| Objective:This paper adopts HPLC fingerprint method to research Marchantia convoluta Gao et Chang from Guangxi and compared with the fingerprints of different origins and different varieties of medicinal materials.which was identified,classified and evaluated by similarity analysis,hierarchical cluster analysis（HCA）,principal component analysis（PCA）.What’s more,in order to control the quality of Marchantia convoluta,we study the spectral effect relationship of total flavonoids with Marchantia convoluta Gao et Chang.Methods:1.The establishment of HPLC fingerprints.:taking Marchantia convoluta Gao et Chanand,other different origins and varieties from Guangxi as research objects.The separation was developed on InertSustain-C18（4.6 mm*250 mm,5μm）by gradient elution with acetonitrile-water（bothcontaining 0.1%phosphoric acid）as mobile phase at a flow rate of 1.0 m L·min-1,the column temperature at 30℃and detection wavelength at 270 nm.and the reference fingerprints were established.2.Extraction,purification and content determination of total flavonoids from.Marchantia convoluta Gao et Chang in Guangxi The total brass extract was purified by extraction silica gel column chromatography.The content of total flavonoids from 11 different regions of Guangxi was determined by Ultraviolet Spectrophotometry with 1%triethylamine-methanol colorimetry and apigenin as control at 275 nm.3.Establishing fingerprint of the total flavonoids by HPLC:Chromatography:Inert Sustain C18 column（250mm x 4.6mm,5um）,ultraviolet detector detector,acetonitrile（A）-0.1%phosphoric acid（B）as mobile phase,injection volume,column temperature,25 degrees,flow rate.With the detection wavelength of 270 nm and the fingerprint.technology of HPLC,the fingerprint of total flavonoids in Radix Scrophulariae was constructed,and the corresponding data of common peak area were obtained by similarity analysis and cluster analysis.4 Pharmacodynamic study on Anti-Acute liver injury:140 male rats were randomly divided into 14 groups:solvent group,model group,bifendate dropping pill control group(3mg.kg-1),total flavonoids group(10Omg.kg-1)from 11 different regions of Guangxi,with 10 rats in each group.The solvent group and model group were intragastric administration of distilled water according to 1ml/100g,Bifendate Pills group to 3mg.Kg-1 crude drug Ig,different batches of total flavones of Marchantia convoluta extract(dose:100mg.kg-1 intragastric administration.After 10 days of administration,AST and ALT were detected as indicators to investigate the Anti-Acute liver injury activity of total flavonoids from Dioscorea fortunei.The Anti-Acute liver injury effects of total flavonoids from 11 different regions of Guangxi were obtained.5.Study on the Spectrum-Effect relationship of total flavonoids from Marchantia convoluta Gao et Chang against acute liver:The common peak data of each fingerprint was standardized,and the gray correlation analysis method was used to correlate the peak information of each chromatographic peak with the efficacy index to screen the acute liver injury.The chromatographic peaks closely related to the function;using SPSS21.0,using linear regression analysis method,using the fingerprint peak information and pharmacodynamic data of 8 batches of herbs to establish the spectral effect relationship model,and the accuracy and reliability of the remaining 3 batches of sample data verification models.Results:1.The established fingerprints of the medicinal herbs from Marchantia convoluta Gao et in Guangxi and and other land-based medicines is stable and reliable.The peak No.5 is identified as apigenin.The similarity of 30 samples of medicinal herbs from Guangxi was more than 0.8,and the composition is relatively stable.The results of cluster analysis and principal component analysis divide the medicinal materials into four categories,and the analysis results are consistent.Cluster analysis showed that A2,A4,A6,A7,A8,A10,A11,B2,B3,B6,B7,B8,B9,B10,B11,C1,C3,C5,C6 and C8 from Guangxi origin were more similar than those from other three origins.Principal component analysis extracts four main components,the.contribution rate of eigenvalue lambda 1 is 58.993%,the contribution rate of lambda 2 is 21.957%,the contribution rate of lambda 3 is 8.654%,the contribution rate of lambda 4 is 5.010%,and the cumulative contribution rate of the first four eigenvalues is 94.606%.The cumulative contribution rate of the first four eigenvalues is 94.606%.The information of the original samples is almost retained,of which the first is the main component.The score has the largest eigenvalue.2.The extraction and purification results of 11 batches of medicinal materials from different places in Guangxi showed that they may be affected by climate,geographical environment and harvesting time.Among them,A5（Huanjiang County of Hechi City）and A9（Pingnan County of Guiping City）had the highest rate of extracting,while A3（Guigang City）had the lowest rate of extracting.Ultraviolet spectrophotometry was used to determine the content of total flavonoids from Marchantia convoluta Gao et.The results showed that the content of total flavonoids in each batch of Marchantia convoluta Gao et was more than 70%.3.The results of 11 batches of total flavonoid extracts from different origins in Guangxi by HPLC showed that the fingerprint was constructed.The fingerprint method was stable and reproducible.A total of 11 common peaks were calibrated,which visually displayed the internal multi-component chemical peak information of total flavonoids from Marchantia convoluta Gao et.Cluster analysis shows that all samples can be divided into three categories,namely the first A4,the second A6,A7,A11,and the third A1,A2,A3,A5,A8,A9,A10.The similarity is higher than the other two categories,which is consistent with the similarity analysis.In the whole fingerprint,the main characteristics of fingerprints from 1,3,4,8,9,11 play an important role in the quality of samples.Other batches constitute the secondary characteristics of fingerprints and have little contribution to fingerprints.It can be seen that the content of total flavonoids in different batches is different.4.The 11 batches of Marchantia convoluta Gao et from different places in Guangxi have anti-acute liver injury effects.The values of AST and ALT in all drug-administered groups were reduced to different extents,and there was significant difference compared with the model group（P<0.05 or P<0.01）,so AST and ALT can be used as screening pharmacodynamic correlation peaks and establishing pharmacodynamic basis for mathematical models.However,the pharmacodynamic effects of different samples of origin are different,which may be due to the difference in the total flavonoid content of the medicinal materials.5.The peak information of HPLC peaks of 11 batches of cotton samples from different producing areas in Guangxi was correlated with the pharmacological action data of acute liver injury.The correlation coefficient between each chromatographic peak and drug efficacy was greater than 0.7,and the fingerprint was established.The characteristic peak was positively correlated with the anti-acute liver injury effect of total flavonoids of F.chinensis,indicating that apigenin is the basis of anti-acute liver injury activity of total flavonoids of P.sinensis,and it has a great correlation with its efficacy;The relationship between the total flavonoids of Didi Qiang and the spectrum of acute liver injury.The equation is reasonable and effective,and the prediction of the efficacy against acute liver injury is good.The absolute value of the relative error of the prediction results is less than 10%.The comprehensive quality analysis of the medicinal materials has important theoretical significance for the quality analysis of the money.Practical significance.Conclusion:The fingerprints and comparisons of Marchantia convoluta Gao et produced in Guangxi and other land-based medicines were established,and the relationship between the spectrum and effect was expounded.It supplemented the systematic research on the pharmacology of medicinal materials in Guangxi,and provided its evaluation system of"component"-"peak"-"efficiency".It measured the quality of its medicinal materials in three dimensions and provided an experimental basis for the improvement of its quality standard.To lay the foundation for the development and utilization of Marchantia convoluta Gao et produced by Guangxi. |
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