Method for determining galactooligosaccharide content in milk powder, rice powder or milk and rice powder and application thereof

A technology of galacto-oligosaccharides and galactose, which is applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of interfering GOS quantitative detection, effective separation of difficult compounds, poor stability of amino columns, etc., and achieves low detection limit, accurate High intensity and less interference

Inactive Publication Date: 2016-07-27
INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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  • Abstract
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Problems solved by technology

However, there are many ingredients in health food, especially milk powder with high lactose content, rice flour and the enzymatic hydrolysis products of their mixtures, such as glucose and galactose, which have various optical isomers, all of which are large polar compounds. approaching, seriously interfered with the quantitative detection of GOS
Therefore, conventional separation liquid phase columns are difficult to achieve effective separation of such compounds, and the stability of the amino column generally used to separate large polar compounds is poor
[0005] In addition, for samples with high lactose and galactose content, the peak eluting time of lactose and kestose is similar, which affects the peak eluting of lactose; according to the detection principle, if the lactose content in the sample is too high, that is, the For dairy products with a content ratio of less than 1:10, such as formula milk powder, the detection error will increase, and this shortcoming cannot be compensated by the AOAC method

Method used

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  • Method for determining galactooligosaccharide content in milk powder, rice powder or milk and rice powder and application thereof
  • Method for determining galactooligosaccharide content in milk powder, rice powder or milk and rice powder and application thereof
  • Method for determining galactooligosaccharide content in milk powder, rice powder or milk and rice powder and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment one: selection and optimization of detection method

[0041] 1. Sample pretreatment method

[0042] (1) Extraction and removal of heteropolysaccharide interference: Weigh 0.5g sample into a 50mL centrifuge tube, add 10mL water, adjust the pH to 4-5 with acetic acid solution, dissolve and shake for 5min; then add 40mL ethanol, shake for 5-10min , centrifuged at 10000r / min, filtered with quantitative filter paper to obtain a clear filtrate and rotary evaporated to dryness.

[0043] (2) Removal of lactose interference: Add 50 mL of isopropanol-ethanol mixed organic solvent with a volume ratio of 4:1 to the concentrate obtained in the step (1), vortex for 5 minutes, filter, and take the filter residue. Repeat five times, and concentrate the filter residue to dryness under low pressure to obtain a concentrate.

[0044] (3) Prepare the test solution before enzymolysis: dilute the concentrate obtained in the step (2) to 5 ml with water to obtain the test sample solu...

Embodiment 2

[0069] Embodiment two: the content determination of galacto-oligosaccharide in milk powder and rice flour

[0070] 1. Standard stability research

[0071] Check the standard stock solution for three months, and compare the response values ​​of the old and new working curves. The results showed that the standard stock solution was stable for three months.

[0072] 2. Methodological investigation

[0073] (1) Linear range and linear regression equation

[0074] By adding different concentration levels of galactose standard and lactose standard to the blank sample, the standard curves of galactose and lactose were drawn respectively, and the mass concentration of galactose and lactose was linear with the peak area within the linear range of 500mg / kg-8000mg / kg The relationship is good, and its linear regression equation and correlation coefficient are shown in Table 2. Figure 13 is the standard curve for galactose, Figure 14 Standard curve for galactose.

[0075] Table 2 ...

Embodiment 3

[0083] Example three: an application of XAmide liquid phase column to measure galactooligosaccharides in milk powder and rice flour

[0084] Using the measurement method established in the above two examples, 19 kinds of milk powder and 16 kinds of rice flour samples on the market were detected, and the contents of galactose and lactose before and after enzymolysis were respectively obtained, and the formula GOS=k*(GB-GA- GL) Calculate the content of galacto-oligosaccharides in the detection sample. Among them, GB in the formula is the content of galactose after enzymatic hydrolysis; GA is the content of free galactose; GL is the content of galactose produced by enzymatic hydrolysis of lactose, which is the content of lactose divided by the conversion factor 1.9; k is the conversion of galactose into GOS Conversion coefficient, k=(180+162n) / (180*n), the coefficient is generally 1.49. Table 4 shows the test results of different brands of milk powder.

[0085] Table 5 is the t...

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Abstract

The invention discloses a method for determining the galactooligosaccharide content in milk powder, rice powder or milk and rice powder and application thereof.The method adopts an XAmide amide column and adopts ethanol and isopropyl alcohol according to the ratio of 1 to 4 in the pretreatment process of a sample to be detected to efficiently reduce lactose interference in the sample to be detected by being mixed with an organic solvent, the detection conditions are optimized, glucose, galactose and their epimers in an enzyme hydrolysis product are effectively separated, a linear regression equation is established, and the galactooligosaccharide content in the sample to be detected is accurately determined.The method is simple and easy to operate, the pretreatment time and costs are greatly reduced, a linear range is wide, and the method has the advantages of being low in interference, low in detection limit, high in accuracy and the like.

Description

technical field [0001] The invention belongs to the field of health food detection, and in particular relates to a method for measuring the content of galactooligosaccharides in milk powder, rice flour and milk rice flour by using an Xamide amide chromatographic column and an application thereof. Background technique [0002] Galactooligosaccharides (GOS) is a kind of active oligosaccharides, using lactose as a substrate, under the action of β-galactosidase with galactosyl transfer activity, it first hydrolyzes lactose into galactose and glucose , and then transfer galactose to galactose molecules to generate galactooligosaccharides. In general, the molecular structure of galactooligosaccharides is that 1 to 7 galactosyl groups are connected to galactose or glucose molecules, that is, Gal-(Gal) n - Glc / Gal (n is 0-6), molecular weight about 300-2000. Galactooligosaccharide is a functional oligosaccharide with natural properties, which can promote the proliferation of intes...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/08
CPCG01N30/02G01N30/08
Inventor 韦晓群易蓉王志元郑璇刘青杜斯航翁文川吴志航黄维龙张玉文梁瑞婷张英鹏
Owner INSPECTION & QUARANTINE TECH CENT OF GUANGDONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU
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