CN103675127A - Method for distinguishing flavor substance in edible mushroom through combination of headspace gas chromatography-mass spectrometer and electronic nose - Google Patents
Method for distinguishing flavor substance in edible mushroom through combination of headspace gas chromatography-mass spectrometer and electronic nose Download PDFInfo
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Abstract
The invention discloses a method for distinguishing a flavor substance in an edible mushroom through combination of a headspace gas chromatography-mass spectrometer and an electronic nose. According to the method, the headspace gas chromatography-mass spectrometer and the electronic nose are combined to collect volatile flavor substances of different edible mushrooms, and the similarity of the edible mushrooms is judged according to principal component analysis and a radar map so as to distinguish different mushrooms. The method can be used to rapidly and accurately distinguish the different edible mushrooms to improve the accuracy, the scientificity and the authority of distinguishing the edible mushrooms.
Description
Technical field
The invention belongs to edible fungi volatile flavor substance and distinguish field, relate to particularly and a kind ofly utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to distinguish method for distinguishing to edible fungi flavor substances.
Background technology
Edible fungi is the macro fungi that a class has high fructification, is commonly called as mushroom or gill fungus, has the kinds such as mushroom, flat mushroom, pleurotus eryngii, Hericium erinaceus, dictyophora phalloidea.Edible fungi has very high nutritive value, is rich in protein, B family vitamin and some polysaccharide, nucleic acid isoreactivity composition, has the health-care effects such as the body immunity of enhancing, antitumor activity, reducing blood lipid.From nutrition angle, edible fungi has been concentrated the gun-ho characteristic of food, is proposed as one of ten large healthy food.
Volatile ingredient in mushroom is of a great variety, mainly comprises containing eight carbon volatile compounds, sulfocompound and aldehydic acid ketone ester class etc.The mushroom of different cultivars, different parts and different developmental phases, each is variant for its contained flavor component.Zheng Jianxian
[1]once applied steam distillation extracting and analyzed through GC-MS, from fragrant mushroom umbrella part and shank, identify respectively 64 kinds and 42 kinds of aroma compounds, and find the two difference little on main taste compound forms, for the comprehensive utilization of mushroom from now on provides good theoretical foundation.Mushroom essence is most important aroma compound in mushroom, yet because its stability is not high, easily decomposes, and decomposition product comprises dimethyl disulfide and NSC 97324, so its analysis content is lower.Volatile compound containing eight carbon, also be that the most important flavor substances of edible fungi is as 1-OCOL etc., they are all that having strong mushroom wind sulfocompound can affect the fragrance of mushroom body integral body conventionally by the enzymatic of linoleic acid essence fat oxidation, are most important fragrance sources in mushroom.Sulfocompound can affect the fragrance of mushroom body integral body conventionally, is most important fragrance source in mushroom.The fragrance of edible fungi is not the result that single compound embodies, but by numerous component interactions, the mutual effect of balance.
Electronic Nose is that a plurality of chemical sensors (abbreviation gas sensor array) and the suitable mode identification method that do as one likes can overlap each other forms, and can identify the bionics instrument of simple and complicated smell.Because smell constituent is complicated, with single gas sensor, be only to evaluate gaseous mass, Electronic Nose detects gas based on gas sensor array.It and general chemistry analytical instrument, as differences such as chromatograph, spectrometer, capillary electrophoresis apparatus, what obtain is not the qualitative and quantitative result of certain or a few compositions in sample, but gives the Global Information of volatile ingredient in sample, also claims " fingerprint " data.This is the same with the nose of humans and animals, " smelling " be the overall breath of object.The present invention adopts is FOX 4000 types that French Alpha Mo Si company produces, this detection system is by 18 metal oxide sensors, each sensor is different to the response characteristic of gas, the metal semiconductor sensitive layer being embodied in sensor is adsorbed onto after detected gas, its conductivity changes, thereby pass out different influence values
The escaping gas of edible fungi research is at present mainly also to carry out the analysis of quantitative and qualitative analysis by the method for gas chromatography-mass spectrography, and the characteristic flavor substance of different edible fungis carries out quantitative test.But existing, this kind of analytical approach cannot carry out to the whole flavor characteristic of edible fungi volatile matter the technical matters of concentrated expression.
list of references
[1], Zheng Jianxian. the separation of Fujian mushroom flavor substance and evaluation [J]. edible fungi of china, 1995,14 (6): 3-6.
