CN111398193A - Method for rapidly detecting protein content in grain for wine brewing - Google Patents
Method for rapidly detecting protein content in grain for wine brewing Download PDFInfo
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- CN111398193A CN111398193A CN202010254937.7A CN202010254937A CN111398193A CN 111398193 A CN111398193 A CN 111398193A CN 202010254937 A CN202010254937 A CN 202010254937A CN 111398193 A CN111398193 A CN 111398193A
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Abstract
The invention discloses a method for rapidly detecting protein content in grain for wine brewing, which relates to the technical field of wine brewing and comprises the steps of S1, grinding a sample to be detected to 150-180 mu m, accurately weighing a sample to be detected with a certain mass, adding concentrated sulfuric acid and hydrogen peroxide for digestion, cooling a digested mixture, S2, taking a product obtained by processing 0.5m L through S1, adding 9.5m L alkaline solution, shaking up, taking a product obtained by processing 0.5m L through S2, adding 9.5m L water, adding 0.4ml of alkaline Nashin reagent, shaking up, standing for 10 minutes in a dark place, measuring absorbance at a wavelength of 430nm, and S4, calculating the content of the protein content according to a detection result by means of a standard working curve.
Description
Technical Field
The invention relates to the technical field of wine brewing, in particular to a method for quickly detecting the content of protein in grains for wine brewing.
Background
Protein is one of important indexes in conventional grain detection projects, and the current commonly used national standard method adopts concentrated sulfuric acid-catalyst digestion on an electric heating or infrared digestion furnace and Kjeldahl method titration. However, the method has the following defects: the operation steps are complicated, the digestion time of the concentrated sulfuric acid-catalyst on an electric heating or infrared digestion furnace is long, generally 4-6 hours are needed, the reagent dosage is large, and the energy consumption is high; there is a great impact on the environmental pollution and the health of the operators. Therefore, a method for measuring the protein content in the grain for brewing wine with high efficiency, accuracy, rapidness, good repeatability and good reproducibility is urgently needed. The method can be applied to detection departments and production enterprises at all levels, and the problem of rapid quantitative detection of the protein is solved.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a method for rapidly detecting the content of protein in grain for wine brewing, provides a method which is efficient, accurate, rapid, simple and convenient to operate, good in repeatability and reproducibility for the determination of protein in grain for wine brewing, and fills the technical blank of rapid determination of protein in grain for wine brewing.
The method comprises the steps of S1, grinding a sample to be detected to 150-180 mu m, accurately weighing the sample to be detected with a certain mass, adding concentrated sulfuric acid and hydrogen peroxide for digestion, cooling a digested mixture, S2, adding 9.5m L to a product obtained by processing 0.5m L through S1, adding 9.5m L alkaline solution, shaking up, S3, adding 9.5m L to a product obtained by processing S2, adding 9.5m L water, adding 0.4ml of alkaline Nashin reagent, shaking up, standing for 10 minutes in a dark place, measuring absorbance at a wavelength of 430nm, and S4, calculating the content of the protein according to a detection result by means of a standard working curve.
Preferably, in step S4, the standard working curve is prepared by accurately transferring 0.1m L, 0.2m L, 0.3m L, 0.4m L, 0.5m L, and 0.6m L of a standard solution with a nitrogen content of 10 mg/L to a 25m L colorimetric tube, adding water to 10m L, adding 0.4m L of an alkaline nash reagent, shaking, standing in dark for 10 minutes, measuring absorbance at a wavelength of 430nm, and preparing the standard working curve with the absorbance as abscissa and the corresponding nitrogen content as ordinate.
Preferably, in step S4, the calculation formula of the protein content is:
in the formula: w-protein content (% or g/100 g); c-nitrogen content (ug) from a standard working curve; k-nitrogen coefficient to protein, rice: 5.95, wheat: 5.83, corn, sorghum: 6.25; m-sample mass (g); 500-dilution factor.
Preferably, in the step S1, the weighed mass of the sample to be detected is 0.0500-0.2500g, the sample to be detected is placed in a digestion tank, and the volume of the digestion tank is 50-70m L.
Preferably, in step S1, the sample to be tested is grain for brewing wine; the sample to be tested comprises but is not limited to one or more of rice, sticky rice, wheat, sorghum and corn.
Preferably, in step S1, the concentrated sulfuric acid is analytically pure concentrated sulfuric acid, and the addition amount of the concentrated sulfuric acid (analytically pure) is 3m L.
Preferably, in step S1, the hydrogen peroxide is analytically pure hydrogen peroxide, and the addition amount of the hydrogen peroxide (analytically pure) is 3m L. preferably, in step S1, the digestion is carried out in a microwave rapid digestion instrument.
Preferably, in step S1, the cooling is performed in a constant temperature cooling water tank, the water temperature is controlled to be-5 to-6 ℃, and the cooling time is 35 minutes.
