Study On Extraction Technology And Quantitative And Qualitative Analysis Methods Of Blueberry Anthocyanins

In order to further develop and utilize the health value of blueberries,especially the utilization value of the natural water-soluble pigment anthocyanins in blueberries,this subject studied the best extraction process of anthocyanins of blueberries through orthogonal experiments and the determination method of anthocyanins content of blueberries Focused on research,optimized the measurement conditions of the p H differential method,and established a high-performance liquid chromatography method for determining the relative content of blueberry anthocyanins,and combined with high-performance liquid chromatography-mass spectrometry to further confirm and anthocyanin substances Identification,and finally on the basis of blueberry anthocyanin research,a preliminary analysis of the quality characteristics of blueberry,also provides reference for the application research and quality evaluation of blueberry breeding,processing,development and other aspects.1)Using blueberries as raw materials,the effects of material-liquid ratio,ultrasonic time,ethanol volume fraction,and ultrasonic temperature on the extraction of blueberry anthocyanins were studied through single factor experiments and orthogonal experiments.The test results show that the optimal extraction conditions of blueberry anthocyanins are:material-liquid ratio 1:20(g/m L),ultrasonic time 50 min,ethanol volume fraction 80%(0.1%hydrochloric acid),ultrasonic temperature 50 ?.Under this optimal extraction condition,the anthocyanin yield of the extract is higher.2)The p H differential method in the determination method of anthocyanin content was optimized.Through the single factor test of the dilution factor and buffer time of the blueberry and wine sample solutions,the p H differential method for determining the total anthocyanin content in the blueberry and wine was determined The best determination conditions,under this condition,the anthocyanin content of blueberry brilliant,round blue and Baldwin were determined to be 1.19 mg / g,1.72 mg / g,2.20 mg / g;blueberry wine Lan Ji,Lan Fei,The content of anthocyanins in Lanjue was 0.005 mg / g,0.013 mg / g,0.014 mg / g.3)Established a method for the determination of the relative content of anthocyanins in blueberries by high performance liquid chromatography.With the main component substance,delphinidin-3-O-glucoside as the target,the relative content of blueberry anthocyanins was determined.By optimizing the elution conditions,the anthocyanin substances in the anthocyanin extract were separated well.Through the inspection of the standard curve linearity,repeatability,precision,spike recovery,detection limit and quantitation limit test,it was found to be 1.0 ? 100?g / m L concentration range,good linearity(r> 0.9999),and good repeatability and precision,RSD <3%,spike recovery is92.61%,RSD is 0.75%,detection limit is 1.0mg / kg,The limit of quantification is 5.0mg /kg,indicating that this method is suitable for the determination of the relative content of anthocyanins in blueberries,and also provides a reference for the qualitative and quantitative determination of other single anthocyanin components and total anthocyanin content.By this method,the contents of Delphinium-3-O-glucoside in the brilliant blueberry,round blue,and Baldwin were determined to be 25 mg / kg,750 mg / kg,and40 mg / kg,respectively.4)Using high performance liquid chromatography-mass spectrometry to further qualitatively confirm the anthocyanin constituents of blueberry,in the multi-reaction detection mode(MRM),by optimizing the anthocyanin substances Feiyan-3-O-glucoside and vector The mass spectrometry conditions of chrysanthemum-3-O-glucoside are used for the qualitative confirmation of anthocyanin substances.The blueberry anthocyanin extract was preliminarily identified under the conditions of first-level mass spectrometry and second-level mass spectrometry by combining literature references and the mass-to-herb ratio(m / z)of fragment ions,and speculated that the anthocyanins in blueberry The substance may contain delphinium-3-galactoside,delphinium-3-glucoside,cornflower-3-galactoside,cornflower-3-glucoside,morning glory pigment-3-galactose Glycoside,morning glory pigment-3-glucoside,paeoniflorin pigment-3-glucoside,mallow pigment-3-glucoside,mallow pigment-3-glucoside.5)In order to study the quality characteristics of blueberries,physical indicators such as blueberry fruit shape,average single fruit quality,fruit shape index,chemical indicators such as Ca,Fe,Zn,Se,Mn,Cu in mineral nutrients and blueberry wine The alcohol level,glucose,mineral elements and other indicators were analyzed.Combined with the above analysis of blueberry anthocyanin content,the brilliant blueberry fruit is large and heavy,suitable for fresh eating,while the round blue fruit is small and single fruit is light,but the mineral elements are rich in nutritional value,and the anthocyanin content is high,suitable for making blueberry wine,etc.Blueberry processing products and development research,Baldwin blue berry fruit is large,and the content of mineral elements is also high,both fresh food and processing,Baldwin blue berry anthocyanin content is the highest,has an important role in the research of anthocyanins.The alcohol content of the three blueberry wines was not much different,ranging from 10.3 ° to 11.4 °.The blue glucose content of blueberry blueberry wine was the highest,which was significantly different from that of Lanjue and Lanfei wine.