Expression Of Banlangen Genealogy Based On Target Component Deletion And Its Application In Screening Drug Substance

Objective:In the view of spectrum-effect relationship, the new pattern for efficient components recognition and quality control of Isatidis Radix by component "deletion/increment" strategy was established rudimentarily. Methods:Optimal select activity antibacterial position of Isatidis Radix by Cylinder-Plate Method which Escherichia coli[AG100A] as its Biological Model, Optimal select Rabbits erythrocyte agglutination activity position from Isatidis Radix. Establish the HPLC fingerprint of 10 batches of activity position which from different habits, and used mathematical statistical methods such as cluster analysis, principal component analysis and radar chart to study the spectral efficiency correlation; Using TLC method, semi-preparative HPLC method to delete the target components from the strongest anti-E.coli[AG100A] and erythrocyte agglutination activity position; Determination IC50 of target components and its negative components of anti-E.coli[AG100A] position with agar dilution method, Determination IC50 of target components and its negative components of erythrocyte agglutination activity position with MTT method; Using the equivalent curve analysis research the relationgship between target components and its negative components, one target components and another.. Explored the content of the component group of key efficacy limited range by gradually "incremental" target composition to the negative samples, and determinated the strongest antibacterial and antiviral activity of the effective components. Determination the contents of activity components with HPLC-DAD,HPLC-ELSD and acid-base titration.Results:Organic acid site was the strongest antimicrobial active site of Isatidis Radix. Spectroscopy-effect relationship found six peaks of organic acid fractions were closely related with antibacterial activity, there have two peaks which were benzoic acid, salicylic acid with the reference standard, and the remaining peaks to be further identified; Six target consitituents were deleted by separating the antibacterial activity sample from Isatidis Radix using thin-layer chromatography(TLC), the six constituents, which were marked as X+, were A+(benzoic acid), B+(syringic acid), C+(unknown constituents), D+(salicylic acid), E+(unknown constituents), F+(unknown constituents), The IC50 of target components and its negative components like these:A+-F+:50.56mg/mL,168.88 mg/mL,147.24mg/mL,1123 mg/mL,182.77mg/mL,220.66mg/mL; A-～F-:200.87mg/mL,97.24 mg/mL,100.30mg/mL,167.08mg/mL,101.71mg/mL,98.29mg/mL; T(total organic acids in parallel samples):96.65mg/mL.the sequence of antibacterial activities of the total organic acids in parallel samples, the six target constituents and its negative samples were A+>total organic acids in parallel samples>B->F->C->D+>E->C+>D->B+>E+>A->F+. The result of equivalent curve analysis shows that benzoic acid and salicylic acid exhibit cooperativity, syringic acid and benzoic acid exhibit additivity, syringic acid and salicylic acid exhibit additivity, benzoic acid, salicylic acid are the main effective components when Isatidis Radix antibacterial. The n-butanol extracts had the best agglutination effect on red blood cells of the rabbit. The study on the spectrum-activity relationship of the extracts found that the 2nd and 11th common peak have close relationship with agglutination effect, the structure of the two common peaks need further identified. Epigoitrin and FG were deleted by separating from n-butanol extracts by the meas of semi-preparative HPLC. IC50 of epigoitrin is 239.87mg/ml, FG is 458.38 mg/ml, the negative component of epigoitrin is 1928.25mg/ml, the negative component of FG is 1636.49mg/ml, the n-butanol extracts in parallel sample is 1130.41 mg/ml. The result of equivalent curve analysis shows that epigoitrin and FG exhibit cooperativity. Anti-virus active of the increment samples of epigoitrin and FG showed that quantity of them play the antivirus action as follows:the lowest content of epigoitrin is 82.5μg/mL, the higest is 289.7μg/mL and the optimum is 225.0μg/mL; the lowest content of FG is138.2μg/mL, the higest is 618.9μg/mL and the optimum is 450.0μg/mL.Conclusion:A pattern based on the component of Isatidis Radix "deletion/increment" strategy was preliminary established for screening the key efficient components and quality control in the paper. The key components of antibacterial and anti-viral were found and their contents limitation were established. These results provided a pragmatic pattern and some useful study ideas for efficient components recognition and quality control of TCD.