Preferably, in step S2, the alkaline solution is a sodium hydroxide solution, and the concentration of the sodium hydroxide solution is 0.1 mol/L.
The method for rapidly detecting the protein content in the grain for brewing provided by the invention adopts microwave micro digestion, uses superior pure sulfuric acid and hydrogen peroxide solution to catalyze and digest a sample, digests organic matters into solution of inorganic substances, accurately and quantitatively adds a color developing reagent for color development, and accurately determines by using a spectrum principle.
The invention has the beneficial effects that: (1) the full-closed microwave digestion is realized, the operation is simple and easy, and the repeatability and the reproduction are good; (2) the dosage of the reagent is less, the cost is low and the pollution is less; (3) the single sample is measured for only 45 minutes, and the real quick detection is realized; (4) the batch detection can be realized, the detection working efficiency is high, and the operation cost of the laboratory of the enterprise and public institution is saved.
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In order to more clearly illustrate the detailed description of the invention or the technical solutions in the prior art, the drawings that are needed in the detailed description of the invention or the prior art will be briefly described below.
FIG. 1 is a standard operating curve.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The following examples are only for illustrating the technical solutions of the present invention more clearly, and therefore are only examples, and the protection scope of the present invention is not limited thereby.
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains.
Example 1
A method for rapidly detecting the content of protein in grain for brewing wine comprises the following steps:
s1, grinding a sample (sorghum) to be detected to 150-;
s2, taking a product obtained by treating 0.5m L with S1, adding 9.5m L of sodium hydroxide solution with the concentration of 0.1 mol/L, and shaking up;
s3, taking a product obtained by treating 0.5m L with S2, adding 9.5m L of water, adding 0.4ml of alkaline Nashin reagent, shaking up, standing for 10 minutes in a dark place, and measuring the absorbance at the wavelength of 430 nm.
S4: and calculating the content of the protein content according to the result obtained by detection by means of a standard working curve.
In step S4, the standard working curve is specifically produced by the steps of:
respectively and accurately transferring standard solutions with the nitrogen content of 10 mg/L, namely 0.1m L, 0.2m L, 0.3m L, 0.4m L, 0.5m L and 0.6m L into a 25m L colorimetric tube, adding water to 10m L, adding an alkaline Nashin reagent, namely 0.4m L, shaking uniformly, standing in a dark place for 10 minutes, and measuring the absorbance at the wavelength of 430 nm.
The absorbance (Abs) was used as the abscissa and the corresponding nitrogen content (ug) was used as the ordinate to prepare a standard working curve, as shown in fig. 1.
The nitrogen content of the sample (sorghum) to be detected is found to be 1.06ug from a standard working curve (figure 1) according to the absorbance value measured by the sample (sorghum), and the protein content is calculated to be 6.62 percent by using a calculation formula of the protein content and taking k as 6.25.
Example 2
A method for rapidly detecting the content of protein in grain for brewing wine comprises the following steps:
s1, grinding a sample (rice) to be detected to 150-;
s2, taking a product obtained by treating 0.5m L with S1, adding 9.5m L of sodium hydroxide solution with the concentration of 0.1 mol/L, and shaking up;
s3, taking a product obtained by treating 0.5m L with S2, adding 9.5m L of water, adding 0.4ml of alkaline Nashin reagent, shaking up, standing for 10 minutes in a dark place, and measuring the absorbance at the wavelength of 430 nm.
S4: and calculating the content of the protein content according to the result obtained by detection by means of a standard working curve.
In step S4, the standard working curve is specifically produced by the steps of:
respectively and accurately transferring standard solutions with the nitrogen content of 10 mg/L, namely 0.1m L, 0.2m L, 0.3m L, 0.4m L, 0.5m L and 0.6m L into a 25m L colorimetric tube, adding water to 10m L, adding an alkaline Nashin reagent, namely 0.4m L, shaking uniformly, standing in a dark place for 10 minutes, and measuring the absorbance at the wavelength of 430 nm.
The absorbance (Abs) was used as the abscissa and the corresponding nitrogen content (ug) was used as the ordinate to prepare a standard working curve, as shown in fig. 1.
The nitrogen content of the sample (sorghum) to be detected is found to be 1.16ug from a standard working curve (figure 1) according to the absorbance value measured by the sample (sorghum), and the protein content is calculated to be 6.88 percent by using a calculation formula of the protein content and taking k as 5.95.
Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; such modifications and substitutions do not depart from the spirit and scope of the present invention, and they should be construed as being included in the following claims and description.
