CN105891375A - Method using ultra-performance convergence chromatography to fast detect chiral ethyl lactate in Baijiu - Google Patents
Method using ultra-performance convergence chromatography to fast detect chiral ethyl lactate in Baijiu Download PDFInfo
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- CN105891375A CN105891375A CN201610447206.8A CN201610447206A CN105891375A CN 105891375 A CN105891375 A CN 105891375A CN 201610447206 A CN201610447206 A CN 201610447206A CN 105891375 A CN105891375 A CN 105891375A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a method using ultra-performance convergence chromatography to fast detect chiral ethyl lactate in Baijiu and belongs to the technical field of detection. The method is characterized in that the chiral ethyl lactate includes L-ethyl lactate and D-ethyl lactate; a Baijiu sample is dried by blowing through nitrogen after being extracted by ethyl acetate and dissolved with methanol, a special ACQUITY UPC2Trefoil CEL1 chiral chromatographic column of 2.5 micrometers is used as the separation column, and carbon dioxide (A) plus methanol (B) are used as the flowing phase to perform gradient elution; the sample is detected by a diode array detector after being separated by ultra-performance convergence chromatography (UPC2). The method is simple and fast and capable of accurately qualitatively and quantitatively detecting the chiral ethyl lactate in Baijiu, and a scientific basis is provided for the accurate determination and fast detection of the two kinds chiral ethyl lactate in Baijiu.
Description
Technical field
The present invention relates to detection technique field, specifically one utilizes ultra high efficiency to close phase chromatograph series diode array detector
The most quickly the method for chirality ethyl lactate (Pfansteihl ethyl ester, D-ALPHA-Hydroxypropionic acid ethyl ester) in detection Chinese liquor, belongs to analysis technical field.
Background technology
Along with the development of analysis and detection technology, particularly efficient capillary gas chromatogram, liquid chromatograph and makings, liquid matter
The development of multiple techniques and application, in conjunction with the sense organ taste of Chinese liquor, the kind verifying framework ingredient in Chinese liquor and content, research
Its compositing characteristic, it is possible to progressively open the secret of all kinds of famous brand of wine " peculiar flavour ".Numerous research worker are attempting effort employing
State-of-the-art technology removes to identify micro constitutent even trace constituent in the Chinese liquor not recognized, and makes a determination district with the style characteristic to Chinese liquor
Not.
Ethyl lactate is the framework ingredient in Chinese liquor, is important one in the big ester of Chinese liquor 4, and content is higher, is about
40-200mg/100mL, it constitutes giving off a strong fragrance, paste flavor, meter Xiang with ethyl hexanoate, ethyl acetate, ethyl n-butyrate. and bata-phenethyl alcohol
And the main aroma composition of mixed-flavouring liquor.In aromatic Chinese spirit, require to reduce the content of ethyl lactate, but content be too low,
Vinosity is boring;Too high levels then increases the astringent taste of wine.From the point of view of molecular structure, ethyl lactate contains a chiral carbon atom, deposits
At a pair i.e. Pfansteihl ethyl ester of chiral enantiomer and D-ALPHA-Hydroxypropionic acid ethyl ester.Chirality correspondence body compound has identical molecular formula, molecule
Between atom connect order the most identical, the simply molecule Atom difference that puts in order in space.The physics of enantiomer
Character and chemical property are the most identical, but most important feature is physiological potency performance difference, and the most left-handed nicotine is than dextrorotation cigarette
The toxicity of alkali is bigger.In Chinese liquor, Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester may cause different sensory responses, at Chinese liquor wind
Taste and mouthfeel aspect play very important effect.Therefore, Pfansteihl ethyl ester and the content of D-ALPHA-Hydroxypropionic acid ethyl ester and ratio in researching white spirit
Example is extremely important.
Summary of the invention
It is an object of the invention to provide one utilizes ultra high efficiency to close the most quickly detection of chromatography series diode array detector
The method of chirality ethyl lactate (Pfansteihl ethyl ester, D-ALPHA-Hydroxypropionic acid ethyl ester) in Chinese liquor, it is intended to realize chirality ethyl lactate inspection in Chinese liquor
That surveys accurately judges, quickly detects.