Summary of the invention
The object of the invention is cannot to the whole flavor characteristic of edible fungi volatile matter, carry out the technical matters of concentrated expression in order to solve above-mentioned gas chromatography-mass spectrography method, and provide a kind of, utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to distinguish method for distinguishing to edible fungi flavor substances.The method is to utilize headspace gas chromatography-GC-MS and Electronic Nose to gather the volatile matter of different edible fungis, by principal component analysis (PCA) in edible fungi (PCA) and local flavor fingerprint, be that radar map is judged the similarity between edible fungi, and then differentiate different edible fungis.The method can fundamentally improve accuracy, the science and authoritative that edible fungi is identified.
Know-why of the present invention
The gas that tested edible fungi essential oil sample volatilizes acts on Electronic Nose sensor array, the conductivity that causes each sensor changes, this changes the kind with each sensor specificity sensitive gas, concentration is relevant, this mutual relationship can be used as demarcate sample information according to " a kind of gas is presented on before a sensor cover specifically, sensor converts electric signal to gas input, a plurality of sensors have just formed the response spectra of sensor array to this smell to a kind of response of gas, every kind of gas all can have its characteristic response, according to the characteristic response of multisensor, just can distinguish the kind concentration of gas.
Chemical composition in edible fungi essential oil is very complicated, and different edible fungis has different flavor characteristics, and flavor characteristic material, has specificity.And the Volatile flavor components of edible fungi of the same race keeps relative stability, therefore, the characteristic spectrum of research edible fungus flavor thing and volatile ingredient qualitative, quantitative, and with this collection of illustrative plates and volatile constituent differentiate edible fungi method be practicable.That is to say, because different edible fungi volatile constituents is different, under certain condition, with head-space sampler, collect the volatile and semi-volatile compounds in edible fungi, detection analysis through gas chromatograph-mass spectrometer and Electronic Nose, will obtain diverse volatile ingredient table and local flavor finger-print (being radar map).
Technical scheme of the present invention
A kind ofly by headspace gas chromatography-mass spectrum and Electronic Nose coupling technique, volatile flavor substance in edible fungi is distinguished to method for distinguishing, utilize headspace gas chromatography-GC-MS and Electronic Nose to analyze the volatile matter of different edible fungis, by volatile ingredient table, principal component analysis (PCA) and local flavor fingerprint analysis figure, judge the similarity between edible fungi, and then differentiate that the whole flavor characteristic of the volatile matter of different edible fungis comprehensively analyzes.
Above-mentioned a kind ofly utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to the edible fungi flavor substances method for distinguishing that reflects, specifically comprise the following steps:
1), edible fungi volatile essential oil is prepared
Get commercially available different dry edible mushroom to be identified, respectively at 80 ℃ of dry 4h, mechanical disintegration, crosses 40 mesh sieves, under the condition that is then 1:10 in solid-to-liquid ratio respectively, uses extraction by steam distillation 2.5h, obtains different edible fungi essential oil samples;
Described edible fungi is mushroom, grifola frondosus, flat mushroom, pleurotus eryngii, bolete, Ji Songrong, grey mushroom (abalonelike), agrocybe or Hericium erinaceus etc., in the preferred embodiment of the invention, only with mushroom, grifola frondosus, describe, but do not limit the application of method of the present invention in other edible fungis;
