CN110514757A - A kind of method of Volatile flavor components in fast resolving white wine - Google Patents
A kind of method of Volatile flavor components in fast resolving white wine Download PDFInfo
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Abstract
The invention discloses a kind of methods of Volatile flavor components in fast resolving white wine, belong to Analysis Technique For Flavor Substances In Food field.HS-SPME-Arrow technical application in the detection of white wine volatile component, is combined with GC × GC-TOFMS and develops a kind of higher detection method of sensitivity, be suitable for the detection of volatile component in white wine after optimizing to method by the present invention.A large amount of Wine Samples are parsed by constructed method, construct volatile component database (Baijiu Flavor Library) in white wine, can be used cooperatively with complete two-dimentional instrument, substitute existing commercialization database.The method optimized and database are combined, rapidly and accurately Volatile flavor components in white wine can be detected, convenient for the confirmation of fragrance whole in unknown wine sample and the quick excavation of key sapidity ingredient.
Description
Technical field
The present invention relates to a kind of methods of Volatile flavor components in fast resolving white wine, belong to Analysis Technique For Flavor Substances In Food
Field.
Background technique
China white wine is one of Spirit most ancient in the world, has thousands of years developing histories in China, is Chinese biography
The a part for culture of uniting.Due to a variety of Particular crafts such as use multiple-microorganism is inoculated with naturally, prolonged solid state fermentations, bring up
China white wine unique and complex flavor characteristic.The volatile component type that white wine is included is more, quantity is big, the fragrance of white wine
It is generally codetermined by thousands of kinds of volatile compounds, the flavor substance with flavoring function is to determine white wine whole style feature
Key, it is significant in terms of process optimization, quality monitoring, product, largely determine consumer's
Consumption choice.However it is also not comprehensive to the understanding of white wine volatile component at present, flavor function is still unintelligible.
In white wine volatile component analyze common pre-treating method include liquid-liquid extraction (LLE), Solid Phase Extraction (SPE),
Headspace solid-phase microextraction (HS-SPME) etc., they respectively have advantage and disadvantage, wherein solid phase micro-extraction technique, are integrated with extraction and concentration
Two steps, save time and labour, and good separating effect can be also played in the presence of matrix interference effect, but is adsorbed
Ability is affected by coated fiber.And HS-SPME-Arrow this novel solid phase micro extraction technology (using novel extraction head)
Appearance, bigger adsorption area can be provided, compared with traditional SPME fiber, micro-analysis in more samples can be adsorbed
Object greatly shortens the extraction time of sample, improves detection sensitivity, provides more high-throughput sample pre-treatments scheme.
Traditional one-dimensional gas-chromatography is unable to satisfy point of this kind of complex sample volatile component of white wine since separating degree is limited
Analysis, comprehensive two dimensional gas chromatography/flight time mass spectrum (GC × GC-TOFMS) technology have been developed in recent years analysis method, phase
Than one-dimensional gas chromatograph, peak capacity, sensitivity and separating degree have significant advantage, efficiently solve traditional one-dimensional chromatography point
Existing total outflow phenomenon is separated out, is the effective means for analyzing complex sample.But due to GC × GC-TOFMS technology high resolution, spirit
The technical advantage that sensitivity is high, peak capacity is big, a sample analysis tends to obtain thousands of in the analysis of white wine volatile component
A chromatographic peak brings huge challenge to the identification work of volatile component in white wine.GC × GC-TOFMS analysis is thousands of
The confirmation of parsing one by one of chromatographic peak needs to expend great manpower, and the comprehensive of volatile component, precise Identification need in a wine sample
The time for expending for 1~2 week leads to sample analysis inefficiency.The main reason for causing the above problem, which is a lack of, to be directed to
Property structural database magnanimity chromatographic peak that GC × GC-TOFMS is parsed carry out effective comparing identification.It is commercialized at present
Structural Identification database mainly has National Institute of Standards and Technology (National Institute of Standards
And Technology, NIST) canonical reference mass spectrometry database and Wiley chemline and mass spectral database.Based on these wide spectrums
The comparison retrieval of database often provides multiple possible Structural Identifications as a result, accuracy is low, to the further of qualification result
Confirmation process is cumbersome, time-consuming huge.
