CN109917046A - A kind of detection method of Yunnan truffle aroma substance - Google Patents
A kind of detection method of Yunnan truffle aroma substance Download PDFInfo
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- CN109917046A CN109917046A CN201910298890.1A CN201910298890A CN109917046A CN 109917046 A CN109917046 A CN 109917046A CN 201910298890 A CN201910298890 A CN 201910298890A CN 109917046 A CN109917046 A CN 109917046A
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Abstract
The invention discloses a kind of detection methods of Yunnan truffle aroma substance, which is characterized in that the aroma substance in extraction truffle；Aroma substance is tested and analyzed using internal standard method for gas chromatography technology；Smelling is carried out to truffle using the method for subjective appreciation, establishes multiple sensory attributes；Filter out crucial aroma substance；The aroma substance in truffle is investigated using Partial Least Squares and the correlation between sensory attribute identifies the characteristic perfume substance in truffle according to the correlation of aroma substance and note.The present invention gives a mark to the analysis and sensory evaluation of the characteristic perfume substance progress qualitative, quantitative of the fresh truffle of same place of production different cultivars to not note branch using headspace solid-phase microextraction and gas chromatography combined with mass spectrometry and sense organ smelling, data are analyzed by chemo metric software simultaneously, establish truffle aroma substance and sense organ model.The present invention carries out the analysis of qualitative, quantitative to the Volatile infochemicals in truffle, filters out and constitutes the critical aroma substance of truffle.
Description
Technical field
The present invention relates to a kind of detection methods of Yunnan truffle aroma substance, belong to technical field of analysis and detection.
Background technique
Truffle, also known as ferfas are a kind of symbiotic effects that oneself life cycle is strictly completed by other biological, generally
Silk be grown in plant root, establish exotrophic mycorrhiza, form fructification.External many researchers have centainly truffle kind
Research, so far, from the truffle of different cultivars discovery more than 200 kind of volatile organic compounds.Flavour of food products section
Worker is ground to describe the fragrance of truffle are as follows: it is sexy, there is temptation, soil fragrance, Moschus taste, acid and unique garlic are fragrant.
Although different truffle kinds has different smells, it is eight carbonizations that it is biggish, which to occupy specific gravity, for content in volatile flavor
Object is closed, specifically includes that 1-OCOL, 3- octanol, 2- octen-1-ol, 1- octene -3- ketone, 3- octanone etc..Some Containing Sulfurs
Object is closed, although content is smaller, very big effect is played to the flavor contribution of truffle, such as: dimethyl disulfide, dimethyltrisulfide
Ether, 1,2,4- tri- tiacyclopentanes, thiophene derivant etc., it is considered to be the character flavor compound of truffle.
The characteristics of gas chromatography (GC) is separating capacity strong, high sensitivity, quantitative quasi-, equipment and easy to operate, is suitable for
In doing multi-component quantitative analysis.The characteristics of mass spectrography (MS) is that distinguishing ability is strong, high sensitivity, fast response time, is suitable for
The Qualitative Identification of pure compound.Mass spectrography can carry out effective qualitative analysis, but just show to the analysis of complex organic compound
It obtains helpless；And chromatography is that one kind efficiently separates analysis method to organic compound, particularly suitable for organising
The quantitative analysis of object is closed, but qualitative analysis is then relatively difficult.Therefore, the effective of the two combines one progress of offer complicated organic
The efficient qualitative and quantitative analysis tool of compound, i.e., present " gas chromatograph-mass spectrometer ".Currently, common SPME combination GC-MS energy
The simply and efficiently volatile flavor component in extractive analysis truffle.
Summary of the invention
The technical problems to be solved by the present invention are: providing a kind of detection method of Yunnan truffle aroma substance, this method
Detailed analysis can be carried out to the aroma substance of truffle, provide reference for the aroma substance preferably using truffle.
