CN104849390B - A kind of method of applying component in the qualitative white wine of comprehensive two dimensional gas chromatography-flight time mass spectrum coupling - Google Patents
A kind of method of applying component in the qualitative white wine of comprehensive two dimensional gas chromatography-flight time mass spectrum coupling Download PDFInfo
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Abstract
The invention discloses a kind of method of applying component in the qualitative white wine of comprehensive two dimensional gas chromatography-flight time mass spectrum coupling, first by deionized water, wine sample to be measured is diluted to final alcohol content 10～15%vol, draw 8～10mL dilution wine sample to 20mL head space bottle, add 1～3gNaCl, then use the component in headspace solid-phase microextraction enrichment wine sample, finally adopt the coupling of comprehensive two dimensional gas chromatography-flight time mass spectrum to measure. Comprehensive two dimensional gas chromatography adopts two different chromatographic columns of polarity to carry out orthogonal separation to component in wine, and under the effect of modulator, enter flight time mass spectrum and complete detection, sample pre-treatments mode of the present invention is simple, analytical equipment has the feature of high flux, high sensitivity and high accuracy, solve white wine complicated components, difficult separation, difficulty problem qualitatively, provide an effective approach for further dissecting white wine micro constitutent.
Description
Technical field
The present invention relates to a kind of method of analytical test component, especially a kind of method of analyzing component in white wine.
Background technology
White wine is the distinctive a kind of Spirit of China, is large Spirit (brandy Brandy, whiskey of the world eightWhisky, vodka Vodka, gin Gin, Rum Rum, mescal Tequila, sake sake, China white wineSpirit) one of, obtain through distillation after making wine unstrained spirits or fermentation by starch or saccharine material.
Fragrance is the first impressions of people to white wine, is an extremely important factor weighing white wine quality. But along withWhite wine component there are differences, and white wine presents different fragrance. The micro constitutent kind containing in China's conventional solid-state fermentation white wineThousands of, how the classification of component, can be divided into alcohols, acids, ester class, aldehydes, ketone, ethers and heterocyclic etc.Comprehensively these components are carried out qualitatively, this is to our researching white spirit local flavor, investigation white wine risk and hazard, backstepping microbial metabolismApproach, improvement White wine brewing process all have very important effect.
In recent years, along with the continuous renewal of Instrumental Analysis equipment, analysis level improves constantly, and white wine component analysis equipmentThrough developing into gas chromatography mass spectrometry, LC-MS from gas-chromatography, liquid chromatogram before, the composition that can detect from white wine also fromBefore tens kinds are developed into present hundreds of kind. But due to the complexity of Chinese conventional solid-state fermentation white spirit process, causeIn white wine, still exist a lot of micro constitutent in unknown state. Therefore select simple, fast and efficient white wine pre-treatment sideMethod, and the component of analyzing in white wine by the analytical equipment with high flux, high sensitivity and high accuracy remains ChinaThe important topic of conventional solid-state fermentation method liquor flavor research.
SPME (solid-phasemicroextraction, SPME) technology is to rise the nineties in 20th centuryNovel sample pre-treatments and beneficiation technologies are that a kind of centralized procurement sample, extraction, concentrated and sample introduction are in the solvent-free sample of oneMicro-extraction new technology. Compared with solid phase extraction techniques, SPME operation is simpler, carries more conveniently, and operating cost also moreAdd cheap; Overcome in addition that the SPE rate of recovery is low, the susceptible to plugging shortcoming in adsorbent duct. Therefore become currently usedOne of method being most widely used in Sample Pretreatment Technique.
Comprehensive two dimensional gas chromatography (ComprehensiveTwo-dimensionalGasChromatography, GC ×GC) be the method occurring the beginning of the nineties, it is that separating mechanism difference is had to two separate chromatographic columns with series systemBe combined into two-dimensional gas chromatography, trap and transmission effect again through plaing of modulator, each separation through first chromatographic column is heated up in a steamerDivide and all need to be introduced into modulator, after focusing on, deliver to second chromatographic column again with pulse mode, all components are from secondChromatographic column enters detector, and signal, after data system is processed, obtains taking post 1 retention time as the first abscissa, when post 2 retainsBetween be the second abscissa, signal strength signal intensity is ordinate three chromatograms or profile diagram. Comprehensive two dimensional gas chromatography and one dimension chromatogramCompare, have that resolution ratio is high, peak capacity is large, highly sensitive, qualitative reliability is a little strong, very applicable analysis is as China's traditionThe substance system of this complicated components of white wine. Comprehensive two dimensional gas chromatography instrument and the time of-flight mass spectrometer full two-dimensional gas forming of arranging in pairs or groupsPhase chromatogram-flight time mass spectrum combined instrument (GC × GC-TOFMS) can be described as at present the most powerful gas of analytic function in the worldMatter equipment.
