CN107505407A - It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content - Google Patents
It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content Download PDFInfo
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- CN107505407A CN107505407A CN201710592118.1A CN201710592118A CN107505407A CN 107505407 A CN107505407 A CN 107505407A CN 201710592118 A CN201710592118 A CN 201710592118A CN 107505407 A CN107505407 A CN 107505407A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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Abstract
Especially a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content the present invention relates to white wine composition detection technical field, this method includes dilute fermented grain, extracting, centrifuging, diluting again, filtering, and finally goes up liquid chromatogram and is analyzed.By the optimization of the diluting condition to the dosage of fermented grain, the amount of being diluted with water, leaching mode, centrifugal condition and centrifuged supernatant and liquid phase analysis condition etc., lactic acid in the fermented grain based on liquid-phase chromatographic analysis technology, acetic acid qualitativing quantitative measuring method are established.This method has the advantages of quick, easy, high sensitivity, is determined simultaneously for lactic acid, acetic acid in white wine fermented grain and provides an approach.
Description
Technical field
It is especially a kind of to determine lactic acid, acetic acid in white wine fermented grain simultaneously the present invention relates to white wine composition detection technical field
The method of content.
Background technology
White wine has the developing history of thousands of years, is indispensable in China's cultural continuity as the distinctive wine kind in China
A part, taken the course of its own in world's strong drink class product, be one of major class Spirit of the world six.
Lactic acid, acetic acid are organic acids important in white wine, the number of its content not only the taste to white wine and rear taste have compared with
It is big to influence, and whether in evolution normal, therefore, detect lactic acid, second in white wine and fermented grain if directly reflecting fermented grain
The content of acid all has very important significance to vast wine brewing worker and hook tune worker, while also has weight to improving product quality
Want directive function.
In the prior art, mainly there is gas phase direct injected chromatogram for the detection method of lactic acid in white wine, acetic acid content
Method, esterification treatment indirect sample introduction chromatography, colorimetric method, physico-chemical analysis and chromatography synthesis, high performance liquid chromatography.Such as Feng
The lactic acid and acetic acid (Feng Xiangdong high effective liquid chromatography for measuring white wine in high performance liquid chromatography method measure white wine are employed eastwards
Middle lactic acid and acetic acid content [J] brew wine science and technology, 2009 (5):115-116.).However, fermented grain is different from the matrix of white wine, fermented grain
Matrix in sample is extremely complex, containing substantial amounts of protein, carbohydrate, lipid, pigment and many small molecular organic acids etc., this
A little matrix can all influence the extraction and detection of lactic acid, acetic acid, and appearance is unstable when detecting the chromatogram of lactic acid, acetic acid and is mixed with miscellaneous
Peak, its appearance can not be judged well.
At present by liquid chromatogram be used for white wine fermented grain in lactic acid, acetic acid measure have not been reported, also do not occur clearly
Lactic acid, the detection method of acetic acid in a kind of white wine fermented grain that can be simultaneously quick, accurate, easy.
The content of the invention
In order to solve the above technical problems, present invention offer is a kind of while determines lactic acid in white wine fermented grain, the side of acetic acid content
Method, to ensure properly reasonably to control lactic acid, acetic acid content during brewed spirit.
The present invention determines lactic acid in white wine fermented grain simultaneously, the method for acetic acid content includes fermented grain dilution, extraction, centrifugation, mistake
Filter and liquid-phase chromatographic analysis, described liquid-phase chromatographic analysis are to carry out qualitative and quantitative analysis to lactic acid, acetic acid.
It is achieved particular by following technical scheme：
It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content, it is characterised in that to comprise the following steps：
1) fermented grain sample dilutes；
2) fermented grain sample extracts；
3) fermented grain leaching liquor centrifuges；
4) the centrifuged supernatant dilution of fermented grain leaching liquor；
5) supernatant liquid filtering after diluting；
6) liquid-phase chromatographic analysis.
Wherein, in step 1), the dilution of described fermented grain sample is according to quality by fermented grain sample and ultra-pure water：Volume ratio is
1:(1~5) is mixed.As preferred embodiment, by fermented grain sample and water according to quality：Volume ratio is 1:2 are mixed
Close.
