CN111307973A - Method for releasing combined-state aroma substances of kiwi fruit juice - Google Patents

Method for releasing combined-state aroma substances of kiwi fruit juice Download PDF

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CN111307973A
CN111307973A CN202010155276.2A CN202010155276A CN111307973A CN 111307973 A CN111307973 A CN 111307973A CN 202010155276 A CN202010155276 A CN 202010155276A CN 111307973 A CN111307973 A CN 111307973A
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周元
傅虹飞
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Northwest A&F University
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Abstract

The invention relates to a method for releasing combined-state aroma substances of kiwi fruit juice, which comprises the following steps of 1) selecting kiwi fruits, namely selecting kiwi fruit fruits with good maturity, regular fruit types and no damage, 2) juicing, namely peeling fresh kiwi fruits, cutting into pieces, juicing and removing fruit residues to obtain kiwi fruit juice, 3) performing enzymolysis, namely adding β -glucosidase into the kiwi fruit juice, performing heat preservation and enzymolysis, wherein the adding amount of β -glucosidase is 4-12U/g of fruit juice, the enzymolysis temperature is 20-50 ℃, the enzymolysis time is 0.5-12h, and 4) analyzing, namely taking an enzymolyzed fruit juice sample, analyzing the influence of enzymolysis treatment on the composition and content of the aroma substances of the kiwi fruit juice by an electronic nose and solid-phase microextraction gas chromatography-mass spectrometry combined technology, and determining the release degree of the combined-state aroma substances.

Description

Method for releasing combined-state aroma substances of kiwi fruit juice
Technical Field
The invention relates to the technical field of fruit juice processing technologies, in particular to a method for releasing combined-state aroma substances of kiwi fruit juice.
Background
Kiwi fruit is popular with consumers due to its unique taste and rich nutrition. The planting area and the yield of Chinese kiwi fruits are the first in the world, the main product consumed in the domestic market at present is still fresh kiwi fruits, and the variety and the quality of the kiwi fruit deep-processed products are far from those of common fruits such as apples and grapes. Although products such as kiwi fruit juice, kiwi fruit wine and the like are already in the domestic market, the kiwi fruit juice and the kiwi fruit wine have thin flavor and poor aroma quality. The kiwi fruit juice is one of kiwi fruit processed products, maintains and effectively improves the types and the content of aroma components, and is a technical problem to be solved urgently.
At present, two ways for improving the aroma of fruit processing products are mainly provided, one is to reduce the loss of the aroma of the fruits during the processing and storage processes as much as possible, and the other is to release potential aroma substances in the fruits as much as possible. The content of the combined aroma precursor in the fruit, which is considered as a potential aroma source, is 10 times that of the free aroma. Since the bound aroma precursor itself is non-odorous and non-volatile or non-volatile, in order for these potential aroma precursor molecules to function, it is necessary to release volatile aroma substances by unbinding the bound groups. The enzymolysis technology is considered to be one of methods for treating the precursor of the combined-state aroma substances and releasing potential volatile aroma substances, at present, researches on releasing the combined-state aroma substances of kiwi fruits by the enzymolysis technology are rarely reported, and the method for releasing the combined-state aroma substances of kiwi fruit juice is still blank.
Disclosure of Invention
The invention aims to solve the technical problems in the background art, β -glucosidase from different sources is adopted to treat the aroma precursor substance in the combined state in the fresh kiwi fruit juice, and the change of the composition and the content of the aroma substance of the kiwi fruit before and after the enzymolysis treatment is analyzed by an electronic nose (E-nose) and solid phase micro-extraction gas chromatography-mass spectrometry (SPME-GC-MS) to clarify the release degree of the aroma precursor substance in the combined state, namely to effectively release the aroma substance with volatility.
The technical solution of the invention is as follows: the invention relates to a method for releasing combined aroma substances of kiwi fruit juice, which is characterized by comprising the following steps: the method comprises the following steps:
1) selecting fruits: selecting kiwi fruits with good maturity, regular fruit type and no damage;
2) juicing: peeling fresh kiwi fruits, cutting into pieces, squeezing to obtain juice, and removing fruit residues to obtain kiwi fruit juice;
3) performing enzymolysis, namely adding β -glucosidase into kiwi fruit juice, and performing heat preservation and enzymolysis, wherein β -glucosidase is selected from enzyme I, enzyme II and enzyme III, the addition amount of β -glucosidase is 4-12U/g fruit juice, the enzymolysis temperature is 20-50 ℃, and the enzymolysis time is 0.5-12 h;
4) and (3) analysis: taking an enzymolysis result juice sample, analyzing the influence of enzymolysis treatment on the composition and the content of aroma substances of the kiwi fruit juice by adopting an electronic nose and an SPEM-GC-MS, and determining the release degree of the combined aroma precursor substances.