In addition, at present database can only all provide Structural Identification as a result, GC × GC-TOFMS analysis Wine Sample often
The qualification result of several hundred thousands of kinds of compounds can be obtained, how to excavate from such huge compound identification result has wind
The component of function of odor is the huge difficult problem that current GC × GC-TOFMS technical application faces.Now more commonly used method be by
GC-O gas-chromatography-electronic nose combined system hears the fragrant aroma component found in wine sample, but this is also required to devote a tremendous amount of time,
Higher requirement is proposed to operator, be easy to cause laboratory technician tired.
Summary of the invention
[technical problem]
Volatile component type is more in white wine, quantity is big, and conventional pre-treating method sensitivity is limited, it is difficult to which complete detection arrives
All volatile components；The confirmation of parsing one by one that GC × GC-TOFMS analyzes thousands of chromatographic peaks needs to expend great people
Power, comprehensive, the precise Identification of volatile component need to expend the time in 1~2 week in a wine sample, and sample analysis is caused to imitate
Rate is low, and the database being lack of pertinence carries out effective compare and identifies.
[technical solution]
To solve the above-mentioned problems, the invention proposes a kind of method of Volatile flavor components in fast resolving white wine,
The method of the present invention utilizes the database of special foundation, and according to HS-SPME-Arrow pretreatment technology and GC × GC-TOFMS phase
In conjunction with method, can fast and accurately detect to obtain the Volatile flavor components in Wine Sample, it is convenient and efficient, be easy to push away
Extensively.
Specifically, the technical solution of the present invention is as follows: in a kind of fast resolving white wine Volatile flavor components method, it is described
Method the following steps are included:
(1) it is volatilized using HS-SPME-Arrow pre-treating method to the volatile compound and half in Wine Sample to be measured
Compound is extracted；
(2) it is extracted using comprehensive two dimensional gas chromatography/flight time mass spectrum GC × GC-TOFMS analysis HS-SPME-Arrow
The volatile compound and half volatile compound arrived；
(3) the TOFMS data of experiment acquisition are subjected to data processing by work station, automatic identification signal-to-noise ratio is greater than 50 color
Automatic integration deconvolution is carried out after spectral peak, by the white wine volatile flavor group of the retention index of each component and ms fragment and building
(Baijiu Flavor Library) is compared in divided data library, and Volatile flavor components in Wine Sample to be measured can be obtained
Qualification result；
Wherein, the white wine Volatile flavor components database (Baijiu Flavor Library) is by with lower section
What formula obtained:
Step 1: 12 big aromatic white spirit finished wines and former wine are chosen, using HS-SPME-Arrow, the micro- extraction of head space solid phase
Take more kinds of HS-SPME, liquid-liquid extraction LLE, Solid Phase Extraction SPE, magnetic agitation sorption extraction SBSE pre-treating methods to above-mentioned white
Volatile compound and half volatile compound in wine sample are extracted；
Step 2: the volatile compound and partly wave that a variety of pretreatment technologies are obtained by extraction are analyzed using GC × GC-TOFMS
Compound is sent out, data are acquired；
Step 3: the white wine volatile component of precise Identification is obtained using multistage identification strategy: testing the TOFMS number of acquisition
Data processing is carried out according to by TOFMS software workstation, automatic identification signal-to-noise ratio carries out automatic integration solution after the chromatographic peak greater than 50
Convolution sum mass spectral database compares；Level-one evaluation and screening mass spectrum similarity is greater than 700, and possibility is greater than 4000 compound；Second level mirror
It is fixed that matching result is proofreaded one by one by artificial；Three-level identification is confirmed based on retention index (RI) comparing, finally in difference
2959 kinds of white wine volatile components have been identified in aromatic white spirit；
Step 4: the essential information of white wine volatile component described in collection step three, and by white wine volatile flavor group
Divided data library, which is added in the database of work station, can be obtained white wine Volatile flavor components database (Baijiu Flavor
Library)。
In one embodiment of the invention, the work station is LECO companyWork station.
In one embodiment of the invention, in HS-SPME-Arrow pretreatment process, by Wine Sample or to be measured white
The alcoholic strength of wine sample is diluted to 1-15%vol, it is preferable that 2%vol.
In one embodiment of the invention, in HS-SPME-Arrow pretreatment process, Wine Sample or white wine to be measured
Addition ionic strength adjustor is needed in sample, the ionic strength adjustor is NaCl, and additive amount is that 0.125~0.5g/mL is dilute
Wine sample after releasing.
