A method for analyzing the saccharification capacity of brewing Daqu
By preparing the Daqu sample solution and conducting blank and formal tests, recording the volume of glucose consumption and calculating the saccharification force value, the problem of inaccurate data in traditional methods is solved, and a highly accurate saccharification force analysis is achieved.
Patent Information
- Application Number
- CN202211020781.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-08-24
- Publication Date
- 2024-12-10
- Estimated Expiration
- 2042-08-24
AI Technical Summary
The traditional method of saccharification of Daqu is affected by a variety of factors, resulting in inaccurate data and prone to deviations.
The accuracy of the data is ensured by using methods including preparing the large quintillus sample solution, conducting blank and formal tests, recording the volume of glucose consumed, and calculating the saccharification force values through specific formulas.
By comparing the data independently measured two times, the absolute difference does not exceed 10% of the arithmetic average, the accuracy of the glycation force data is improved and the impact in traditional manual titration methods is reduced.
Smart Images
Figure QLYQS_1 
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of saccharification power analysis, in particular to a method for analyzing the saccharification power of brewing koji. Background Art
[0002] Saccharification refers to the process of decomposing starch into sweet products by adding water. It is the main process in the manufacturing process of starch sugars and the main process of many intermediate products in the fermentation process of koji. The method of saccharification depends on the required sweetness of the product and the corresponding physical and chemical properties. It is basically divided into three categories: acid method, enzyme method, and acid-enzyme combined method.
[0003] When analyzing and detecting the saccharification power of brewing Daqu, the traditional analysis and detection method is manual titration. However, the determination of the manual titration method is affected by many factors such as titration speed, evaporation amount, stirring speed, heating power and endpoint judgment, which can easily cause a certain deviation in the measured data of brewing Daqu and cause inaccurate saccharification power values. Therefore, a technical personnel in this field has proposed a method for analyzing the saccharification power of brewing Daqu. Summary of the invention
[0004] In view of the shortcomings of the prior art, the present invention provides a method for analyzing the saccharification power of brewing Daqu, which solves the problem that the saccharification power values are inaccurate and prone to deviations due to many influencing factors in traditional analysis methods.
[0005] To achieve the above objectives, the present invention is implemented by the following technical scheme: A method for analyzing the saccharification capacity of brewing Daqu, comprising the following experimental analysis steps:
[0006] S1: Prepare a Daqu sample solution, place the Daqu sample in a beaker, then add an acetic acid-sodium acetate buffer solution thereto, then add water to the beaker, then stir the solution in the beaker and put it into a constant temperature water bath, finally filter the solution in the beaker, collect the filtrate, and obtain the Daqu sample solution for use;
[0007] S2: Determine the Daqu sample liquid, and perform two test operations, namely, a blank test and a formal test, to obtain the glucose consumption volume V1 and V2 in each test respectively;
[0008] S3: result analysis and calculation, the consumed glucose data V1 and V2 obtained in step S2 are put into the calculation formula for calculation;
[0009] S4: Obtain the saccharification capacity value.
[0010] Preferably, the specific steps of preparing the Daqu sample liquid in S1 are as follows: according to the measured water content of the Daqu sample, weigh 10 g of the absolute dry sample, accurate to 0.001 g, put it into a 250 mL beaker, add 20 mL of acetic acid-sodium acetate buffer solution, then add water, stir evenly with a glass rod, make the volume to 200 mL, then place the beaker in a constant temperature water bath at 35°C for 1 hour, filter, collect the filtrate, and obtain the Daqu sample liquid for use.
[0011] Preferably, the specific operation steps of the blank experiment in step S2 are to add 25.0 mL of soluble starch solution into a test tube, then add 5.0 mL of Daqu sample solution, shake well, add 1 mL of 20% NaOH solution, and then draw 5.0 mL as blank solution into a 150 mL conical flask containing 5.0 mL of Fehling's solution A and B respectively, add 50 mL of water, shake well, and then heat it to boiling, keep the solution in the flask slightly boiling for 2 minutes, add 2 drops of methylene blue indicator, and continue to titrate with glucose standard solution within 1 minute under boiling state until the blue color just disappears as the end point, and record the volume of glucose consumed V1. .