Summary of the invention
The present invention aims to provide a kind of sample pre-treatments mode simply and can detect component in more multi-component qualitative white wineMethod, in order to overcome existing white wine component analysis method length consuming time, to detect the less deficiency of composition,
The method of component in the qualitative white wine of the present invention's application comprehensive two dimensional gas chromatography-flight time mass spectrum coupling, its featureBe: first by deionized water, wine sample to be measured being diluted to final alcohol content is 10～15%, then uses headspace solid-phase microextractionComponent in enrichment wine sample to be measured, then use comprehensive two dimensional gas chromatography-flight time mass spectrum coupling qualitative determination wine sample to be measuredMiddle component. Comprise the following steps:
A, dilution process: by deionized water, wine sample to be measured being diluted to final alcohol content volume ratio is 10～15%;
B, headspace solid-phase microextraction: draw 8～10mL dilution wine sample to 20mL head space bottle, add 1～3gNaCl, screwHead space bottle cap; Then after 45～55 DEG C of constant temperature preheating 3～5min, extracting head is inserted in described head space bottle, in 45～55 DEG CConstant temperature extracting 30～40min; The insertion depth of extracting head in described head space bottle is 40～43mm;
C, comprehensive two dimensional gas chromatography-flying time mass spectrum analysis: after headspace solid-phase microextraction finishes, extracting head is insertedTo the injection port of comprehensive two dimensional gas chromatography-flight time mass spectrum, insertion depth 50～54mm, 230～250 DEG C of pyrolysis analyse 3～5min, through comprehensive two dimensional gas chromatography-flying time mass spectrum analysis, Comprehensive analysis results obtains the component in wine sample to be measured.
After the component in automatic headspace solid-phase microextraction adsorption and enrichment wine sample to be measured, be adsorbed in to be measured group in extracting headDivide the mode of analysing by pyrolysis at front injection port to enter in comprehensive two dimensional gas chromatography instrument, component to be measured separates through one dimension chromatographic columnAfter, after modulation /demodulation, entering two-dimensional gas chromatography post and again separate, component to be measured is examined by flight time mass spectrumSurvey.
Further, in step B, extraction adopts headspace solid-phase microextraction technology to carry out, and extracting head is 50/30μ mDVB/CAR/PDMS self-action extracting head.
Further, in step C, the analysis condition of comprehensive two dimensional gas chromatography instrument is as follows:
Chromatographic column adopting GC × GC column system, one dimension chromatographic column is 60m × 0.25mm × 0.25 μ mDB-wax polar column,Two dimension chromatographic column is 2m × 0.18mm × 0.18 μ mRtx-200 middle polarity post, and one dimension chromatographic column and two-dimentional chromatographic column are by hairCapillary column joint connector connects with series system;
230～250 DEG C of injector temperatures; Not shunt mode;
4～6s modulation period (cold spray time 1～2s, hot spray time 1～3s), cooling agent is liquid nitrogen, heat modulation gas isCompressed air, cool tone gas processed is nitrogen;
Carrier gas is He, and flow velocity is 0.9～1.1mL/min;
Column oven heating schedule is: 40～45 DEG C of one dimension column oven initial temperatures, keep 2～4min, then with 1～3 DEG C/The heating rate of min is warming up to 90～100 DEG C, then is warming up to 180～200 DEG C with the heating rate of 4～6 DEG C/min, finally with 6The heating rate of～8 DEG C/min is warming up to 230～250 DEG C, keeps 12～15min;
45～50 DEG C of two-dimensional columns incubator initial temperatures, keep 2～4min, then heat up with the heating rate of 1～3 DEG C/minTo 100～115 DEG C, then be warming up to 185～200 DEG C with the heating rate of 4～6 DEG C/min, finally with the intensification of 6～7 DEG C/minSpeed is warming up to 225～235 DEG C, keeps 12～15min;
Modulation compensated temperature is 10～15 DEG C;
Further, in step C, the analysis condition of flight time mass spectrum is as follows: quality acquisition range 30～400amu, 230～250 DEG C of transmission line temperature, 100～120 full spectrogram/s of frequency acquisition, 200～220 DEG C of ion source temperatures,Detector voltage 1450～1500V;
Advantage of the present invention and good effect are as follows:
The present invention adopts headspace solid-phase microextraction sample pre-treatments mode, be a kind of centralized procurement sample, extraction, concentrated and sample introduction inThe solvent-free sample micro-extraction new technology of one. Compared with solid phase extraction techniques, SPME operation is simpler, carries more squareJust, operating cost is also cheaper, has overcome in addition that the SPE rate of recovery is low, the susceptible to plugging shortcoming in adsorbent duct; With liquidLiquid-liquid extraction method is compared, SPME operation simpler, consuming time shorter, security is higher; In addition, the present invention byGerstelMPS three-in-one full automatic sample pre-treatments platform, has realized the mass processing of sample pre-treatments, has more improvedThe analysis efficiency of sample.
