CN107796690A - 10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods - Google Patents
10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods Download PDFInfo
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- 235000019991 rice wine Nutrition 0.000 title claims abstract description 58
- 238000003556 assay Methods 0.000 title claims abstract description 24
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 90
- 235000014101 wine Nutrition 0.000 claims abstract description 55
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000012634 fragment Substances 0.000 claims abstract description 22
- 238000002835 absorbance Methods 0.000 claims abstract description 20
- 238000004925 denaturation Methods 0.000 claims abstract description 15
- 230000036425 denaturation Effects 0.000 claims abstract description 15
- 230000001476 alcoholic effect Effects 0.000 claims description 23
- 238000000034 method Methods 0.000 claims description 20
- 239000007787 solid Substances 0.000 claims description 19
- ZHNUHDYFZUAESO-UHFFFAOYSA-N Formamide Chemical compound NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 claims description 16
- 239000003795 chemical substances by application Substances 0.000 claims description 12
- 238000002965 ELISA Methods 0.000 claims description 8
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 7
- 239000004202 carbamide Substances 0.000 claims description 7
- 238000001514 detection method Methods 0.000 claims description 7
- 239000012153 distilled water Substances 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 abstract description 4
- 238000012360 testing method Methods 0.000 abstract description 4
- 235000019441 ethanol Nutrition 0.000 description 29
- 108020004414 DNA Proteins 0.000 description 20
- 238000010561 standard procedure Methods 0.000 description 5
- 102000053602 DNA Human genes 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000000643 oven drying Methods 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000002203 pretreatment Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 230000003694 hair properties Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- -1 small molecule carbohydrate Chemical class 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
Classifications
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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
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
Present invention absorbance difference in the case where different water cause 260nm wavelength with denaturation degrees difference in the ethanolic aqueous system of ethanol mass volume ratio using specific double chain DNA fragment, so as to set up a kind of 10 to 12 brand-new degree Non-sugar Solidity in Chinese Rice Wine assay methods.Specific double chain DNA fragment such as SEQ ID NO：Shown in 1, the relational expression between its absorbance x and ethanolic aqueous system reclaimed water and the mass volume ratio y of ethanol is y=29.197x+13.555.The non-sugar solidity assay method of the present invention is applied to the measure of 10 12 degree of yellow rice wine, has good accuracy and reliability, safer without long time drying, and easy to operate rapid, required wine sample amount is seldom, and testing cost is cheap.
Description
Technical field
The present invention relates to non-sugar solidity detection method, it is easy to be fast to spend yellow rice wine non-sugar solidity for more particularly to one kind 10 to 12
Fast assay method.
Background technology
Yellow rice wine is the characteristic wine kind in China, its bright color, give off a strong fragrance, sweetness is delicious, flavor is mellow, and containing abundant
Amino acid, small molecule carbohydrate, organic acid, vitamin and trace element etc..Yellow rice wine is because its is nutritious, alcoholic strength is low and has
There are unique flavor taste and effect and liked deeply by broad masses of the people.Non-sugar solidity is that non-in addition to sugar is waved in wine sample
The general designation of hair property component, its accounting very little in yellow rice wine is generally less than 3%.Although content is low, non-sugar solidity acts on very
Greatly, it has been largely fixed color and luster, flavour, nutrition and effect of yellow rice wine, is that one of yellow rice wine quality grade evaluation is important
Index.According to national standard GB/T 13662-2008《Yellow rice wine》Middle parameter, same type of Non-sugar Solidity in Chinese Rice Wine content is higher,
The quality of yellow rice wine is better, taste is better.
In existing yellow rice wine national standard the non-sugar solidity content in wine sample, this method are determined using 4h oven drying methods
Time-consuming, and quality is not up to constant weight after wine sample drying 4h being present, is unfavorable for quick measure, accurate measurement and batch
Detection.In addition, also have the wine sample non-sugar solidity rapid assay methods established based near infrared spectrum, but this method needs
By means of valuable equipment, and need to expend higher testing cost.Therefore need badly at present foundation accurately and reliably, it is easy quick
And economical and practical yellow rice wine non-sugar solidity assay method.
Ethanol content is alcoholic strength and total sugar content is the index that must survey of yellow rice wine, and the wine sample quality of unit volume can be held very much
Easily weighed, if can now measure the gross mass of ethanolic aqueous system in wine sample, wine sample quality subtracts ethanolic aqueous system quality and subtracted again
Total reducing sugar is gone to can be obtained by non-sugar solidity quality.The present invention is using specific double chain DNA fragment in different water and ethanol mass body
Denaturation degrees difference causes under 260nm wavelength absorbance difference in the ethanolic aqueous system of product ratio, a kind of complete so as to set up
10 to 12 new degree Non-sugar Solidity in Chinese Rice Wine assay methods, have accurately and reliably, it is easy to be quick and economical and practical the characteristics of, fit
Actual demand is detected together in brewery.