Wherein, in step 2), the extraction of described fermented grain sample is by the way of vortex oscillation extraction.Specifically, whirlpool
The time of rotation vibration extraction is 5~20min；Preferably, the vortex oscillation extraction time 10min of described fermented grain sample.Through examination
Checking is real, and after vortex oscillation extraction 10min, extraction effect is without significant difference.
Wherein, in step 3), described fermented grain leaching liquor centrifugal condition is 4 DEG C, rotating speed 10000-13000rpm of temperature, when
Between 5~20min.As preferred embodiment, described fermented grain leaching liquor centrifugal condition is 4 DEG C, rotating speed 12000rpm of temperature,
Time 10min.
Wherein, in step 4), the dilution of described centrifuged supernatant is according to volume by the supernatant after centrifugation and ultra-pure water
Than for 1:(10~60) are mixed；As preferred embodiment, supernatant is 1 according to volume ratio with ultra-pure water：(20~
40)；According to volume ratio it is 1 by supernatant and ultra-pure water as preferred embodiment:30 are mixed.
Wherein, in step 6), described liquid phase chromatogram condition is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent
ZORBX RRHT SB-C18 chromatographic columns；0.10%~0.20% phosphoric acid solution is as mobile phase A, and 100% methanol is as mobile phase
B, gradient elution；The column temperature of chromatographic column is 20~50 DEG C, 210~280nm of Detection wavelength, 0.1~1.0mL/min of flow velocity, sample introduction
Measure as 5~20 μ L.
Condition of gradient elution is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
As preferred embodiment, during liquid-phase chromatographic analysis, using 0.12% phosphoric acid solution as mobile phase A, 100% first
Alcohol is 30 DEG C, Detection wavelength 214nm as Mobile phase B, gradient elution column temperature, and flow velocity 0.2 is mL/min, and sample size is 5 μ L.
In addition, the standard curve preparation method in the present invention is：Lactic acid, the acetic acid of finite concentration gradient are prepared with ultra-pure water
The aqueous solution, using lactic acid, acetic acid peak area as ordinate, lactic acid, acetic acid concentration are the standard curve that abscissa is drawn.Lactic acid, second
Acid has good linear relationship (R in surveyed concentration range2>0.99), lactic acid, acetic acid test limit (LOD) are in fermented grain
300.00mg/Kg, to the relative standard deviation of same sample replication 3 times all within 10%, to the breast of 5 concentration knowns
Acid, the rate of recovery of acetic acid solution are 88.0%~107.0%.Show the present invention method can accurately, reliably determine wine
Lactic acid, acetic acid content in unstrained spirits.
Beneficial effects of the present invention are embodied in：
The present invention is by fermented grain dosage, amount of water, extraction, centrifugation, diluting condition and liquid-phase chromatographic analysis condition etc.
Optimization, establish lactic acid in the fermented grain based on liquid-phase chromatographic analysis technology, acetic acid qualitativing quantitative measuring method.This method has
Quickly, the advantages of simplicity, high sensitivity, determined simultaneously for lactic acid, acetic acid in white wine fermented grain and provide an approach.
Lactic acid, acetic acid analysis method in existing fermented grain are mainly that acid number titration method surveys total acid.Acid number titration method is exclusive
Property is poor, error is big, cannot be distinguished by the species and content of organic acid in acidulant.It is of the invention compared to existing analysis method
Advantage is that large batch of sample pre-treatments can be carried out simultaneously, can effectively distinguish the lactic acid in fermented grain, acetic acid and be determined
Amount, sensitivity reach ppm ranks, and whole detection process only needs 13min and can persistently detected.The method can be analyzed simultaneously
Go out the content of lactic acid in fermented grain, acetic acid, detection is quick, the degree of accuracy is high, detectable concentration scope is wide.
The analysis method of the present invention is applied to the fermented grain of any fermentation stage, such as goes out cellar for storing things or goes out rice steamer fermented grain.
