CN111505197A - Method for detecting protein content in food - Google Patents
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- CN111505197A CN111505197A CN202010467434.8A CN202010467434A CN111505197A CN 111505197 A CN111505197 A CN 111505197A CN 202010467434 A CN202010467434 A CN 202010467434A CN 111505197 A CN111505197 A CN 111505197A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- G01N31/002—Determining nitrogen by transformation into ammonia, e.g. KJELDAHL method
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N31/00—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
- G01N31/16—Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods using titration
Abstract
The invention discloses a method for detecting protein content in food, which belongs to the technical field of food detection and comprises the following steps of S1, mixing a sample with a catalyst and digestion liquid, heating, boiling, placing in an ultrasonic oscillator, ultrasonically oscillating, S2, distilling the sample solution obtained in the step S1 by a Kjeldahl method, and S3, titrating the distillate by 0.1 mol/L of hydrochloric acid solution, and recording the dosage of the hydrochloric acid solution when the color changes.
Description
Technical Field
The invention relates to the technical field of food detection, in particular to a method for detecting protein in food.
Background
With the continuous improvement of the life quality of people and the increasing emphasis on body health, the requirements of consumers on the quality aspects of food such as palatability, nutrition and the like are also improved. Proteins are the material basis of life, are important components constituting cell tissues of living bodies, and are also important nutritional indexes in food. The method for determining the content of the protein in the food has great significance for evaluating the nutritional value of the food, reasonably developing and utilizing food resources and the like, can provide data for reasonable diet, master the variation of the nutritional value and the quality of the food, and ensure the requirements of different people on the protein. The method for measuring protein in food comprises the following steps: kjeldahl method, salicylic acid colorimetric method, biuret colorimetric method, Folin-phenol reagent method, ultraviolet absorption method, etc. The most common method at present is the Kjeldahl method, which comprises the steps of digestion, distillation and titration, has the advantages of wide application range, higher sensitivity, better recovery rate, no need of expensive instruments and the like, but is time-consuming to operate, for example, the digestion time of a high-fat-assistant high-protein sample needs more than 5 hours. Therefore, in terms of detecting aging, the kjeldahl method is increasingly difficult to meet the market demand.
Patent document No. CN108205045A discloses a method for detecting the protein content in regenerated protein fibers. Weighing a certain amount of regenerated protein fibers which are dried to constant weight, putting the regenerated protein fibers into a digestion tube, adding a certain amount of copper sulfate, potassium sulfate and concentrated sulfuric acid, digesting for a period of time in a digestion furnace with constant temperature, cooling, testing on an automatic Kjeldahl azotometer, and converting by a set protein conversion coefficient to obtain the content of the protein.
The patent document with publication number CN104122367A discloses a rapid detection technology for protein content in liquid starch, which comprises (1) digesting by sucking 10 ml of liquid corn starch uniformly stirred by a pipette, transferring into a 500 ml Kjeldahl flask or a 300 ml nitrogen-fixing test tube, adding 10 ml of copper sulfate, potassium sulfate mixed catalyst and 25 ml of concentrated sulfuric acid and 3 glass beads in turn, (2) distilling, after cooling the decomposed liquid, washing a glass funnel and a bottle neck with distilled water, diluting to 200 ml, transferring the Kjeldahl flask onto a distilling rack, connecting a 500 ml conical flask as a receiver at the lower end of a condensing tube, (3) titrating the liquid in the conical flask with 0.1 mol/L of sulfuric acid standard solution, so that the solution is changed from blue-green to grey-purple, namely, to obtain an end point, (4) simultaneously performing a blank test, and (5) calculating that X = (V1-V0) × C × 0.014.014 0.014 × 6.25.25/[ × (1-X1) ] × 100.
Although the two methods improve the defects of complex operation, long test period and the like of the conventional Kjeldahl method to a certain extent, the digestion heating time is still long, and needs to be further improved.