Preferably, β -glucosidase used in step 3) is derived from microbial glycosidase I, microbial glycosidase II and plant glycosidase III, which are all commercially available.
Preferably, in step 3), β -glucosidase was added in an amount determined according to enzyme activity measured by mixing 100mg enzyme with 5mL of 40mM p-nitrophenyl- β -D-glucopyranoside solution and 4mL of 1M citrate-phosphate buffer (pH 4.0), incubating at 40 ℃ for 30min, adding 2mL of 3M sodium carbonate solution to stop the reaction, and colorimetric reading at 405nm with enzyme activity expressed as U/mg enzyme.
Preferably, in step 3), the enzymolysis temperature can be controlled by water bath, gas bath or jacket heat preservation, and the enzymolysis temperature is based on the central temperature of the fruit juice.
Preferably, in the step 4), the method for detecting the aroma composition by the electronic nose comprises the following steps: diluting 1g of the kiwi fruit juice sample subjected to enzymolysis by 5 times by adding pure water, collecting data every second with the inlet flow rate of an electronic nose being 300mL/min for 60s, and cleaning for 300 s; each group is six parallel; table 1 shows the characteristics of the 10 sensors of the electronic nose.
TABLE 1 characteristics of electronic nose sensor
Sensor serial number Sensor name Feature(s)
S1 W1C Sensitive to aromatic substances
S2 W5S High sensitivity and sensitivity to nitrogen oxides
S3 W3C Sensitive to ammonia and aromatic compounds
S4 W6S Is mainly sensitive to hydrogen
S5 W5C Sensitive to alkanes and aromatics
S6 W1S Sensitive to methane, similar to sensor No. 8
S7 W1W Is sensitive to terpenes
S8 W2S Sensitive to alcohols, partially aromatic compounds
S9 W2W Sensitive to aromatic compounds and organic sulfides
S10 W3S Is sensitive to alkanes
Preferably, in the step 4), the method for detecting the aroma components by SPME-GC-MS comprises the following steps: taking 5.0g of the kiwi fruit juice sample after enzymolysis, adding 2.5mL of saturated sodium chloride solution and 10 microliters of cyclohexanone internal standard (0.5mg/mL), preheating at 45 ℃ for 2.5min, extracting and adsorbing for 45min, and analyzing for 5min at GC 250 ℃; a gas chromatography column using DB-1MS (60 m.times.0.25 mm, 0.25 μm); helium flow rate is 1.5mL/min, no split flow; GC temperature setting: maintaining at 40 deg.C for 2min, increasing to 120 deg.C at 6 deg.C/min for 5min, increasing to 200 deg.C at 8 deg.C/min for 2min, and increasing to 250 deg.C at 10 deg.C/min for 8 min; an EI ionization source is adopted by MS, the electron energy is 70eV, the ion source temperature is 230 ℃, and the scanning range is 30.00-500.00 m/z; and (3) the test result is searched and compared with a standard spectrogram in an NIST mass spectrum library to determine the aroma components, and the content of the aroma components is calculated according to the response value of the internal standard cyclohexanone added in the sample.
Preferably, in the step 4), the detected electronic nose is combined with a SPEM-GC-MS detection technology to analyze the change of the composition and the content of the aroma substances, and PCA and O2PLS analysis are carried out to determine main substances influencing the detection of the change of the aroma response value of the kiwi fruit juice by the electronic nose.
Preferably, the step 4) is followed by a step 5) of determining the optimal process parameters for releasing the combined-state aroma substances according to the analysis result, wherein the optimal process parameters comprise the addition amount of the microbial source β -glucosidase II of 8U/g fruit juice, the enzymolysis temperature of 35 ℃ and the enzymolysis time of 4 hours.
Preferably, in the step 1), the hardness range of the kiwi fruit is 5.0-20.0N, the weight range of a single fruit is 50-150g, and the fruit shape is full.
Preferably, in the step 2), the kiwi fruits are peeled, the bitter and astringent feeling is reduced, and the method for removing the pomace after juicing comprises filtering, extruding or centrifuging.