To solve the above-mentioned problems, the present invention provides a kind of detection method of Yunnan truffle aroma substance, feature exists
In, comprising the following steps:
Step 1): with the aroma substance in the method extraction truffle of headspace solid-phase microextraction；
Step 2): the aroma substance that step 1) obtains is tested and analyzed using internal standard method for gas chromatography technology, to truffle
In aroma substance carry out qualitative and quantitative analysis；
Step 3): smelling is carried out to truffle using the method for subjective appreciation, establishes multiple sensory attributes；
Step 4): calculating the fragrance energy value of the aroma substance detected, filters out the key that energy value is greater than rated value
Aroma substance；
Step 5): using Partial Least Squares investigate truffle in crucial aroma substance and sensory attribute between it is related
Property, establish correlative relationship between note corresponding to the aroma substance in the truffle of Partial Least Squares model and sensory attribute
Dependent variable be " crucial aroma substance ", independent variable is " sensory attribute ", related to sensory attribute according to crucial aroma substance
Property, further verify crucial aroma substance characteristic perfume contribution in truffle.
Preferably, the truffle in the step 1) uses the black pine of Yunnan Nanhua County autonomous region, the Yi nationality, distributed over Yunnan, Sichuan and Guizhou, Yunnan kahikatea
The false recessed truffle of dew or Yunnan.
Preferably, the step 1) specifically: after cleaning the truffle of fresh picking, select that head is uniform in size, color
Similar truffle is ground into truffle mud, is put into -8 DEG C of refrigerators and is sealed, for use；Fresh truffle sample is put into head space
In solid phase microextraction bottle, internal standard compound is added, then 30min is balanced at (45 ± 1) DEG C, with the good solid-phase micro-extracting device of aging
Headspace extraction 30min, then extraction equipment is inserted into GC sample injector, 5min is desorbed under 250 DEG C not shunt mode.
It is highly preferred that the internal standard compound uses concentration for the o-dichlorohenzene of 100mg/L.
Preferably, in the step 2) gas-chromatography parameter are as follows: chromatographic column use HP-INNOWAX (60m × 0.25mm
× 0.25 μm) chromatographic column；N2For carrier gas, carrier gas flux 1.8mL/min, input mode is not shunt, and keeps 3min；Injection port
Temperature is 250 DEG C, temperature program: 50 DEG C of reservations 3min are warming up to 100 DEG C with the rate of 10 DEG C/min, reservation 5min, with 3 DEG C/
Min is warming up to 140 DEG C, retains 10min, rises to 200 DEG C with 2 DEG C/min, retains 5min；Using EI ion source, ionization voltage is
70eV, ion source temperature are 230 DEG C；Interface temperature is 250 DEG C, using full scan mode, mass scan range are as follows: m/z=20
~350.
Preferably, the step 3) specifically: sense organ smelling group chooses multiple senses by 10 experienced member compositions
Official's attribute, sensory evaluation personnel carry out sensory evaluation using fragrance of the ten point system to truffle to every kind of sensory attribute, wherein 0 indicates
Indicate that fragrance is moderate without fragrance, 5,10 indicate aroma strength highest, everyone repeats every kind of sensory attribute of truffle to evaluate 1 respectively
~3 times, and each appraisal result is recorded, finally it is averaged.
It is highly preferred that the sensory attribute is divided into 7 kinds of notes, respectively mushroom fragrance, nut taste, rotten vegetable leaf taste, fat
Green fragrance, roast potato taste, flower sweet fragrance and sulphur sample musty.Strong mushroom fragrance is 1-OCOL, 3- octanone, 3- octanol
Deng eight carbon alcohol ketone odor characteristic present；Nut note is by a large amount of 2 methyl butyraldehyde, the performance of 3- methylbutyraldehyd；Rotten Chinese cabbage
Dimethyl sulfide has the taste of rot dish leaf and corn in note；Hexanal, aldehyde C-9, capraldehyde and anti-, trans- 2,4 decadienals
Fatty note and green fragrant note are then provided；The fragrance of roast potato is embodied by 3 methylthiol propyl alcohol and 3- methylthiopropionaldehyde；Benzene
Acetaldehyde, benzyl carbinol, methyl para-cresol are then with rose, and spending for Yilan is fragrant and sweet；Double methyl mercapto methane, double disulfides etc. then have
Pungent sulphur sample smell.
Preferably, the step 4) specifically: utilize formula: fragrance energy value=aroma substance concentration/aroma substance threshold
Value calculates the fragrance energy value for the volatile compound that gaschromatographic mass spectrometry detected, and filters out the key that energy value is greater than 1
Aroma substance.
