CN103645148A - Method and reagent for detecting nutrition ingredient in food - Google Patents
Method and reagent for detecting nutrition ingredient in food Download PDFInfo
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- CN103645148A CN103645148A CN201310616197.7A CN201310616197A CN103645148A CN 103645148 A CN103645148 A CN 103645148A CN 201310616197 A CN201310616197 A CN 201310616197A CN 103645148 A CN103645148 A CN 103645148A
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Abstract
The invention belongs to the field of food detection, and relates to a method and a reagent for detecting a nutrition ingredient in food. The reagent for detecting the nutrition ingredient in the food is prepared from the following components: 100mmol/L of a solvent, 300-50000U/L of endopeptidase, 2000-80000U/L of exopeptidase, 200-1000U/L of subtilisin from bacillus subtilis, 4000-30000 U/L of amino acid dehydrogenase, 0.2-0.5mol/L of ammonium sulfate and 0.2-0.5mol/L of glycerin. The reagent for detecting the nutrition ingredient in the food, provided by the invention, is long in storage time and convenient to use, is capable of effectively detecting the content of true protein, and prevents a nonprotein nitrogen-containing compound from interfering the measurement of the content of the protein, thereby avoiding the result deviation caused in detection of the content of the protein by a Kjeldahl method, and being more accurate in testing.
Description
Technical field
The invention belongs to food inspection field, relate to a kind of detection method of food nutrition composition and detect reagent.
Background technology
Food is the elementary necessaries that people live, and the number of protein content in foodstuff is closely bound up with people's health.The content of Protein in Food not only represents the quality of the quality of food, and is being related to the health of human body, and therefore, the content of measuring Protein in Food is not only the Important Action of check food quality, is also to guarantee the healthy important means of people simultaneously.At present, national food hygienic standard also makes explicit provisions to the protein content in Partial Food especially animal breast and animal dairy products.Due to protein determination difficulty comparatively, and protein in food is unique source of nitrogen in human body, and therefore, traditional method adopts the content of the content indirect determination protein of measuring nitrogen more.At present, protein measuring adopts Kjeldahl's method more, the method Ye Shi China measures the standard method of protein content, its applicability is strong, reproducible, but the maximum defect that this method exists is exactly that the result recording has not only comprised protein, but also urea nitrogen and inorganic amine salt etc. have been comprised containing nitrogen compound, thus, caused many lawless persons or enterprise to illegitimately make exorbitant profits, as their adding in food containing nitrogen compound cheapnesss such as three chlorocyanamides and urea, to pretend to be protein, like this serious harm people's health, the infant of especially edible dairy products, therefore, provide a kind of easy to detect, result accurately protein detection method is most important.
Summary of the invention
For solving the above-mentioned technical matters existing in prior art, the object of this invention is to provide a kind of detection method of food nutrient composition and detect reagent, can detect rapidly the protein content in food, get rid of the interference of non-protein nitrogenous constituents to testing result.
Technical scheme of the present invention is as follows:
A test agent for food nutrient composition, is made by the component that comprises following content:
Solvent 100mmol/L
Endopeptidase 300 ~ 50000U/L
Exopeptidase 2000 ~ 80000U/L
Subtilopeptidase A 200 ~ 1000U/L
Amino acid dehydrogenase 4000 ~ 30000U/L
Ammonium sulfate 0.2 ~ 0.5mol/L
Glycerine 0.2 ~ 0.5mol/L.
Described solvent is selected from: one or more in 2-amino-2-methyl-I-propyl alcohol damping fluid, 2-amino-2-methyl-I-propyl alcohol damping fluid, diethanolamine buffer, glycylglycine damping fluid or boric acid-borate buffer solution.
Preferred described solvent is that 2-amino-2-methyl-I-propyl alcohol damping fluid, 2-amino-2-methyl-I-propyl alcohol damping fluid, three kinds of damping fluids of diethanolamine buffer are the potpourri of 1:1:2 in molar ratio.
Described endopeptidase is N-endopeptidase, and described exopeptidase is C-exopeptidase; The ratio of endopeptidase and exopeptidase is 1U/L:(1.2 ~ 3) U/L.
