was evaluated.2.The damp type ulcerative colitis(UC)model of rats were copied by 2,4,6-trinitro-benzene-sulfonic acid(TNBS)which combined with high fat and high carbohydrate diet,the pharmacodynamic informations were obtained by comprehensively evaluating effect of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine on the basis of indexes including disease activity index(DAI),colon mucosa damage index(CMDI),pathology observation,IL-4,IL-10,TNF-α,IFN-γcontents of rats serum and colon.3.The relative areas of constituents which were represented by main common peaks from fingerprint chromatogram of Gegen Qinlian Decoction and the pharmacodynamic informations were correlated by the method of principal component analysis and grey relational analysis and the correlation between components and efficiency of Gegen Qinlian Decoction was determined.Results:1.The UPLC-MS/MS fingerprint chromatograms of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine were established and components which were represented by 11 common peaks from fingerprint chromatogram of Gegen Qinlian Decoction,which were determined by relative retention time and relative peak areas,were qualitatively identified based on using the reference substances which combined with mass fragmentation rule.The result of the similarity showed that the UPLC-MS/MS fingerprint chromatograms of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine reached the standard of fingerprint establishment.2.The result of study on pharmacodynamics showed that high dose groups of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine could reduce DAI score and CMDI score,impove colonic mucosa hyperaemia,edema,inflammatory cell infiltration,increase anti-inflammatory cytokine contents including IL-4,IL-10 and decrease pro inflammatory cytokine contents including TNF-a,IFN-y of rats serum and colon in varying degrees,which had statistical significance compared with the model group(P<0.05).Among all the high dose groups,the efficiency of Gegen Qinlian Decoction is the most significant(P<0.01).3.The result of study on spectrum-effect relationship showed that the correlation between components of puerarin,daidzin,coptisine,epiberberine,j atrorrhizine hydrochloride,baicalin,berberine hydrochloride,palmatine hydrochloride,wogonoside,baicalein and ammonium glycyrrhizinate which could represent the whole information of Gegen Qinlian Decoction in major degree and efficiency all above 0.5,the constituents of Gegen Qinlian Decoction and efficiency had a certain correlation which revealed that it was the result from combined action of multicomponent group of Gegen Qinlian Decoction.Conclusion:It is the first time to build the UPLC-MS/MS fingerprint chromatograms of Gegen Qinlian Decoction,separated prescriptions and single herbal medicine.At the same time,Gegen Qinlian Decoction,separated prescriptions and single herbal medicine can improve the inflammation condition of damp type ulcerative colitis(UC)model of rats in varying degrees and the efficiency of Gegen Qinlian Decoction is the most significant because of the result from combined action of multicomponent group.To sum up,it lays the foundation of study on the quality control and evaluation,pharmacodynamic material basis,compatibility regularity and preparation technology of traditional Chinese herbal compounds. |
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