(2), the different edible fungi essential oil samples of step (1) gained carried out to headspace gas chromatography-mass spectrophotometry measure, detailed process is as follows:
1. get, respectively 6～12ml edible fungi essential oil sample and put into 15ml sampling receptacle, and be placed in 30～60 ℃ of water-baths, SPME extracting head (65 μ m PDMS/DVB, Supelco company) head space Static Adsorption 30 ~ 60min;
2., gas chromatographic sample introduction mouth that SPME extracting head is inserted in headspace gas chromatography-GC-MS resolves, 200～280 ℃ of resolution temperatures, the parsing time is 1～4min, then carries out the mensuration of different edible fungi volatile ingredients;
Gas chromatography condition of work in above-mentioned headspace gas chromatography-GC-MS:
HP-INNOWAX (30m * 25mm * 25um) capillary column; Carrier gas is He, and flow is 0.5～2ml/min, and injection port adopts not shunt mode; Temperature programme: 200～280 ℃ of injection ports, 40 ℃ of post initial temperature, stop 2～5min, with 5 ℃/min, rise to 150 ℃, stop 1min, then with 5～20 ℃/min, rise to 220 ℃, finally retain 1～4min; Mass spectrum condition of work: 250～300 ℃ of mass spectrometer interface temperature, current system is EI, 210～250 ℃ of ion source temperatures, level Four bar 150e, quality of scanning scope is 30～500u;
3., after machine examination rope and MAINLIB, NISTDEMO, 3 standard spectrum storehouses of WILLEY match as calculated by the mass spectrometric data obtaining in the mensuration process of above-mentioned different edible fungi volatile ingredient, the principal ingredient of different edible fungi volatile ingredients is carried out to qualitative and quantitative analysis;
(3), different edible fungi essential oil samples is carried out to Electronic Nose analysis, detailed process is as follows:
1., the different edible fungi essential oil sample 0.1～0.4g of step (1) gained is put into respectively to 10ml sampling receptacle;
2., sampling receptacle is placed in to Electronic Nose automatic sampler, under normal temperature, the sample introduction syringe needle of Electronic Nose is inserted to head space absorption in sampling receptacle, adsorption time 700～1000s, 40～60 ℃ of adsorption temps, stirring rate 500r/min;
3. the gas that, the gas sensor in Electronic Nose air chamber distributes different edible fungi essential oil samples detects 60～160s, lag time 400～800s, collect the data that gas sensor array gathers, the data that gather are processed, obtain Electronic Nose local flavor fingerprint analysis figure;
(4), by step (2) 3. the principal ingredient of the different edible fungi volatile ingredient of gained carry out the result of qualitative and quantitative analysis and Electronic Nose local flavor fingerprint analysis figure and the principal component analysis (PCA) figure of the 3. gained in step (3) comprehensively analyze, when recording the detailed volatile ingredient of different edible fungis, can obtain the whole flavor characteristic of different edible fungi volatile matter, thereby realize the discriminating of the flavor substance in different edible fungis.
Beneficial effect of the present invention
The present invention a kind ofly utilizes headspace gas chromatography-GC-MS and Electronic Nose coupling to distinguish method for distinguishing to edible fungi flavor substances, owing to having adopted headspace gas chromatography-GC-MS and Electronic Nose coupling technique, therefore when recording the detailed volatile ingredient of edible fungi, the whole flavor characteristic of edible fungi volatile matter can be obtained, thereby solved headspace gas chromatography-mass spectrometric hyphenated technique, the technical matters of concentrated expression cannot be carried out to the whole flavor characteristic of edible fungi volatile matter.Further, different according to the characteristic volatile component of different types of edible fungi, difference with the local flavor finger-print of its volatile ingredient integral body, can differentiate quickly and accurately and judge different edible fungi samples, the different chemical material corresponding according to each Electronic Nose sensor, can roughly determine the volatile species of edible fungi.