In one embodiment of the invention, in HS-SPME-Arrow pretreatment process, extraction time be 35~
50min, preferably 45min.
In one embodiment of the invention, HS-SPME-Arrow pre-treatment concrete operation step are as follows: by Wine Sample
It is diluted to 1~6%vol with ultrapure water, the wine sample and 1~4g sodium chloride after taking 8mL to dilute are placed in 20mL ml headspace bottle, after sealing
HS-SPME operation is carried out by multi-functional automatic sample handling system MPS2, Wine Sample balances 5~10min at 35~6050 DEG C,
35~50min is extracted, revolving speed is set as 400~1000r/min.
In one embodiment of the invention, the operating parameter of the GC × GC-TOFMS are as follows:
Chromatographic condition: chromatographic column uses capillary column, one-dimensional chromatographic column DB-FFAP, Two way chromatograms column Rxi-17Sil MS；
After the temperature program of one-dimensional post case is 45 DEG C of holding 3min of initial temperature, 230 DEG C are warming up to 5 DEG C/min and keeps 10min, two
The temperature whole process for tieing up post case keeps 5~10 DEG C higher than one-dimensional baking oven；Modulation compensated temperature be 15~20 DEG C, modulation period be 2~
8s；Carrier gas is helium, flow rate of carrier gas 1mL/min；
Mass Spectrometry Conditions: ion source voltage 70eV, ion source temperature are 200~250 DEG C, and transmission line temperature is 240 DEG C, inspection
Survey device voltage is 1430V；Acquisition quality number range is 35~400amu, and frequency acquisition is 50~200spectra/s.
In one embodiment of the invention, purity > 99.9995% of the helium.
In one embodiment of the invention,
In one embodiment of the invention, described to confirm it is to calculate mass spectral results check and correction identification based on RI comparing
Compound RI, further in document report compound identical chromatographic conditions under RI Value Data be compared, screen RI
It is worth compound of the deviation less than 30 as qualification result.
In one embodiment of the invention, the calculating of the RI is completed by ChromaTOF software.
In one embodiment of the invention, the essential information includes physical chemistry information, mass spectrometry parameters, flavor function
It can information；The physical chemistry information include they Chinese name, English name, No. CAS, molecular formula, molecular weight, fusing point, boiling point,
The parameters such as polarity；The mass spectrometry parameters include one-dimensional retention time, two-dimentional retention time, similarity, anti-similarity, possibility,
The parameters such as retention index, quota ion and peak area；The flavor functional information includes the letter such as parameters such as flavor description, threshold value
Breath.
It is that the present invention obtains the utility model has the advantages that
The present invention by HS-SPME-Arrow technical application white wine volatile component detection in, with GC × GC-TOFMS phase
In conjunction with a kind of higher detection method of sensitivity is developed, more suitable for the detection of volatile component in white wine.The present invention passes through
Constructed method parses a large amount of Wine Samples, and building has obtained volatile component database in white wine, can be with complete two
Dimension instrument is used cooperatively, and substitutes existing commercialization database.The method optimized and database are combined, it can be quick and precisely
Ground detects Volatile flavor components in white wine, confirmation and key sapidity ingredient convenient for fragrance whole in unknown wine sample
Quick excavation, method is easy, has a good application prospect.
Detailed description of the invention
Fig. 1: Volatile flavor components method flow diagram in a kind of fast resolving white wine of the invention.
HS-SPME-Arrow-GC × the GC-TOFMS and HS-SPME-GC × GC-TOFMS of Fig. 2 embodiment 2 analyze white wine
Two-dimentional contour chromatogram and 4 class compound two dimension appearance time distribution maps, wherein (a) be HS-SPME-Arrow-GC ×
The two-dimentional contour chromatogram of GC-TOFMS analysis white wine；(b) it is analyzed in white wine for HS-SPME-Arrow-GC × GC-TOFMS
Four class compound two dimension appearance time distribution maps；(c) the two-dimentional contour color of white wine is analyzed for HS-SPME-GC × GC-TOFMS
Spectrogram；(d) four class compound two dimension appearance time distribution maps in white wine are analyzed for HS-SPME-GC × GC-TOFMS.