Traditional one dimension gas-chromatography can only adopt a root chromatogram column, chromatographic column or polarity or nonpolar, and full two-dimensional gasPhase chromatogram adopts GC × GC dual column system, first presses molecular polarity size when the component to be measured in sample is passed through one dimension polarity chromatographic columnSeparate, then press molecular size separation through the nonpolar chromatographic column of two dimension, in like manner, component to be measured also can first be pressed molecular size and be separated, thenPress molecular polarity and separate, thereby make component to be measured realize orthogonal separation. Therefore the inventive method is applicable to multi-component compartment analysis,There is absolute advantage for analyzing component in white wine;
Headspace solid-phase microextraction technology and comprehensive two dimensional gas chromatography-flight time mass spectrum are applied to analysis white wine by the present inventionIn the research of component, its result is accurately and reliably, highly sensitive, the disposable 1000 lot of trace compositions that detect of energy in white wine,The component detecting in white wine considerably beyond other analytical methods.
Brief description of the drawings
Fig. 1 is comprehensive two dimensional gas chromatography-flight time mass spectrum total ion current figure of component in embodiment wine sample;
Fig. 2 is comprehensive two dimensional gas chromatography-flight time mass spectrum exterior view of component in embodiment wine sample;
Fig. 3 is comprehensive two dimensional gas chromatography-flight time mass spectrum profile diagram of component in embodiment wine sample.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail:
Instrument: LecoPegasus4D comprehensive two dimensional gas chromatography-time of-flight mass spectrometer (GC × GC-TOFMS); Be equipped withAgilent7890B two-dimensional gas chromatography instrument, GerstelMPS (GerstelGmbH, MulheimanderRuhr,Germany) automatic sampler.
Reagent: absolute ethyl alcohol, chromatographically pure; NaCl, analyzes pure.
Wine sample to be measured is diluted to final alcohol content 13%vol by deionized water; Draw dilution after wine sample 9mL inIn 20mL head space bottle, and add 2gNaCl, screw head space bottle cap, adopt full-automatic SPME and full-automatic input mode to enterThe absorption of row sample and parsing. Online SPME condition is: extracting head is 50/30 μ mDVB/CAR/PDMS self-action extractionGet head, sample incubation temperature is 50 DEG C; Hatching balance 4min; The degree of depth that extracting head is inserted head space bottle is 42mm, and extraction time is37min, when sample introduction, extracting head is inserted the degree of depth 52mm of injection port.
Instrument condition:
One dimension chromatographic column is 60m × 0.25mm × 0.25 μ mDB-wax polar column;
Two dimension chromatographic column is 2m × 0.18mm × 0.18 μ mRtx-200 middle polarity post;
Carrier gas: He, flow velocity 1mL/min;
Shunt mode: do not shunt;
Sampling condition: 240 DEG C of injection port resolution temperatures, resolve time 4min, 240 DEG C of injector temperatures;
Column oven heating schedule is: 40 DEG C of one dimension column oven initial temperatures, keep 3min, then with the intensification of 2 DEG C/minSpeed is warming up to 95 DEG C, then is warming up to 190 DEG C with the heating rate of 5 DEG C/min, is finally warming up to the heating rate of 7 DEG C/min230 DEG C, keep 14min;
45 DEG C of two-dimensional columns incubator initial temperatures, keep 3min, are then warming up to 100 DEG C with the heating rate of 2 DEG C/min, thenBe warming up to 195 DEG C with the heating rate of 5 DEG C/min, be finally warming up to 235 DEG C with the heating rate of 7 DEG C/min, keep 14min;
Modulation compensated temperature is 15 DEG C.
6s modulation period (cold spray time 1s, hot spray time 2s), cooling agent is liquid nitrogen, heat modulation gas is compressed air,Cool tone gas processed is nitrogen;
Mass spectrum condition: mass detector is flight time mass spectrum, 210 DEG C of EI ion source temperatures, ionization voltage :-70v, passes240 DEG C of defeated line temperature, adopt full scan mode, quality acquisition range 30-400amu, and 100 full spectrogram/s of frequency acquisition, detectDevice voltage 1450v;
Data processing: the data obtained is processed automatically through the work station of Pegasus4D again, qualitative spectrum library used isNIST/EPA/NIHVersion2.0. The data obtained is again through further processing and obtain final result. Result shows, utilizes thisBright method detects more than 1600 and plants micro constitutent in wine sample to be measured, and concrete outcome (only lists main 350 in table 1 in tableKind).