Disclosure of Invention
In view of this, the present invention provides a method for detecting protein content in food.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a method for detecting the protein content in food comprises the following steps:
step S1: mixing the sample with a catalyst and a digestion solution, heating to boil, and placing in an ultrasonic oscillator for ultrasonic oscillation;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method;
and step S3, titrating the distillate with 0.1 mol/L hydrochloric acid solution, and recording the dosage of the hydrochloric acid solution when the color changes.
Further, in step S1, the catalyst is a mixture of copper sulfate, potassium sulfate, a 98% sulfuric acid solution, and mercury oxide, where the ratio of copper sulfate: potassium sulfate: sulfuric acid solution with a concentration of 98%: the mercury oxide is prepared from the following components in parts by weight: 7.5-8:35-40:0.07-0.1.
Further, in step S1, the digestion solution is a hydrogen peroxide solution with a mass fraction of 35%.
Further, in step S1, the sample: catalyst: the weight portion ratio of the digestion solution is 1: 12-15:20-25.
Further, in the step S1, the time of the ultrasonic oscillation is 15-30 min.
Further, in the step S1, the boiling time is 8-10 min.
The protein is used as an important physicochemical index in food, the content of the protein in the food is accurately measured, the understanding of the quality of the food has very important significance, data can be provided for reasonable diet, the change of the nutritional value and the quality of the food can be mastered, and the requirements of different people on the protein can be ensured. The protein determination method is a plurality of methods, and the protein detection method adopted by the inventor is a classical constant Kjeldahl method which is one of the most accurate methods for determining total organic nitrogen and simpler and more convenient to operate. The method has the advantages of wide application range, high sensitivity, high recovery rate, no need of expensive instruments and the like, but the operation is time-consuming, and if high lipid is helpful and the digestion time of a high protein sample is more than 5 hours. To speed up the detection efficiency, the digestion process is modified by those skilled in the art. For example, patent publication No. CN107356702A discloses a method for detecting crude protein in castor seeds or castor cake, which comprises adding copper sulfate, potassium sulfate, and concentrated sulfuric acid into a sample to be detected, and mixing them uniformly to make the sample completely soaked by the concentrated sulfuric acid; and adding hydrogen peroxide, heating, fully digesting, and detecting the content of crude protein in the obtained digestion solution. Zhanghua, rapid protein detection and method improvement discussion [ J ], Chinese science and technology journal database science, 2018,1:129, aiming at long digestion time of the Kjeldahl method, a protein mixed digestion solution and a mixed catalyst are designed to improve the digestion speed. The sample is heated and digested together with concentrated sulfuric acid and a catalyst, so that protein is decomposed, carbon and hydrogen are oxidized into carbon dioxide and water to escape, and organic nitrogen in the sample is converted into ammonia and sulfuric acid to be combined into ammonium sulfate. Then, alkali is added for distillation, ammonia is distilled out, and after absorption with boric acid, titration is carried out with a standard hydrochloric acid or sulfuric acid solution. The protein content can be calculated according to the standard acid consumption. However, these studies are improved, complicated in operation and long in digestion time by heating. To overcome the above disadvantages, there are many methods for rapidly determining proteins, such as: biuret method, ultraviolet spectrophotometry, dye binding method, salicylic acid colorimetry, refraction method, optical rotation method, near infrared spectroscopy and the like. However, these methods have different limitations in terms of flexibility, application range, etc., and are susceptible to interference from environmental factors and their own factors. Therefore, a new digestion method is needed to be researched, so that the accuracy of the kjeldahl method detection is ensured, the digestion time is shortened, and the detection efficiency is improved.
The invention has the beneficial effects that: the sample, the digestion solution and the catalyst are heated and digested together to decompose organic matters in the food, wherein carbon and hydrogen are oxidized into carbon dioxide and water to escape, nitrogen in the protein is converted into ammonia and combined with sulfuric acid to generate ammonium sulfate to be left in the sulfuric acid, then alkali is added according to a Kjeldahl method for distillation to evaporate the ammonia, the ammonia is absorbed by boric acid and titrated by sulfuric acid or hydrochloric acid standard titration solution, and the consumption of the standard titration solution is multiplied by a coefficient, namely the protein content.