The method of utilizing above-mentioned food nutrient composition test agent instrument nutritional labeling, comprises the following steps:
(1) test agent is divided equally for three equal parts, in first part, add testing sample, be mixed with the testing sample solution that concentration is 30 ~ 50g/L, in second part, add with the first duplicate samples in the pure protein of equal in quality be mixed with the first comparative solution with first part of same concentrations, in the 3rd part of test agent, add the test agent of equal in quality in the first duplicate samples as the second comparative solution;
(2) adopt same condition to carry out while stirring microwave dispersion the testing sample solution making in step (1), the first comparative solution and the second comparative solution;
(3) adopt scattered three kinds of solution absorbance under 540nm in absorbance tester testing procedure (2) over time, to record respectively δ testing sample solution, δ first comparative solution δ the second comparative solution;
(4) protein content in testing sample is (δ testing sample solution-δ the second comparative solution)/(δ the first comparative solution-δ the second comparative solution).
Preferably, in described step (2), in three kinds of solution, add respectively the NaOH of same amount, then stir the NaOH that the addition of NaOH is 0.3 ~ 0.5g/L.
Stirring condition in described step (2) is specially, and first solution is stirred to 1 ~ 3min, then carries out while stirring microwave dispersion, continues to stir 2 ~ 5min.
Probe temperature in described step (3) is 25 ~ 30 ℃.
Absorbance tester described in step (3) is UV, visible light spectrophotometer or automatic clinical chemistry analyzer.
Compared to the prior art, beneficial effect of the present invention is:
1. detection reagent of the present invention is can storage time long, easy to use, can effectively detect the content of true protein, avoided the interference of non-protein nitrogenous constituents to protein content determination, thereby the result error having caused while having avoided triumphant formula nitriding to detect protein content, it is more accurate to test.
2. in the mensuration process of content in food, the compositional selecting of test agent and concrete content are extremely important, the present invention, by selecting suitable solvent and the enzyme matching with it, rationally regulates the content proportioning between each component, makes the test agent in the present invention have higher activity.Wherein endopeptidase can be by protein molecule inner cut-out, forms the peptide of molecular weight, exopeptidase from the free amine group of protein molecule or the end of carboxyl one by one by peptide bond hydrolysis, and the amino acid that dissociates; Each component interaction makes to test sample can meet the needs that absorbance tester is tested.
3. test solution guarantees that with comparative solution the content of each composition is identical, makes test more accurate, adds NaOH to make the environment of solution be alkalescence simultaneously in test solution and comparative solution, is conducive to improve the efficiency of various enzymes in test solution.
Embodiment:
Below in conjunction with embodiment, the present invention will be further described:
Embodiment 1
A test agent for food nutrient composition, is made by the component that comprises following content:
Solvent 100mmol/L
Endopeptidase 4000U/L
Exopeptidase 6000U/L
Subtilopeptidase A 800U/L
Amino acid dehydrogenase 5000U/L
Ammonium sulfate 0.3mol/L
Glycerine 0.5mol/L
Described solvent is glycylglycine damping fluid or boric acid-borate buffer solution;
Described endopeptidase is N-endopeptidase, and described exopeptidase is C-exopeptidase; The ratio of endopeptidase and exopeptidase is 1U/L:3U/L;
The method of utilizing above-mentioned food nutrient composition test agent test food nutrient composition, comprises the following steps:
(1) test agent is divided equally for three equal parts, in first part, add testing sample (soymilk), be mixed with the testing sample solution that concentration is 50g/L, in second part, add with the first duplicate samples in the pure protein of equal in quality be mixed with the first comparative solution with first part of same concentrations, in the 3rd part of test agent, add the test agent of equal in quality in the first duplicate samples as the second comparative solution;
(2) will in the testing sample solution making in step (1), the first comparative solution and the second comparative solution, add respectively the NaOH of 0.5g/L, then adopt same condition to carry out while stirring microwave dispersion; In whipping process, first solution is stirred to 3min, then carry out while stirring microwave dispersion, continue to stir 5min;
(3) adopt scattered three kinds of solution absorbance under 540nm in automatic clinical chemistry analyzer testing procedure (2) over time, to record respectively δ testing sample solution, δ first comparative solution δ the second comparative solution;
(4) protein content in testing sample is (δ testing sample solution-δ the second comparative solution)/(δ the first comparative solution-δ the second comparative solution);
Protein content by test testing sample milk powder is 16.5%.