Claims (10)
1. A method for rapidly detecting protein content in grain for wine brewing is characterized by comprising the steps of S1, grinding a sample to be detected to 150-180 mu m, accurately weighing the sample to be detected with a certain mass, adding pure concentrated sulfuric acid and hydrogen peroxide for digestion, cooling a digested mixture, S2, adding 9.5m L into a product obtained by processing 0.5m L through S1, adding 9.5m L alkaline solution, shaking uniformly, S3, adding 9.5m L into a product obtained by processing 0.5m L through S2, adding 0.4ml of alkaline Nashin' S reagent, shaking uniformly, standing for 10 minutes in a dark place, measuring absorbance at a wavelength of 430nm, and S4, calculating the content of protein according to a detection result by means of a standard working curve.
2. The method for rapidly detecting the protein content in the grain for wine brewing according to claim 1, wherein in the step S4, the preparation of the standard working curve specifically comprises the steps of accurately transferring a standard solution with the nitrogen content of 10 mg/L, namely 0.1m L, 0.2m L, 0.3m L, 0.4m L, 0.5m L and 0.6m L, into a 25m L colorimetric tube, adding water to 10m L, adding an alkaline Nashin reagent, namely 0.4m L, shaking up, standing in a dark place for 10 minutes, measuring the absorbance at the wavelength of 430nm, taking the absorbance as the abscissa and the corresponding nitrogen content as the ordinate, and preparing the standard working curve.
3. The method for rapidly detecting the protein content in the grains for brewing wine as recited in claim 1, wherein in step S4, the calculation formula of the protein content is as follows:
in the formula: w-protein content (% or g/100 g); c-nitrogen content (ug) from a standard working curve; k-nitrogen coefficient to protein, rice: 5.95, wheat: 5.83, corn, sorghum: 6.25; m-sample mass (g); 500-dilution factor.
4. The method for rapidly detecting the protein content in the grain for brewing wine as recited in claim 1, wherein in step S1, the weighed mass of the sample to be detected is 0.0500-0.2500g, the sample to be detected is placed in a digestion tank, and the volume of the digestion tank is 50-70m L.
5. The method for rapidly detecting the protein content in the brewing grain according to claim 1, which is characterized in that: in the step S1, the sample to be detected is grain for brewing wine; the sample to be detected comprises one or more of rice, sticky rice, wheat, sorghum and corn.
6. The method for rapidly detecting the protein content in the grain for brewing wine as recited in claim 1, wherein in step S1, the concentrated sulfuric acid is analytically pure concentrated sulfuric acid, and the addition amount of the concentrated sulfuric acid is 3m L.
7. The method for rapidly detecting the protein content in the grain for brewing wine as recited in claim 1, wherein in step S1, said hydrogen peroxide is analytically pure hydrogen peroxide, and the addition amount of said hydrogen peroxide is 3m L.
8. The method for rapidly detecting the protein content in the brewing grain according to claim 1, which is characterized in that: in step S1, the digestion is performed in a microwave rapid digestion apparatus.
9. The method for rapidly detecting the protein content in the brewing grain according to claim 1, which is characterized in that: in step S1, the cooling is carried out in a constant temperature cooling water tank, the water temperature is controlled to be-5 to-6 ℃, and the cooling time is 35 minutes.
10. The method for rapidly detecting protein content in grain for brewing wine as recited in claim 1, wherein in step S2, said alkaline solution is sodium hydroxide solution, and the concentration of said sodium hydroxide solution is 0.1 mol/L.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101354387A (en) * | 2008-09-24 | 2009-01-28 | 北京市大兴区疾病预防控制中心 | Method for determining protein, sodium, kalium, calcium, magnesium in prescription nutrition food |
| CN104165886A (en) * | 2014-08-01 | 2014-11-26 | 湖北富邦科技股份有限公司 | Method for determining grain protein |
| CN104316524A (en) * | 2014-11-25 | 2015-01-28 | 中国农业科学院麻类研究所 | Method for determining content of crude protein in fiber plant products for fodder |
| CN109805270A (en) * | 2019-01-28 | 2019-05-28 | 江南大学 | A method of low albumen millet is produced using two-way solid state fermentation |
-
2020
- 2020-04-02 CN CN202010254937.7A patent/CN111398193A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101354387A (en) * | 2008-09-24 | 2009-01-28 | 北京市大兴区疾病预防控制中心 | Method for determining protein, sodium, kalium, calcium, magnesium in prescription nutrition food |
| CN104165886A (en) * | 2014-08-01 | 2014-11-26 | 湖北富邦科技股份有限公司 | Method for determining grain protein |
| CN104316524A (en) * | 2014-11-25 | 2015-01-28 | 中国农业科学院麻类研究所 | Method for determining content of crude protein in fiber plant products for fodder |
| CN109805270A (en) * | 2019-01-28 | 2019-05-28 | 江南大学 | A method of low albumen millet is produced using two-way solid state fermentation |
Non-Patent Citations (1)
| Title |
|---|
| 曹坎涛: "蛋白质测定方法改进", 《河北化工》 * |
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Application publication date: 20200710 |