The present invention solves technical problem and adopts the following technical scheme that
The present invention utilizes ultra high efficiency to close the method for chirality ethyl lactate in phase chromatograph the most quickly detection Chinese liquor, and its feature is:
Described chirality ethyl lactate includes Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester;
Instrument is: be furnished with diode array detector ultra high efficiency close chromatography, ACQUITY UPC2Trefoil CEL1,
2.5 μm chromatographic columns (cellulose 3-(3,5-dimethylphenylcarbamate));Vortex instrument;Liquid-transfering gun;
Described method comprises the steps:
(1) Chinese liquor wine sample to be measured is carried out pre-treatment:
Pipette 5mL Chinese liquor to be measured wine sample in 10mL color comparison tube, add 2mL ethyl acetate, vortex 1min, pipette upper strata
Solution dries up, and uses 2mL methanol to dissolve, injects 2mL sample bottle, treat sample introduction;
(2) set ultra high efficiency close chromatography condition as:
Chromatographic column: ACQUITY UPC2Trefoil CEL1,2.5 μm chromatographic column (cellulose 3-(3,5-dimethylphenylamino formic acid
Ester));Column temperature is 30~50 DEG C;
Sample room temperature: 10~20 DEG C
Flowing phase: A phase is carbon dioxide, and B phase is methanol;Flow velocity is 2~3mL/min;Type of elution is gradient elution;Enter
Sample amount is 1~5 μ L;Detection time is 5min;ABPR pressure is 1885~2200psi;Concretely comprising the following steps of gradient elution:
Time initial, the Volume fraction of mobile phase A and Mobile phase B is 98%:2%;By in this time period of 0min to 3.0min,
The Volume fraction of mobile phase A and Mobile phase B is gradually varied to 96%:4% from 98%:2%;By 3min to 4.0min this
In one time period, the Volume fraction of mobile phase A and Mobile phase B is gradually varied to 95%:5% from 96%:4%;By 4.0min
Within this time period of 5.0min, the Volume fraction of mobile phase A and Mobile phase B is gradually varied to 98% from 95%:5%:
2%.Gradient elution each time change curve all select the curve 6 in instrument.
The condition of diode array detector PDA is set as:
3D passage is 200nm-410nm;
Separating degree is 1.2~2.0nm;
Sampling rate is 20~50 points/second
(3) detection of Chinese liquor wine sample to be measured
After Chinese liquor wine sample to be measured is carried out pre-treatment by the mode of step (1), make UPC by the condition of step (2)2Detection,
Employing external standard method is measured, and carries out qualitative and quantitative with retention time and peak area.Concrete mode is:
(31) drafting of standard curve
Take Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester preparation mass concentration scope works molten at the mixing series standard of 12.5~300 μ g/L
Liquid, then according to the condition set by (2) takes 1.5mL each standard working solution sample introduction, makees ultra high efficiency and closes the detection of phase chromatograph, with
The peak area of determinand mass concentration corresponding to its institute carries out linear regression, it is thus achieved that the linear regression side of two kinds of chirality ethyl lactates
Journey, the curve corresponding to each equation of linear regression, it is the standard curve of corresponding chirality ethyl lactate;
Concrete, equation of linear regression that two kinds of chirality ethyl lactates are corresponding and correlation coefficient r2It is shown in Table 1.
The equation of linear regression of 1: two kind of ethyl lactate of table, correlation coefficient, detection limit, quantitative limit
(32) detection of Chinese liquor wine sample to be measured
After Chinese liquor wine sample to be measured is carried out pre-treatment by the mode of step (1), take 1.5mL and make UPC by the condition of step (2)2
Detection, by using 3D length scanning, it is thus achieved that the chromatogram of testing sample, is carried out according to retention time Chinese liquor wine sample to be measured
Qualitative analysis, then carries out quantitative analysis according to standard curve to Chinese liquor wine sample to be measured.
The inventive method is made following sensitivity test: sensitivity test includes the sensitivity of instrument and the sensitivity of method, instrument
The detection limit of sensitivity instrument represent, the Cmin of the mixed standard solution taking the chirality ethyl lactate of signal to noise ratio >=3 is instrument
Device detection limit;The quantitative limit of the sensitivity method of method represents, takes the chirality ethyl lactate mixed standard solution of signal to noise ratio >=6
Cmin is method quantitative limit.The related data of gained is shown in Table 1.
