CN110879260A - Method for predicting barley malt filtering speed through arabinoxylan content - Google Patents
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Abstract
The invention discloses a method for predicting barley malt filtering speed through arabinoxylan content, and belongs to the technical field of beer production. The molecular exclusion chromatography provided by the invention can separate the arabinoxylan related to the wort filtration speed, and according to a univariate linear equation reflecting the relationship between the filtration speed and the arabinoxylan content with the molecular weight of more than 50,000Da, y is 484.892-0.852x, wherein y represents the filtration speed of barley malt in the industrial production of wort, and x represents the arabinoxylan content with the molecular weight of more than 50000Da, so that the filtration speed of a malt sample in the industrial production can be accurately predicted; the influence degree of the arabinoxylan in the barley malt on the filtration speed of the saccharified wort in the beer production can be predicted in advance, so that the preparation method can be improved conveniently according to different barley in the beer production, thereby reducing unnecessary loss and improving the production efficiency of the beer.
Description
Technical Field
The invention relates to a method for predicting barley malt filtering speed through arabinoxylan content, and belongs to the technical field of beer production.
Background
The filtration speed of wort is one of the most important indexes for brewers, which determines the efficiency of beer production process, and the research on the influence factors of filtration speed is always a hot point in the research of beer industry. The filtration rate of barley malt also has an important influence on the quality of the finished beer: the filtering speed is slow, the saccharification viscosity is high, the contact and the degradation of enzyme and substrate are not facilitated, macromolecular substances such as protein, starch and the like cannot be fully degraded, the extract yield is low, the production time of a single batch of beer is prolonged, and the production cost is increased.
Barley malt is the main raw material for beer brewing. Arabinoxylan is the most important component in the barley endosperm cell wall, belongs to non-starch polysaccharide, and accounts for about 20% of the dry weight of the endosperm cell wall and 4-10% of the total weight of barley seeds. Researches show that wort and finished beer still contain high-concentration arabinoxylan, the highest concentration of the arabinoxylan in 36 kinds of domestic and foreign beers reaches 849mg/L, and high-molecular-weight arabinoxylan aqueous solution has high viscosity and potential possibility of influencing wort viscosity and filtration speed, so that the arabinoxylan aqueous solution has more and more attention of brewers and researchers in recent years.
At present, in domestic beer production and research, the methods for detecting the arabinoxylan content mainly include gas chromatography (Debyser W, Derdelink X G, Delcour J A. Arabinoxylan and arabinyloxyanhydrans activity in barley and salts derivative from the [ J ]. Journal of Cereal Science,1997,26(1):67-74), phenolate method (plum, land, health, see the national sages. determination of pentosan in beer-licheno-muriate method [ J ]. food and fermentation industry, 2003,29(9):35-38) and phloroglucinol method (Kanauchi M, Ishikura W, BamHforr C W. β -glucans and pentosans and xyloglucan production W. 2011. filtration method of xylan content, which is not determined by the above mentioned methods, but by filtration of xylose from the above mentioned methods, the influence of the arabinoxylan content is not determined accurately by the filtration rate of the above-filtration of barley J. filtration rate of xylan, the filtration rate of barley 124. filtration method, the influence of arabinoxylan content is not determined by the above mentioned methods of barley.
Disclosure of Invention
To solve the above problems, the present invention provides a method for predicting barley malt filtration rate by arabinoxylan content. The technical scheme of the invention is as follows:
the invention provides a method for predicting barley malt filtering speed, which comprises the following steps:
(1) measuring the content of arabinoxylan with molecular weight of more than 50,000Da in barley malt protocol wort;
(2) the filtration rate of barley malt in an industrial beer mashing process is predicted from an equation of linear relationship between the content of arabinoxylan with a molecular weight of more than 50,000Da in the wort and the filtration rate of barley malt in an industrial beer mashing process.
In one embodiment of the invention, the wort for detecting the arabinoxylan content with the molecular weight of more than 50,000Da in the step (1) is prepared according to an agreement saccharification method, and the method comprises the following steps of ①, pouring 48-52 g of finely crushed malt of different samples into a saccharification cup with a known weight, adding 200mL of 42-48 ℃ purified water, placing the saccharification cup on an automatic saccharification device, preserving heat at 42-48 ℃ for 25-35 min to prepare mash, ②, heating the mash to 70-72 ℃ at the speed of 0.5-1.5 ℃/min, adding 100mL of 70-72 ℃ purified water, preserving heat at 70-72 ℃ for 55-65 min, cooling the malt mash to 22-26 ℃, adding purified water to enable the total weight of the mash to reach 450.0g, ③, filtering the mash in a 32cm funnel by adopting medium-speed qualitative filter paper, and collecting filtrate.
