Comparative Analysis Of High Performance Liquid Chromatography And Ultra Performance Liquid Chromatography Tandem Time Of Flight Mass Spectrometry Detection Methods Of Vitamin B₁₂ In 4 Kinds Of Infant Formula Milk Powder

Infant formula milk powder is a special dietary food,which plays an important role in the growth and development of infants and young children.At present,infant formula milk powder still has many food safety problems,and the types and contents of nutrients in different products are also different.Vitamin B₁₂is the only water-soluble vitamin containing metal elements.It is one of the essential vitamins for the human body.It is also an important indicator to measure the nutrition of infant formula milk powder.Therefore,it is necessary to accurately determine the content of vitamin B₁₂in infant formula milk powder.In this study,high performance liquid chromatography and ultra high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry were established to detect the content of vitamin B₁₂in infant formula milk powder.The purpose is to establish a simple,efficient and rapid detection of infant formula milk powder.The main research content and results of the analysis method of vitamin B₁₂content in the spore are as follows:1.Optimize the pretreatment process of infant formula milk powderUsing infant formula milk powder as the raw material,the sample solution is subjected to protein precipitation experiments,p H adjustment,ultrasonic extraction,membrane filtration and frozen centrifugation,etc.,and then the pretreatment process of infant formula milk powder is optimized,and finally effective extraction is achieved Vitamin B₁₂purpose in the sample solution.2.Establish a method for detecting vitamin B₁₂in infant milk powder and milk powder by high performance liquid chromatographyThe experiment established an analytical method for HPLC to detect vitamin B₁₂in infant formula milk powder.This method selects Athena C₁₈-120 A(4.6 mm×250mm,5.0?m)as the chromatographic column,the mobile phase is water(75%)+methanol(25%),the flow rate is 0.5 m L/min,and the elution method is isocratic.For elution,the injection volume was 45?L,the column temperature was 40?,and the wavelength was 361 nm.The sample is pre-treated and passed through a 0.22?m filter membrane and measured on the machine.The results show that the standard solution of vitamin B₁₂has a good linearity in the concentration range of 0.05?2.0?g/m L,the correlation coefficient R²is 0.9998,the peak time of vitamin B₁₂is 17.006min,the average recovery rate is 91.3%,and the detection limit is 0.6?g/100g.It shows that the method can be applied to detect the content of vitamin B₁₂in infant formula milk powder.3.Establish an ultra-high performance liquid chromatography quadrupole time of flight mass spectrometry method for the detection of vitamin B₁₂in infant formula milk powderThe experiment established a UPLC-Q-TOF-MS quantitative analysis method for vitamin B₁₂in infant formula milk powder.This method uses Eclise Plus C₁₈(2.1mm×50 mm,1.8?m)chromatographic column,methanol(25%)+water(75%)ratio isometric elution,injection volume 5?L,flow rate 0.3 m L/min,column temperature45?.Scanning in positive ion mode,the scanning range is 100?1600 m/z,and the reference ions are 121.0509 m/z and 922.0098 m/z.The mass-to-charge ratio of the vitamin B₁₂target ion was found to be 689.28 m/z by MS mode MS MS scanning,and the time was 0.663 min.Then,the vitamin B₁₂in the standard solution was quantitatively analyzed by Target MS/MS quantitative analysis mode.The results showed that the vitamin B₁₂standard The solution has a good linear relationship in the range of 2-50 ng/m L,the correlation coefficient R²is 0.9985,and the detection limit is 0.2?g/100g,indicating that this method can be applied to the detection of vitamin B₁₂content in infant formula milk powder.4.HPLC and UPLC-Q-TOF-MS detection of vitamin B₁₂in infant formula milk powder and comparative analysis of methodsThis study established the HPLC method and UPLC-Q-TOF-MS method to detect the content of vitamin B₁₂in infant formula milk powder,as well as the comparative analysis between the two methods and the microbiological method,sample pretreatment process,detection limit and sample solution Comparative analysis of the peak time of vitamin B₁₂shows that the two methods are compared with the microbiological method,while reducing the complicated and tedious experimental steps and improving the detection efficiency.The detection limit of UPLC-Q-TOF-MS method and the peak time of vitamin B₁₂in the sample solution.Due to the HPLC method,the UPLC-Q-TOF-MS method is suitable for the detection of infant formula milk powder with low vitamin B₁₂content,but HPLC The method has good repeatability,convenience and simplicity,and strong maneuverability.It is also suitable for the determination of vitamin B₁₂content in common infant formula milk powder.