The inventive method is made following accuracy and repeatability experiment: select same Wine Sample as blank sample after pre-treatment
Product, are divided into 3 parts, are separately added into hybrid standard working solution and carry out recovery testu, calculate the response rate;Choose 1 Chinese liquor sample
Product process 6 according to same pre-treating method, test respectively, by calculating the scope of its relative standard deviation (RSD)
Carry out the repeatability of discriminatory analysis method.The accuracy response rate of method represents, is shown in Table 2;The repeatability of method mark relatively
Quasi-deviation (RSD) represents, is shown in Table 3.Can be seen that the response rate 80~120%, RSD < 10%.
The recovery of standard addition experiment of 2: two kinds of ethyl lactates of table
The repeatability experiment of 3: two kinds of ethyl lactates of table
Advantage for present invention is as follows:
1, the present invention establishes one and utilizes hands in ultra high efficiency conjunction phase chromatograph series diode array detector the most quickly detection Chinese liquor
The method of property ethyl lactate, it is possible to exactly the ethyl lactate in Chinese liquor is carried out qualitative, quantitative, in Chinese liquor two kinds of chirality breasts
The accurately judgement of acetoacetic ester, quickly detection provide scientific basis.
2, ultra high efficiency of the present invention close phase chromatograph series diode array detector simple and quick, accurately and reliably, reproducible.
3, the present invention special chiral column ACQUITY UPC2Trefoil CEL1,2.5 μm (cellulose 3-(3,5-3,5-dimethylphenyl
Carbamate)) and CO2The selection of+methanol as mobile phase has reached excellent separation to the separation of in Chinese liquor two kinds of chirality ethyl lactates
Effect.
4, the pre-treatment operating procedure of the present invention is few, thus reduces experimental error and improve the accuracy of data.
5, the detection method of the present invention is environmental protection " green " technology.The main flowing phase carbon dioxide used in analysis comes
From in the recovery carbon dioxide of other industry release, the carbon dioxide that uses in experiment will not produce new greenhouse gases again.Use
During the method, the modifying agent (methanol) that each sample introduction is consumed is only 0.9mL, compared with similar detection method, reduces 95%
Organic solvent usage amount.
6, the present invention is by reducing the use of laboratory consumable, and then can be cost-effective.
Accompanying drawing explanation
Fig. 1 be concentration be the spectrogram of 12.5 μ g/L standard working solution.
Detailed description of the invention
For illustrating the understanding of the present invention, below in conjunction with embodiment, the present invention is further elaborated.
Chirality ethyl lactate in Chinese liquor is detected by the present embodiment as follows:
Instrument is: be furnished with diode array detector ultra high efficiency close chromatography, ACQUITY UPC2Trefoil CEL1,
2.5 μm chromatographic columns (cellulose 3-(3,5-dimethylphenylcarbamate));Vortex instrument;Liquid-transfering gun;
Concretely comprise the following steps:
(1) Chinese liquor wine sample to be measured is carried out pre-treatment:
Pipette 5mL Chinese liquor to be measured wine sample in 10mL color comparison tube, add 2mL ethyl acetate, vortex 1min, pipette upper strata molten
Liquid dries up, and uses 2mL methanol to dissolve, injects 2mL sample bottle, treat sample introduction;
(2) set ultra high efficiency close chromatography condition as:
Chromatographic column: ACQUITY UPC2Trefoil CEL1,2.5 μm chromatographic columns, chromatographic column filler is cellulose 3-(3,5-diformazans
Base carbanilate);Column temperature is 45 DEG C;
Sample room temperature: 15 DEG C
Flowing phase: A phase is carbon dioxide, and B phase is methanol;Flow velocity is 2.5mL/min;Type of elution is gradient elution;Enter
Sample amount is 2 μ L;Detection time is 5min;ABPR pressure is 2000psi;Concretely comprising the following steps of gradient elution: time initial,
The Volume fraction of mobile phase A and Mobile phase B is 98%:2%;By in this time period of 0min to 3.0min, mobile phase A
It is gradually varied to 96%:4% from 98%:2% with the Volume fraction of Mobile phase B;By this time period of 3min to 4.0min
In, the Volume fraction of mobile phase A and Mobile phase B is gradually varied to 95%:5% from 96%:4%;By 4.0min to 5.0min
In this time period, the Volume fraction of mobile phase A and Mobile phase B is gradually varied to 98%:2% from 95%:5%.Gradient
The change curve in eluting each stage all selects the curve 6 in instrument.