Embodiment 2
A test agent for food nutrient composition, is made by the component that comprises following content:
Solvent 100mmol/L
Endopeptidase 300U/L
Exopeptidase 2000U/L
Subtilopeptidase A 200U/L
Amino acid dehydrogenase 4000U/L
Ammonium sulfate 0.2mol/L
Glycerine 0.2mol/L
Described solvent is that 2-amino-2-methyl-I-propyl alcohol damping fluid, 2-amino-2-methyl-I-propyl alcohol damping fluid, three kinds of damping fluids of diethanolamine buffer are the potpourri of 1:1:2 in molar ratio;
Described endopeptidase is N-endopeptidase, and described exopeptidase is C-exopeptidase; The ratio of endopeptidase and exopeptidase is 1U/L:1.2U/L;
The method of utilizing above-mentioned food nutrient composition test agent test food nutrient composition, comprises the following steps:
(1) test agent is divided equally for three equal parts, in first part, add testing sample (baby milk), be mixed with the testing sample solution that concentration is 30g/L, in second part, add with the first duplicate samples in the pure protein of equal in quality be mixed with the first comparative solution with first part of same concentrations, in the 3rd part of test agent, add the test agent of equal in quality in the first duplicate samples as the second comparative solution;
(2) will in the testing sample solution making in step (1), the first comparative solution and the second comparative solution, add respectively the NaOH of 0.3g/L, then adopt same condition to carry out while stirring microwave dispersion; In whipping process, first solution is stirred to 1min, then carry out while stirring microwave dispersion, continue to stir 2min;
(3) adopt scattered three kinds of solution absorbance under 540nm in UV, visible light spectrophotometer testing procedure (2) over time, to record respectively δ testing sample solution, δ first comparative solution δ the second comparative solution;
(4) protein content in testing sample is (δ testing sample solution-δ the second comparative solution)/(δ the first comparative solution-δ the second comparative solution);
Protein content by test testing sample milk powder is 18.2%.
Embodiment 3
A test agent for food nutrient composition, is made by the component that comprises following content:
Solvent 100mmol/L
Endopeptidase 50000U/L
Exopeptidase 80000U/L
Subtilopeptidase A 1000U/L
Amino acid dehydrogenase 30000U/L
Ammonium sulfate 0.5mol/L
Glycerine 0.5mol/L
Described solvent is 2-amino-2-methyl-I-propyl alcohol damping fluid;
Described endopeptidase is N-endopeptidase, and described exopeptidase is C-exopeptidase; The ratio of endopeptidase and exopeptidase is 1U/L:3U/L;
The method of utilizing above-mentioned food nutrient composition test agent test food nutrient composition, comprises the following steps:
(1) test agent is divided equally for three equal parts, in first part, add testing sample (soymilk), be mixed with the testing sample solution that concentration is 50g/L, in second part, add with the first duplicate samples in the pure protein of equal in quality be mixed with the first comparative solution with first part of same concentrations, in the 3rd part of test agent, add the test agent of equal in quality in the first duplicate samples as the second comparative solution;
(2) will in the testing sample solution making in step (1), the first comparative solution and the second comparative solution, add respectively the NaOH of 0.5g/L, then adopt same condition to carry out while stirring microwave dispersion; In whipping process, first solution is stirred to 3min, then carry out while stirring microwave dispersion, continue to stir 5min;
(3) adopt scattered three kinds of solution absorbance under 540nm in automatic clinical chemistry analyzer testing procedure (2) over time, to record respectively δ testing sample solution, δ first comparative solution δ the second comparative solution;
(4) protein content in testing sample is (δ testing sample solution-δ the second comparative solution)/(δ the first comparative solution-δ the second comparative solution);
Protein content by test testing sample milk powder is 16.5%.
Above-described embodiment is only the preferred embodiment of the present invention, and should not be construed as limitation of the invention, and those skilled in the art are according to announcement of the present invention, and not departing from the improvement that category of the present invention makes and revise all should be within protection scope of the present invention.
Claims (10)
1. a test agent for food nutrient composition, is characterized in that, by the component that comprises following content, is made:
Solvent 100mmol/L
Endopeptidase 300 ~ 50000U/L
Exopeptidase 2000 ~ 80000U/L
Subtilopeptidase A 200 ~ 1000U/L
Amino acid dehydrogenase 4000 ~ 30000U/L
Ammonium sulfate 0.2 ~ 0.5mol/L
Glycerine 0.2 ~ 0.5mol/L.
2. the test agent of a kind of food nutrient composition according to claim 1, it is characterized in that, described solvent is selected from one or more in 2-amino-2-methyl-I-propyl alcohol damping fluid, 2-amino-2-methyl-I-propyl alcohol damping fluid, diethanolamine buffer, glycylglycine damping fluid or boric acid-borate buffer solution.