Brief description of the drawings
Fig. 1 is to determine lactic acid in white wine fermented grain, the high performance liquid chromatography of acetic acid content method simultaneously in the embodiment of the present invention 1
Figure.
Fig. 2 is that the present invention determines the different dilution extractions of lactic acid in white wine fermented grain, the fermented grain sample of acetic acid content method simultaneously
Influence of the ratio to analytical effect.
Fig. 3 is that the present invention determines lactic acid in white wine fermented grain simultaneously, the different leaching modes of acetic acid content method are imitated to analysis
The influence of fruit.
Fig. 4 is that the present invention determines lactic acid in white wine fermented grain, the different vortex oscillation times of acetic acid content method to dividing simultaneously
Analyse the influence of effect.
Fig. 5 is that the present invention determines lactic acid in white wine fermented grain simultaneously, the different centrifugal conditions of acetic acid content method are imitated to analysis
The influence of fruit.
Fig. 6 be the present invention simultaneously determine lactic acid in white wine fermented grain, acetic acid content method different centrifugations after supernatant
Influence of the diluting condition to analytical effect.
Fig. 7 is that the present invention determines lactic acid in white wine fermented grain, the different gradients of acetic acid content method to analysis simultaneously
The influence of effect.
Embodiment
Limited technical scheme is further, but required with specific embodiment below in conjunction with the accompanying drawings
The scope of protection is not only limited to made description.
Material and facility selects explanation in the embodiment of the present invention：
Lactic acid, acetic acid, phosphoric acid are purchased from Sigma Co., USA；Ethanol and methanol (chromatographically pure) are purchased from TEDIA companies of the U.S..
Ultra Performance Liquid Chromatography instrument (Agilent 1290Series), 1290FLD fluorescence detectors, assay balance
(METTLER TOLEDO XP205), pure water meter (Ai Kepu RM-220), centrifuge (FDAC), Talboys digital displays are more
Tubular type vortex mixer (U.S.), KQ-500DA Kunshans ultrasonic instrument (Kunshan Shu Mei).
Embodiment 1 is a kind of while determines lactic acid in white wine fermented grain, the method for acetic acid content
Its step is including in the dilution of fermented grain sample, the extraction of fermented grain sample, the centrifugation of fermented grain leaching liquor, the centrifugation of fermented grain leaching liquor
The dilution of clear liquid, the filtering of dilution and liquid-phase chromatographic analysis.
Specifically, by fermented grain sample and water according to quality：Volume ratio is 1:1 is mixed, and then vortex oscillation extracts
10min.After extraction terminates, 20min is centrifuged under conditions of 4 DEG C of temperature, rotating speed 11000rpm.Centrifugation terminates, after taking centrifugation
Supernatant is 1 according to volume ratio with ultra-pure water:30 are mixed.Finally carry out liquid-phase chromatographic analysis.
The condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT
SB-C18 chromatographic columns；0.12% phosphoric acid solution carries out gradient elution, gradient as mobile phase A, 100% methanol as Mobile phase B
Elution requirement is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 5 μ L.
Embodiment 2 is a kind of while determines lactic acid in white wine fermented grain, the method for acetic acid content
Its step is including in the dilution of fermented grain sample, the extraction of fermented grain sample, the centrifugation of fermented grain leaching liquor, the centrifugation of fermented grain leaching liquor
The dilution of clear liquid, the filtering of dilution and liquid-phase chromatographic analysis.
Specifically, by fermented grain sample and water according to quality：Volume ratio is 1:2 carry out mixing vortex oscillation extraction 15min.Leaching
After proposing end, 10min is centrifuged under conditions of 4 DEG C of temperature, rotating speed 12000rpm.Centrifugation terminates, take centrifugation after supernatant with
Ultra-pure water is 1 according to volume ratio:30 are mixed.Finally carry out liquid-phase chromatographic analysis.
The condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT
SB-C18 chromatographic columns；0.14% phosphoric acid solution carries out gradient elution, gradient as mobile phase A, 100% methanol as Mobile phase B
Elution requirement is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 10 μ L.