The condition of diode array detector PDA is set as:
3D passage is 200nm-410nm;
Separating degree is 1.8nm;
Sampling rate is 20 points/second;
(31) drafting of standard curve
Take Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester preparation mass concentration be 12.5 μ g/L, 50 μ g/L, 100 μ g/L, 200 μ g/L,
The mixing series standard working solution of 300 μ g/L, enters then according to the condition set by (2) takes each standard working solution of 1.5mL
Sample, makees ultra high efficiency and closes phase chromatograph detection (spectrogram of 12.5 μ g/L standard working solution is as shown in Figure 1), with the peak area of determinand
Mass concentration corresponding to its institute carries out linear regression, it is thus achieved that the equation of linear regression of two kinds of chirality ethyl lactates, each linear regression
Curve corresponding to equation, is the standard curve of corresponding chirality ethyl lactate;The appearance time of Pfansteihl ethyl ester is 1.24min;
The appearance time of D-ALPHA-Hydroxypropionic acid ethyl ester is 1.58min.
(32) detection of Chinese liquor wine sample to be measured
After Chinese liquor wine sample to be measured is carried out pre-treatment by the mode of step (1), take 1.5mL and make UPC by the condition of step (2)2
Detection, by using 3D length scanning, it is thus achieved that the chromatogram of testing sample, is carried out according to retention time Chinese liquor wine sample to be measured
Qualitative analysis, then carries out quantitative analysis according to standard curve to Chinese liquor wine sample to be measured.
Calculate the ratio of the content of Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester and be analyzed, it is possible to obtain the size of ratio between two is with white
The relation of wine taste mouthfeel.
Claims (3)
1. one kind utilizes ultra high efficiency conjunction phase chromatograph quickly to detect the method for chirality ethyl lactate in Chinese liquor, it is characterised in that:
Described chirality ethyl lactate includes Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester;
Instrument is: the ultra high efficiency being furnished with diode array detector closes chromatography;Vortex instrument;Liquid-transfering gun;
Described method comprises the steps:
(1) Chinese liquor wine sample to be measured is carried out pre-treatment:
Pipette 5mL Chinese liquor to be measured wine sample in 10mL color comparison tube, add 2mL ethyl acetate, vortex 1min, pipette upper strata
Solution dries up, and uses 2mL methanol to dissolve, injects 2mL sample bottle, treat sample introduction;
(2) set ultra high efficiency close chromatography condition as:
Chromatographic column: ACQUITY UPC2 Trefoil CEL1,2.5 μm chromatographic columns, chromatographic column filler is cellulose 3-(3,5-diformazans
Base carbanilate);Column temperature is 30~50 DEG C;
Sample room temperature: 10~20 DEG C
Flowing phase: A phase is carbon dioxide, and B phase is methanol;Flow velocity is 2~3mL/min;Type of elution is gradient elution;Enter
Sample amount is 1~5 μ L;Detection time is 5min;ABPR pressure is 1885~2200psi;
The condition of diode array detector PDA is set as:
3D passage is 200nm-410nm;
Separating degree is 1.2~2.0nm;
Sampling rate is 20~50 points/second;
(3) detection of Chinese liquor wine sample to be measured
After Chinese liquor wine sample to be measured is carried out pre-treatment by the mode of step (1), make UPC by the condition of step (2)2Detection,
Employing external standard method is measured, and carries out qualitative and quantitative with retention time and peak area.