The content of the invention
It is an object of the invention to provide 10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods.
The technical solution adopted for the present invention to solve the technical problems is that 10 to 12 degree yellow rice wine non-sugar solidities are easy to be quick
The key step of assay method is as follows：
（1）In yellow rice wine Quality Detection, first determining its alcoholic strength v according to existing national standard, this must survey index, while weighed 1L wine
Sample mass M o, determine the temperature of wine sample and inquire about ethanol density p accordingly.
（2）320ul room temperatures yellow rice wine wine sample to be measured is pipetted into the ELISA Plate of sky, the agent of 40ul denaturations is added and 40ul is special
Determine double chain DNA fragment and stand 4min after fully mixing, while set control wells to add 40ul denaturations for 320ul wine samples to be measured
Agent adds 40ul distilled water, and wine sample to be measured and control make even row survey three times, ELISA Plate are placed in ELIASA after sample process is good.
（3）Set absorbance of the ELIASA program under 260nm wavelength in determination sample hole and control wells, then sample
The mean light absorbency that mean light absorbency subtracts control is the actual absorbance x of specific double chain DNA fragment in sample.X is substituted into
The mass volume ratio y of wine sample reclaimed water and ethanol can be then calculated in relational expression y=- 29.197x+13.555, then total solid in wine sample
Content M=Mo-10 × ρ × v-10 × y × v namely M=Mo-10 ×（ρ+y）× v, and then non-sugar solidity content m=M-Mt, Mt are
Total sugar content.
Wherein, step（1）Described in yellow rice wine refer to that National Standard Method measure alcoholic strength v is more than or equal to 10.0 and is less than or equal to
12.0 yellow rice wine；The alcoholic strength v is that yellow rice wine Quality Detection must survey index, and just to have originally directly to use；The 1L wine sample
Mass M o assay method is：When determining wine sample alcoholic strength by National Standard Method, the appearance equipped with wine sample is weighed after measuring 100ml wine samples
Measuring bottle weight, then it is 100ml wine sample weight again that the weight, which deducts spatial capacity bottle, and 1L wine sample mass M o are can obtain through conversion, its
Unit is g/L；The wine sample temperature is normal room temperature, it is preferable that control is between 30.0 ± 5.0 DEG C.
Wherein, step（2）Described in denaturation agent be 3% mass volume fraction urea and 3% volume volume fraction
Formamide mixed aqueous solution, i.e., per 100ml denaturation agent in the urea containing 3g and 3ml formamide；Described is specific
Double chain DNA fragment, its sequence such as SEQ ID NO：Shown in 1, concentration 16umol.
Wherein, step（3）Described in mean light absorbency refer to the arithmetic mean of instantaneous values of three parallel absorbances；It is described total
Solid content M=Mo-10 × ρ × v-10 × y × v, wherein 10 × ρ × v, which is ethanol density, is multiplied by alcoholic strength i.e. 100ml
The alcohol volume content of wine sample is multiplied by with 10, it is possible to obtains the ethanol quality in 1L wine samples；And 10 × y × v is ethanol volume
The mass volume ratio of water and ethanol is multiplied by multiplied by with 10, it is possible to obtain the water quality in 1L wine samples, then 10 ×（ρ+y）× v is real
It is exactly the quality sum of ethanolic aqueous system in 1L wine samples on border；Therefore total solids content M=Mo-10 ×（ρ+y）× v, namely 1L
It is then the total solids content in 1L wine samples that wine sample gross mass, which subtracts ethanolic aqueous system quality therein, and its unit is g/L；And then
Non-sugar solidity content subtracts total sugar content namely m=M-Mt equal to total solids content, and total sugar content Mt is the inspection of yellow rice wine quality
That surveys must survey index, and just having originally directly to use, and its unit is g/L.
The present invention screens its sequence of specific double chain DNA fragment such as SEQ ID NO：Shown in 1, and it is disclosed in 10-
Ultraviolet absorptivity constructs on this basis with the relation between the water and the mass volume ratio of ethanol in system in 12 degree of yellow rice wine
The easy rapid assay methods of 10-12 degree yellow rice wine non-sugar solidities.The non-sugar solidity assay method of the present invention is applied to 10-12
The measure of yellow rice wine is spent, there is good accuracy and reliability, easy to operate rapid, institute safer without long time drying
Need wine sample amount seldom, and testing cost is cheap, is suitable for such yellow rice wine non-sugar solidity measure practice in brewery.