Embodiment 3 is a kind of while determines lactic acid in white wine fermented grain, the method for acetic acid content
Its step is including in the dilution of fermented grain sample, the extraction of fermented grain sample, the centrifugation of fermented grain leaching liquor, the centrifugation of fermented grain leaching liquor
The dilution of clear liquid, the filtering of dilution and liquid-phase chromatographic analysis.
Specifically, by fermented grain sample and water according to quality：Volume ratio is 1:2 carry out mixing vortex oscillation extraction 10min.Leaching
After proposing end, 10min is centrifuged under conditions of 4 DEG C of temperature, rotating speed 13000rpm.Centrifugation terminates, take centrifugation after supernatant with
Ultra-pure water is 1 according to volume ratio:30 are mixed.Finally carry out liquid-phase chromatographic analysis.
The condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT
SB-C18 chromatographic columns；0.18% phosphoric acid solution carries out gradient elution, gradient as mobile phase A, 100% methanol as Mobile phase B
Elution requirement is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 5 μ L.
Influence of the different dilution extraction ratio of the fermented grain sample of embodiment 4 to analytical effect
Its step is including in the dilution of fermented grain sample, the extraction of fermented grain sample, the centrifugation of fermented grain leaching liquor, the centrifugation of fermented grain leaching liquor
The dilution of clear liquid, the filtering of dilution and liquid-phase chromatographic analysis.
Specifically, take 2g, 4g to go out to store and fermented grain and go out rice steamer fermented grain respectively, by its with water according to quality：Volume ratio is 1：(1~
5) mixed respectively, fermented grain dosage and amount of water are optimized, remaining condition is same as Example 1.
As a result the volume ratio of display such as table 2 and table 3, fermented grain and water is 1:Symmetrical peak shape, RSD are can obtain under (1~5)
Also in tolerance interval, it is contemplated that the inhomogeneity of solid-state sample, and RSD differences are smaller when sample is 2g and 4g, most
Determine that fermented grain dosage is 4g eventually.In terms of the consideration of dilution factor, it is also necessary to take into account operation and lactic acid, the acetic acid marks such as follow-up centrifugation
The range of linearity of directrix curve.To sum up, as preferred embodiment, it is 2 times of fermented grain quality to select amount of water, i.e. 4g fermented grains add
8mL ultra-pure waters are diluted extraction.
The lactic acid of table 2
The acetic acid of table 3
Influence of the different leaching mode of embodiment 5 to analytical effect
Its step is including in the dilution of fermented grain sample, the extraction of fermented grain sample, the centrifugation of fermented grain leaching liquor, the centrifugation of fermented grain leaching liquor
The dilution of clear liquid, the filtering of dilution and liquid-phase chromatographic analysis.
Specifically, in the case of time identical (15min), static, ultrasonic (frequency 40KHz, power is respectively adopted
500W, 25 DEG C of constant temperature) and 3 kinds of leach extraction methods progress of vortex oscillation (rotating speed 2500rpm, turnover diameter 3.6mm, power 75W)
Fermented grain sample extracts, and remaining condition is same as Example 2.
Influence static, ultrasonic and that 3 kinds of leach extraction methods of vortex oscillation are to analysis result, experimental result is as shown in figure 3, adopt
It is maximum with vortex vibration peak area, it is best to show that vortex oscillation mixes leaching mode effect.
Influence of the different vortex oscillation time of embodiment 6 to analytical effect
It is determined that leaching mode with vortex oscillation it is best on the basis of, investigated the vortex oscillation time again to extracting effect
Influence, except vortex oscillation, remaining step are same as Example 2.Experimental result as shown in figure 4, vortex oscillation after 10min,
The peak area no significant difference of acetic acid and ethanol, it is optimal to improve analysis efficiency, vibration 10min or so.
Influence of the different centrifugal condition of embodiment 7 to analytical effect
In the case where other conditions are same as Example 2, peak face of the different centrifugation times to lactic acid and acetic acid has been investigated
Long-pending influence.Experimental result is as shown in Figure 5, the results showed that, centrifugation time is in 10~20min, to the peak face of lactic acid and acetic acid
Product is without much influences, and in order to save experimentation, centrifugation time may be selected within 10min.