Method the most according to claim 1, is characterized in that: the method for the gradient elution described in step (2) is:
Time initial, the Volume fraction of mobile phase A and Mobile phase B is 98%:2%;By 0min to 3.0min, mobile phase A
It is gradually varied to 96%:4% from 98%:2% with the Volume fraction of Mobile phase B;By 3min to 4.0min, mobile phase A
It is gradually varied to 95%:5% from 96%:4% with the Volume fraction of Mobile phase B;By 4.0min to 5.0min, mobile phase A
It is gradually varied to 98%:2% from 95%:5% with the Volume fraction of Mobile phase B.
Method the most according to claim 1, it is characterised in that: the concrete mode of step (3) is:
(31) drafting of standard curve
Take Pfansteihl ethyl ester and D-ALPHA-Hydroxypropionic acid ethyl ester preparation mass concentration scope works molten at the mixing series standard of 12.5~300 μ g/L
Liquid, then according to the condition set by (2) takes 1.5mL each standard working solution sample introduction, makees ultra high efficiency and closes the detection of phase chromatograph, with
The peak area of determinand mass concentration corresponding to its institute carries out linear regression, it is thus achieved that the linear regression side of two kinds of chirality ethyl lactates
Journey, the curve corresponding to each equation of linear regression, it is the standard curve of corresponding chirality ethyl lactate;
(32) detection of Chinese liquor wine sample to be measured
After Chinese liquor wine sample to be measured is carried out pre-treatment by the mode of step (1), take 1.5mL and make UPC by the condition of step (2)2
Detection, by using 3D length scanning, it is thus achieved that the chromatogram of testing sample, is carried out according to retention time Chinese liquor wine sample to be measured
Qualitative analysis, then carries out quantitative analysis according to standard curve to Chinese liquor wine sample to be measured.
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Cited By (8)
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CN106814155A (en) * | 2017-03-10 | 2017-06-09 | 国家烟草质量监督检验中心 | The chiral analysis of nicotine close phase chromatographic tandem mass spectrography in a kind of tomato |
CN106841454A (en) * | 2017-03-10 | 2017-06-13 | 国家烟草质量监督检验中心 | The chiral analysis of nicotine close phase chromatographic tandem mass spectrography in a kind of bolete |
CN106896173A (en) * | 2017-03-10 | 2017-06-27 | 国家烟草质量监督检验中心 | The chiral analysis of nicotine close phase chromatographic tandem mass spectrography in a kind of electronic cigarette liquid |
CN106908541A (en) * | 2017-03-10 | 2017-06-30 | 国家烟草质量监督检验中心 | The chiral analysis of nicotine close phase chromatographic tandem mass spectrography in a kind of cigarette mainstream flue gas |
CN108303482A (en) * | 2018-02-13 | 2018-07-20 | 江南大学 | A kind of method of extracting and developing and volatility bitter taste substance in identification white wine |
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CN113791144A (en) * | 2021-08-13 | 2021-12-14 | 四川绵竹剑南春酒厂有限公司 | Method for detecting chiral ethyl lactate in white spirit by using high performance gas chromatography |
CN115656385A (en) * | 2022-11-14 | 2023-01-31 | 中轻检验认证有限公司 | Method for detecting content of L-lactic acid and D-lactic acid chiral isomers in white spirit |
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CN108303482A (en) * | 2018-02-13 | 2018-07-20 | 江南大学 | A kind of method of extracting and developing and volatility bitter taste substance in identification white wine |
CN108303482B (en) * | 2018-02-13 | 2019-08-20 | 江南大学 | A kind of method of extracting and developing and volatility bitter taste substance in identification white wine |
CN108535395A (en) * | 2018-03-12 | 2018-09-14 | 安徽古井贡酒股份有限公司 | A method of using 32 kinds of free fatties in UPLC-QTof Rapid Simultaneous Determination health liquors |
CN113791144A (en) * | 2021-08-13 | 2021-12-14 | 四川绵竹剑南春酒厂有限公司 | Method for detecting chiral ethyl lactate in white spirit by using high performance gas chromatography |
CN115656385A (en) * | 2022-11-14 | 2023-01-31 | 中轻检验认证有限公司 | Method for detecting content of L-lactic acid and D-lactic acid chiral isomers in white spirit |
CN115656385B (en) * | 2022-11-14 | 2023-03-21 | 中轻检验认证有限公司 | Method for detecting content of L-lactic acid and D-lactic acid chiral isomers in white spirit |
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