Brief description of the drawings
Fig. 1 is the standard curve that the present invention 10 to 12 spends yellow rice wine non-sugar solidity measure new method, is related to specific double-stranded DNA
Absorbance x and ethanolic aqueous system reclaimed water and ethanol of the fragment under 260nm mass volume ratio y relation.Y in figure=-
29.197x+13.555 be the softwares of SPSS 23.0 fitting x and y relational expression, R2=0.9986 expression is determining for matched curve
Determine coefficient.
Embodiment
Below in conjunction with specific embodiment, the present invention is expanded on further.
The present invention is additionally added urea and the formamide mixing of optimized certain concentration in wine sample to be measured and DNA systems
Liquid, it is easy to primarily to making DNA fragmentation be in one in the environment of denaturation, so that the ethanolic aqueous system quality based on this
Assay method has higher sensitivity.The specific double chain DNA fragment such as SEQ ID NO that present invention screening and optimization obtain：1 institute
Show, it has been investigated that its water and second in the absorbance and system under 260nm in 10 to 12 alcoholic strengthes this narrow scopes
Alcohol mass ratio can realize linear fit.System of the present invention and under the conditions of, less than 10 degree of alcoholic strength is still insufficient to allow specific
Double chain DNA fragment is denatured, and more than 12 degree of alcoholic strength has been enough to make specific double chain DNA fragment be denatured completely, therefore the present invention
The standard curve of offer is only applicable to the non-sugar solidity measure of the yellow rice wine between 10-12 degree.Because second alcohol and water accounts for master in wine
Body, solid content is very low therefore it influences very little to absorbance of the system in 260nm, and uv-absorption maximum wavelength exists
260nm specific double chain DNA fragment at concentrations up to 16umol, on the other hand on the one hand can improve detection sensitivity can arrange
Except ambient interferences.In addition, it can be use up as control there is provided the wine sample for only adding denaturation agent and being not added with specific double chain DNA fragment
Influence of the background to absorbance may be eliminated.In the detection of drinks product quality, AAS is conventional means, can be surveyed
Surely colourity, methanol and specific amino acids etc. are included, therefore spectrophotometer is conventional equipment.
Embodiment 1
Embodiment 1 is that the present invention 10 to 12 spends yellow rice wine non-sugar solidity simplicity rapid assay methods reclaimed water and ethanol mass volume ratio
The making of bioassay standard curve.
（1）Artificial ethanolic aqueous system is prepared
Measure 10.00ml, 10.40ml, 10.80ml, 11.20ml, 11.60ml and 12.00ml absolute ethyl alcohol respectively with pipettor
In 6 100ml volumetric flasks, volumetric flask be placed on assay balance be correspondingly separately added into 90.00g, 89.60g, 89.20g,
88.80g, 88.40g and 88.00g distilled water, cover tightly capacity bottle cap and overturn fully mixing repeatedly, be stored at room temperature 10min, it is final to obtain
It is quality/ethanol volume of water to different water and ethanol mass volume ratio（It is corresponding be respectively 9.000,8.615,8.259,
7.929th, 7.621 and 7.333）Artificial ethanolic aqueous system.
（2）Artificial ethanolic aqueous system pre-treatment
320ul room temperatures artificial ethanolic aqueous system to be measured is pipetted respectively with liquid-transfering gun into the common ELISA Plate of sky, first adds 3% matter
The urea of volume fraction and the formamide mixed aqueous solution 40ul of 3% volume volume fraction are measured as denaturation agent, add as
SEQ ID NO：Specific double chain DNA fragment shown in 1（16umol）40ul, then total system is 400ul, is stood after fully mixing
4min.It should be noted that the urea of 3% mass volume fraction and the formamide of 3% volume volume fraction refer to 100ml distilled water
In dissolved with 3g urea and 3ml formamide, specific double chain DNA fragment is synthesized by Shanghai Sheng Gong bioengineering Co., Ltd
Complementary single strand mixed in equal amounts form.
Control wells are set to add the above-mentioned denaturation agent of 40ul to add 40ul distilled water for the artificial ethanolic aqueous systems of 320ul simultaneously,
I.e. control is to substitute specific double chain DNA fragment with distilled water, and each artificial ethanolic aqueous system and corresponding control make even row three times,
ELISA Plate is placed in ELIASA after sample preparation is good（The Multiskan GO of Thermo companies of the U.S.）In.