[0012] Preferably, the specific operating steps of the sample determination experiment in step S2 are: adding 25.0 mL of soluble starch solution into another test tube, adding 5.0 mL of Daqu sample solution, shaking, placing in a 35°C warm water bath, accurately timing, saccharifying for 1 hour, adding 1 mL of 20% NaOH solution, and then absorbing 5.0 mL of saccharified liquid into a 150 mL conical flask containing 5.0 mL of Fehling's solution A and 5.0 mL of Fehling's solution B, adding 50 mL of water, shaking, and then heating it to boiling, and keeping the solution in the flask slightly boiling for 2 minutes, adding 2 drops of methylene blue indicator, and continuing to titrate with glucose standard solution within 1 minute under boiling state until the blue color just disappears as the end point, and recording the volume V2 of glucose consumed.
[0013] Preferably, the saccharification power calculation formula in step S3 is: In the formula: X is the saccharification power of the sample, in U; V1 is the volume of the glucose standard solution consumed when titrating the blank, in milliliter (mL); V2 is the volume of the glucose standard solution consumed when titrating the sample, in milliliter (mL); 2.5 is the mass of glucose contained in each milliliter of glucose standard solution, in milligram (mg); 30 is the total volume of the saccharification mixture (soluble starch solution plus Daqu sample solution), in milliliter (mL); 0.25 is the mass of 5 mL Daqu sample solution equivalent to Daqu, in gram (g); 5 is the volume of the saccharification liquid absorbed during titration, in milliliter (mL).
[0014] Preferably, the calculation result in step S4 is retained to one decimal place, and under repeatability conditions in step S4, the absolute difference between two independent measurement results shall not exceed 10% of the arithmetic mean.
[0015] Preferably, the specific preparation steps of the Fehling's solution A are first weighing 69.28g of copper sulfate (CuS04.5H20), then dissolving it with water and diluting it to 1000mL; finally obtaining Fehling's solution A.
[0016] Preferably, the specific preparation steps of the Fehling's solution liquid B are first weighing 346g of potassium sodium tartrate and 100g of sodium hydroxide, then dissolving and diluting to 1000mL with water, then shaking the aqueous solution, and finally filtering the shaken aqueous solution to obtain Fehling's solution liquid B.
[0017] Preferably, the heating device used after the mixed solution prepared in the blank experiment and the formal experiment in step S2 is shaken is an electric furnace, and the mixed solution prepared in the blank experiment and the formal experiment is heated to boiling by the electric furnace.
[0018] Preferably, the pH value of the acetic acid-sodium acetate buffer solution added to the Daqu sample solution prepared in step S1 is 4.6.
[0019] The present invention provides a method for analyzing the saccharification power of brewing Daqu. It has the following beneficial effects:
[0020] The present invention conducts two different experiments, namely a blank experiment and a formal experiment, to record and count the glucose consumption volume under different experimental conditions, then calculates the saccharification power data through a specific formula, and then retains the calculated saccharification power data to one decimal place, and finally compares the data of the two independent measurements, and the absolute value of the measured data shall not exceed 10% of the arithmetic mean, so as to obtain relatively accurate saccharification power data, and the measurement is not affected by many factors in the traditional manual drip method, and the data accuracy is high. DETAILED DESCRIPTION
[0021] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.
[0022] Example:
[0023] The embodiment of the present invention provides a method for analyzing the saccharification capacity of brewing Daqu, comprising the following experimental analysis steps:
[0024] S1: Prepare a Daqu sample solution, place the Daqu sample in a beaker, then add an acetic acid-sodium acetate buffer solution thereto, then add water to the beaker, then stir the solution in the beaker and put it into a constant temperature water bath, finally filter the solution in the beaker, collect the filtrate, and obtain the Daqu sample solution for use;
[0025] S2: Determine the Daqu sample liquid, and perform two test operations, namely, a blank test and a formal test, to obtain the glucose consumption volume V1 and V2 in each test respectively;
[0026] S3: result analysis and calculation, the consumed glucose data V1 and V2 obtained in step S2 are put into the calculation formula for calculation;
[0027] S4: Obtain the saccharification capacity value.
[0028] Sampling method of Daqu test sample: one block is drawn from each of the four corners and the center of the top layer and the bottom layer of the Quku block stacking, and a total of 10 blocks are drawn, and the sensory quality of the 10 blocks is tested respectively. After the sensory test is completed, the 10 Qu samples are crushed, the Qu powder is mixed evenly, and then the samples are sieved, and the small samples are reserved for testing the overall quality indicators.