Influence of the supernatant diluting condition to analytical effect after the different centrifugation of embodiment 8
Specifically, by fermented grain sample and water according to quality：Volume ratio is 1:1 is mixed, and then vortex oscillation extracts
10min.After extraction terminates, 10min is centrifuged under conditions of 4 DEG C of temperature, rotating speed 12000rpm.Centrifugation terminates, after taking centrifugation
Supernatant is 1 according to volume ratio with ultra-pure water:(10~60) are mixed.Finally carry out liquid-phase chromatographic analysis.
The condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT
SB-C18 chromatographic columns；0.12% phosphoric acid solution carries out gradient elution, gradient as mobile phase A, 100% methanol as Mobile phase B
Elution requirement is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 5 μ L.
Experimental result is as shown in Figure 6, the results showed that, supernatant after centrifugation is 1 according to volume ratio with ultra-pure water:(20~
40) when being mixed, the synthesis such as peak area, peak type, separating degree and rate of recovery of lactic acid and acetic acid sees it is relatively good, Neng Gouyou
One good analytical effect, wherein 1:30 effects are best, and the peak area of acetic acid and lactic acid all reaches maximum.Supernatant with it is ultrapure
Water volume ratio hour, lactic acid, the separating degree of acetic acid and peak shape are poor；When supernatant and ultrapure water volume ratio are big, lactic acid, acetic acid
The rate of recovery is relatively low.
Influence of the different gradient of embodiment 9 to analytical effect
Specifically, by fermented grain sample and water according to quality：Volume ratio is 1:2 are mixed, and then vortex oscillation extracts
10min.After extraction terminates, 10min is centrifuged under conditions of 4 DEG C of temperature, rotating speed 12000rpm.Centrifugation terminates, after taking centrifugation
Supernatant is 1 according to volume ratio with ultra-pure water:30 are mixed.Finally carry out liquid-phase chromatographic analysis.
The condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT
SB-C18 chromatographic columns；0.10,0.12% is respectively adopted, 0.20% phosphoric acid solution is as mobile phase A, and 100% methanol is as flowing
Phase B, carries out gradient elution, and condition of gradient elution is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 5 μ L.
Experimental result is as shown in Figure 7, the results showed that, when phosphoric acid solution concentration is 0.12% in mobile phase A, lactic acid and acetic acid
Analytical effect it is best.When phosphoric acid solution concentration is relatively low or higher, peak shape, peak area and the guarantor of lactic acid and acetic acid can be all influenceed
Stay the time.
The linear and detection line of the analysis method of embodiment 10
The analysis method of organic acid mainly has：Acid-base titration, thin-layered chromatography, gas chromatography, the chromatography of ions and
High performance liquid chromatography etc..High performance liquid chromatography (HPLC) is a kind of analytical technology for integrating separation and determining, sensitive
Degree is high, and accuracy is good, and specificity is good, and sample pretreatment is simple, can a variety of organic acids of Simultaneous Determination.But in the prior art also
Do not go out to be now able to using high performance liquid chromatography while determine the lactic acid in fermented grain, the pertinent literature or report of acetic acid.Pass through access
Data of literatures and test of many times, it is determined that the condition of liquid-phase chromatographic analysis step is：Chromatographic column uses 4.6 × 50mm, and 1.8
μm Agilent ZORBX RRHT SB-C18 chromatographic columns；0.12% phosphoric acid solution is as mobile phase A, and 100% methanol is as flowing
Phase B, carries out gradient elution, and condition of gradient elution is shown in Table 1.
The mobile phase linear elution condition of table 1
| Time/minute | Mobile phase A/% | Mobile phase B/% |
| 0 | 98 | 2 |
| 4 | 0 | 100 |
| 11 | 98 | 2 |
| 13 | 0 | 100 |
Wherein, 30 DEG C of column temperature, Detection wavelength 214nm, flow velocity 0.2mL/min, sample size are 5~10 μ L.