In one embodiment of the invention, in the step (1), the arabinoxylan with molecular weight more than 50,000Da in the barley malt agreement wort is separated by using molecular exclusion chromatography, and then the content of the arabinoxylan with molecular weight more than 50,000Da is determined by using a phloroglucinol method, wherein the molecular exclusion chromatography comprises the steps of ① calibrating the molecular exclusion chromatography column by using glucans with different molecular weights as calibration molecular weight standards, determining the elution time of each molecular weight standard, and ② passing the wort prepared by the agreement saccharification method through the molecular chromatography column, collecting eluent according to the different elution times determined before, and obtaining the arabinoxylan eluent with molecular weight more than 50,000 Da.
In one embodiment of the present invention, linear regression analysis is used to determine the relationship between the arabinoxylan content of barley malt having a molecular weight greater than 50,000Da and the filtration rate of barley malt in wort production by an industrial beer mashing process, and a linear equation is constructed reflecting the relationship between the arabinoxylan content of barley malt having a molecular weight greater than 50000Da and the filtration rate of barley malt in wort production by an industrial beer mashing process, where y is 484.892-0.852 x.
In one embodiment of the invention, the saccharification process of the industrial beer production in the step (2) comprises the following steps of putting ① 20-30 kg of finely ground malt and 90-110L of 45-50 ℃ tap water into a saccharification pot, preserving heat at 42-50 ℃ for 25-35 min, preserving heat at ② ℃ for 35-45 min, raising the temperature to 70-75 ℃ until starch is completely decomposed, standing the malt mash for 25-40 min, filtering, and collecting wort.
The invention provides application of a method for predicting the wort filtration speed through the content of arabinoxylan in beer production.
In one embodiment of the present invention, the beer includes a draft beer, a dry beer, an ice beer, a low alcohol beer, a non-alcohol beer, a wheat beer, a turbid beer, a fruit and vegetable juice type beer, and a fruit and vegetable taste type beer.
Has the advantages that: the invention provides a method for determining the content of arabinoxylan with molecular weight more than 50,000Da in barley malt agreement wort by adopting a molecular exclusion chromatography, the molecular exclusion chromatography provided by the invention can separate the arabinoxylan related to the filtration speed of the wort, and according to a univariate linear equation reflecting the relationship between the filtration speed and the content of the arabinoxylan with molecular weight more than 50,000Da, y is 484.892-0.852x, wherein y represents the filtration speed of barley malt in the industrial production of the wort, and x represents the content of the arabinoxylan with molecular weight more than 50000Da, the filtration speed of a malt sample in the industrial production can be accurately predicted; the influence degree of the arabinoxylan in the barley malt on the filtration rate of the saccharified wort in beer production can be predicted in advance. The detection result can help a brewer to predict the filtration speed of the barley malt in the batch for preparing the wort by adopting an industrial beer saccharification process in advance, so that the preparation method can be improved conveniently according to different barley in the beer production, thereby reducing unnecessary loss and improving the production efficiency of beer.
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FIG. 1 is a typical chromatogram for the determination of the content of arabinoxylans of different molecular weights by size exclusion chromatography.
Detailed Description
EXAMPLE 1 calibration of size exclusion chromatography columns
The size exclusion column was Sephacryl S-300HR from GE, column size 26mm by 1,000mm in inside diameter by bed height.
Dextran (Dextran) having molecular weights of 21,000Da,55,000Da,123,000Da,500,000Da,760,000Da, respectively, was used as a molecular weight standard for calibration. The loading concentration of the molecular weight standard is 400mg/L, and the loading amount is 10 mL.
The mobile phase was 0.05mol/L NaCl and the flow rate was 100 mL/h. And collecting the eluent by using an automatic fraction collector, collecting 5mL of the eluent in each tube, determining the total sugar content in each tube of the eluent by adopting an anthrone-sulfuric acid method, and determining the elution time of each molecular weight standard substance, thereby completing the calibration of the molecular exclusion chromatographic column.
Example 2 preparation of malt protocol for barley mashing and size exclusion chromatography for the determination of arabinoxylan content of different molecular weights in wort
(1) Preparation of barley malt protocol wort
① pouring 50.0g of finely pulverized malt of different samples into a known weight of saccharifying cup, adding 200mL of purified water at 46 deg.C, placing on an automatic saccharifying device, and keeping the temperature at 45 deg.C for 30 min;
② heating the mash to 70 deg.C at a rate of 1 deg.C/min, adding 100mL 70 deg.C purified water, maintaining at 70 deg.C for 60min, cooling the malt mash to about 25 deg.C, and adding purified water to make the total weight of mash reach 450.0 g;
③ the filtrate was collected by filtration through a 32cm funnel using medium speed qualitative filter paper.