[0029] The sensory quality standards of Daqu are shown in Table 1.
[0030] Table 1: Daqu sensory quality standards
[0031]
[0032]
[0033] The sensory quality of the third round of Daqu is shown in Table 2:
[0034] Table 2: Sensory quality report of the third round of Daqu in 2021
[0035] Serial number Sample No. Tracking number Level 1 Level 2 Level 3 1 21090730808 80％ 20％ 2 21090530909 80％ 10％ 10％ 3 21090331010 70％ 30％ 4 21083031313 80％ 10％ 10％ 5 21082631717 60％ 30％ 10％ 6 21082831554 80％ 10％ 10％ 7 21082320255 80％ 20％ 8 21082120356 90％ 10％ 9 21081920457 80％ 20％ 10 21090131458 70％ 30％
[0036] The complete quality indicators of the third round of Daqu are shown in Table 3.
[0037] Table 3: Report on the quality indicators of the third round of Daqu in 2021
[0038]
[0039]
[0040] The experimental principle of this analytical method is that the saccharifying amylase in Daqu can hydrolyze starch to produce glucose. Under specified conditions, the sample hydrolyzes the a-1,4-glucosidic bonds from the non-reducing end of the starch in sequence to produce glucose, and the amount of glucose generated is determined by the Fehling method to indicate the saccharifying power.
[0041] The specific steps for preparing the Daqu sample liquid in S1 are as follows: according to the measured water content of the Daqu sample, weigh 10 g of the absolute dry sample, accurate to 0.001 g, put it into a 250 mL beaker, add 20 mL of acetic acid-sodium acetate buffer solution, then add water, stir evenly with a glass rod, make the volume to 200 mL, then place the beaker in a constant temperature water bath at 35°C for 1 hour, filter, collect the filtrate, and obtain the Daqu sample liquid for use.
[0042] Before the blank experiment and the formal experiment begin, the Fehling's solution to be used must be prepared. The specific preparation steps of Solution A and Solution B in the Fehling's solution are as follows. After the Fehling's solution is prepared, it will be set aside.
[0043] The prepared Fehling solution was used for calibration. The calibration was divided into two steps: preliminary experiment and formal experiment. The specific steps of the preliminary experiment were as follows: 5.0 mL of Fehling's solution A and B were taken into a 150 mL conical flask, 50 mL of water was added, shaken well, heated to boiling on an electric stove, and titrated with the prepared glucose standard solution under boiling state. When the blue color of the solution was about to disappear, two drops of methylene blue indicator were added, and the titration was continued until the blue color just disappeared as the end point, and the volume of the consumed glucose standard solution was recorded;
[0044] First-level and second-level sample index report (physical and chemical)
[0045]
[0046] First-level and second-level sample index report (microorganisms)
[0047]
[0048] The specific operation steps of the formal experiment are as follows: take 5.0 mL of Fehling's A and B solutions respectively into a 150 mL conical flask, add 50 mL of water and 1 mL less glucose standard solution than the preliminary test, shake well, heat to boiling on an electric stove, and keep it boiling for 2 minutes, add two drops of methylene blue indicator, titrate with glucose standard solution to the end point within 1 minute under boiling state, and record the total volume V of glucose standard solution consumed;
[0049] Acetic acid-sodium acetate buffer solution (pH 4.6), weigh 164g of anhydrous sodium acetate (CH3COONa), dissolve in water, add 114mL of glacial acetic acid, and dilute to 1000mL with water. The pH of the buffer solution should be calibrated with an acidometer;
[0050] Soluble starch solution (20g / L): Take 2g of soluble starch dried at 100℃~105℃ for 2h, accurate to 0.001g, mix with water to make a paste, pour in 70mL boiling water with constant stirring, stir and boil for 2min until completely transparent, cool to room temperature, transfer completely to a 100mL volumetric flask and make up to volume. This solution is prepared and used immediately;
[0051] Glucose standard solution (2.5g / L): weigh 2.5g of anhydrous glucose dried at 103℃~105℃ to constant weight, accurate to 0.0001g, dissolve in water, and make up to 1000mL. This solution needs to be prepared on the same day;
[0052] Sodium hydroxide solution (mass fraction 20%), weigh 20g of sodium hydroxide into a beaker, add 80mL of water to dissolve, set aside, and prepare a constant temperature water bath for later use;
[0053] After completing the above experimental preparations, you can conduct the saccharification power analysis experiment of brewing Daqu.