In the digestion process, the sample, the catalyst and the digestion solution are heated and boiled to decompose protein, and carbon and hydrogen are oxidized into carbon dioxide and water to escape. Then the conversion of organic nitrogen into ammonia and the combination of sulfuric acid into ammonium sulfate is accelerated by ultrasonic oscillation. The digestion method provided by the invention reduces the heating time in the digestion process in the Kjeldahl method, reduces the energy consumption and improves the digestion efficiency. The ultrasonic wave has strong capability of digesting the sample, utilizes ultrasonic energy to improve the reaction rate, and the frequency is 15-20 KHz.
The hydrogen peroxide solution is decomposed to accelerate the oxidation of organic matters. The catalyst is formed by mixing copper sulfate, potassium sulfate, 98% sulfuric acid solution and mercury oxide, wherein the copper sulfate and the potassium sulfate increase the digestion temperature and accelerate the decomposition of organic matters, and the mercury oxide and hydrogen peroxide have synergistic effect to accelerate the oxidation of the organic matters and shorten the digestion time.
The method disclosed by the invention combines ultrasonic waves and a Kjeldahl method, can quickly and accurately measure the content of protein in food, is almost the same as the result measured by the Kjeldahl method, is high in accuracy and speed, and has positive popularization significance.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention are clearly and completely described below. It is to be understood that the embodiments described are only a few embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the described embodiments of the invention, are within the scope of the invention.
Example 1
The embodiment provides a method for detecting the content of protein in eggs, which comprises the following steps:
step S1: boiling ovum gallus Domesticus, removing shell, mashing with tissue mashing machine, mixing the sample with catalyst and digestion solution, heating to boil, and placing in ultrasonic oscillator for ultrasonic oscillation at frequency of 15-20 KHz; the catalyst is formed by mixing copper sulfate, potassium sulfate, sulfuric acid solution with the concentration of 98 percent and mercury oxide, wherein the copper sulfate: potassium sulfate: sulfuric acid solution with a concentration of 98%: the mercury oxide is prepared from the following components in parts by weight: 7.5: 38: 0.07;
the digestion solution is a hydrogen peroxide solution with the mass fraction of 35 percent;
sample preparation: catalyst: the weight portion ratio of the digestion solution is 1: 12: 20;
the time of ultrasonic oscillation is 15 min; boiling for 9 min;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method; connecting a Kjeldahl flask with a distillation device, plugging a bottle opening tightly, and inserting the lower end of a condensation pipe below the liquid level of an absorption bottle (50 parts by weight of 2% boric acid solution and 5 drops of mixed indicator are pre-filled in the absorption bottle); loosening the clamp, adding 40% sodium hydroxide solution through a funnel until the solution in the bottle becomes dark blue or generates black precipitate, clamping the clamp, heating and distilling until ammonia is completely evaporated, lifting the lower end of the condensing tube away from the liquid level, washing the tube opening with distilled water, and continuously distilling for 1 minute to stop heating; wherein, the mixed indicator is a methyl red bromocresol green mixed indicator: mixing 1 part of 0.1% methyl red ethanol solution and 5 parts of 0.1% bromocresol green ethanol solution at the moment of use;
and step S3, directly titrating the distillate with 0.1 mol/L hydrochloric acid standard solution until the color of the distillate is changed from blue to reddish, namely, the end point, recording the consumption of the hydrochloric acid solution, simultaneously making a reagent blank (except that no sample is added, the operation is completely the same from digestion to digestion), and recording the volume of the hydrochloric acid standard solution consumed in the blank experiment.
Protein (%) = C × (V1-V2) × m in sampleNitrogen is present in×100/{1000×F×m×(1-mWater (W))}
In the formula: c-concentration of hydrochloric acid Standard solution, N
V1Titration sampleThe volume of the hydrochloric acid standard solution is consumed when the product absorbs the solution, m L;
V2consumption of hydrochloric acid standard solution volume upon titration of the blank absorption solution, m L;
m-sample mass, g;
mnitrogen is present in-molar mass of nitrogen, 14.01 g/mol;
mwater (W)-sample moisture content,%;
f-nitrogen is converted to a coefficient of protein.
