Simple and quick determination method of alcoholic strength of wine with alcoholic strength of 1 to 10 degree

A technology for rapid determination and alcohol content, which is applied in the direction of color/spectral characteristic measurement and analysis through chemical reaction of materials, etc. It can solve the problems of unsatisfactory rapid detection development, long detection process, poor stability, etc., and achieve low price , good accuracy and reliability, the effect of less wine sample

Inactive Publication Date: 2017-12-19

黄海霞

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AI-Extracted Technical Summary

Problems solved by technology

The entire detection process is time-consuming, has many steps, and poo...

Method used

The present invention also adds the urea and the formamide mixed solution of optimized specific concentration in wine sample to be tested and DNA system, mainly in order to make DNA fragment be in an environment that is easy to denaturation, thereby making ethanol assay method have more High sensitivity. The short double-stranded DNA system provided by the present invention includes 2 DNA fragments, and its melting temperature increases according to the increasing order of SEQ ID NO: 1-2. Therefore, with the increase of alcohol content, the types and types of denatured double-stranded DNA in the system will increase. The amount also increases, so there is a positive correlation between absorbance and alcohol. In addition, the denaturation of DNA is achieved within a narrow range, so screening and condition optimization are required, so that the boundaries of DNA fragments with different melting temperatures in the DNA system are superimposed and continued under specified conditions. In the narrow range of ethanol, a mild denaturant (within 10ml) and ultra-short DNA fragments (less than 15bp), it is found t...

Abstract

The alcoholic strength is a necessary index for testing an alcohol product, but at present, an accurate, reliable, simple, convenient, fast and economical and practical alcoholic strength determination method is in shortage. A new alcoholic strength determination method is established by utilizing the phenomenon that a DNA (Deoxyribonucleic Acid) fragment is unlinked in an ethanol system so as to cause the change of absorbancy of the DNA fragment under 260nm wave length. According to the method provided by the invention, aiming at wine with the alcoholic strength of 1 to 10 degree, an appropriate short double-chain DNA fragment mixing system is selected as shown in SEQ ID NO:1-2, and a standard curve equation of the absorbancy x and the alcohol strength y of a wine sample, which is that y is equal to 8.8599x-0.5236 (y is larger than or equal to 1 and smaller than or equal to 10) is established. The alcoholic strength determination method provided by the invention is applicable to determining the wine with the alcoholic strength of 1 to 10 degree, has favorable accuracy and reliability, and is safer without the need for distilling, simple, convenient and fast to operate, less in required wine sample amount, low in price, and suitable for determination practice of the alcohol strength of the wine with the alcoholic strength of 1 to 10 degree in a winery.

Application Domain

Analysis by subjecting material to chemical reactionColor/spectral properties measurements

Technology Topic

ChemistryStandard curve +6

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Examples

Experimental program(4)

Example Embodiment

[0013] Example 1

[0014] Example 1 is the preparation of a standard curve for the determination of alcohol in the simple and rapid method for determining the alcohol content of 1 to 10 alcoholic liquors of the present invention.

[0015] (1) Manual preparation of wine samples of 1 to 10 degrees

[0016] Use the corresponding pipette to take 0.0, 1.0, 3.0, 5.0, 7.0, 9.0, 10.0ml of absolute ethanol into a 100ml volumetric flask, continuously add distilled water and repeatedly shake and stand, and finally set the volume to 100ml, and let it stand at room temperature In 10 minutes, artificial 1 to 10 alcohol samples with different alcohol levels are obtained. 0.00 is the extreme point, representing the ethanol-free system.

[0017] (2) Manual pre-processing of wine samples from 1 to 10 degrees

[0018] Use a pipette to transfer 400ul of the artificial wine sample (30.0℃) to be tested from 1 to 10 degrees to an empty ordinary ELISA plate, and first add 50ul of a mixed aqueous solution of 2% by mass urea and 3% by volume of formamide As the initial denaturant, add 50ul of the short double-stranded DNA fragment mixing system shown in SEQ ID NO: 1-2 (each fragment concentration is 10μmol/l), then the total system will be 500ul, mix well and let stand still 5min. It should be noted that 2% by mass volume fraction of urea and 3% volume by volume fraction of formamide means that 2g of urea and 3ml of formamide are dissolved in 100ml of distilled water. The short double-stranded DNA fragment is produced by Shanghai Shenggong Biological Engineering Co., Ltd. Synthetic complementary single strands are mixed in equal amounts.