3. the test agent of a kind of food nutrient composition according to claim 2, it is characterized in that, preferred described solvent is that 2-amino-2-methyl-I-propyl alcohol damping fluid, 2-amino-2-methyl-I-propyl alcohol damping fluid, three kinds of damping fluids of diethanolamine buffer are the potpourri of 1:1:2 in molar ratio.
4. the test agent of a kind of food nutrient composition according to claim 2, is characterized in that, described endopeptidase is N-endopeptidase, and described exopeptidase is C-exopeptidase.
5. the test agent of a kind of food nutrient composition according to claim 4, is characterized in that, described endopeptidase and the ratio of exopeptidase are 1U/L:(1.2 ~ 3) U/L.
6. adopt the method for the test agent instrument nutritional labeling of arbitrary described food nutrient composition in claim 1 ~ 5, it is characterized in that, comprise the following steps:
(1) test agent is divided equally for three equal parts, in first part, add testing sample, be mixed with the testing sample solution that concentration is 30 ~ 50g/L, in second part, add with the first duplicate samples in the pure protein of equal in quality be mixed with the first comparative solution with first part of same concentrations, in the 3rd part of test agent, add the test agent of equal in quality in the first duplicate samples as the second comparative solution;
(2) adopt same condition to carry out while stirring microwave dispersion the testing sample solution making in step (1), the first comparative solution and the second comparative solution;
(3) adopt scattered three kinds of solution absorbance under 540nm in absorbance tester testing procedure (2) over time, to record respectively δ testing sample solution, δ first comparative solution δ the second comparative solution;
(4) protein content in testing sample is (δ testing sample solution-δ the second comparative solution)/(δ the first comparative solution-δ the second comparative solution).
7. the method for a kind of instrument nutritional labeling according to claim 6, it is characterized in that, in described step (2), in three kinds of solution, add respectively the NaOH of same amount, then stir the NaOH that the addition of NaOH is 0.3 ~ 0.5g/L.
8. the method for a kind of instrument nutritional labeling according to claim 6, is characterized in that, the stirring condition in described step (2) is specially, and first solution is stirred to 1 ~ 3min, then carries out while stirring microwave dispersion, continues to stir 2 ~ 5min.
9. the method for a kind of instrument nutritional labeling according to claim 6, is characterized in that, the probe temperature in described step (3) is 25 ~ 30 ℃.
10. the method for a kind of instrument nutritional labeling according to claim 6, is characterized in that, the absorbance tester described in step (3) is UV, visible light spectrophotometer or automatic clinical chemistry analyzer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310616197.7A CN103645148A (en) | 2013-11-28 | 2013-11-28 | Method and reagent for detecting nutrition ingredient in food |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310616197.7A CN103645148A (en) | 2013-11-28 | 2013-11-28 | Method and reagent for detecting nutrition ingredient in food |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030026794A1 (en) * | 2001-07-31 | 2003-02-06 | Howard Fein | Selective enzyme treatment of skin conditions |
| US20030170867A1 (en) * | 1996-12-23 | 2003-09-11 | Young David Michael | Novel thermostable proteolytic enzymes and uses thereof in peptide and protein synthesis |
| CN101477129A (en) * | 2008-12-19 | 2009-07-08 | 苏州艾杰生物科技有限公司 | Protein measuring method and protein detection reagent kit |
| CN102298025A (en) * | 2010-06-25 | 2011-12-28 | 苏州艾杰生物科技有限公司 | Method for determining glycine and glycine determination kit |
| CN102735664A (en) * | 2012-06-18 | 2012-10-17 | 中国科学院化学研究所 | Potassium ion concentration detection method |
-
2013
- 2013-11-28 CN CN201310616197.7A patent/CN103645148A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030170867A1 (en) * | 1996-12-23 | 2003-09-11 | Young David Michael | Novel thermostable proteolytic enzymes and uses thereof in peptide and protein synthesis |
| US20030026794A1 (en) * | 2001-07-31 | 2003-02-06 | Howard Fein | Selective enzyme treatment of skin conditions |
| CN101477129A (en) * | 2008-12-19 | 2009-07-08 | 苏州艾杰生物科技有限公司 | Protein measuring method and protein detection reagent kit |
| CN102298025A (en) * | 2010-06-25 | 2011-12-28 | 苏州艾杰生物科技有限公司 | Method for determining glycine and glycine determination kit |
| CN102735664A (en) * | 2012-06-18 | 2012-10-17 | 中国科学院化学研究所 | Potassium ion concentration detection method |
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