[0054] The specific operation steps of the blank experiment in step S2 are as follows: add 25.0mL of soluble starch solution into a test tube, add 5.0mL of Daqu sample solution, shake well, add 1mL of 20% NaOH solution, and then absorb 5.0mL as blank solution into a 150mL conical flask containing 5.0mL of Fehling's solution A and 5.0mL of Fehling's solution B, add 50mL of water, shake well, and then heat it to boiling, keep the solution in the flask slightly boiling for 2min, add 2 drops of methylene blue indicator, and continue to titrate with glucose standard solution within 1min under boiling state until the blue color just disappears as the end point, and record the volume of glucose consumed V1.
[0055] The specific operation steps of the sample determination experiment in step S2 are: add 25.0mL of soluble starch solution into another test tube, then add 5.0mL of Daqu sample solution, shake well, place in a 35℃ warm water bath, accurately time, saccharify for 1h, add 1mL of 20% NaOH solution, then pipette 5.0mL of saccharified solution into a 150mL conical flask containing 5.0mL of Fehling's solution A and B, add 50mL of water, shake well, then heat it to boiling, and keep the solution in the flask slightly boiling for 2min, add 2 drops of methylene blue indicator, and continue to titrate with glucose standard solution within 1min under boiling state until the blue color just disappears as the end point, and record the volume of glucose consumed V2.
[0056] The formula for calculating the saccharification power in step S3 is: In the formula: X is the saccharification power of the sample, in U; V1 is the volume of the glucose standard solution consumed when titrating the blank, in milliliter (mL); V2 is the volume of the glucose standard solution consumed when titrating the sample, in milliliter (mL); 2.5 is the mass of glucose contained in each milliliter of glucose standard solution, in milligram (mg); 30 is the total volume of the saccharification mixture (soluble starch solution plus Daqu sample solution), in milliliter (mL); 0.25 is the mass of 5 mL Daqu sample solution equivalent to Daqu, in gram (g); 5 is the volume of the saccharification liquid absorbed during titration, in milliliter (mL).
[0057] The calculation result in step S4 is retained to one decimal place. Under repeatability conditions in step S4, the absolute difference between two independent measurement results shall not exceed 10% of the arithmetic mean.
[0058] This analytical method uses two different experiments, a blank experiment and a formal experiment, to record and count the volume of glucose consumption under different experimental conditions, and then calculates the saccharification power data through a specific formula. The calculated saccharification power data is then retained to one decimal place. Finally, by comparing the data from the two independent measurements, the absolute value of the measured data must not exceed 10% of the arithmetic mean, thereby obtaining relatively accurate saccharification power data. The measurement is not affected by many factors in the traditional manual drip method, and the data accuracy is high.
[0059] The specific preparation steps of Fehling's solution A are as follows: first weigh 69.28 g of copper sulfate (CuS04.5H20), then dissolve it in water and dilute it to 1000 mL; finally, Fehling's solution A is obtained.
[0060] The specific preparation steps of Fehling's solution liquid B are as follows: first weigh 346 g of potassium sodium tartrate and 100 g of sodium hydroxide, then dissolve and dilute to 1000 mL with water, then shake the aqueous solution, and finally filter the shaken aqueous solution to obtain Fehling's solution liquid B.
[0061] After the mixed solution prepared in the blank experiment and the formal experiment in step S2 is shaken, the heating device used is an electric furnace. The mixed solution prepared in the blank experiment and the formal experiment is heated to boiling by the electric furnace. When heating the mixed solution, other heating devices can be used instead. The specific device can be replaced according to the specific device at the experimental site.
[0062] The pH value of the acetic acid-sodium acetate buffer solution added to the Daqu sample solution prepared in step S1 is 4.6. Precise control of the pH value of the acetic acid-sodium acetate buffer solution facilitates subsequent experiments and can also improve the accuracy of subsequent saccharification test data.
[0063] The Daqu tracking sampling and Daqu crushing sample preparation were completed. A total of 8 Daqu samples were drawn from the 4 Daqu libraries. The testing of the 8 samples has been completed. The test results are detailed in Tables 8 and 9.
[0064] Table 8: Daqu test data report (physical and chemical)
[0065]
[0066]
[0067]
[0068] Table 9: Daqu test data report (microorganisms)
[0069]
[0070] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.
