CN114184697A - Research method for interaction of sweetener and key aldehyde aroma substances in passion fruit - Google Patents
Research method for interaction of sweetener and key aldehyde aroma substances in passion fruit Download PDFInfo
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
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- G—PHYSICS
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- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
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Abstract
The invention discloses a method for researching interaction between a sweetener and key aldehyde aroma substances in passion fruit, which comprises the following steps: analyzing key aldehyde aroma substances in the passion fruit juice, extracting volatile substances in the passion fruit juice by SPME combined with SBSE, analyzing by GC-MS, identifying the key aroma substances by GC-O combined with OAV, and selecting the key aldehyde substances A from the key aroma substances as an object for researching interaction; detecting respective ultraviolet-visible absorption spectrums of the blank group sweetener B solution, the mixed solution of the sweetener B and the key aldehyde substance A at different temperatures by using an ultraviolet-visible spectrophotometer; the type of interaction force between the key aldehyde substance A and the sweetener B is judged by calculating thermodynamic parameters under a binary system. The method is simple and effective, has intuitive and reliable result and wide applicability, and provides theoretical basis for the blending of the essence of the medium-low sugar juice or beverage and the regulation and control of the flavor quality in actual production.
Description
Technical Field
The invention relates to a method for researching interaction between aroma substances and non-nutritive sweetener components in fruit juice or beverage products, in particular to a method for analyzing interaction and mechanism between key aldehyde aroma substances and sweeteners in passion fruit juice based on instrument analysis (GC-O-MS and ultraviolet spectrum) and thermodynamics, and belongs to the field of food flavor chemical analysis.
Background
Passion fruit, also known as passion fruit, belongs to the genus Passiflora, is the largest of the Passifloraceae family, and is primarily native to tropical America. Passion fruit is often eaten as fresh fruit or juice and is popular for its organoleptic properties-it has exotic sentiments, floral and fruity notes. The passion fruit is also called as the king of fruit juice because the fruit juice has fragrant smell, sweet and sour taste and pleasant color. The passion fruit has high nutritive value and has multiple functional characteristics of anti-inflammation, antioxidation, antianxiety and the like. For the consumption of passion fruit juice, the impact of aroma and taste complement each other, because the flavor of the juice depends on a delicate balance between other tastes such as sweetness and sourness and aroma. Wherein the volatile part of the substances responsible for the aroma plays an important role in the overall acceptability. At the same time, once the juice has a strong acid taste, water, sugar or high intensity sweeteners should be added to provide a palatable juice. For decades, sugar has been the primary sweetener in human diets and accounts for a large portion of the daily energy intake. However, excessive intake of such high calorie foods causes various complex conditions such as caries, diabetes, obesity and the like as negative problems. From the demand end, the pursuit of internal health and external beauty, the demand of reducing sugar (sucrose) for consumers is continuously promoted, low sugar/sugar-free products are gradually preferred to be selected in the product consumption, and the trend of reducing sugar is gradually increased. This makes the non-nutritive sweeteners playing a vital role as food additives.
Thaumatin, a type of sweet protein sweetener, also known as thaumatin, tastes like sucrose and also acts as a good flavor enhancer. These two properties (sweetener and flavor enhancer) also make this additive for food more important. Meanwhile, the protein can interact with aroma substances in the product, and can absorb or release flavor compounds due to environmental changes, so that the whole flavor is influenced. It is therefore necessary to understand the mechanism of interaction with flavor compounds in fruit juice products, as it not only modulates flavor, but also improves the organoleptic properties of the juice. The invention aims to analyze the interaction mechanism of key aldehyde aroma substances and sweeteners in passion fruit juice by combining instrument analysis and thermodynamics, provides a method for researching the mechanism between the sweeteners and the aroma substances in fruit juice and beverage formulas, and provides a theoretical basis for blending low-sugar fruit juice or beverage essence and regulating and controlling flavor quality in actual production.
Disclosure of Invention
The technical problem to be solved by the invention is as follows: provides a method for researching the interaction of the sweetener and key aldehyde aroma substances in the passion fruit juice.
In order to solve the technical problems, the invention provides a method for researching the interaction between a sweetener and key aldehyde aroma substances in passion fruit, which comprises the following steps:
step 1): analyzing key aldehyde aroma substances in the passion fruit juice, extracting volatile substances in the passion fruit juice by SPME combined with SBSE, analyzing by GC-MS, identifying the key aroma substances by GC-O (AEDA) combined with OAV, and selecting the key aldehyde substances A as a target for researching interaction;
step 2): detecting respective ultraviolet-visible absorption spectrums (observing the absorption of the sweetener) of the blank group sweetener B solution, the mixed solution of the sweetener B and the key aldehyde substance A at different temperatures by using an ultraviolet-visible spectrophotometer; the type of interaction force between the key aldehyde substance A and the sweetener B is judged by calculating thermodynamic parameters under a binary system.