The present invention is using headspace solid-phase microextraction and gas chromatography combined with mass spectrometry and sense organ smelling to same place of production difference product
The characteristic perfume substance of the fresh truffle of kind carries out the analysis of qualitative, quantitative and gives a mark to sensory attribute, living using fragrance
Force value filters out crucial aroma substance and analyzes data by chemo metric software simultaneously, establishes truffle key aroma substance and sense
The model of official's attribute further verifies the characteristic perfume contribution of crucial aroma substance.The present invention is to the volatile aroma in truffle
Substance carries out the analysis of qualitative, quantitative, filters out and constitutes the critical aroma substance of truffle.The present invention is directed to understand different cultivars
The difference of truffle flavor components provides certain reference for the flavor research of domestic truffle, and also the Related product for truffle flavor is opened
Hair provides technical parameter and guidance.
Detailed description of the invention
Fig. 1 is the total ion current figure of fresh black pine；
Fig. 2 is the total ion current figure of fresh white truffle；
Fig. 3 is the total ion current figure of the fresh recessed truffle of vacation；
Fig. 4 is the radar map of three different cultivars truffle sensory attributes；
Fig. 5 is the correlation analysis figure using Partial Least Squares to aroma substance and sensory attribute crucial in truffle.
Specific embodiment
In order to make the present invention more obvious and understandable, hereby with preferred embodiment, and attached drawing is cooperated to be described in detail below.
Embodiment
A kind of detection method of Yunnan truffle aroma substance, comprising the following steps:
1, material and instrument and equipment
1.1 material
The black pine (T1) for originating from Yunnan Nanhua County autonomous region, the Yi nationality, distributed over Yunnan, Sichuan and Guizhou is selected, Yunnan kahikatea reveals (T2), the false recessed truffle in Yunnan
(T3) truffle of three different cultivars is tested
1.2 instrument and equipment
7890A-5973C type GC-MS combined instrument, Agilent Technologies (China) Co., Ltd, water-bath, blender, low temperature
Refrigerator
2, experimental method and content
The pre-treatment of 2.1 truffles
It goes after cleaning the truffle of fresh picking, selects that head is uniform in size, and truffle similar in color is put into JYL-C051
In type blender, it is ground into truffle mud, -8 DEG C of refrigerators is put into and is sealed, for use.
2.2 GC-MS detection
2.2.1 headspace solid-phase microextraction: the fresh truffle sample of 2.0g is accurately weighed, 20mL headspace solid-phase microextraction bottle is put into
In, add 5 μ L internal standard compounds (100mg/L o-dichlorohenzene), balance 30min at (45 ± 1) DEG C, is filled with the good solid phase microextraction of aging
Headspace extraction 30min is set, then extraction equipment is inserted into GC sample injector, desorbs 5min under 250 DEG C not shunt mode；
2.2.2 HP-INNOWAX (60m × 0.25mm × 0.25 μm) chromatographic column, N GC condition: are used2For carrier gas, flow is
1.8mL/min, input mode are not shunt, and keep 3min.Injector temperature is 250 DEG C, temperature program: 50 DEG C of reservation 3min,
100 DEG C are warming up to the rate of 10 DEG C/min, retains 5min, 140 DEG C is warming up to 3 DEG C/min, retains 10min, with 2 DEG C/min
200 DEG C are risen to, 5min is retained.Using EI ion source, ionization voltage 70eV, ion source temperature is 230 DEG C, and interface temperature is
250 DEG C, using full scan mode (mass scan range are as follows: m/z=20~350)
2.2.4 the qualitative-and-quantitative method of fragrance (volatile component): a. is using NIST08 and Wiley spectrum library to isolating
Peak is retrieved, through the retention time of n-alkane (C4-C30) under the same conditions, based on KOVATS retention index formula
Calculate the retention index (RI) of volatile materials.The retention index being calculated is matched with Wiley7n.l database, it is qualitative
Obtained effective volatile materials carries out quantitative calculating by internal standard method.It carries out 3 repetitions to test, Aromatic Matter Contents data
Take 3 laboratory mean values.
Using subjective appreciation method to truffle carry out smelling the step of in, sense organ smelling group by 10 it is experienced at
Member's composition, sensory evaluation personnel carry out sensory evaluation using fragrance of the ten point system to truffle to every kind of sensory attribute, wherein 0 indicates
Indicate that fragrance is moderate without fragrance, 5,10 indicate aroma strength highest, everyone repeats 7 kinds of sensory attributes of truffle to evaluate 1 respectively
~3 times, and each appraisal result is recorded, finally it is averaged.