The inventive method is applicable to the qualitative determination of component in white wine
Subordinate list: the main component table in this routine wine sample
Claims (4)
1. a method of applying component in the qualitative white wine of comprehensive two dimensional gas chromatography-flight time mass spectrum coupling, is characterized in thatFirst by deionized water, wine sample to be measured being diluted to final alcohol content is 10～15%, then treats with headspace solid-phase microextraction enrichmentSurvey the component in wine sample, then use the component in comprehensive two dimensional gas chromatography-flight time mass spectrum coupling qualitative determination wine sample to be measured;
The analysis condition of comprehensive two dimensional gas chromatography is:
Chromatographic column adopting GC × GC column system, one dimension chromatographic column is 60m × 0.25mm × 0.25 μ mDB-wax polar column, two dimensionChromatographic column is 2m × 0.18mm × 0.18 μ mRtx-200 middle polarity post, and one dimension chromatographic column and two-dimentional chromatographic column are passed through capillaryColumn joint connector connects with series system;
230～250 DEG C of injector temperatures; Not shunt mode;
Modulation period 4～6s, wherein cold spray time 1～2s, hot spray time 1～3s, cooling agent is liquid nitrogen, heat modulation gas is for pressingContracting air, cool tone gas processed is nitrogen;
Carrier gas is He, and flow velocity is 0.9～1.1mL/min;
Column oven heating schedule is: 40～45 DEG C of one dimension column oven initial temperatures, keep 2～4min, then with 1～3 DEG C/minHeating rate be warming up to 90～100 DEG C, then be warming up to 180～200 DEG C with the heating rate of 4～6 DEG C/min, finally with 6～8DEG C/heating rate of min is warming up to 230～250 DEG C, keeps 12～15min;
45～50 DEG C of two-dimensional columns incubator initial temperatures, keep 2～4min, are then warming up to the heating rate of 1～3 DEG C/min100～115 DEG C, then be warming up to 185～200 DEG C with the heating rate of 4～6 DEG C/min, finally with the intensification speed of 6～7 DEG C/minRate is warming up to 225～235 DEG C, keeps 12～15min;
Modulation compensated temperature is 10～15 DEG C.
2. method according to claim 1, is characterized in that comprising the following steps:
A, dilution process: by deionized water, wine sample to be measured being diluted to final alcohol content is 10～15%;
B, headspace solid-phase microextraction: draw 8～10mL dilution wine sample to 20mL head space bottle, add 1～3gNaCl, screw head spaceBottle cap; Then after 45～55 DEG C of constant temperature preheating 3～5min, extracting head is inserted in described head space bottle, in 45～55 DEG C of constant temperatureExtraction 30～40min; The insertion depth of extracting head in described head space bottle is 40～43mm;
C, comprehensive two dimensional gas chromatography-flying time mass spectrum analysis: after headspace solid-phase microextraction finishes, extracting head is inserted into entirelyThe injection port of two-dimensional gas chromatography-flight time mass spectrum, insertion depth 50～54mm, 230～250 DEG C of pyrolysis are analysed 3～5min, warpComprehensive two dimensional gas chromatography-flying time mass spectrum analysis, Comprehensive analysis results obtains the component in wine sample to be measured.
3. method according to claim 1 and 2, is characterized in that: extracting head used is 50/30 μ mDVB/CAR/PDMSSelf-action extracting head.
4. method according to claim 1 and 2, is characterized in that: the analysis condition of flight time mass spectrum is:
Quality acquisition range 30～400amu, 230～250 DEG C of transmission line temperature, frequency acquisition is 100～120 full spectrogram/s,200～220 DEG C of ion source temperatures, detector voltage 1450～1500V.
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| CN107389823A (en) * | 2017-07-31 | 2017-11-24 | 江苏省环境监测中心 | The method of SPME analysis of off-flavors system detectio pickles factory uncontrollable discharge smell substance |
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| CN103018370A (en) * | 2012-12-21 | 2013-04-03 | 江苏今世缘酒业股份有限公司 | Method for judging important flavor substance of liquor |
| CN103969391A (en) * | 2014-05-23 | 2014-08-06 | 江南大学 | Method for determining volatile components in millet wine |
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Effective date of registration: 20150915 Address after: 236820 Gujing Town, Qiaocheng District, Anhui City, Bozhou Applicant after: ANHUI RUISIWEIER TECHNOLOGY CO., LTD. Address before: 236826 Anhui Province, Qiaocheng District of Bozhou City Gujing Town Industrial Park Applicant before: Gujinggong Spirits Co., Ltd., Anhui Prov. Applicant before: ANHUI RUISIWEIER TECHNOLOGY CO., LTD. |
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