Study On The Chemical Composition And Quality Evaluation Of Red Millet Rice Wine

As a characteristic product of Nanyang City,Henan Province,red millet rice wine is rich in nutrients and has many functions such as heart protection,anti-oxidation,anti-aging,lowering blood pressure and cholesterol.However,there is no research on Nanyang red millet rice wine.In this thesis,three kinds of red rice wines,Xinye red millet red koji rice wine[CRW-XY（RK）],Xinye red millet wheat koji rice wine（CRW-XY）,and Neixiang red millet wheat koji rice wine（CRW-NX）are used as researches objects,comparing Dengzhou glutinous rice wheat koji rice wine（CRW-DZ）and Shaoxing glutinous rice wheat koji rice wine（CRW-SX）,studying the chemical composition,nutrient composition,quality evaluation and myocardial protection of red millet rice wine.The study consists of five parts:1.Chemical composition analysis.The methanol extract from concentration of CRW-XY（RK）was separated and purified by silica gel column chromatography,ODS,Sephadex LH-20,recrystallization and precipitation,and identified by nuclear magnetic resonance NMR.17 compounds,p-hydroxyphenethyl alcohol（1）,ferulic acid（2）,caffeic acid（3）,erythritol（4）,catechol（5）,catechin（6）,syringic acid（7）,rutin（8）,quercetin（9）,β-sitosterol（10）,proline（11）,protocatechuic acid（12）,epicatechin（13）,chlorogenic acid（14）,aspartic acid（15）,lysine（16）andα-naphthol（17）,were obtain from it,which were all isolated from red millet yellow wine for the first time.2.Nutrient composition analysis.Referring to national standards,nutrient compositions of rice wine were studied,such as the amino acids,vitamins B,minerals,proteins,polysaccharides,reducing sugars and total acids.The results showed that the contents of alanine,leucine,γ-aminobutyric acid,K,Na and Ca were higher in red millet rice wine.The content of vitamins B in red millet rice wine（111.1μg/100g）was higher than that in glutinous rice wine（36.5μg/100g）,and that in wheat koji rice wine（111.1μg/100g）was higher than that in red koji rice wine（24.5μg/100g）.The total acid content of red millet rice wine（9.95 g/L）was higher than that of glutinous rice wine（6.21 g/L）.The contents of protein,reducing sugar and total sugar had no significant correlation with the origin,raw materials and koji of rice wine.On the whole,red millet rice wine is rich in nutrients,and wheat koji rice wine is better than red koji rice wine.3.The establishment of HPLC fingerprints.To establish HPLC fingerprints of 5kinds of rice wine and their total extract and methanol parts,a Kromasil C₁₈ column（250 mm×4.6 mm,5μm）was employed during the experiment with the mobile phase of acetonitrile-0.4%phosphoric acid water in a gradient mode,the column temperature was maintained at 38°C,the detection wavelength was 280 nm,the flow rate was 1.0m L/min,and the injection volume was 10μL.Seven components in rice wine,including gallic acid,protocatechuic acid,catechin,chlorogenic acid,epicatechin,5-hydroxy furfural and resveratrol,were identified by reference substance and UPLC-MS/MS.The comparison found that different rice wines had similar components,but the content was quite different.Among them,the content of chlorogenic acid and resveratrol in red millet rice wine CRW-XY（RK）was higher.4.Content determination.The Prussian blue method and the sodium nitrite-aluminum nitrate-sodium hydroxide method were used to determine the content of total phenolic acids and total flavonoids of rice wine with gallic acid and rutin as reference substances,respectively.The results showed that the content of Xinye（1.530、11.336mg/m L）and Neixiang（1.092、6.101 mg/m L）red millet wheat koji rice wine were higher,followed by red millet red koji rice wine（0.727,4.217 mg/m L）,and Deng Zhou（0.573,2.495 mg/m L）and Shaoxing（0.662,3.320 mg/m L）glutinous rice wheat koji rice wine were lower.It showed that the content of total phenolic acid and total flavonoids in rice wine was related to its raw materials and koji.Red millet rice wine was higher than glutinous rice wine,wheat koji rice wine was higher than red koji rice wine,and the influence of raw materials on it was greater than that of koji.5.Study on myocardial protection.The protective effect of rice wine on myocardium was studied by establishing H₂O₂ oxidative stress model of H9c2cardiomyocytes.Compared with H₂O₂ group,it was found that the total extracts of five kinds of rice wine could significantly increase cell survival rate,reduce leakage rate of LDH and the content of MDA,and significantly increase the activities of SOD and GSH（P<0.05）,indicating that the five kinds of rice wine had protective effects on H₂O₂-induced H9c2 cardiomyocyte damage,of which CRW-XY had the best effect,which was equivalent to resveratrol.Western Blot was used to detect the expression of SIRT1 protein,and the results showed that CRW-XY could significantly up-regulate the level of SIRT1 protein in cardiomyocytes,suggesting that CRW-XY may play an anti-oxidative stress role by up-regulating SIRT1 protein,and thus protect cardiomyocytes.To sum up,this paper conducted a systematic study on red millet rice wine for the first time,which clarified its unique unique functional material basis,provided a scientific basis for the quality evaluation of rice wine and the establishment of quality standard system,and laid the foundation for further improving the quality of red millet rice wine and promoting the development of the rice wine industry.