The Study On Detection Of Alcohol Metabolism Markers Ethyl Glucuronide In Biological Sample Based On GC-MS Method

Objective:Using Gas Chromatography-Mass Spectrometry?GC-MS?,determination of alcohol metabolism markers ethyl glucuronide,optimization of biological sample pretreatment method of urine,blood and hair,optimization of determination conditions of testing instruments,strive for high recovery rate,high degree of sensitivity and specific and accurate detection methods.Methods:First using ethyl glucuronide of determine the retention time of chromatographic peak and standard,and then use blank standard substance is added to the blood,urine and hair form simulated biological samples,blood,urine and hair samples by solid phase extraction?SPE?and liquid-liquid extraction pretreatment,two methods to precipitate protein in biological samples and various other has interference of impurity to the testing results of the experiment,after the extraction of the target using different derivatization reagent for derivatization reaction,the gas chromatography-mass spectrometry,using FULL SCAN,and SIM?ion?pattern detection analysis and qualitative analysis,The internal standard method?EtG-D₅?was used for quantitative analysis.Results:the blood,urine,hair biological sample methodology to investigate the method,standard working curve drawing respectively,?1?ethyl glucuronid in blood samples has a good linear relationship between 0.1³?g/ml concentration range with chromatographic response values?R²=0.9998?,the average recovery range of 95.86¹01.88,accuracy?standard deviation,SD?was 1.52².91,the relative standard deviation?RSD?of 0.51³.45;?2?There was a good linear relationship between 0.1³ g/ml ethyl glucuronide and chromatographic response within the concentration range in urine samples?R²=0.9992?.The average recovery was 98.10¹02.71,SD value was 0.81⁶.65,and RSD value was 0.27⁹.94;?3?There was a good linear relationship between5⁵0ng/mg ethyl glucuronide and the chromatographic response within the concentration range in the hair samples?R²=0.9949?.The average recovery was 75.16⁹9.54,SD value was0.18¹.29,and RSD value was 0.80⁴.91.?4?The derivative effects of BSTFA/TMCS,PFPA and HFBA were compared.BSTFA/TMCS and HFBA were determined as the best derivative reagents,followed by PFPA,according to the size of the matrix peak in the chromatogram and the abundance of the typical ion mass to charge ratio?m/z?in the mass spectrometry.?5?Compare CNWBOND NH₂ pillars,Generik MAX pillars and UCT Clean-Screen ETG pillars of blood,urine and hair three kinds of biological samples by solid phase extraction,comparing three kinds of extraction column extraction recovery of the target for blood samples,Generik MAX pillars and UCT Clean-Screen ETG pillars extraction recovery rate is higher than CNWBOND NH₂ pillars extraction recovery rate,recovery rate difference was statistically significant?P<0.05?;There was no significant difference in EtG recovery rate between,Generik MAX column,UCT Clean-Screen ETG column and CNWBOND NH2 column in urine samples?P>0.05?.For hair samples,the recovery rate of pillars Generik MAX was higher than that of UCT clean-screen ETG pillars and CNWBOND NH₂ pillars,and the difference in recovery rate was statistically significant?P<0.05?.?6?After biological samples were treated with two pretreatment methods,solid phase extraction?SPE?and liquid-liquid extraction,the recovery rate of the target substance was compared,and it was found that the recovery rate of solid phase extraction was significantly higher than liquid-liquid extraction,and the difference in recovery rate was statistically significant?P<0.05?.Conclusion:The method established in this experiment has good sensitivity,accuracy and linear relationship for the detection of ethyl glucuronide in blood,urine and hair of drinkers or alcohol abusers,and the recovery rate is high.It is of good application value to provide reliable reference data for the judicial identification of drinking or drunk driving and the diagnosis and treatment of alcohol abstinence in a short period of time.