The method adopts β -glucosidase to carry out enzymolysis treatment on kiwi fruit juice, analyzes the change of the composition and the content of aroma substances by combining an electronic nose with an SPEM-GC-MS detection technology, analyzes main substances influencing the aroma response value of the kiwi fruit juice detected by the electronic nose through PCA and O2PLS analysis, and determines the release degree of the aroma substances in the combined state of the kiwi fruit juice, and not only can obtain the optimal process parameters of β -glucosidase to carry out enzymolysis treatment on the kiwi fruit juice to release the combined state aroma precursor substances through the technology of combining the electronic nose with the SPEM-GC-MS detection and a scientific quantitative analysis method, but also can determine the positive effects of the enzymolysis treatment on the increase of the types and the total amount of the aroma substances in the kiwi fruit juice.
Drawings
FIG. 1 is a curve of the response values of different sensors of the electronic nose to the aroma components of kiwi fruit juice after enzymolysis;
FIG. 2 shows the variation of the kiwi fruit juice aroma component response values before and after enzymolysis detected by different sensors of the electronic nose;
FIG. 3 is a SPME-GC-MS (detection of total ion flow spectrum of aroma components of kiwi fruit juice before and after enzymolysis);
FIG. 4 is a comparison of the total amount of aroma components of kiwi fruit juice before and after SPME-GC-MS detection of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific examples below:
the method of the embodiment of the invention comprises the following steps:
1) selecting fruits: selecting kiwi fruits with good maturity, regular fruit type and no damage; the hardness range of the kiwi fruit is required to be 5.0-20.0N, the weight range of a single fruit is required to be 50-150g, and the fruit shape is full.
2) Juicing: peeling fresh kiwi fruits, cutting into pieces, squeezing to obtain juice, and removing fruit residues to obtain kiwi fruit juice; the kiwi fruit is peeled, the bitter and astringent feeling is reduced, and the method for removing the fruit residues after juicing comprises filtering, extruding or centrifuging.
3) And (2) performing enzymolysis, namely adding β -glucosidase into kiwi fruit juice, performing heat preservation enzymolysis, wherein β -glucosidase can be selected to be enzyme I, enzyme II and enzyme III, the addition amount of β -glucosidase is 4-12U/g of fruit juice, the enzymolysis temperature is 20-50 ℃, the enzymolysis time is 0.5-12h, the selected β -glucosidase is respectively derived from microbial source glucosidase I, microbial source glucosidase II and plant source glucosidase III, the addition amount of the glycosidase belongs to commercial sources, β -glucosidase is determined according to enzyme activity, the enzyme activity determination method comprises the steps of mixing 100mg of enzyme with 5mL of 40mM p-nitrophenyl- β -D-glucopyranoside solution and 4mL of 1M citric acid-phosphate buffer solution (pH 4.0), incubating for 30min at 40 ℃, adding 2mL of 3M sodium carbonate solution to terminate the reaction, performing colorimetric reading at 405nm, and controlling the enzyme activity by U/mg of enzyme, wherein the temperature can be controlled by adopting a water bath, a gas bath or a heat preservation temperature, and the enzymolysis center is used as a standard for the fruit juice.
4) And (3) analysis: taking an enzymolysis result juice sample, analyzing the influence of enzymolysis treatment on the composition and the content of aroma substances of the kiwi fruit juice through an electronic nose and an SPEM-GC-MS, and determining the release degree of the combined aroma substances. The method for detecting the fragrance composition by the electronic nose comprises the following steps: diluting 1g of the kiwi fruit juice sample subjected to enzymolysis by 5 times by adding pure water, collecting data every second with the inlet flow rate of an electronic nose being 300mL/min for 60s, and cleaning for 300 s; each group is six parallel; table 1 shows the characteristics of the 10 sensors of the electronic nose.