Specific embodiment
Equipment: (2cm, 50/30 μm, Switzerland's Si Tesi analysis instrument has HS-SPME-Arrow extracting head DVB/CAR/PDMS
Limit company), comprehensive two dimensional gas chromatography-time of-flight mass spectrometer (GC × GC-TOFMS,4D, LECO company, the U.S.),
System includes the tetra- hot modulator of spout twin-stage of 7890B gas-chromatography (Agilent company of the U.S.) and LECO；One-dimensional chromatographic column DB-
FFAP (60m × 0.25mm × 0.25 μm, Agilent company of the U.S.), Two way chromatograms column Rxi-17Sil MS (1.5m × 0.25mm
× 0.25 μm, Restek company, the U.S.)；Multi-functional automatic sample handling system MPS2 (German Gerstel company)；Ultrapure water instrument
Mili-Q (Millipore Corp., the U.S.)；Ultrasonic washing instrument SB-25-12D (NingBo XinZhi Biology Science Co., Ltd).
Chromatographic condition: chromatographic column uses capillary column, and one-dimensional chromatographic column is DB-FFAP, and Two way chromatograms column is Rxi-17Sil
MS；After the temperature program of one-dimensional post case is 45 DEG C of holding 3min of initial temperature, 230 DEG C are warming up to 5 DEG C/min and is kept
10min, the temperature whole process of two-dimentional post case keep 5 DEG C higher than one-dimensional post case；Modulation compensated temperature be 20 DEG C, modulation period 4s；
Carrier gas is helium, flow rate of carrier gas 1mL/min；
Mass Spectrometry Conditions: ion source voltage 70eV, ion source temperature are 230 DEG C, and transmission line temperature is 240 DEG C, detector
Voltage is 1430V；Acquisition quality number range is 35-400amu, frequency acquisition 100spectra/s.
Wine Sample: different flavor white wine finished wine and former wine representative in the whole country are chosen, contains ten
The standard wine sample of two kinds of typical odor types, amounts to 113 kinds.
Reagent: chromatographic grade ethyl alcohol (Sigma-Aldrich), n-alkane C5~C30 (U.S. Sigma-
Aldrich), sodium chloride (Chinese traditional Chinese medicines Solution on Chemical Reagents in Shanghai company).
The foundation of 1 white wine volatile flavor database of embodiment
It is white to choose different flavor representative in the whole country for the building of Volatile flavor components database in white wine
Wine finished wine and former wine contain the standard wine sample of 12 kinds of typical odor types, amount to 113 kinds.
Step 1: by more kinds of pretreatment modes of HS-SPME-Arrow, HS-SPME, LLE, SPE, SBSE in white wine
Volatile compound is extracted, as far as possible volatile compound and half volatile compound all in acquisition white wine；
Step 2: detecting the volatile component being obtained by extraction using GC × GC-TOFMS technology, acquires data；
Step 3: the white wine volatile component of precise Identification is obtained using multistage identification strategy.Test the TOFMS number of acquisition
According to by LECO companyWork station carries out data processing, and automatic identification signal-to-noise ratio carries out after the chromatographic peak greater than 50
Automatic integration deconvolution and mass spectral database (NIST 2014 and Weliy9 database) compare；Level-one evaluation and screening mass spectrum similarity is big
In 700, possibility is greater than 4000 compound；Second level identification carries out mass spectral results check and correction to matching result one by one by artificial；；
Three-level identification is compared based on the RI (RI lit) of the substance in RI (RI cal) calculated to compound under experiment condition and document
Right, group of the difference within 30 is divided into relatively reliable authenticating compound；Finally obtain 2959 white wine volatility of secured identification
Component.
Step 5: the essential information for the volatile component that secured identification goes out in every a sample, including structural information are collected:
Their mass spectrometry parameters are added in white wine Volatile flavor components database；Physical chemistry information: including their molecule
The parameters such as amount, fusing point, boiling point, polarity, are added in database one by one；Flavor functional information: including their fragrance description, threshold
The parameters such as value confirm the compound in white wine with flavor function, construct white wine Volatile flavor components database, and will be white
Wine Volatile flavor components database is added toIn workstation database.