2.2.5 it filters out the method for crucial aroma substance: utilizing formula: fragrance energy value=aroma substance concentration/fragrance
Substance threshold value calculates the fragrance energy value for the volatile compound that gaschromatographic mass spectrometry detected, and filters out energy value greater than 1
Crucial aroma substance.
3, results and discussion
3.1 fresh black pine (T1) is obtained by the above method aroma substance it is as shown in table 1.
Table 1
Fig. 1 is the total ion current figure of fresh black pine (T1).
3.2 is as shown in table 2 by the aroma substance of above method acquisition fresh white truffle (T2).
Table 2
Fig. 2 is the total ion current figure of fresh white truffle (T2).
3.3 the fresh recessed truffle of vacation (T3) is obtained by the above method aroma substance it is as shown in table 3.
Table 3
Fig. 3 is the total ion current figure of the fresh recessed truffle of vacation (T3).
3.4 carry out smelling to truffle using the method for subjective appreciation, and choose 7 kinds of sensory attributes (mushroom, nut, rotten dishes
Leaf, fat is green fragrant, and roast potato spends fragrant and sweet and sulphur sample musty) being described property of truffle is analyzed, perfumer beats different notes
Divide situation as shown in table 4.
Table 4
By table 4 as it can be seen that the corresponding sensory attribute marking of the truffle fragrance of different cultivars is all different, explanation can pass through sense
The method of official's evaluation identifies the odor characteristic of truffle.
Fig. 4 is the radar map of the sensory attribute of three kinds of different cultivars truffles.
3.5 from the truffle of three different cultivars, calculates fragrance energy value, picks out 24 kinds that fragrance energy value is greater than 1
Crucial aroma substance, as shown in table 5.
Table 5
Fig. 5 be using Partial Least Squares to key aroma substance A1-A24 (Y-axis) in truffle and sensory attribute (X-axis) into
Row correlation analysis.PC1 indicates X-axis (7 kinds of sensory attributes), and PC2 indicates Y-axis (24 crucial aroma substance), the contribution rate of X variable
Contribution rate for 90% and Y variable is 83%, shows the information that Partial Least Squares model can preferably explain that sample provides.
Conclusion: the volatile component in three different cultivars truffles is divided using gas chromatography-mass spectrography (GC-MS)
From identification.The result shows that as shown in table 1-3, detecting 32,27 and 34 kind of substance respectively in T1, T2 and T3.Calculate all inspections
The fragrance energy value of volatile materials out picks out the crucial aroma substance that 24 kinds of energy values are greater than 1.Utilize offset minimum binary
Method carries out correlation analysis to crucial aroma substance and sensory attribute, further verifies crucial aroma substance to truffle characteristic perfume
Contribution.Dimethyl sulfide, 3- methylbutyraldehyd, dimethyl disulfide, 3- octanone, double methyl mercapto methane, octanal, 1- octene -3-
Ketone, 1-OCOL and phenylacetaldehyde contain higher concentration and higher fragrance energy value in uantitative analytical, to truffle
Fragrance contribution it is larger.Generally speaking, the fragrance of Yunnan truffle has more the fragrance of a flower, mushroom tone and fragrant and sweet, this is also that Yunnan truffle is special
Some fragrance.By studying to domestic truffle fragrance deeply, comprehensively, truffle product with Chinese characteristics is further developed,
Chinese truffle market is changed from original original exploitation to intensive processing type.
The present invention provides not only the detection method of truffle characteristic perfume, and also the analysis result based on GC-MS is to truffle perfume
Gas carries out sensory evaluation and sensory attribute, is carried out with correlation of the method for data statistics to crucial aroma substance and sensory attribute
Analysis.