Claims
1. A method for analyzing the saccharification capacity of brewing Daqu, characterized in that: The experimental analysis steps include the following: S1: Prepare a Daqu sample solution, place the Daqu sample in a beaker, then add an acetic acid-sodium acetate buffer solution thereto, then add water to the beaker, then stir the solution in the beaker and put it into a constant temperature water bath, finally filter the solution in the beaker, collect the filtrate, and obtain the Daqu sample solution for use; S2: Determine the Daqu sample solution, and perform two test operations, namely a blank test and a formal test, to obtain the glucose volumes V1 and V2 consumed in each test respectively; the specific operation steps of the blank experiment in the S2 step are to add 25.0 mL of soluble starch solution into a test tube, and then add 5.0 mL of Daqu sample solution, shake well, add 1 mL of 20% NaOH solution, and then absorb 5.0 mL as a blank solution into a 150 mL conical flask containing 5.0 mL of Fehling's solution A and B respectively, add 50 mL of water, shake well, and then heat it to boiling, keep the solution in the bottle slightly boiling for 2 minutes, add 2 drops of methylene blue indicator, and continue to titrate with glucose standard solution within 1 minute under boiling state until the blue color just disappears as the end point, and record the volume of glucose consumed V1; the specific operation steps of the sample determination experiment in the S2 step are to add 25.0 mL of soluble starch solution into another test tube, and then add 5.0 mL Daqu sample solution, shake well, place in a 35℃ warm water bath, accurately time, saccharify for 1h, add 1mL of 20% NaOH solution, pipette 5.0mL of saccharified solution into a 150mL conical flask containing 5.0mL of Fehling's solution A and B, add 50mL of water, shake well, then heat it to boiling, and keep the solution in the flask slightly boiling for 2min, add 2 drops of methylene blue indicator, continue to titrate with glucose standard solution within 1min under boiling state until the blue color just disappears as the end point, and record the volume of glucose consumed V2; S3: Result analysis and calculation, the consumed glucose data V1 and V2 obtained in step S2 are brought into the calculation formula for calculation; the saccharification power calculation formula in step S3 is: , where: X is the saccharification power of the sample, in U; V1 is the volume of the glucose standard solution consumed when titrating the blank, in milliliters (mL); V2 is the volume of the glucose standard solution consumed when titrating the sample, in milliliters (mL); 2.5 is the mass of glucose contained in each milliliter of glucose standard solution, in milligrams (mg); 30 is the total volume of the saccharification mixture, the soluble starch solution plus the koji sample solution, in milliliters (mL); 0.25 is the mass of 5 mL of the koji sample solution equivalent to the koji, in grams (g); 5 is the volume of the saccharification solution absorbed during titration, in milliliters (mL); S4: Obtain the saccharification capacity value.
2. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: The specific steps for preparing the Daqu sample liquid in S1 are as follows: according to the measured water content of the Daqu sample, weigh 10 g of the absolute dry sample, accurate to 0.001 g, put it into a 250 mL beaker, add 20 mL of acetic acid-sodium acetate buffer solution, then add water, stir evenly with a glass rod, and make the volume to 200 mL. Then, place the beaker in a constant temperature water bath at 35°C for 1 hour, filter, collect the filtrate, and obtain the Daqu sample liquid for use.
3. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: The calculation result in step S4 is retained to one decimal place. In step S4, under repeatability conditions, the absolute difference between two independent measurement results shall not exceed 10% of the arithmetic mean.
4. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: The specific preparation steps of the Fehling's solution A liquid are as follows: first weigh 69.28 g of CuS04.5H20, then dissolve it in water and dilute it to 1000 mL; finally, Fehling's solution A liquid is obtained.
5. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: The specific preparation steps of the Fehling's solution liquid B are as follows: firstly weigh 346g of potassium sodium tartrate and 100g of sodium hydroxide, then dissolve and dilute to 1000mL with water, then shake the aqueous solution, and finally filter the shaken aqueous solution to obtain Fehling's solution liquid B.
6. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: After the mixed solution prepared in the blank experiment and the formal experiment in step S2 is shaken, the heating device used is an electric furnace, and the mixed solution prepared in the blank experiment and the formal experiment is heated to boiling by the electric furnace.
7. The method for analyzing the saccharification power of brewing Daqu according to claim 1, characterized in that: The pH value of the acetic acid-sodium acetate buffer solution added to the Daqu sample solution prepared in step S1 is 4.6.