Example 2
This example provides a method for detecting protein content in oat bran, comprising the following steps:
step S1: mixing oat bran with a catalyst and a digestion solution, heating to boil, and placing in an ultrasonic oscillator for ultrasonic oscillation at the frequency of 15-20 KHz; the catalyst is formed by mixing copper sulfate, potassium sulfate, sulfuric acid solution with the concentration of 98 percent and mercury oxide, wherein the copper sulfate: potassium sulfate: sulfuric acid solution with a concentration of 98%: the mercury oxide is prepared from the following components in parts by weight: 8: 39: 0.1;
the digestion solution is a hydrogen peroxide solution with the mass fraction of 35 percent;
sample preparation: catalyst: the weight portion ratio of the digestion solution is 1: 15: 25;
the time of ultrasonic oscillation is 15 min; boiling for 9 min;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method; connecting a Kjeldahl flask with a distillation device, plugging a bottle opening tightly, and inserting the lower end of a condensation pipe below the liquid level of an absorption bottle (50 parts by weight of 2% boric acid solution and 5 drops of mixed indicator are pre-filled in the absorption bottle); loosening the clamp, adding 40% sodium hydroxide solution through a funnel until the solution in the bottle becomes dark blue or generates black precipitate, clamping the clamp, heating and distilling until ammonia is completely evaporated, lifting the lower end of the condensing tube away from the liquid level, washing the tube opening with distilled water, and continuously distilling for 1 minute to stop heating; wherein, the mixed indicator is a methyl red bromocresol green mixed indicator: mixing 1 part of 0.1% methyl red ethanol solution and 5 parts of 0.1% bromocresol green ethanol solution at the moment of use;
and step S3, directly titrating the distillate with 0.1 mol/L hydrochloric acid standard solution until the color of the distillate is changed from blue to reddish, namely, the end point, recording the consumption of the hydrochloric acid solution, simultaneously making a reagent blank (except that no sample is added, the operation is completely the same from digestion to digestion), and recording the volume of the hydrochloric acid standard solution consumed in the blank experiment.
Protein (%) = C × (V1-V2) × m in sampleNitrogen is present in×100/{1000×F×m×(1-mWater (W))}
In the formula: c-concentration of hydrochloric acid Standard solution, N
V1Consumption of hydrochloric acid standard solution volume upon titration of sample absorption solution, m L;
V2consumption of hydrochloric acid standard solution volume upon titration of the blank absorption solution, m L;
m-sample mass, g;
mnitrogen is present in-molar mass of nitrogen, 14.01 g/mol;
mwater (W)-sample moisture content,%;
f-nitrogen is converted to a coefficient of protein.
Example 3
The embodiment provides a method for detecting the content of protein in milk, which comprises the following steps:
step S1: mixing and heating the sample milk, a catalyst and a digestion solution to boil, and placing the mixture in an ultrasonic oscillator for ultrasonic oscillation, wherein the frequency is 15-20 KHz; the catalyst is formed by mixing copper sulfate, potassium sulfate, sulfuric acid solution with the concentration of 98 percent and mercury oxide, wherein the copper sulfate: potassium sulfate: 30% strength sulfuric acid solution: the mercury oxide is prepared from the following components in parts by weight: 7.6: 35: 0.07;
the digestion solution is a hydrogen peroxide solution with the mass fraction of 35 percent;
sample preparation: catalyst: the weight portion ratio of the digestion solution is 1: 12: 20;
the time of ultrasonic oscillation is 17 min; boiling for 8 min;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method; connecting a Kjeldahl flask with a distillation device, plugging a bottle opening tightly, and inserting the lower end of a condensation pipe below the liquid level of an absorption bottle (50 parts by weight of 2% boric acid solution and 5 drops of mixed indicator are pre-filled in the absorption bottle); loosening the clamp, adding 40% sodium hydroxide solution through a funnel until the solution in the bottle becomes dark blue or generates black precipitate, clamping the clamp, heating and distilling until ammonia is completely evaporated, lifting the lower end of the condensing tube away from the liquid level, washing the tube opening with distilled water, and continuously distilling for 1 minute to stop heating; wherein, the mixed indicator is a methyl red bromocresol green mixed indicator: mixing 1 part of 0.1% methyl red ethanol solution and 5 parts of 0.1% bromocresol green ethanol solution at the moment of use;
and step S3, directly titrating the distillate with 0.1 mol/L hydrochloric acid standard solution until the color of the distillate is changed from blue to reddish, namely, the end point, recording the consumption of the hydrochloric acid solution, simultaneously making a reagent blank (except that no sample is added, the operation is completely the same from digestion to digestion), and recording the volume of the hydrochloric acid standard solution consumed in the blank experiment.