The invention firstly uses SBSE method to extract the aroma substances in the passion fruit juice and combines the result with the SPME extraction result. Each method has its limitations, and the combination of the results of the various methods can more completely evaluate the key aroma compounds of the passion fruit juice.
Preferably, the identification method of the key aroma substances in the step 1) is as follows: an aroma substance with OAV more than or equal to 1 is taken as a component contributing to the whole aroma, and the component contributes more to the whole aroma when the OAV value is larger, namely a key aroma substance. Not all volatile compounds analyzed by GC-MS are aroma-sensitive and require GC-O screening, but not all aromas are essential. Therefore, the Flavor Dilution (FD) value is calculated by AEDA detection, the FD value reflects the contribution degree of each compound to the flavor, and higher FD value indicates greater contribution to the overall flavor. AEDA, however, does not take into account the matrix effect and further validation was performed by introducing the OAV method, which is equal to the ratio of the concentration of volatile components in the food to their detection threshold in the food, and is capable of analyzing the contribution of a certain aroma substance to the overall aroma.
Preferably, the concentration of the sweetener B is firstly fixed in the step 2), and the absorbance A corresponding to the maximum ultraviolet absorption wavelength at the concentration is measured0(ii) a Mixing key aldehyde substances A with a plurality of concentration points (generally four concentration points are selected, and the concentration points can be selected within a reasonable absorption value range according to specific experimental conditions) with a sweetener B with a fixed concentration, and determining the light absorption value A corresponding to the maximum absorption wavelength of the sweetener B at different temperatures; and (3) solving the binding constants among molecules by adopting a Lineweave-Burk double reciprocal curve and solving the binding constants at different reaction temperatures:
in the formula, C represents the molar concentration of the small molecule compound, K is a binding constant, and a and b are constants respectively.
Preferably, the thermodynamic parameters of the interaction between the key aldehyde substance a and the sweetener B in step 2) are determined by Van't Hoff equation:
in the formula, K is a binding constant when the temperature is T, R is a gas constant, and Delta H and Delta S are enthalpy change and entropy change respectively; drawing InK to 1/T, and obtaining entropy change delta H and entropy change delta S by the slope and intercept of the straight line; then calculating the free energy change deltaG of the combination reaction by the following formula:
ΔG＝ΔH-T·ΔS。
more preferably, the judgment method of the type of the interaction force between the key aldehyde substance A and the sweetener B is as follows: when the delta S is more than 0 and the delta H is more than 0, hydrophobic acting force exists between the delta S and the delta H; when the delta S is less than 0 and the delta H is less than 0, an oxygen bond and van der Waals force exist between the two; when Δ S > 0 and Δ H < 0, there is an electrostatic interaction between the two. And analyzing the related parameters to determine the main driving force of the interaction process of the two.
The invention researches the interaction and mechanism of the sweetener and key aldehyde aroma substances in the passion fruit juice, utilizes SPME to combine SBSE to extract volatile substances in the passion fruit juice, identifies the key aldehyde aroma substances in the passion fruit juice by AEDA combining OAV, and obtains the interaction research object by screening. Meanwhile, the combination mode and the interaction force type between the sweetener and the key aldehyde aroma substances are judged by utilizing ultraviolet spectrum and calculating thermodynamic parameters. The research on the modification of the product formula, the flavor control and the sense retention of the interaction provides theoretical research. The method combines the instrument analysis and the thermodynamic calculation, makes up the problem of the vacancy of the research method for the interaction of the aroma substances and the sweetening agents, and has the advantages of visual and reliable result, simple and feasible research method and wide applicability.
Drawings
FIG. 1 is a graph of the UV absorption of 0.1g/L thaumatin;
FIG. 2 is a graph of the UV absorption spectra of mixed solutions of different concentrations of furfural (increasing concentrations from a to e) and thaumatin at 25 ℃;
FIG. 3 is a linear regression plot of binding constants for furfural and thaumatin (25 ℃, 30 ℃, 37 ℃);
FIG. 4 is the Van't Hoff equation for furfural interaction with thaumatin.
Detailed Description
In order to make the invention more comprehensible, preferred embodiments are described in detail below with reference to the accompanying drawings.