④ A10 mL sample of the collected wort was applied to a size exclusion chromatography column with a mobile phase of 0.05mol/L NaCl and a flow rate of 100mL/h and the eluate was collected from an autosegregation apparatus.
(2) Method for determining content of arabinoxylan in each sample by phloroglucinol method
Preparing a color developing agent: 1g of phloroglucinol is dissolved by 5mL of absolute ethyl alcohol, and then 110mL of glacial acetic acid, 1mL of 17.5g/L glucose and 2mL of concentrated hydrochloric acid are sequentially added, and the mixture is shaken up and is prepared for use.
Respectively taking 2mL of the collected liquid from each tube, placing the collected liquid in a test tube with a plug scale, adding 10mL of phloroglucinol color developing agent, oscillating the collected liquid uniformly, accurately reacting the liquid in a boiling water bath for 25min, cooling the liquid to room temperature, measuring an OD value at 552nm, calculating the content of the arabinoxylan in each tube, and respectively calculating the content of the arabinoxylan with the molecular weight of more than 5,000, more than 50,000, more than 500,000 and more than 1000,000 according to the peak-off time of a molecular weight standard product obtained after the molecular exclusion chromatography calibration, wherein the results are shown in Table 1. A typical chromatogram for the determination of the content of arabinoxylans of different molecular weights in wort by size exclusion chromatography is shown in FIG. 1.
TABLE 1 arabinoxylan content of different molecular weight size in barley malt samples
Example 3 correlation analysis of the content of arabinoxylan with different molecular weights in barley malt and the filtration rate of wort for simulated industrial production
The preparation method of the wort for simulating the industrial beer saccharification process comprises the following steps:
25.0kg of finely pulverized malt → 100L of tap water at 46 ℃ are put into a mash kettle (the volume of the mash kettle is 200L) → 30min holding at 45 ℃ C. → 65 ℃ C. → 40min holding at 65 ℃ C./min raising the temperature of the mash at 1 ℃/min to 72 ℃ C. → 5min apart for iodine test until color development is complete → malt mash is pumped into a filter tank, and the mash is left stand for 30min → wort is filtered by a sieve plate at the bottom of the filter tank, and clear wort is collected. The filtration rate (V) is expressed as the volume of wort collected per unit time (in mL/min). The filtration rate of wort for simulating the industrial beer mashing process and the arabinoxylan content in the wort with different molecular weight according to the agreement in example 2 are shown in Table 2.
TABLE 2 arabinoxylan content and wort filtration rate for simulated industrial beer saccharification process
Pearson correlation analysis of the arabinoxylan content of different molecular weights in the barley malt agreement wort as determined in example 2 and the wort filtration rate as determined in the simulated industrial beer production of example 3 was performed using the SPSS19.0 software and the results are shown in Table 3.
TABLE 3 analysis of the correlation between the molecular weight of arabinoxylan and the filtration rate
Note: n is the number of samples; *: significant correlation at the 0.05 level; **: significant correlation was found at the 0.01 level.
The results in table 3 show that the correlation coefficient of arabinoxylan content with molecular weight greater than 50,000Da with the filtration rate of the saccharified wort simulating industrial beer production is at most 0.789, which is a significant negative correlation (P <0.001), with significance results of 0.000. It is demonstrated that the arabinoxylan with molecular weight of more than 50,000Da segment is the main reason for influencing the filtration rate of the malt agreement, therefore, the filtration rate of the saccharified wort of barley malt in the industrial beer production can be predicted by detecting the content of the arabinoxylan with molecular weight of more than 50,000Da segment in the malt agreement wort of barley.
Example 4 Single Linear equation for the filtration Rate of malt mash and the arabinoxylan content with molecular weight above 50000Da in simulated Industrial beer production
The linear regression analysis in SPSS19.0 is adopted to further analyze the relationship between the filtration speed of the wort prepared by the simulated industrialized beer saccharification process and the content of the arabinoxylan with the molecular weight of more than 50000Da so as to construct a unitary linear equation capable of reflecting the relationship between the two. In linear regression analysis, the independent variable is the filtration rate and the dependent variable is the arabinoxylan content with a molecular weight of more than 50000 Da. The results of the linear regression analysis are shown in Table 4.
TABLE 4 model summary of Linear regression analysis
a, predicting variables.
TABLE 4 model Overall, model fitness R20.623, the adjusted model fitness R2The value is 0.598, which indicates that the filtration speed of wort in the simulated industrialized saccharification can be explained by the model, and the fitting degree is higher.
TABLE 5 analysis of variance table of regression model
a: and predicting variables.
From the results of the analysis of variance test of the regression model in table 5, the statistic F was 24.770 and the P value was 0.000, demonstrating that the model is significant.
TABLE 6 coefficientsa
a. The dependent variable V.