Accompanying drawing explanation
The edible fungi mushroom that in Fig. 1, embodiment 1, step (2) headspace gas chromatography-mass spectrometry records and the principal ingredient analysis chart of grifola frondosus volatile ingredient, as can be seen from Figure 1 the cumulative proportion in ANOVA of major component 1 and major component 2 surpasses 90%, major component 1 is 51.72%, major component 2 is 48.28%, substantially most information of edible fungi volatile ingredient have been represented, therefore get major component 1 and major component 2 represents all volatile ingredients of edible fungi that headspace gas chromatography-mass spectrometry records;
In Fig. 2, embodiment 1, step (3) utilizes Electronic Nose the flavor substance of mushroom and grifola frondosus to be analyzed to the Electronic Nose local flavor finger-print of gained, as can be seen from Figure 2 the overall flavor substance of grifola frondosus and mushroom is identical, but both responses are different in individual individual sensor; 1 mushroom wherein, 2 grifola frondosus;
Fig. 3, step in embodiment 1 (3) utilizes Electronic Nose the major component of the mushroom edible fungi volatile ingredient different with grifola frondosus to be carried out to the analysis chart of qualitative and quantitative analysis, wherein 1 is grifola frondosus, 2 is mushroom, as can be seen from Figure 3 utilize Electronic Nose to analyze the major component of the volatile ingredient of mushroom and grifola frondosus, the variance accumulation contribution rate of finding major component 1 just reaches 91.976%, illustrate that major component 1 has represented most information of edible fungi volatile ingredient substantially, in the X direction at major component 1 place, mushroom and grifola frondosus distance differ seldom, illustrate that difference is little.
Embodiment
Below by specific embodiment, also by reference to the accompanying drawings the present invention is further set forth, but do not limit the present invention.
Utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to the edible fungi flavor substances method for distinguishing that reflects, specifically comprise the following steps:
(1), preparation of samples:
Get respectively commercially available mushroom to be identified and grifola frondosus dry product, 80 ℃ of dry 4h, mechanical disintegration, crosses 40 mesh sieves, under the condition that is then 1:10 in solid-to-liquid ratio respectively, uses extraction by steam distillation 2.5h, obtains respectively the essential oil sample of mushroom and grifola frondosus;
(2), respectively the essential oil sample of the mushroom of step (1) gained and grifola frondosus is carried out to head space-gas chromatography-mass spectrometry analysis and measures, detailed process is as follows:
1. get, respectively the essential oil sample of 8ml mushroom and grifola frondosus and put into 15ml sampling receptacle, and be placed in 50 ℃ of water-baths, SPME extracting head (65 μ m PDMS/DVB, Supelco company) head space Static Adsorption 40min;
2., gas chromatographic sample introduction mouth that SPME extracting head is inserted in headspace gas chromatography-GC-MS resolves, 250 ℃ of resolution temperatures, the parsing time is 2min, then carries out the mensuration of mushroom and grifola frondosus volatile ingredient;
Above-mentioned headspace gas chromatography-GC-MS (Agilent 7890A gas chromatograph; Agilent 5975C mass spectrometer) gas chromatography condition of work in:
HP-INNOWAX (30m * 25mm * 25um) capillary column; Carrier gas is He, and flow is 1.0ml/min, and injection port adopts not shunt mode; Temperature programme: 250 ℃ of injection ports, 40 ℃ of post initial temperature, stop 3min, with 5 ℃/min, rise to 150 ℃, stop 1min, then with 10 ℃/min, rise to 220 ℃, finally retain 2min;
Mass spectrum condition of work: 280 ℃ of mass spectrometer interface temperature, current system is EI, 230 ℃ of ion source temperatures, level Four bar 150e, quality of scanning scope is 40～400u;
3., by the mass spectrometric data obtaining in the mensuration process of above-mentioned different edible fungi mushroom and grifola frondosus volatile ingredient, machine examination rope and MAINLIB, NISTDEMO, 3 standard spectrum storehouses of WILLEY match as calculated, respectively the principal ingredient of mushroom and grifola frondosus volatile ingredient is carried out to qualitative and quantitative analysis, result as shown in Figure 1, from Fig. a kind, can find out, the cumulative proportion in ANOVA of major component 1 and major component 2 surpasses 90%, major component 1 is 51.72%, major component 2 is 48.28%, has substantially represented most information of edible fungi volatile ingredient.Therefore get all volatile ingredients of edible fungi that major component 1 and major component 2 replace GC-MS to record, main volatile compounds in mushroom is sulfocompound, wherein 1,2,4-tri-thiacyclopentane content are higher, and in the volatile constituent of grifola frondosus, contain hardly sulfocompound;