Further to confirm the reliability and applicability of the inventive method, inventor is evaluated method：With ultrapure
Water prepares serial lactic acid, acetic acid standard liquid, is carried out according to above-mentioned condition, using lactic acid, acetic acid peak area as ordinate, lactic acid,
Acetic acid concentration is abscissa, draws standard curve.Wherein, the concentration of standard solution gradient of lactic acid and acetic acid is 0.25,0.50,
1.00,2.00,4.00%v/v.
Experimental result is shown in Table 4, and this method has good linear relationship (R in measurement range2>0.99).With signal to noise ratio
It is standard equal to 3, the test limit (LOD) of lactic acid, acetic acid under the conditions of experiment with computing.
Table 4
| Material title | R2 | n | The range of linearity (mg/L) | Linear equation | LOD(mg/L) |
| Lactic acid | 0.999 | 6 | 10-1000 | Y=0.5303x-0.1921 | 10.0 |
| Acetic acid | 0.999 | 6 | 10-1000 | Y=0.3285x+27.205 | 15.0 |
Determined under above-mentioned analysis condition, calculate the rate of recovery and relative standard deviation (RSD) of this method, the results are shown in Table 5.
Table 5
The result of table 4 and table 5 shows that this method is accurate, reliable, can meet that lactic acid in fermented grain, acetic acid content are quickly surveyed
Fixed demand.
Detection method of the present invention can be examined suitable for any white wine fermented grain while lactic acid, acetic acid content
Survey.
It is important to point out that, the technical scheme that above example and experimental example are only limitted to invention is further explained herein
Stating and illustrate so that those skilled in the art can more accurately understand the invention thinking and operation scheme of the present invention, and
It is not the further limitation to the present invention, non-protruding essential characteristics that those skilled in the art make on this basis and non-
The improvement of marked improvement, belong to the protection category of the present invention.
Claims (10)
- It is 1. a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content, it is characterised in that to comprise the following steps：1) fermented grain sample dilutes；2) fermented grain sample extracts；3) fermented grain leaching liquor centrifuges；4) centrifuged supernatant dilutes；5) dilution filters；6) liquid-phase chromatographic analysis；Wherein, in step 4), it by supernatant and ultra-pure water according to volume ratio is 1 that the dilution of described centrifuged supernatant, which is,:10~60 Mixed；Wherein, in step 6), chromatogram is using 0.10%~0.20% phosphoric acid solution as mobile phase A, and 100% methanol is as mobile phase B。
- 2. as claimed in claim 1 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 1) in, the dilution of described fermented grain sample is according to quality by fermented grain sample and ultra-pure water：Volume ratio is 1:1~5 is mixed；It is excellent Selection of land, by fermented grain sample and ultra-pure water according to quality：Volume ratio is 1:2 are mixed.
- 3. as claimed in claim 1 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 2) in, the extraction of described fermented grain sample is to vibrate to extract by vortex.
- 4. as claimed in claim 3 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that is vortexed Vibration extraction time is 5~20min；Preferably, vortex oscillation extraction time is 10min.
- 5. as claimed in claim 1 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 3) in, described fermented grain leaching liquor centrifugal condition is 4 DEG C, 10000~13000rpm of rotating speed, 5~20min of time of temperature.
- 6. as claimed in claim 5 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that fermented grain Leaching liquor centrifugal condition is 4 DEG C, rotating speed 12000rpm, time 10min of temperature.
- 7. as claimed in claim 1 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 4) in, it by supernatant and ultra-pure water according to volume ratio is 1 that the dilution of described centrifuged supernatant, which is,:20~40.
- 8. as claimed in claim 7 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 4) in, it by supernatant and ultra-pure water according to volume ratio is 1 that the dilution of described centrifuged supernatant, which is,:30 are mixed.
- 9. as claimed in claim 1 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that step 6) in, described liquid phase chromatogram condition is：Chromatographic column uses 4.6 × 50mm, 1.8 μm of Agilent ZORBX RRHT SB-C18 Chromatographic column；0.12% phosphoric acid solution is as mobile phase A, and 100% methanol is as Mobile phase B, gradient elution；The column temperature of chromatographic column is 20~50 DEG C, 210~280nm of Detection wavelength, 0.1~1.0mL/min of flow velocity, sample size is 5~20 μ L.