Claims (8)
1. a kind of detection method of Yunnan truffle aroma substance, which comprises the following steps:
Step 1): with the aroma substance in the method extraction truffle of headspace solid-phase microextraction；
Step 2): the aroma substance that step 1) obtains is tested and analyzed using internal standard method for gas chromatography technology, in truffle
Aroma substance carries out qualitative and quantitative analysis；
Step 3): smelling is carried out to truffle using the method for subjective appreciation, establishes multiple sensory attributes；
Step 4): calculating the fragrance energy value of the aroma substance detected, filters out the crucial fragrance that energy value is greater than rated value
Substance；
Step 5): the crucial aroma substance in truffle and the correlation between sensory attribute are investigated using Partial Least Squares, really
Between note corresponding to aroma substance and sensory attribute in the truffle of vertical Partial Least Squares model correlative relationship because
Variable is " crucial aroma substance ", independent variable is " sensory attribute ", according to the correlation of crucial aroma substance and sensory attribute, into
One step demonstrate,proves crucial aroma substance characteristic perfume contribution in truffle.
2. the detection method of truffle aroma substance in Yunnan as described in claim 1, which is characterized in that in the step 1)
Truffle is using the black pine of Yunnan Nanhua County autonomous region, the Yi nationality, distributed over Yunnan, Sichuan and Guizhou, the false recessed truffle of Yunnan kahikatea dew or Yunnan.
3. the detection method of truffle aroma substance in Yunnan as described in claim 1, which is characterized in that the step 1) is specific
Are as follows: it after cleaning the truffle of fresh picking, selects that head is uniform in size, truffle similar in color, is ground into truffle mud, put
Enter -8 DEG C of refrigerators to be sealed, for use；Fresh truffle sample is put into headspace solid-phase microextraction bottle, internal standard compound is added, then
30min is balanced at (45 ± 1) DEG C, is inserted into the good solid-phase micro-extracting device headspace extraction 30min of aging, then by extraction equipment
GC sample injector desorbs 5min under 250 DEG C not shunt mode.
4. the detection method of truffle aroma substance in Yunnan as claimed in claim 3, which is characterized in that the internal standard compound is using dense
Degree is the o-dichlorohenzene of 100mg/L.
5. the detection method of truffle aroma substance in Yunnan as described in claim 1, which is characterized in that gas phase in the step 2)
The parameter of chromatography are as follows: chromatographic column uses HP-INNOWAX (60m × 0.25mm × 0.25 μm) chromatographic column；N2For carrier gas, carrier gas stream
Amount is 1.8mL/min, and input mode is not shunt, and keeps 3min；Injector temperature is 250 DEG C, temperature program: 50 DEG C of reservations
3min, is warming up to 100 DEG C with the rate of 10 DEG C/min, retains 5min, is warming up to 140 DEG C with 3 DEG C/min, retains 10min, with 2
DEG C/min rises to 200 DEG C, retain 5min；Using EI ion source, ionization voltage 70eV, ion source temperature is 230 DEG C；Interface temperature
Degree is 250 DEG C, using full scan mode, mass scan range are as follows: m/z=20~350.
6. the detection method of truffle aroma substance in Yunnan as described in claim 1, which is characterized in that the step 3) is specific
Are as follows: sense organ smelling group chooses multiple sensory attributes, sensory evaluation personnel are to every kind of sense organ by 10 experienced member compositions
Attribute carries out sensory evaluation using fragrance of the ten point system to truffle, wherein 0 indicates no fragrance, 5 indicate that fragrance is moderate, and 10 indicate fragrant
Gas intensity highest, everyone repeats to evaluate 1~3 time to every kind of sensory attribute of truffle respectively, and records each appraisal result, finally
It is averaged.
7. the detection method of truffle aroma substance in Yunnan as claimed in claim 6, which is characterized in that the sensory attribute is divided into
7 kinds of notes, respectively mushroom fragrance, nut taste, rotten vegetable leaf taste, the green fragrance of fat, roast potato taste, flower sweet fragrance and sulphur sample are mould
Taste.
8. the detection method of truffle aroma substance in Yunnan as described in claim 1, which is characterized in that the step 4) is specific
Are as follows: formula: fragrance energy value=aroma substance concentration/aroma substance threshold value is utilized, calculates what gaschromatographic mass spectrometry detected
The fragrance energy value of volatile compound filters out the crucial aroma substance that energy value is greater than 1.
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| CN114578001A (en) * | 2022-03-14 | 2022-06-03 | 厦门市燕之屋丝浓食品有限公司 | Sensory quality evaluation method for high-quality bowl-packaged cubilose |
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