The present invention identifies China's allusion quotation by using various samples pretreatment technology combination GC × GC-TOFMS network analysis
Volatile flavor components in 113 kinds of white wine finished wines of type odor type and former wine, analytically going out is more than to identify altogether in 10,000 chromatographic peaks
2959 kinds of volatile component in white wine out summarize mass spectroscopy structural authentication information, the physics of 2959 kinds of volatile components in white wine
Chemical information, and volatile component flavor function synthesized in white wine is analyzed, establish volatile flavor in fast resolving white wine
Volatile component standard mass spectrometry database and include that 905 Characterization of Odor-active Compounds form in the method for component, first white wine
White wine Volatile flavor components database.
The selection of 2 pre-treating method of embodiment
(1) using HS-SPME-Arrow or HS-SPME pre-treating method to the volatile compound in Wine Sample to be measured
It is extracted with half volatile compound；Wine Sample is diluted to 2.0%vol with ultrapure water, the wine sample and 3g after taking 8mL to dilute
Sodium chloride is placed in 20mL ml headspace bottle, after the sealing of hollow magnetic metal cover with PTEE blue silica gel dottle pin by it is multi-functional from
Dynamic sampling system MPS2 carries out HS-SPME operation, and Wine Sample balances 5min at 50 DEG C, extracts 45min, and revolving speed is set as
400r/min。
(2) HS-SPME-Arrow and HS- is detected using comprehensive two dimensional gas chromatography/flight time mass spectrum GC × GC-TOFMS
The volatile compound and half volatile compound that SPME is obtained by extraction；Primary data analysis is completed by ChromaTOF software, root
Volatile component is identified according to white wine volatile flavor database constructed by embodiment 1.
The two-dimentional contour chromatogram of two kinds of pre-treating methods treated Wine Sample is as shown in Figure 2.According to esters, alcohol
Class and acids three classes compound are divided into 3 regions, it is seen then that HS-SPME-Arrow method has one to volatile compound in white wine
Fixed extraction advantage, extracting head adsorption area is bigger, in same extraction time, can more adsorb the volatilization in white wine
Property component, be very suitable to extract, therefore, the present invention selects method of the HS-SPME-Arrow technology as pre-treatment.
The selection of alcoholic strength after the dilution of 3 Wine Sample of embodiment
Different alcohol dilution 0.5%vol, 1.0%vol, 2.0%vol, 5.0%vol, 10.0%vol is selected to treat
The volatile component surveyed in Wine Sample is detected, detecting step are as follows:
(1) it is volatilized using HS-SPME-Arrow pre-treating method to the volatile compound and half in Wine Sample to be measured
Compound is extracted；Wine Sample is diluted to 0.5%vol, 1.0%vol, 2.0%vol, 5.0% with ultrapure water respectively
Vol or 10.0%vol, take 8mL dilute after wine sample and 3g sodium chloride be placed in 20mL ml headspace bottle, with PTEE blue silica gel every
HS-SPME operation is carried out by multi-functional automatic sample handling system MPS2 after the hollow magnetic metal cover sealing of pad, Wine Sample exists
5min is balanced at 50 DEG C, extracts 45min, and revolving speed is set as 400r/min.
(2) it is extracted using comprehensive two dimensional gas chromatography/flight time mass spectrum GC × GC-TOFMS detection HS-SPME-Arrow
The volatile compound and half volatile compound arrived；Primary data analysis is completed by ChromaTOF software, according to embodiment 1
Constructed white wine volatile flavor database identifies volatile component.
Statistic of classification is carried out to qualification result according to compound property, as shown in table 1.As it can be seen that 5%vol and 10%vol wine
The compound numbers that precision sample identification goes out are considerably less than the other three experiment.The quantity of esters, alcohols and carbonyls exists
It differs greatly in different samples.This is because higher (main ester type compound concentration is equal for many ester type compound concentration in white wine
In 100mg/L or more), and ester type compound is more volatile, and the absorption of HS-SPME-Arrow method is more, therefore chromatography peak intensity
It is higher.Carbonyls is close with ester type compound property simultaneously, and the distributed areas in two-dimensional chromatogram are close, therefore low dense
The esters and carbonyls of degree may be subjected to the interference of high concentration ester type compound chromatographic peak, influence qualitative and quantitative analysis.