（3）Artificial ethanolic aqueous system reclaimed water and ethanol mass volume ratio assay method standard curve
Absorbance of the ELIASA program under 260nm wavelength in determination sample hole and control wells is set, then the average suction of sample
Luminosity（Three parallel to be averaged）Subtract the mean light absorbency of control（Three parallel to be averaged）Specific double-stranded DNA as in sample
The actual absorbance x of fragment, as shown in table 1.
Absorbance x and corresponding ethanolic aqueous system reclaimed water and ethanol mass volume ratio y scatterplot are observed, discovery is in
Existing linear trend（Fig. 1）, it is y=- 29.197x+13.555 to be fitted to obtain relational expression, and R2=0.9986 shows linear relationship
Well, then this is ethanolic aqueous system reclaimed water and ethanol mass volume ratio assay method standard curve.
The ethanolic aqueous system reclaimed water of table 1 and ethanol mass volume ratio standard curve data
| Mean light absorbency x | Water and ethanol mass volume ratio y |
| 0.156 | 9.000 |
| 0.170 | 8.615 |
| 0.180 | 8.259 |
| 0.194 | 7.929 |
| 0.203 | 7.621 |
| 0.213 | 7.333 |
Embodiment 2
Embodiment 2 is the reality that the non-sugar solidity for the yellow rice wine wine sample for being 10 to mark alcoholic strength using new method of the present invention is carried out
Measure.
（1）Select commercially available Fang County Swim juice yellow rice wine（Shiyan valley head liquid yellow rice wine Co., Ltd produces）, body mark alcoholic strength is 10
Degree.Need to determine alcoholic strength again for the sake of accurate, therefore this yellow rice wine is determined using alcoholic strength meter method based on existing yellow rice wine national standard
Alcoholic strength is 10.1.Through weighing and converting, the quality of this yellow rice wine of 1L is 1027.462g.Wine sample temperature is 30 DEG C, is existed through looking into ethanol
Density at a temperature of this is 0.781g/ml.
（2）Yellow rice wine wine sample pre-treatment to be measured
320ul yellow rice wine wine samples to be measured are pipetted into the common ELISA Plate of sky with liquid-transfering gun, first add the urine of 3% mass volume fraction
Element and the formamide mixed aqueous solution 40ul of 3% volume volume fraction are added such as SEQ ID NO as denaturation agent：1 institute
The specific double chain DNA fragment shown（16umol）40ul, then total system is 400ul, and 4min is stood after fully mixing.
Control wells are set to add 40ul denaturation agent to add 40ul to distill aqueous systems for 320ul yellow rice wine wine samples to be measured simultaneously（Always
Volume is also 400ul）, each yellow rice wine wine sample to be measured and corresponding control make even row three times, put ELISA Plate after sample preparation is good
In ELIASA（The Multiskan GO of Thermo companies of the U.S.）In.
（3）Yellow rice wine wine sample non-sugar solidity measure to be measured
Absorbance of the ELIASA program under 260nm wavelength in determination sample hole and control wells is set, then the average suction of sample
Luminosity（Three parallel to be averaged）Subtract the mean light absorbency of control（Three parallel to be averaged）Specific double-stranded DNA as in sample
Actual absorbance x=0.163 ± 0.001 of fragment, the relation substituted between water and ethanol mass volume ratio y and absorbance x
Formula is y=- 29.197x+13.555, calculates water and ethanol mass volume ratio y=8.796, as shown in table 2.Then total solid in wine sample
Thing content M=Mo-10 × ρ × v+10 × y × v is M=1027.462-10 × 0.781 × 10.1-10 × 8.796 × 10.1, through meter
Calculate as 60.185g/L.
The inventive method empirical value of table 2
| Yellow rice wine | Alcoholic strength | Ethanol density/g × ml-1 | 1L wine sample quality/g | The mass volume ratio of water and ethanol | Total solid/g × L of the inventive method measure-1 | Total solid/g × L of National Standard Method measure-1 | Deviation % |
| Swim juice yellow rice wine | 10.1 | 0.781 | 1027.462 | 8.796 | 60.185 | 59.869 | 0.53 |
In addition, it is 59.869g/L to determine total solids content according to the 4h oven drying methods of national standard.If the total solid measured with National Standard Method
On the basis of thing content, then the inventive method measures numerical value and its deviation are 0.53%（Total solid that the inventive method measures with
The difference of national standard measured value accounts for the percentage of national standard measured value）, meet the measurement accuracy of total solid.Finally again by total solid
Content subtracts the content that total sugar content obtains non-sugar solidity.Due to being to take commercially available finished wine to survey, its total sugar content is only marked
Scope is noted, its total reducing sugar is 15.1-40g/L if half-dry type yellow rice wine.In factory in practice, there is the specific measure numerical value of total reducing sugar, can
Directly bring use.