TABLE 1 characteristics of electronic nose sensor
Sensor serial number Sensor name Feature(s)
S1 W1C Sensitive to aromatic substances
S2 W5S High sensitivity and sensitivity to nitrogen oxides
S3 W3C Sensitive to ammonia and aromatic compounds
S4 W6S Is mainly sensitive to hydrogen
S5 W5C Sensitive to alkanes and aromatics
S6 W1S Sensitive to methane, similar to sensor No. 8
S7 W1W Is sensitive to terpenes
S8 W2S Sensitive to alcohols, partially aromatic compounds
S9 W2W Sensitive to aromatic compounds and organic sulfides
S10 W3S Is sensitive to alkanes
The method for detecting the aroma components by SPME-GC-MS comprises the following steps: taking 5.0g of the kiwi fruit juice sample after enzymolysis, adding 2.5mL of saturated sodium chloride solution and 10 microliters of cyclohexanone internal standard (0.5mg/mL), preheating at 45 ℃ for 2.5min, extracting and adsorbing for 45min, and analyzing for 5min at GC 250 ℃; a gas chromatography column using DB-1MS (60 m.times.0.25 mm, 0.25 μm); helium flow rate is 1.5mL/min, no split flow; GC temperature setting: maintaining at 40 deg.C for 2min, increasing to 120 deg.C at 6 deg.C/min for 5min, increasing to 200 deg.C at 8 deg.C/min for 2min, and increasing to 250 deg.C at 10 deg.C/min for 8 min; an EI ionization source is adopted by MS, the electron energy is 70eV, the ion source temperature is 230 ℃, and the scanning range is 30.00-500.00 m/z; and (3) the test result is searched and compared with a standard spectrogram in an NIST mass spectrum library to determine the aroma components, and the content of the aroma components is calculated according to the response value of the internal standard cyclohexanone added in the sample.
The change of the composition and the content of the aroma substances is analyzed by combining the electronic nose with a SPEM-GC-MS detection technology, and the main substances influencing the detection of the change of the aroma response value of the kiwi fruit juice by the electronic nose are determined through PCA and O2PLS analysis.
And 5) determining the optimal process parameters of the release of the combined aroma substances according to the analysis result, wherein the addition amount of β -glucosidase II is 8U/g of fruit juice, the enzymolysis temperature is 35 ℃, and the enzymolysis time is 4 hours.
Example 1
The method comprises the steps of taking slow-fragrance kiwi fruits as raw materials, taking kiwi fruits with the hardness range of 5.0-10.0N, the weight range of single fruits of 50-150g and cylindrical fresh fruits of 5kg, peeling, cutting into blocks, juicing, filtering to remove fruit residues, uniformly mixing the kiwi fruits, taking 200g of kiwi fruits, adding β -glucosidase I according to the using amount of 4U/g, carrying out heat preservation for 2 hours in water bath at 35 ℃, shaking once every 30min during the process, and immediately carrying out electronic nose and SPME-GC-MS measurement after enzymolysis is finished.
Through electronic nose detection analysis, as shown in fig. 1, 5 sensors (S1, S3, S4, S5 and S9) are insensitive to volatile substances in kiwi juice, and the sensor response value is between 1.00 and 1.20. The response values of the sensors S7 and S2 both decrease slowly after increasing to the highest value and finally gradually tend to be stable, the response values of the sensors S6, S8 and S10 increase slowly and gradually tend to be stable after increasing to the highest value, wherein the response value is highest when the concentration of a substance represented by the number S7 (sensitive to terpenes) is higher, the response value of the sensor S2 (high sensitivity and sensitive to nitrogen oxides) is next to that of the sensor S6 (sensitive to methane, similar to the number 8 sensor), the response value of the sensor S8 (alcohols, partial aromatic compounds) and the response value of the sensor S10 (sensitive to alkanes) are subsequent.
The fragrance change of the kiwi fruit juice subjected to enzymolysis in example 1 is shown in fig. 2, and the response values of 4 sensors of the electronic nose are obviously increased (S1 aromatics, S6 alkanes, S7 terpenes and S8 alcohols); the response values of 5 sensors did not change significantly (S3 ammonia, S4 hydrogen, S5 alkanes, S9 organosulfides, S10 alkanes); there was a slight decrease in the response of 1 sensor (S2 nitroxide). Example 1 is shown to be effective in releasing aroma precursor materials of aromatic, alkane, terpene and alcohol types that may be present in kiwi juice.
The changes of the aroma substances of the kiwi fruit juice after the SPME-GC-MS detection and the enzymolysis are shown in the table 2. 41 substances are obtained from a sample after enzymolysis, wherein the sample comprises 7 alcohols, 9 aldehydes, 4 acids, 6 terpenes, 6 ketones, 2 furans, 6 olefin alkanes and 1 ester, and main volatile compounds comprise: cis-3-hexen-1-ol, trans-2-hexen-1-ol, hexanol, trans-2-hexenal, 1, 8-cineole and methylheptenone, and the like.
