The Identification Technology Research After Drinking Rde Wine

Objectives1. To find the characteristic substances in the red wine that have differential significance.2. To establish the method of UPLC-MSn for the characteristic substances in the red wine.3. To establish the method of extraction and purification for the characteristic substances inthe red wine, and compare the different extraction and purification methods of recovery tochoose the optimal method.4. To testing all kinds of fermented wine (including beer, wine, and liquor) and beverages,to confirm the selected substances can be used to identify the different wines and beverages, andprovide scientific basis for samples taken, detection, result analysis and forensic identification insome cases that happened after drinking.5. After detecting of the biological samples (including the saliva, urine and blood) had thecharacteristics substances or not, we can have a conclusion whether the examined people hadhave drunk the red wine.Methods1.Through literature study to know the different trace substances in fermented wine as wellas the characteristic substances in red wine ,and this paper selected quinine acid and resveratrolas the characteristic substances in red wine.2. After pretreatment of samples, using Waters Oasia@MAX 3cc ( 200mg ) solid phaseextraction column to extracted and purified the samples,then using Acquity UPLC @ BEH C181.7 m column as the liquid chromatographic column, electrospray ionization negative ion ( ESI-)on samples UPLC-MSn test.3. Selected red wine, liquor, beer and beverages as samples, detected the content ofquinine acid and resveratrol after extracted and purified the samples .4. After drinking various fermented wine, through the detection of characteristic substancesof red wine in human biological samples (blood, urine, saliva), to determine whether drinkingred wine.Results1. Take the peak area of quinine acid and resveratrol in blank blood as Y-axis separately, andthe concentration of quinine acid and resveratrol as X-axis to built the working curve. Thequinine acid typical equation of working curve was: y =100X-2.5167,γ=0.9999, n =6, the linearrange was1.25μg/ml -20.0μg/ml , the fixed lower quantity was1.01μg/ml. The resveratrol typicalequation of working curve was: y =400.19X-45.579,γ=0.9998, n =6, the linear range was1.25μg/ml -20.0μg/ml , the fixed lower quantity was0.83μg/ml.2. This method was applicable to the samples of blood, saliva and urine3. After the detection of red wine, liquor, beer and beverage, the quinine acid and resveratrolwere contained in red wine, but didn’t, contain in Red star Erguotou and Blue Ribbon beer. In thebeverages, Beauty source fruit had resveratrol but didn’t have quinine acid. Unified grapes didn’thave quinine acid or resveratrol.4. After drinking red wine, the samples of blood, saliva, urine didn’t have quinine acid orresveratrol.Conclusions1. Using an Acquity UPLC @ BEH C18 1.7μm chromatographic column ,the shape of peakof quinine acid and resveratrol had a good symmetry that was better than using the HILICchromatographic column.so this paper’s method was using anAcquity UPLC BEH C18 1.7μmcolumn. The established method after UPLC-MSn optimization, the test result was proved to beaccurate, sensitive, good reproducibility and to achieve qualitative and quantitative analysisrequirements.2. In this study, MAX column and C18 column were used as solid phase extraction column,parallel operation, found that the MAX column C18 column with high recovery and goodreproducibility. This paper uses the MAX column for solid phase extraction column to extractand purify samples . Using solid phase extraction to extract and purify the sample does notrequire derivatization processing ,so this method can be used for rapid detection of biologicalsamples.3. In this study, samples for extraction and purification by solid phase extraction technology,the quinine acid and resveratrol had a high recovery, especially suitable for UPLC-MSn methodwhich needs samples clean, less impurity. Combined with the high sensitivity of the LTQ XLlinear ion trap mass spectrometry, can detect the trace characteristics of substances in thesamples.4.Different types of red wines containing different amounts of quinine acid andresveratrol,,but the Red Star Erguotou and the Blue Ribbon beer have not been detected both,indicating that the method established in this paper can be applied to distinguish the red wine,liquor, and beer in the fermented wine5. The content of quinine acid and resveratrol are different in the red wine , cause the grapeshave different regions and different varieties.6. Unified grapes has grape juice, but cannot detected have quinine acid and resveratrol,may be the form of quinine acid and resveratrol were affected by the additives in the beverage. 7. In this study, after the volunteers drinking red wine, samples of blood, urine, saliva werenot detected have quinine acid and resveratrol. The reason may be the red wine had less quinicacid and resveratrol, and the volunteers drunk less, so the content of characteristic substances inthe human body was less than the instrument can detect the smallest concentration; or may bedue to when the quinine acid and the resveratrol came into the human body rapidly dissolve inthe blood and into metabolites leaded to the instrument could not detect them.