And wine sample alcoholic strength is reduced, increase extension rate, effectively reduces high concentration ester type compound to micro esters and carbonyl compound
The qualitative analysis of object influences.In 1%vol and 2%vol sample, sulfide and terpenes both volatile micro chemical combination
Object quantity is apparently higher than other samples, this may be reduced with high concentrations of compounds content, and the competition of HS-SPME-Arrow extracting head is inhaled
Attached decrease is related.To sum up, when white wine be diluted to alcoholic strength be 2%vol when, both reduced white wine middle and high concentration component chromatographic peak
Overload to it is total outflow component qualitative, quantitative influence, while also reduce HS-SPME-Arrow method middle and high concentration component with it is low
The competitive Adsorption of concentration microcomponent facilitates extraction and the qualitative and quantitative analysis of low concentration component.Therefore, the present invention selects
Alcoholic strength after Wine Sample dilution is 2%vol.
HS-SPME-Arrow-GC × GC-TOFMS of volatile component analyzes and identifies result in the different dilution wine samples of table 1
The selection of the extraction time of 4 HS-SPME-Arrow pre-treatment of embodiment
The alcohol of Wine Sample is diluted to 2%vol, 35,40,45 and 50min of extraction time, according to 2 institute of embodiment
12 kinds of " amount micro-perfume is big " volatile materials peak areas are as shown in table 2 in the method detection Wine Sample stated, it can be seen that with extraction
The increase of time, the peak area logarithms of 12 kinds of volatile materials are in a metastable variation, when 45min, big portion
The peak area of compound is divided to be in the larger value, when extraction time increases to 50min, most of volatile materials is (in addition to acetic acid
Pentyl ester and caproic acid) peak area decrease.Comprehensively consider, the extraction for the HS-SPME-Arrow pre-treatment that the present invention selects
Time is 45min.
2 extraction time of table influences 12 kinds of volatile materials peak areas
When 5 HS-SPME-Arrow pre-treatment of embodiment, the selection of sample ionic strength additive amount
Inorganic salts are added in Wine Sample to increase the distribution coefficient that polar compound enters coating using salting-out effect,
The efficiency of extraction is improved, the inorganic salts that the present invention selects are NaCl, and the additive amount of inorganic salts is respectively 1,2,3 or 4g, by white wine
The alcohol of sample is diluted to 2%vol, extraction time 45min, according to method described in embodiment 3 (after Wine Sample dilution
Alcoholic strength is 2%vol) 12 kinds of " amount micro-perfume is big " volatile materials peak areas are detected in Wine Sample with the change of NaCl additional amount
Change, the results are shown in Table 3, it is found that when NaCl additive amount is 3g, compound chromatographic peak area is maximum, when continuing addition, peak
Area declines instead, and extends the time for reaching balance, comprehensively considers, it is 3g as ion that the present invention, which selects NaCl additive amount,
Intensity adjustment agent.
3 NaCl additive amount of table influences 12 kinds of volatile materials peak areas
The selection of the heating rate of chromatography in 6 GC of embodiment × GC-TOFMS detection process
Heating rate in the temperature program of GC × GC-TOFMS is respectively 4,5,6,7 DEG C/min, remaining condition and implementation
The operating method of example 3 is consistent (alcoholic strength after Wine Sample dilution is 2%vol).The correspondence detected under different heating rates
Compound numbers it is as shown in table 4, it can be seen that be rapidly heated and will lead to peak big, the compound amounts detected that separate difficulty
It is few；The heating rate of 5 DEG C/min can detecte more compound, when heating rate continues slack-off, the chemical combination that detects
Object quantity increase it is unobvious, and also result in analysis time extension, comprehensively consider, select the temperature program of 5 DEG C/min as
The operating parameter of the method for the present invention.
HS-SPME-Arrow-GC × GC-TOFMS of volatile component analyzes and identifies result under the different temperature programs of table 4
Detection of the embodiment 7 to volatile flavor substance in Wine Sample to be measured
(1) it is volatilized using HS-SPME-Arrow pre-treating method to the volatile compound and half in Wine Sample to be measured
Compound is extracted: Wine Sample being diluted to 2.0%vol with ultrapure water, the wine sample and 3g sodium chloride after taking 8mL to dilute are set
In 20mL ml headspace bottle, pass through multi-functional automatic sampling system after the hollow magnetic metal cover sealing with PTEE blue silica gel dottle pin
The MPS2 that unites carries out HS-SPME operation, and Wine Sample balances 5min at 50 DEG C, extracts 45min, and revolving speed is set as 400r/min.