In example 1, the content of 41 aroma substances in the kiwi fruit juice after enzymolysis is increased to different degrees compared with the juice sample without enzymolysis, wherein 6 substances (2-propylheptanol, phenethyl alcohol, 2-ethylhexanoic acid, benzoic acid, β -pinene and trans-2- (2-pentenyl) furan) are added, and 12 substances (3,7, 11-trimethyl-2, 6, 10-dodecen-1-ol, decanal, acetic acid, octanoic acid, 4-terpenol, cymene, gamma-terpinene, 1-oct-3-one, geranylacetone, 2-ethylfuran, trans-2-decene and trans-2-hexenol acetate) are added in multiple times, the change of the aroma substances is shown in fig. 4 in the example 1 compared with the juice sample after enzymolysis and the juice sample without enzymolysis, the total amount of the kiwi fruit juice aroma substances after enzymolysis is increased by 24.15% compared with the juice sample without enzymolysis, wherein the content of the acid compounds is increased by 1277.34%, the content of the esters compounds is increased by 890.51%, the compounds is increased by 341.41.36%, and the contents of the alkanes compounds are increased by 13.83%.
Example 2
The method comprises the steps of taking slow-fragrance kiwi fruits as raw materials, taking kiwi fruits with the hardness range of 10.0-14.0N, taking the weight range of 60-100g of single fruits and 5kg of cylindrical fresh fruits, peeling, cutting into blocks, juicing, removing pomace through extrusion, taking 200g of kiwi fruit juice after the kiwi fruit juice is uniformly mixed, adding β -glucosidase II according to the amount of 8U/g, carrying out heat preservation for 4 hours in a water bath at 35 ℃, shaking once every 30min during the process, and immediately carrying out electronic nose and SPME-GC-MS measurement after enzymolysis is finished.
The aroma change of kiwi fruit juice subjected to enzymolysis in the process of example 2 is shown in fig. 2, and the response values of 4 sensors of the electronic nose are obviously increased (S1 aromatics, S6 alkanes, S7 terpenes and S8 alcohols); the response values of 5 sensors did not change significantly (S3 ammonia, S4 hydrogen, S5 alkanes, S9 organosulfides, S10 alkanes); there was a slight decrease in the response of 1 sensor (S2 nitroxide). Example 2 is shown to be effective in releasing aroma precursor materials of aromatic, alkane, terpene and alcohol types that may be present in kiwi juice.
Example 2 compared with the non-enzymolyzed juice, 41 aroma compounds are increased in content to a different extent, wherein 7 additional substances (2-propylheptanol, phenethyl alcohol, 2-ethylhexanoic acid, benzoic acid, β -pinene, 2-heptanone, trans-2- (2-pentenyl) furan) are added in multiple, 14 additional substances (3,7, 11-trimethyl-2, 6, 10-dodecen-1-ol, trans-2-nonenal, decanal, acetic acid, octanoic acid, 4-terpene alcohol, cymene, γ -terpinene, α -piperonyl terpene, 1-octan-3-one, geranylacetone, 2-ethylfuran, trans-2-decene, trans-2-hexenol acetate) are added in multiple, the change of aroma compounds is shown in fig. 4, compared with the enzymolyzed juice and the non-enzymolyzed juice sample, the total amount of the enzymolysis kiwi juice is shown in example 2, the aroma compounds content of 3648.54% of the total amount of the kiwi juice and the non-enzymolyzed juice is increased by 28.39%, the aroma compounds are increased by 6739.26%, and the hydrocarbon compounds are added by 10.58%.
Example 3
The method comprises the steps of taking slow-fragrance kiwi fruits as raw materials, taking kiwi fruits with the hardness range of 14.0-20.0N, taking the weight range of 60-90g of single fruits and 5kg of cylindrical fresh fruits, peeling, cutting into blocks, juicing, removing pomace through centrifugation, taking 200g of kiwi fruit juice after the kiwi fruit juice is uniformly mixed, adding β -glucosidase III according to the amount of 8U/g, keeping the temperature of a gas bath at 45 ℃ for 8 hours, shaking once every 30min during the process, and immediately carrying out electronic nose and SPME-GC-MS determination after enzymolysis is finished.