- 10. as claimed in claim 9 determine lactic acid in white wine fermented grain, the method for acetic acid content simultaneously, it is characterised in that described Condition of gradient elution be：
Time/minute Mobile phase A/% Mobile phase B/% 0 98 2 4 0 100 11 98 2 13 0 100 Column temperature is 30 DEG C, Detection wavelength 214nm, flow velocity 0.2mL/min, and sample size is 5 μ L.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201710592118.1A CN107505407A (en) | 2017-07-19 | 2017-07-19 | It is a kind of while determine lactic acid in white wine fermented grain, the method for acetic acid content |
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| CN109239223A (en) * | 2018-10-12 | 2019-01-18 | 江苏力凡胶囊有限公司 | A kind of test method for Capsules acetic acid |
| CN110426481A (en) * | 2019-09-16 | 2019-11-08 | 亚太星原农牧科技海安有限公司 | The method of lactic acid content and the measurement extracting method of prepare liquid in a kind of measurement fermented bean dregs |
| CN110441256A (en) * | 2019-08-16 | 2019-11-12 | 四川省绵阳市丰谷酒业有限责任公司 | The near-infrared spectral analytical method of fermented grain |
| CN112063573A (en) * | 2020-08-27 | 2020-12-11 | 贵州茅台酒股份有限公司 | Formula and culture method of lactobacillus isolation medium |
| CN114062561A (en) * | 2021-11-29 | 2022-02-18 | 江南大学 | Method for simultaneously detecting lactic acid, acetic acid and derivative ester thereof in fen-flavor fermented grains based on high performance liquid chromatography technology |
| CN114107434A (en) * | 2020-08-31 | 2022-03-01 | 贵州茅台酒股份有限公司 | Method for evaluating lactic acid production capacity of Daqu in high throughput |
| CN114965731A (en) * | 2022-03-22 | 2022-08-30 | 贵州茅台酒股份有限公司 | Method for detecting pectin in fermented grains by using ultra-high performance liquid chromatography-quadrupole electrostatic field orbit trap |
| CN115032298A (en) * | 2022-05-26 | 2022-09-09 | 宜宾五粮液股份有限公司 | Method and device for detecting content of yellow water flavor substances in fermentation process in cellar |
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| CN109239223A (en) * | 2018-10-12 | 2019-01-18 | 江苏力凡胶囊有限公司 | A kind of test method for Capsules acetic acid |
| CN110441256A (en) * | 2019-08-16 | 2019-11-12 | 四川省绵阳市丰谷酒业有限责任公司 | The near-infrared spectral analytical method of fermented grain |
| CN110426481A (en) * | 2019-09-16 | 2019-11-08 | 亚太星原农牧科技海安有限公司 | The method of lactic acid content and the measurement extracting method of prepare liquid in a kind of measurement fermented bean dregs |
| CN112063573A (en) * | 2020-08-27 | 2020-12-11 | 贵州茅台酒股份有限公司 | Formula and culture method of lactobacillus isolation medium |
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| CN114107434A (en) * | 2020-08-31 | 2022-03-01 | 贵州茅台酒股份有限公司 | Method for evaluating lactic acid production capacity of Daqu in high throughput |
| CN114062561A (en) * | 2021-11-29 | 2022-02-18 | 江南大学 | Method for simultaneously detecting lactic acid, acetic acid and derivative ester thereof in fen-flavor fermented grains based on high performance liquid chromatography technology |
| CN114965731A (en) * | 2022-03-22 | 2022-08-30 | 贵州茅台酒股份有限公司 | Method for detecting pectin in fermented grains by using ultra-high performance liquid chromatography-quadrupole electrostatic field orbit trap |
| CN114965731B (en) * | 2022-03-22 | 2023-08-15 | 贵州茅台酒股份有限公司 | Method for detecting pectin in fermented grains by ultra-high performance liquid chromatography-four-level rod electrostatic field track trap |
| CN115032298A (en) * | 2022-05-26 | 2022-09-09 | 宜宾五粮液股份有限公司 | Method and device for detecting content of yellow water flavor substances in fermentation process in cellar |
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