Metabonomics Based On LC-MS To Study Volatile Oils Of Different Processed Products From Angelica Sinensis On Rat Model Of Acute Inflammatory Paw Edema

LC-Q/TOF-MS was used for analysis platform, through the rat model of acute inflammatory paw edema induced by carrageenan was established, volatile oils from charred AS(C-ASVO) and from parched AS with alcohol(J-ASVO), soil(T-ASVO),and sesame oil( Y-ASVO) were respectively used to treat rats with acute inflammation, and evaluate the anti-inflammatory effect of angelica naphtha.rats plasma and inflammatory leachate as the biological samples, biological sample pretreatment method and chromatographic separation conditions were applied to discuss, and Quality control(QC) was used to evaluate samples.the strategy of metabonomics research was adopted to investigate the anti-inflammatory mechanism of volatile Oils of Different Processed Products from Angelica Sinensis, SIMCA-p13.0 analysis software was taken to get the differences metabolites, last metabolic pathways of different metabolites was built.In order to provide new ideas and new methods for the overall effect and material base of traditional Chinese medicine, the acute inflammation mechanism of volatile oils of different processed products from Angelica Sinensis on rat urine metabonomics was studied as a whole. The main research conclusions are as follows:1. LC-Q/TOF-MS analysis platform was adopted to get the samples of plasma and inflammatory leaching liquid and the chromatographic separation conditions were optimized.The results showed that methanol was used to deproteinization,which was more suitable for the sample of LC-Q/TOFMS determination.Chromatographic separation conditions as follows: the flow rate was 0.4 m L/min; column temperature was 35℃; mobile phase composition A: ultrapure water(containing 0.1% formic acid), B: acetonitrile(including 0.1% formic acid); Gradient elution procedure was:5% B in 0-1 min,5% changed to 20% of B in 1-6 min, and B linear change change from 20% to 50% in 6-9 min.B linear change change from 50% to 95% in 13-15 min,and kept 95% B about 13-15 min. Sample quantity is 4μL, automatic injector temperature of 4℃. QC samples were used to evaluate the confirmed sample pretreatment methods and the chromatographic separation conditions.The resultsshowed that QC samples of plasma and inflammatory lixivium were all got good separation, the peak shape was symmetrical and the baseline was stable.In the positive ion detection mode, the plasma and inflammatory components average detection rate after calculated were respectively 93.628% and 90.1535%, relative standard deviation RSD were respectively 5.784% and 2.197%.In the negative ion detection mode, the plasma and inflammatory components average detection rate after calculated were respectively 95.628% and 90.2981%, relative standard deviation RSD were respectively 4.162% and 6.4708%. It was suggested that the selective chromatographic separation conditions could effectively make the target sample of plasma and inflammatory leach liquor components were well separated, analysis platform repeatability was good.Which could be used for analyzing all the samples in the experiment, and the datas could be also used in multi-dimensional statistical analysis of metabonomics further analysis.2. LC-Q/TOF-MS was used to test the metabolites of plasma and inflammatory leach liquor in control group, model group, C-VOAS group, J-VOAS group, T-VOAS group and Y-VOAS group.34, 34 metabolites were respectively identified and speculated from the plasma and inflammatory leaching liquid. Principal component analysis and orthogonal partial least squares discriminant analysis were used to analyze plasma and inflammatory leachate.The control and the model were found to complete separation, C-VOAS, J-VOAS, T-VOAS and Y-VOAS were closed to the control group, showed that model group built successfully. C-VOAS, J-VOAS,T-VOAS and Y-VOAS has a certain role on the rat model of acute inflammatory paw edema induced by carrageenan.Which presented a trend that returned to normal.According to the variable importance projection, combined with T-test results to screen differences metabolites, which found that potential pharmacological effect of C-VOAS was more obvious than any other group, and it could make more difference metabolites back to normal. Metabolomics Pathway Analysis(Met PA) database was used to analyze the differences metabolites of metabolic pathways.3. systematic research was used to analysis the difference metabolites of the plasma and inflammatory leaching liquid and then integrated them, 42 kinds of differencesmetabolites was got in total.Input all of them to Met PA database, built relevent metabolic pathways, further study the acute inflammation mechanism of volatile Oils of different processed products from Angelica Sinensis. Thereby provided a more integrity, more systematically theoretical basis to these studies. Analysis results showed that Angelica Sinensis could restore the fatty acid metabolism disorder by adjusting the Krebs cycle, improve the glucose content,then played an anti-inflammatory effects.The restored metabolites by Angelica Sinensis, combined with speculating the mechanism of carageen glue caused, arachidonic acid, succinic acid, pyruvic acid, glucose, stearic acid, malic acid,which screened from those metabolites has biological significance,and it could be used as biomarks of the rat model of acute inflammatory paw edema induced by carrageenan, and anti-inflammatory effect of C-ASVO was better than T-ASVO, J-ASVO and Y-ASVO.