The aroma change of the kiwi fruit juice subjected to enzymolysis in the process of example 3 is shown in fig. 2, and the response values of 4 sensors of the electronic nose are obviously increased (S1 aromatics, S6 alkanes, S7 terpenes and S8 alcohols); response values of 4 sensors were not significantly changed (S3 ammonia, S4 hydrogen, S5 alkanes, S9 organosulfur); there was a slight decrease in the 2 sensor response values (S2 nitroxide, S10 alkane). Example 3 is shown to be effective in releasing aroma precursor materials of aromatic, alkane, terpene and alcohol types that may be present in kiwi juice.
Example 3 compared with the non-enzymatic hydrolyzed juice, 39 aroma substances are increased in content to different degrees, wherein 7 new substances (2-propylheptanol, phenethyl alcohol, 2-ethylhexanoic acid, benzoic acid, β -pinene, 2-heptanone, trans-2- (2-pentenyl) furan) are added, and 5 (3,7, 11-trimethyl-2, 6, 10-dodecen-1-ol, acetic acid, octanoic acid, 2-ethylfuran, trans-2-hexenol acetate) are added in multiple, compared with the enzymatic hydrolyzed juice and the non-enzymatic hydrolyzed juice, the aroma substance changes are shown in fig. 4. in example 3, the total amount of the aroma substances of the enzymatic hydrolyzed kiwi juice is increased by 4.93% compared with the non-enzymatic hydrolyzed juice, wherein the content of the ester compounds is increased by 522.53%, the content of the furan compounds is increased by 347.46%, the content of the acid compounds is increased by 278.76%, the content of the ketone compounds is increased by 14.51%, the content of the terpene compounds is increased by 13.70%, the content of the olefin compounds is increased by 3.82%, and the alcohol compounds is increased by 2.71%.
Table 2 shows the composition and content of kiwi fruit juice aroma before and after enzymatic hydrolysis in SPME-GCMS for three examples:
table 2 SPME-GC-MS detection of change of aroma substances of kiwi fruit juice before and after enzymolysis
Figure BDA0002403582280000091
Figure BDA0002403582280000101
The results of the correlation coefficient analysis result shown in table 3 show that the response value of the electronic nose sensor S2 is in positive correlation with 1-pentene-3-one (| r | >0.7, FDR <0.01), and the response values of the electronic nose sensor S6, S7, S8, and S10 are in positive correlation with 9 substances (i r | >0.7, FDR <0.01) such as β -pinene and trans-2- (2-pentenyl) furan, except acetic acid and geranyl acetone, which indicates that the changes of the substances can be used for predicting the response value of the electronic nose.
TABLE 3 correlation coefficient of volatile substances measured by SPME-GC-MS and response value of electronic nose sensor
Figure BDA0002403582280000111
In the 3 embodiments, the aroma content in the kiwi fruit juice after enzymolysis is increased, which shows that the enzymolysis method can effectively promote the release process of the combined state precursor substance to volatile aroma micromolecules, the difference of the embodiments is that the combined state aroma precursor substance in the kiwi fruit juice has different release degrees, the embodiment 2 has the best effect, and the specific process parameters comprise that fresh kiwi fruits with the fruit hardness of 10.0-14.0N are selected, peeled, cut into blocks and squeezed, the fruit residues are removed through squeezing, β -glucosidase II is adopted to carry out enzymolysis treatment on the kiwi fruit juice, the adding amount of enzyme is 8U/g, the kiwi fruit juice is subjected to water bath enzymolysis for 4 hours at 35 ℃, and the kiwi fruit juice is shaken every 30 minutes.
In conclusion, all detection results of the electronic nose and the SPME-GC-MS show that the kiwi fruit juice combined aroma substance is released, can be used for enhancing typical aroma in kiwi fruit juice and related products, and has a positive effect on improving the quality of kiwi fruit products.
The above embodiments are only specific embodiments disclosed in the present invention, but the scope of the present invention is not limited thereto, and the scope of the present invention disclosed in the present invention should be subject to the scope of the claims.

Claims (9)

1. A method for releasing combined-state aroma substances of kiwi fruit juice is characterized by comprising the following steps: the method comprises the following steps:
1) selecting fruits: selecting kiwi fruits with good maturity, regular fruit type and no damage;
2) juicing: peeling fresh kiwi fruits, cutting into pieces, squeezing to obtain juice, and removing fruit residues to obtain kiwi fruit juice;
3) performing enzymolysis, namely adding β -glucosidase into kiwi fruit juice, and performing enzymolysis under heat preservation, wherein β -glucosidase I, II and III derived from microorganisms or plants are respectively selected, and the addition amount of β -glucosidase is 4-12U/g of fruit juice, the enzymolysis temperature is 20-50 ℃, and the enzymolysis time is 0.5-12 h;
4) and (3) analysis: taking an enzymolysis result juice sample, analyzing the influence of enzymolysis treatment on the composition and the content of aroma substances of the kiwi fruit juice by an electronic nose and solid-phase microextraction gas chromatography-mass spectrometry combined technology, and determining the release degree of the combined aroma substances.