Embodiment 3
Embodiment 3 is the reality that the non-sugar solidity for the yellow rice wine wine sample for being 11 to mark alcoholic strength using new method of the present invention is carried out
Measure.
Select commercially available Hei Tanxie villages high-grade Shaoxing wine yellow rice wine（Shaanxi Qinyang Changsheng Winery Co., Ltd. produces）, body mark alcoholic strength be
11 degree.Need to determine alcoholic strength again for the sake of accurate, therefore this yellow rice wine is determined using alcoholic strength meter method based on existing yellow rice wine national standard
Alcoholic strength be 11.2.Through weighing and converting, the quality of this yellow rice wine of 1L is 1070.548g.Wine sample temperature is 30 DEG C, through looking into ethanol
Density at this temperature is 0.781g/ml.
It is measured with reference to embodiment 2, actual absorbance x=0.188 ± 0.001, correlation values refer to table 3.If with national standard
On the basis of the total solids content that method measures, then the inventive method measures numerical value and its deviation are 0.20%, meet total solid
Measurement accuracy.
The inventive method empirical value of table 3
| Yellow rice wine | Alcoholic strength | Ethanol density/g × ml-1 | 1L wine sample quality/g | The mass volume ratio of water and ethanol | Total solid/g × L of the inventive method measure-1 | Total solid/g × L of National Standard Method measure-1 | Deviation % |
| Thank to village's yellow rice wine | 11.2 | 0.781 | 1070.548 | 8.066 | 79.684 | 79.523 | 0.20 |
In summary, non-sugar solidity assay method of the invention is applied to the measure of 10-12 degree yellow rice wine, has well accurate
Property and reliability；It is safer without long time drying compared with national standard 4h oven drying methods, operate 10min and simple, convenient and rapid；In addition
Required wine sample amount is seldom, and testing cost is very cheap less than 1 yuan/sample, is suitable for such yellow rice wine non-sugar solidity in brewery and surveys
Fixed practice.
Sequence table
<110>Chen Rukai
<120>10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 10
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 1
catgcaacga 10
Claims (3)
1.10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods, it is characterised in that the yellow rice wine non-sugar solidity is easy
Rapid assay methods mainly comprise the following steps：
（1）In yellow rice wine Quality Detection, first determining its alcoholic strength v according to existing national standard, this must survey index, and v needs to meet
10.0≤v≤12.0, while weighed 1L wine sample mass M o, determine the temperature of wine sample and inquire about ethanol density p accordingly；
（2）320ul room temperatures yellow rice wine wine sample to be measured is pipetted into the ELISA Plate of sky, the agent of 40ul denaturations is added and 40ul is specific double
Chain DNA fragment simultaneously fully stands 4min after mixing, while sets control wells to add 40ul denaturation agent to add for 320ul wine samples to be measured
40ul distilled water, wine sample to be measured and control make even row survey three times, and ELISA Plate is placed in ELIASA after sample process is good；
（3）Set absorbance of the ELIASA program under 260nm wavelength in determination sample hole and control wells, then sample is averaged
The mean light absorbency that absorbance subtracts control is the actual absorbance x of specific double chain DNA fragment in sample, and x is substituted into relation
The mass volume ratio y of wine sample reclaimed water and ethanol can be then calculated in formula y=- 29.197x+13.555, then total solids content in wine sample
M=Mo-10×（ρ+y）× v, and then non-sugar solidity content m=M-Mt, Mt are total sugar content.
2. 10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods according to claim 1, it is characterised in that step
Suddenly（2）Described in specific double chain DNA fragment, its sequence such as SEQ ID NO：Shown in 1, concentration 16umol.
3. 10 to 12 degree yellow rice wine non-sugar solidity simplicity rapid assay methods according to claim 1, it is characterised in that step
Suddenly（2）Described in denaturation agent be 3% mass volume fraction urea and 3% volume volume fraction formamide mixing it is water-soluble
Liquid.
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| CN101216419A (en) * | 2007-12-28 | 2008-07-09 | 陈郁 | Method for quickly detecting yellow wine quality index |
| CN102759607A (en) * | 2012-07-23 | 2012-10-31 | 福州大学 | Method for rapidly determining non-sugar solid in yellow rice wine |
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2017
- 2017-11-06 CN CN201711074959.XA patent/CN107796690A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101216419A (en) * | 2007-12-28 | 2008-07-09 | 陈郁 | Method for quickly detecting yellow wine quality index |
| CN102759607A (en) * | 2012-07-23 | 2012-10-31 | 福州大学 | Method for rapidly determining non-sugar solid in yellow rice wine |
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| Title |
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