TABLE 7 residual statisticsa
a. The dependent variable V.
In table 6, the regression coefficient constant is 484.892, the significance is 0.000, and the constant term is significant; the regression coefficient was-0.852, the significance was 0.000, and the significance was obtained. The model is significant in the analysis of variance table of the regression model. The unitary linear equation reflecting the relationship between the filtration rate of wort prepared by the industrial beer saccharification process and the arabinoxylan content with molecular weight of more than 50000Da is as follows: y is 484.892-0.852 x; wherein y represents the filtration rate of barley malt in the commercial production of wort, and x represents the arabinoxylan content.
Example 5
The method for detecting the content of the arabinoxylan with the molecular weight of more than 50000Da in a certain batch of Jiangsu barley malt is the same as that in example 1 according to the steps in example 2, and the result shows that the content of the arabinoxylan with the molecular weight of more than 50000Da is 560mg/L, and the filtering speed is 7.6mL/min according to the calculation of y being 484.892-0.852 x.
When the batch of malt is used for industrial scale beer production in a certain brewery in Jiangsu, the adopted saccharification process is as follows:
8000kg of finely ground malt → 48kL of 46 ℃ tap water are put into a mash kettle (the volume of the mash kettle is 60kL) → the heat preservation is carried out for 30min at 45 ℃ → the mash is heated to 65 → the heat preservation is carried out for 40min at 65 ℃ at the rate of 1 ℃/min → the mash is heated to 72 ℃ → the iodine test is carried out at 5min intervals at the rate of 1 ℃/min until the color development is complete → the malt mash is pumped into a filter tank, the malt mash is stood for 30min → the wort is filtered by a sieve plate at the bottom of the filter tank, and clear wort is collected. The actual filtration rate of the wort was 7.8mL/min, indicating that the prediction equation can predict the filtration rate of barley malt well.
The method can accurately predict the filtering speed of a certain batch of barley malt mash before saccharification is finished, is beneficial to timely adjusting the process, and plays an important role in improving the efficiency of the beer production process.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (10)
1. A method for predicting barley malt filtration rate, comprising the steps of:
(1) detecting the content of arabinoxylan with molecular weight more than 50,000Da in barley malt agreement wort;
(2) the filtration rate of barley malt in an industrial beer mashing process is predicted from an equation of linear relationship between the content of arabinoxylan with a molecular weight of more than 50,000Da in the wort and the filtration rate of barley malt in an industrial beer mashing process.
2. The method of claim 1, wherein the wort for measuring the arabinoxylan content having a molecular weight greater than 50,000Da is prepared according to a protocol saccharification method.
3. The method of claim 2, wherein the agreement saccharification process step is: (1) pouring 48-52 g of finely ground malt of different samples into a saccharifying cup with a known weight, adding 150-250 mL of 42-48 ℃ purified water, placing on an automatic saccharifying device, and preserving heat at 42-48 ℃ for 25-35 min to prepare mash; (2) heating the mash to 70-72 ℃ at the speed of 0.5-1.5 ℃/min, adding 75-150 mL of 70-72 ℃ purified water, preserving the heat at 70-72 ℃ for 55-65 min, and cooling the malt mash to 22-26 ℃.
4. The method of claim 1, wherein the arabinoxylan content of the agreed wort with a molecular weight of more than 50,000Da is separately determined by size exclusion chromatography.
5. The method of claim 4, wherein the size exclusion chromatography step is: (1) calibrating the size exclusion chromatographic column by using dextrans with different molecular weights as calibration molecular weight standards, and determining the elution time of each molecular weight standard; (2) passing the wort prepared by the conventional saccharification method through a molecular chromatographic column, and collecting eluent according to different elution times measured before to obtain arabinoxylan eluent with molecular weight of more than 50,000 Da.
6. The method according to claim 1, wherein the arabinoxylan content with a molecular weight greater than 50,000Da is determined using the phloroglucinol method.
7. The method of claim 1, wherein the relationship between the content of arabinoxylan with a molecular weight of greater than 50,000Da in the wort and the filtration rate of barley malt in the industrial beer mashing process is analyzed by linear regression to construct a linear equation reflecting the relationship.
8. The method of claim 1, wherein the calculation is performed according to the unary linear equation y-484.892-0.852 x; wherein y represents the filtration rate of barley malt in the commercial production of wort and x represents the arabinoxylan content with a molecular weight of more than 50000 Da.
9. Use of the method according to claim 1 in the production of beer.
10. The use according to claim 9, wherein the beer comprises a draft beer, dry beer, ice beer, low alcohol beer, non-alcohol beer, wheat beer, cloudy beer, fruit and vegetable juice type beer, fruit and vegetable flavored type beer.
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Application publication date: 20200313 |