2. The method for releasing the flavor substances in the combined state of kiwi fruit juice according to claim 1, wherein in step 3), the addition amount of β -glucosidase is determined according to the enzyme activity, and the enzyme activity is determined by mixing 100mg enzyme with 5mL of 40mM p-nitrophenyl- β -D-glucopyranoside solution and 4mL of 1M citric acid-phosphate buffer solution (pH 4.0), incubating at 40 ℃ for 30min, adding 2mL of 3M sodium carbonate solution to terminate the reaction, and performing colorimetric reading at 405nm, wherein the enzyme activity is expressed in U/mg enzyme.
3. The method of kiwi fruit juice combined fragrance release according to claim 2, wherein: in the step 3), the enzymolysis temperature can be controlled by adopting water bath, gas bath or jacket heat preservation, and the enzymolysis temperature is based on the central temperature of the fruit juice.
4. The method of kiwi fruit juice combined-state aroma release of claim 3, wherein: in the step 4), the method for detecting the fragrance composition by the electronic nose comprises the following steps: taking 1g of the kiwi fruit juice sample after enzymolysis, adding pure water to dilute the kiwi fruit juice sample by 5 times, collecting data every second with the inlet flow rate of an electronic nose being 300mL/min, collecting time being 60s, cleaning for 300s, and enabling six of each group to be parallel.
5. The method of kiwi fruit juice combined fragrance delivery according to claim 4, wherein: in the step 4), the method for detecting the aroma components by SPME-GC-MS comprises the following steps: taking 5.0g of the kiwi fruit juice sample after enzymolysis, adding 2.5mL of saturated sodium chloride solution and 10 microliters of cyclohexanone internal standard (0.5mg/mL), preheating at 45 ℃ for 2.5min, extracting and adsorbing for 45min, and analyzing for 5min at GC 250 ℃; a gas chromatography column using DB-1MS (60 m.times.0.25 mm, 0.25 μm); helium flow rate is 1.5mL/min, no split flow; GC temperature setting: maintaining at 40 deg.C for 2min, increasing to 120 deg.C at 6 deg.C/min for 5min, increasing to 200 deg.C at 8 deg.C/min for 2min, and increasing to 250 deg.C at 10 deg.C/min for 8 min; an EI ionization source is adopted by MS, the electron energy is 70eV, the ion source temperature is 230 ℃, and the scanning range is 30.00-500.00 m/z; and (3) the test result is searched and compared with a standard spectrogram in an NIST mass spectrum library to determine the aroma components, and the content of the aroma components is calculated according to the response value of the internal standard cyclohexanone added in the sample.
6. The method of delivering kiwi fruit juice combined with aroma according to claim 5, wherein: in the step 4), the detected electronic nose is combined with a SPEM-GC-MS detection technology to analyze the change of the composition and the content of the aroma substances, and main substances influencing the detection of the change of the aroma response value of the kiwi fruit juice by the electronic nose are analyzed through PCA and O2PLS analysis.
7. The method for releasing the combined-state aroma substances of the kiwi fruit juice according to claim 6, wherein the step 4) is followed by the step 5) of determining the optimal technological parameters for releasing the combined-state aroma substances according to the analysis result, wherein the optimal technological parameters comprise the addition amount of the microbial source β -glucosidase II being 8U/g fruit juice, the enzymolysis temperature being 35 ℃ and the enzymolysis time being 4 hours.
8. The method of delivering kiwi fruit juice combined aroma according to any one of claims 1-7, further comprising: in the step 1), the hardness range of the kiwi fruit is required to be 5.0-20.0N, the weight range of a single fruit is required to be 50-150g, and the fruit shape is full.
9. The method of kiwi fruit juice combined fragrance release according to claim 8, wherein: in the step 2), the kiwi fruits are peeled, the bitter and astringent feeling is reduced, and the method for removing the fruit residues after juicing comprises filtering, extruding or centrifuging.
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