(2) it is extracted using comprehensive two dimensional gas chromatography/flight time mass spectrum GC × GC-TOFMS detection HS-SPME-Arrow
The volatile compound and half volatile compound arrived；
(3) by the retention time and ms fragment of each component for detecting GC × GC-TOFMS, with GC × GC-
It scans for comparing in white wine Volatile flavor components database in TOFMS database, precise Identification, which goes out 465 kinds, altogether has wind
The volatile compound of taste feature.
Since the substance in database is obtained by repeatedly screening, accuracy is higher, saves the time that screening is deleted, only
Need instrument analysis that can complete the determination of Volatile flavor components in white wine, substituting 1~2 week of previous needs could be complete
At multiple screening operation, volatile component in white wine is quick and precisely identified；Aroma component in white wine can also be quickly excavated,
Now more commonly used method is that the fragrant aroma component found in wine sample is heard by GC-O electronic nose, when needing to spend a large amount of
Between, higher requirement is proposed to operator, is also easy to cause laboratory technician tired, passes through white wine Volatile flavor components data
Library can be quickly found out the compound with flavor function, and directly obtain the fragrance description and threshold value of related compound, In
The flavor characteristic of unknown wine sample is understood in short time.
Embodiment 8
To same wine sample use respectively conventional qualitative method (when conventional qualitative, one by one screening obtain precise Identification substance it is big
The time in a week is about needed, then is excavated by flavor function of the electronic nose to these substances, it is also necessary to a week
Time) and the method for the embodiment of the present invention 7 analyzed, it is found that and the method for the present invention will be conventional in data processing
Commercialization database replace with white wine Volatile flavor components database constructed by the present invention, it is not only a large amount of to save gradually
Screen the time of confirmation, moreover it is possible to which the fragrance description and threshold value for directly obtaining related compound can understand unknown in a short time
The flavor characteristic of wine sample.
When being analyzed respectively with two methods, what individually be detected using the method for the present invention in 672s is valeric acid isobutyl
Ester, and conventional qualitative method is butyl valerate in the matching result that the time provides, their isomer Information in Mass Spectra each other
Like, but (pass through configuration standard sample by standard items verifying and detected), it is found that isobutyl isovalerate standard items
Appearance time is 672s, and the appearance time of butyl valerate is 748.8s, it is possible thereby to judge, the detection of the method for the present invention is obtained
Result it is more correct, the step of this has benefited from the multistage identification of database of the invention carried out when establishing, also furtherly
Bright this method can accurately excavate the compound in white wine with flavor function, improve the reliability of qualification result.
Although the present invention has been described by way of example and in terms of the preferred embodiments, it is not intended to limit the invention, any to be familiar with this skill
The people of art can do various change and modification, therefore protection model of the invention without departing from the spirit and scope of the present invention
Enclosing subject to the definition of the claims.
Claims (10)
1. a kind of method of Volatile flavor components in fast resolving white wine, which is characterized in that the described method comprises the following steps:
(1) using HS-SPME-Arrow pre-treating method to the volatile compound and half volatilization chemical combination in Wine Sample to be measured
Object is extracted；
(2) it is obtained by extraction using comprehensive two dimensional gas chromatography/flight time mass spectrum GC × GC-TOFMS analysis HS-SPME-Arrow
Volatile compound and half volatile compound；
(3) the TOFMS data of experiment acquisition are subjected to data processing by work station, automatic identification signal-to-noise ratio is greater than 50 chromatographic peak
Automatic integration deconvolution is carried out afterwards, by the white wine Volatile flavor components number of the retention index of each component and ms fragment and building
It is compared according to library, the qualification result of Volatile flavor components in Wine Sample to be measured can be obtained；
Wherein, the white wine Volatile flavor components database obtains in the following manner:
Step 1: 12 big aromatic white spirit finished wines and former wine are chosen, using HS-SPME-Arrow, headspace solid-phase microextraction HS-
More kinds of SPME, liquid-liquid extraction LLE, Solid Phase Extraction SPE, magnetic agitation sorption extraction SBSE pre-treating methods are to above-mentioned Wine Sample
In volatile compound and half volatile compound extracted；
Step 2: the volatile compound and half volatilizationization that a variety of pretreatment technologies are obtained by extraction are analyzed using GC × GC-TOFMS
Object is closed, data are acquired；
Step 3: the white wine volatile component of precise Identification is obtained using multistage identification strategy: test the TOFMS data of acquisition by
TOFMS software workstation carries out data processing, and automatic identification signal-to-noise ratio carries out automatic integration deconvolution after the chromatographic peak greater than 50
It is compared with mass spectral database；The process of level-one identification are as follows: screening mass spectrum similarity is greater than 700, and possibility is greater than 4000 compound；Two
The process of grade identification are as follows: by manually carrying out mass spectral results check and correction to matching result one by one；The process of three-level identification are as follows: based on guarantor
It stays index (RI) comparing to confirm, 2959 kinds of white wine volatile components has finally been identified in different flavor white wine；
Step 4: the essential information of white wine volatile component described in collection step three, and by white wine Volatile flavor components number
Being added in the database of work station according to library can construct to obtain white wine Volatile flavor components database.