(3), mushroom and grifola frondosus essential oil sample are carried out to Electronic Nose (FOX4000, Alpha M.O.S) analysis, detailed process is as follows:
1., the mushroom of step (1) gained and grifola frondosus essential oil sample 0.2g are put into respectively to 10ml sampling receptacle;
2., sampling receptacle is placed in to Electronic Nose automatic sampler, under normal temperature, the sample introduction syringe needle of Electronic Nose is inserted to head space absorption in sampling receptacle, adsorption time 900s, 50 ℃ of adsorption temps, stirring rate 500r/min;
3., the gas that gas sensor in Electronic Nose air chamber distributes mushroom and grifola frondosus essential oil sample respectively detects 120s, lag time 600s, collect the data that gas sensor array gathers, the data that gather are processed, the Electronic Nose local flavor fingerprint analysis figure of gained distinguishes as shown in Figures 2 and 3 with the analysis chart that utilizes Electronic Nose to analyze the major component of the volatile ingredient of mushroom and grifola frondosus, 1 mushroom wherein, 2 grifola frondosus, the local flavor finger-print that the Electronic Nose local flavor fingerprint analysis figure of gained is comprised of the response of Electronic Nose different sensors the volatile flavor substance component of mushroom and grifola frondosus, as can be seen from Figure 2, the overall flavor substance of grifola frondosus and mushroom is identical, but both responses are different in individual individual sensor, the volatile constituent major part that has shown thus grifola frondosus and mushroom is similar, but also having any different property composition between the two, be that some component content in mushroom is than high in grifola frondosus.As can be seen from Figure 3, by Electronic Nose, the major component of the volatile ingredient of mushroom and grifola frondosus is analyzed, the variance accumulation contribution rate of finding first principal component just reaches 91.976%, illustrate that major component 1 has represented most information of edible fungi volatile ingredient substantially, in the X direction at major component 1 place, mushroom and grifola frondosus distance differ seldom, illustrate that difference is little;
(4), by step (2) 3. the major component of the mushroom of the gained edible fungi volatile ingredient different with grifola frondosus carry out the result of qualitative and quantitative analysis and comprehensively analyze from the Electronic Nose local flavor fingerprint analysis figure of 3. gained in step (3) and the result of utilizing Electronic Nose to carry out qualitative and quantitative analysis to the major component of the mushroom edible fungi volatile ingredient different with grifola frondosus, thereby realize the discriminating of the flavor substance in mushroom edible fungis different from grifola frondosus.
By the flavor substance in the above-mentioned mushroom edible fungis different from grifola frondosus that obtain in embodiment 1, contrast, result shows, different edible fungis have different flavor substance features, through head space-gas chromatography-mass spectrum and detection by electronic nose, show as different volatile constituents and local flavor finger-print.By gas chromatography-mass spectrography technology, can obtain detailed edible fungi volatile flavor substance component, and quantitative and qualitative analysis, by the local flavor fingerprint analysis to edible fungi volatile constituent of Electronic Nose, can obtain the general profile of edible fungi volatile constituent on each sensor, the different chemical material corresponding according to each Electronic Nose sensor, can roughly determine the volatile species of edible fungi.
Foregoing is only the basic explanation of the present invention under conceiving, and according to any equivalent transformation that technical scheme of the present invention is done, all should belong to protection scope of the present invention.
Claims (4)
1. one kind is utilized headspace gas chromatography-GC-MS and Electronic Nose coupling to distinguish method for distinguishing to edible fungi flavor substances, it is characterized in that described to utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to carry out discriminating conduct to edible fungi flavor substances be to utilize headspace gas chromatography-GC-MS and Electronic Nose to gather the volatile matter of different edible fungis, by principal component analysis (PCA) and radar map, judge the similarity between edible fungi, and then differentiate different edible fungis;
Wherein principal component analysis (PCA), by a plurality of variablees by linear transformation to select a kind of Multielement statistical analysis method of less number significant variable.
2. as claimed in claim 1ly a kind ofly utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to the edible fungi flavor substances method for distinguishing that reflects, it is characterized in that described edible fungi is mushroom, grifola frondosus, flat mushroom, pleurotus eryngii, bolete, Ji Songrong, grey mushroom (abalonelike), agrocybe or Hericium erinaceus.