Protein (%) = C × (V1-V2) × m in sampleNitrogen is present in×100/{1000×F×m×(1-mWater (W))}
In the formula: c-concentration of hydrochloric acid Standard solution, N
V1Consumption of hydrochloric acid standard solution volume upon titration of sample absorption solution, m L;
V2consumption of hydrochloric acid standard solution volume upon titration of the blank absorption solution, m L;
m-sample mass, g;
mnitrogen is present in-molar mass of nitrogen, 14.01 g/mol;
mwater (W)-sample moisture content,%;
f-nitrogen is converted to a coefficient of protein.
Example 4
The embodiment provides a method for detecting the content of protein in peanut kernels, which comprises the following steps:
step S1: mixing and heating a sample peanut kernel, a catalyst and a digestion solution to boil, and placing the mixture in an ultrasonic oscillator for ultrasonic oscillation, wherein the frequency is 15-20 KHz; the catalyst is formed by mixing copper sulfate, potassium sulfate, sulfuric acid solution with the concentration of 98 percent and mercury oxide, wherein the copper sulfate: potassium sulfate: 30% strength sulfuric acid solution: the mercury oxide is prepared from the following components in parts by weight: 7.8: 40: 0.08;
the digestion solution is a hydrogen peroxide solution with the mass fraction of 35 percent;
sample preparation: catalyst: the weight portion ratio of the digestion solution is 1: 13: 22;
the time of ultrasonic oscillation is 30 min; boiling for 10 min;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method; connecting a Kjeldahl flask with a distillation device, plugging a bottle opening tightly, and inserting the lower end of a condensation pipe below the liquid level of an absorption bottle (50 parts by weight of 2% boric acid solution and 5 drops of mixed indicator are pre-filled in the absorption bottle); loosening the clamp, adding 40% sodium hydroxide solution through a funnel until the solution in the bottle becomes dark blue or generates black precipitate, clamping the clamp, heating and distilling until ammonia is completely evaporated, lifting the lower end of the condensing tube away from the liquid level, washing the tube opening with distilled water, and continuously distilling for 1 minute to stop heating; wherein, the mixed indicator is a methyl red bromocresol green mixed indicator: mixing 1 part of 0.1% methyl red ethanol solution and 5 parts of 0.1% bromocresol green ethanol solution at the moment of use;
and step S3, directly titrating the distillate with 0.1 mol/L hydrochloric acid standard solution until the color of the distillate is changed from blue to reddish, namely, the end point, recording the consumption of the hydrochloric acid solution, simultaneously making a reagent blank (except that no sample is added, the operation is completely the same from digestion to digestion), and recording the volume of the hydrochloric acid standard solution consumed in the blank experiment.
Protein (%) = C × (V1-V2) × m in sampleNitrogen is present in×100/{1000×F×m×(1-mWater (W))}
In the formula: c-concentration of hydrochloric acid standard solution, N;
V1consumption of hydrochloric acid standard solution volume upon titration of sample absorption solution, m L;
V2consumption of hydrochloric acid standard solution volume upon titration of the blank absorption solution, m L;
m-sample mass, g;
mnitrogen is present in-molar mass of nitrogen, 14.01 g/mol;
mwater (W)-sample moisture content,%;
f-nitrogen is converted to a coefficient of protein.