[0019] At the same time, the control hole is set to 400ul artificial 1 to 10 degree wine sample plus 50ul initial denaturant plus 50ul distilled water, that is, the control is a mixed system with distilled water instead of short double-stranded DNA fragments, and each artificial 1 to 10 degree wine sample and the corresponding control are taken Parallel for three times. After the sample is prepared, place the microplate reader in the microplate reader (Multiskan GO from Thermo Company, USA).

[0020] (3) Fitting the standard curve of the alcohol content determination method

[0021] Set up the microplate reader program to measure the absorbance in sample wells and control wells at a wavelength of 260nm, then the average absorbance of the sample (average for three parallels) minus the average absorbance of the control (average for three parallels) is the sample The actual absorbance of the DNA system is shown in Table 1.

[0022] Observe the scatter points of absorbance and the corresponding alcohol content, and find that the six alcohol content points of 1.0, 3.0, 5.0, 7.0, 9.0 and 10.0 show a linear trend (the point of 0.0 alcohol content does not conform to the discarding), and the fitting is obtained The relationship between alcohol y and absorbance x is y=8.8599x-0.5236 (1≤y≤10), and R 2 =0.9998 is very close to 1 indicating a significant linear relationship, then this is the standard curve of the alcohol content determination method.

[0023] Table 1 Alcohol standard curve data

[0024] Average absorbance x

Example Embodiment

[0025] Example 2

[0026] Example 2 is the actual measurement of the alcohol content of a certain 1 to 10 alcohol by using the new method of the present invention.

[0027] (1) Pre-treatment of wine samples to be tested

[0028] Choose RIO Vodka (Rui Ao Weibo, produced by Shanghai Bacchus Wine Co., Ltd.), a commercially available cocktail, and take two bottles (RIO Vodka 1 and RIO Vodka 2) for alcohol determination. Use a pipette to transfer 400ul of the wine sample (30.0℃) to an empty common ELISA plate. First add 50ul of 2% urea and 3% formamide as the initial denaturant. Add 50ul of the short double-stranded DNA fragment mixing system shown in SEQ ID NO: 1-2 (each fragment concentration is 10μmol/l), then the total system will be 500ul, mix well and let stand for 5min.

[0029] At the same time, set the control hole to 400ul wine sample to be tested plus 50ul initial denaturant plus 50ul distilled water system (the total volume is also 500ul), each wine sample to be tested and the corresponding control are taken in parallel three times. After the sample is prepared, the enzyme-labeled plate is placed In the microplate reader (Multiskan GO of Thermo Company, USA).

[0030] (2) Determination of alcohol content of wine samples to be tested

[0031] Set up the microplate reader program to measure the absorbance in sample wells and control wells at a wavelength of 260nm, then the average absorbance of the sample (average for three parallels) minus the average absorbance of the control (average for three parallels) is the sample The actual absorbance of the DNA system is shown in Table 2. Substitute the relationship between alcohol content y and absorbance x as y=8.8599x-0.5236 (1≤y≤10), and calculate the alcohol content result (Table 2). In addition, the density bottle method was used to determine the alcohol content of the wine sample, and the results are shown in Table 2.

[0032] Table 2 Alcohol values determined by the method of the present invention and the density bottle method

[0033] cocktail

[0034] In Table 2, the deviation refers to the alcohol content measured by the density flask method as a reference, and the alcohol content measured by the method of the present invention and the difference thereof account for the percentage of the result of the alcohol meter method. It can be seen from Table 2 that the deviations of the alcohol content of the two bottles of RIO cocktail measured by the method of the present invention and the measurement result of the density bottle method are respectively 0.79% and 0.53%, and both meet the accuracy of the alcohol content determination stipulated by the national standard within 1%.

Example Embodiment

[0035] Example 3

[0036] Example 3 is to use the new method of the present invention to determine the alcohol content of commercial Qingdao high-concentration beer (produced by Tsingtao Brewery Co., Ltd.), which is 1 to 10 degrees alcohol. A total of two bottles (Tsingtao Beer 1 and Tsingtao Beer 2) are taken. . Refer to the operation in Example 2, and the results are shown in Table 3. In addition, at the same time, the alcohol content of this wine is determined by the density bottle method of the current national standard of beer.

[0037] Table 3 Alcohol values determined by the method of the present invention and the density bottle method

[0038] beer

[0039] It can be seen from Table 3 that the deviation of the alcohol content of the two bottles of Tsingtao Beer measured by the method of the present invention and the measurement result of the density bottle method are 0.59% and 0.39%, respectively, which meets the accuracy of the alcohol content determination stipulated by the national standard within 1%.

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good accuracy and reliability

Simple and quick determination method of alcoholic strength of wine with alcoholic strength of 1 to 10 degree

AI-Extracted Technical Summary

Problems solved by technology

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Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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