3. as claimed in claim 1 or 2ly a kind ofly utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to the edible fungi flavor substances method for distinguishing that reflects, it is characterized in that specifically comprising the following steps:
(1), edible fungi volatile essential oil is prepared:
Get commercially available different dry edible mushroom to be identified, respectively at 80 ℃ of dry 4h, mechanical disintegration, crosses 40 mesh sieves, under the condition that is then 1:10 in solid-to-liquid ratio respectively, uses extraction by steam distillation 2.5h, obtains different edible fungi essential oil samples;
(2), the different edible fungi essential oil samples of step (1) gained carried out to headspace gas chromatography-mass spectrophotometry measure, detailed process is as follows:
1. get, respectively 6～12ml edible fungi essential oil sample and put into 15ml sampling receptacle, and be placed in 30～60 ℃ of water-baths, SPME extracting head head space Static Adsorption 30 ~ 60min;
Described SPME extracting head is 65 μ m PDMS/DVB, Supelco company;
2., gas chromatographic sample introduction mouth that SPME extracting head is inserted in headspace gas chromatography-GC-MS resolves, 200～280 ℃ of resolution temperatures, the parsing time is 1～4min, then carries out the mensuration of different edible fungi volatile ingredients;
In above-mentioned headspace gas chromatography-GC-MS, gas chromatography condition of work:
HP-INNOWAX (30m * 25mm * 25um) capillary column; Carrier gas is He, and flow is 0.5～2ml/min, and injection port adopts not shunt mode; Temperature programme: 200～280 ℃ of injection ports, 40 ℃ of post initial temperature, stop 2～5min, with 5 ℃/min, rise to 150 ℃, stop 1min, then with 5～20 ℃/min, rise to 220 ℃, finally retain 1～4min;
Mass spectrum condition of work: 250～300 ℃ of mass spectrometer interface temperature, current system is EI, 210～250 ℃ of ion source temperatures, level Four bar 150e, quality of scanning scope is 30～500u;
3., after machine examination rope and MAINLIB, NISTDEMO, 3 standard spectrum storehouses of WILLEY match as calculated by the mass spectrometric data obtaining in the mensuration process of above-mentioned different edible fungi volatile ingredient, the principal ingredient of different edible fungi volatile ingredients is carried out to qualitative and quantitative analysis;
(3), different edible fungi essential oil samples is carried out to Electronic Nose analysis, detailed process is as follows:
1., the different edible fungi essential oil sample 0.1～0.4g of step (1) gained is put into respectively to 10ml sampling receptacle;
2., sampling receptacle is placed in to Electronic Nose automatic sampler, under normal temperature, the sample introduction syringe needle of Electronic Nose is inserted to head space absorption in sampling receptacle, adsorption time 700～1000s, 40～60 ℃ of adsorption temps, stirring rate 500r/min;
3. the gas that, the gas sensor in Electronic Nose air chamber distributes different edible fungi essential oil samples detects 60～160s, lag time 400～800s, collect the data that gas sensor array gathers, the data that gather are processed, obtain Electronic Nose local flavor fingerprint analysis figure;
(4), by step (2) 3. the principal ingredient of the different edible fungi volatile ingredient of gained carry out the result of qualitative and quantitative analysis and comprehensively analyze from the Electronic Nose local flavor fingerprint analysis figure of 3. gained in step (3) and the result of utilizing Electronic Nose to carry out qualitative and quantitative analysis to the major component of different edible fungi volatile ingredients, thereby realize the discriminating of the flavor substance in different edible fungis.
4. as claimed in claim 3ly a kind ofly utilize headspace gas chromatography-GC-MS and Electronic Nose coupling to the edible fungi flavor substances method for distinguishing that reflects, it is characterized in that described SPME extracting head is 65 μ m PDMS/DVB, Supelco company;
Described headspace gas chromatography-GC-MS, wherein gas chromatograph is Agilent 7890A, mass spectrometer is Agilent 5975C;
Described Electronic Nose is FOX4000, Alpha M.O.S.
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