The content of protein in eggs was determined in control group 1 according to GB 5009.5-2016.
Control group 2 was tested for protein content in oat bran according to GB 5009.5-2016.
Control group 3 the protein content in milk was determined according to GB 5009.5-2016.
The content of protein in peanut is measured according to GB 5009.5-2016 in the control group 4.
Comparative example 1
This comparative example provides a method for measuring the protein content in eggs, similar to example 1, but different from example 1, in this comparative example, the catalyst lacks mercuric oxide.
Comparative example 2
This comparative example provides a method for measuring the protein content in oat bran, which is the same as example 2, but is different from example 2 in that step S1 is not subjected to ultrasonic oscillation and the boiling time is 24 min.
Comparative example 3
This comparative example provides a method for measuring the protein content in eggs, similar to example 3, but different from example 3, in this comparative example, digestion solution was absent.
Comparative example 4
This comparative example provides a method for measuring the protein content in eggs, similar to example 4, but different from example 4, in this comparative example, the sample: catalyst: the weight portion ratio of the digestion solution is 1: 10: 28.
TABLE 1 test results of protein content in examples and comparative examples
As can be seen from Table 1, the protein content of the invention is basically consistent with that of the national standard GB 5009.5-2016, and the invention has the advantages of high food detection accuracy, digestion process only requiring about half an hour, high speed and high efficiency. The mercury oxide is omitted in the comparative example 1, the ultrasonic oscillation is not carried out in the comparative example 2, the digestion solution is omitted in the comparative example 3, the proportion of the sample, the digestion solution and the catalyst is adjusted in the comparative example 4, the measured protein content is low, the adjustment of the components, the content and the method of the invention in the same time is illustrated, the digestion is not completed, and the process and the formula of the invention are good in adaptability.
Finally, the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting, and other modifications or equivalent substitutions made by the technical solutions of the present invention by those of ordinary skill in the art should be covered within the scope of the claims of the present invention as long as they do not depart from the spirit and scope of the technical solutions of the present invention.
Claims (6)
1. A method for detecting the protein content in food is characterized in that: comprises the following steps:
step S1: mixing the sample with a catalyst and a digestion solution, heating to boil, and placing in an ultrasonic oscillator for ultrasonic oscillation;
step S2: distilling the sample solution obtained in the step S1 by a Kjeldahl method;
and step S3, titrating the distillate with 0.1 mol/L hydrochloric acid solution, and recording the dosage of the hydrochloric acid solution when the color changes.
2. The method for detecting the protein content in food according to claim 1, wherein the method comprises the following steps: in the step S1, the catalyst is formed by mixing copper sulfate, potassium sulfate, 98% sulfuric acid solution and mercury oxide, wherein the copper sulfate: potassium sulfate: sulfuric acid solution with a concentration of 98%: the mercury oxide is prepared from the following components in parts by weight: 7.5-8:35-40:0.07-0.1.
3. The method for detecting the protein content in food according to claim 1, wherein the method comprises the following steps: in step S1, the digestion solution is a 35% hydrogen peroxide solution.
4. The method for detecting the protein content in food according to claim 1, wherein the method comprises the following steps: in step S1, sample: catalyst: the weight portion ratio of the digestion solution is 1: 12-15:20-25.
5. The method for detecting the protein content in food according to claim 1, wherein the method comprises the following steps: in the step S1, the time of the ultrasonic oscillation is 15-30 min.
6. The method for detecting the protein content in food according to claim 1, wherein the method comprises the following steps: in the step S1, the boiling time is 8-10 min.
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| CN112485450A (en) * | 2020-11-27 | 2021-03-12 | 四川省农业科学院经济作物育种栽培研究所 | Simple determination method for protein content of vegetable soybean fresh grains |
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