EGCG In Grass Carp Body Metabolism And Its Bacteriostatic Effect On Pathogenic Bacteria

EGCG is isolated from the tea catechins monomers,is tea polyphenols bioactive ingredient.This experiment is to grass carp as experimental object,explore a concentration of 75 mg/kg of EGCG lavage after grass carp,fish plasma and tissue samples at different time points analysis,metabolic rules of catechins in the in addition,we also compare the four types of catechin: EGCG,ECG,EGC,EC in vitro inhibition of grass carp pathogenic bacteria,moisture,bacterium pseudomonas aeruginosa and the effect of citric acid bacillus research.The conclusion is as follows:(1)The establishment of a grass carp plasma and tissue samples before treatment.Were accurately draw 0.6 mL plasma,2 mL of liver and serum in 10 mL with a plug in the centrifuge tube,are joined to 40 ?L 20% of Vc and 200 ?L 0.05%Na2-EDTA solution,after blending,to join the ethyl acetate exactly 4 mL,oscillation 5 min,6000 r/min high-speed centrifugal 5 min,take out all the upper organic phase;Add 2 ml ethyl acetate extraction repeats,merging two centrifugal extraction organic phase on the bottom of the tip of the glass tube,the weak nitrogen flow in 45 ? water bath,to join in plasma samples of 500 ?L10 % acetonitrile-water solution,dissolve the oscillation,18000 r/min speed centrifugal after 5 min,learned that 20 muL supernatant fluid sample analysis.Liver tissue extract wave after working with uL 500 10% acetonitrile-water solution,then add 0.5 mL n-hexane to fat,after the oscillation,18000 r/min speed centrifugal after 5 min,abandon all to n-hexane layer,0.5 Ml n-hexane repeat to fat,lower speed centrifugal after 20 m?L sample analysis.(2)Using HPLC analysis EGCG levels in the blood,liver,according to the standard curve for content determination,make the EGCG absorption metabolism in serum and liver of concentration-time curves.After lavage for 15 min,30 min,45 min,60 min,120 min and 300 min six point to collect fish blood samples.By contrast with EGCG standard,blank sample,15-300 min after lavage in serum can detect EGCG prototype,and 15-45 min time range,increasing the concentration of EGCG in plasma,15 to 45 min over time,the concentration increasing,the maximum concentration at the time of 45 min,Tmax is 45 min,the largest concentration of Cmax is 512.8068 ?g/mL,under the concentration,calculate the relative bioavailability of 5.57%,60-300 min then slow down.(3)In the same way,15 min to 300 min in time within the scope of the liver samples also can detect the EGCG prototype,within the scope of 15 to 60 min time over time,EGCG concentration increasing,lavage after 60 minegcg maximum concentration Cmax434.1476 ?g/mL,Tmax is 60 min.(4)The use of UPLC ESI-Orbitrap-MS/MS analysis of sample group testing,the results showed that 15-300 min time range inner in serum,liver and intestinal tissue samples can detect EGCG prototype,in time for 30 min of liver samples detected the existence of the ECG,60-300 min time range in serum,liver and intestinal tissue samples were detected the presence of ECG,EGCG metabolites.(5)Through a series of increasing concentration of the solution of four kinds of bacteria liquid bacteriostatic test research shows that four kinds of catechin on four kinds of bacterium escherichia coli 44102,citric acid coli,pseudomonas aeruginosa and heavy moisture all have different degrees of bacterium antibacterial effect,low concentration of liquid(0.05-0.4 mM)inhibitory effect belongs to low sensitivity;Concentration in the solution(0.4-2.0 mM)inhibition of ZhongMin;A high concentration of liquid(2.0-10 mm)inhibitory effect belongs to gao min.0.05 mM for MIC(minimum inhibitory concentration),when the solution concentration increased from 0.2 mM to 4.0 mM,the bacteriostatic circle was bigger,increase the size of the bacteriostatic effect is more obvious;When the solution concentration increased from 4.0 mM to 10.0 mM,the bacteriostatic circle small,bacteriostatic effect was enhanced with a narrow,bacteriostatic effect gradually to limit.Overall,with the increase of solution concentration,bacteriostatic circle diameter on the rise,namely the bacteriostatic intensity increases with the increase of solution concentration.The same concentration of solution on different bacteria inhibition.EGCG in three kinds of pathogenic bacteria addicted to moisture,citric acid bacterium coli and pseudomonas aeruginosa bacteriostatic effect is stronger than the nonpathogenic bacteriae.coli 44102,among them,the citric acid bacillus bacteriostatic effect was strongest.And,EGCG bacteriostatic effect is stronger than the rest of the three kinds of catechin,EGCG > EGC > ECG > EC.