2. the method for Volatile flavor components in a kind of fast resolving white wine according to claim 1, which is characterized in that
In HS-SPME-Arrow pretreatment process, the alcoholic strength of Wine Sample or Wine Sample to be measured is diluted to 1-15%vol.
3. the method for Volatile flavor components in a kind of fast resolving white wine according to claim 2, which is characterized in that
In HS-SPME-Arrow pretreatment process, the alcoholic strength of Wine Sample or Wine Sample to be measured is diluted to 2%vol.
4. according to claim 1~3 in a kind of any fast resolving white wine Volatile flavor components method, feature
It is, in HS-SPME-Arrow pretreatment process, needs to add ionic strength into Wine Sample or Wine Sample to be measured adjusting
Agent, the ionic strength adjustor are NaCl, and additive amount is the wine sample after 0.125~0.5g/mL dilution.
5. the method for Volatile flavor components, feature in a kind of fast resolving white wine according to any one of claims 1 to 4
It is, in HS-SPME-Arrow pretreatment process, extraction time is 35~50min.
6. according to claim 1~5 in a kind of any fast resolving white wine Volatile flavor components method, feature
It is, HS-SPME-Arrow pre-treatment concrete operation step are as follows: Wine Sample is diluted to 1~15%vol with ultrapure water, is taken
Wine sample and 1~4g sodium chloride after 8mL dilution are placed in 20mL ml headspace bottle, pass through multi-functional automatic sample handling system MPS2 after sealing
HS-SPME operation is carried out, Wine Sample balances 5~10min at 35~60 DEG C, extracts 35~50min, and revolving speed is set as 400
~1000r/min.
7. according to claim 1~6 in a kind of any fast resolving white wine Volatile flavor components method, feature
It is, the operating parameter of the GC × GC-TOFMS are as follows:
Chromatographic condition: chromatographic column uses capillary column, one-dimensional chromatographic column DB-FFAP, Two way chromatograms column Rxi-17Sil MS；It is one-dimensional
After the temperature program of post case is 45 DEG C of holding 3min of initial temperature, 230 DEG C are warming up to 5 DEG C/min and keeps 10min, two-dimensional columns
The temperature whole process of case keeps 5~10 DEG C higher than one-dimensional baking oven；Modulation compensated temperature is 15~20 DEG C, and modulation period is 2~8s；It carries
Gas is helium, flow rate of carrier gas 1mL/min；
Mass Spectrometry Conditions: ion source voltage 70eV, ion source temperature are 200~250 DEG C, and transmission line temperature is 240 DEG C, detector
Voltage is 1430V；Acquisition quality number range is 35~400amu, and frequency acquisition is 50~200spectra/s.
8. according to claim 1~7 in a kind of any fast resolving white wine Volatile flavor components method, feature
It is, it is described to confirm it is the RI for calculating the compound of mass spectral results check and correction identification based on RI comparing, further and in document
RI Value Data under report compound identical chromatographic conditions is compared, and screens compound of the RI value deviation less than 30 as identification
As a result.
9. according to claim 1~8 in a kind of any fast resolving white wine Volatile flavor components method, feature
It is, the essential information includes physical chemistry information, mass spectrometry parameters and flavor functional information.
10. the method for Volatile flavor components is in liquor determination in a kind of any fast resolving white wine of claim 1~9
Application in field.
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