Examples
A method for researching interaction of key aldehyde aroma substances in sweetener and passion fruit juice based on instrument analysis and thermodynamic calculation comprises the following specific steps:
1. SPME: a fruit juice sample (6 g) was accurately weighed into a 20mL headspace bottle, 20. mu.L of 2-octanol (400mg/kg) was added as an internal standard, and then the headspace bottle was heated in a water bath at 40 ℃ for 30 min. The SPME needle was inserted into the headspace of the headspace bottle, the fiber (75 μm CAR/PDMS) was extended to expose the sample headspace, placed 1cm from the sample liquid level and the volatiles were extracted for 40 minutes. The fibers were thermally desorbed at the inlet of the gas chromatograph at 250 ℃ for 5 minutes for gas chromatograph-mass spectrometer (GC-MS) analysis. SBSE: PDMS coated stir bars of 10mm/0.5mm ACAR/PDMS specification were used. A6 g sample of fruit juice, 20. mu.L of 2-octanol (400mg/kg) as an internal standard and a stir bar were placed in a 20mL sealed vial. The stir bar was used to stir at 600rpm for 40min at 40 ℃. After extraction, the PDMS stir bar was removed from the sample vial, (the stir bar was removed from the solution, rinsed with distilled water, dried with lint-free paper, and immediately transferred to a pyrolysis pipette.) dried with filter paper and inserted into a glass pyrolysis pipette for desorption and analysis using a GC-MS system. And then analyzed by a GC-O instrument: using passion fruit matrix as a solvent and adding 1:4, 1:16, 1: 32,1:64,1:256,1:1024,1: 2048 and diluting the concentrated solution. The person smelling fragrance continuously smells the fragrance materials close to the upper part of the smell mouth, the peak time of the smelled fragrance materials and the description of the fragrance materials are recorded until no fragrance materials are smelled under a certain dilution gradient, and the highest dilution concentration of each fragrance material which can be smelled is recorded and recorded as FD value. The quantitative analysis of the odoriferous active compounds with FD factors of greater than or equal to 16 was carried out. All detected aromatic active compounds were quantified by establishing a standard curve. The abscissa is the ratio of the concentration of the compound to the internal standard, and the ordinate is the peak area of the compound to the internal standard. The contribution of each odor to the overall aroma is evaluated by the odor activity value, which is measured as the ratio of the concentration of each compound to its detection threshold in water, the OAV corresponding to each aroma is calculated, and the aroma with retention OAV ≧ 1 is the key aroma.
2. Selecting key aldehydes in all aroma substances with OAV more than or equal to 1, and selecting furfural from the aldehydes as a research object for interaction with a sweetener (thaumatin). Determining thaumatin concentration of 0.1g/L in a mixed solution system as the concentration for researching interaction, and sequentially determining that the molar concentration of furfural in the system is 3.62 multiplied by 10-4，6.04×10-4，9.66×10-4，1.21×10- 3mol/L. Mixing furfuralThe mixed solution with thaumatin is equilibrated at 25 deg.C, 30 deg.C and 37 deg.C for 10 min. The uv-visible absorption spectrum of the solution was determined using a uv-visible spectrophotometer. Taking a corresponding blank reagent as a reference, carrying out full scanning on 0.1g/L of the thaumatin solution, wherein the scanning wavelength range is 190-400nm to confirm the maximum absorption wavelength of thaumatin (figure 1), and then measuring the light absorption value of the mixed solution system. And then, obtaining the binding constant between molecules by adopting a Lineweave-Burk double reciprocal curve. The interaction and structural change between furfural and thaumatin can be judged by the change of the absorption value under the maximum absorption wavelength of thaumatin under different concentrations of furfural (figure 2).
3. The binding constants at different reaction temperatures were determined (FIG. 3) and then determined by the Van't Hoff equation
Where K is the binding constant at temperature T and R is the gas constant 8.314) to determine the thermodynamic parameters of the interaction between furfural and thaumatin (table 2). The enthalpy change Δ H and entropy change Δ S were obtained by plotting InK versus 1/T and calculating the slope and intercept of the straight line (FIG. 4). Then, the free energy change Δ G ═ Δ H-T · Δ S of the binding reaction at each temperature was calculated from these two data. From Table 2, it can be seen that the furfural and thaumatin system Δ G<0, indicating that the reaction process between the two is spontaneous. The values of Δ H and Δ S were 29.985 kJ. mol, respectively-1>0 and 170.29J mol-1·K-1>And 0, according to a judgment rule, indicating that the main driving force of the interaction process of the two is hydrophobic acting force.
TABLE 2 binding constants K for furfural systems and related thermodynamic parameters
The method for researching the interaction of the sweetener and the key aldehyde aroma substances in the passion fruit juice based on the instrumental analysis and the thermodynamic calculation is simple and rapid to operate, and visual and reliable in result.
Claims (5)
1. A method for researching interaction between a sweetener and key aldehyde aroma substances in passion fruit is characterized by comprising the following steps:
step 1): analyzing key aldehyde aroma substances in the passion fruit juice, extracting volatile substances in the passion fruit juice by SPME combined with SBSE, analyzing by GC-MS, identifying the key aroma substances by GC-O combined with OAV, and selecting the key aldehyde substances A from the key aroma substances as an object for researching interaction;
step 2): detecting respective ultraviolet-visible absorption spectrums of the blank group sweetener B solution, the mixed solution of the sweetener B and the key aldehyde substance A at different temperatures by using an ultraviolet-visible spectrophotometer; the type of interaction force between the key aldehyde substance A and the sweetener B is judged by calculating thermodynamic parameters under a binary system.
2. The method for studying interaction between sweetener and key aldehyde aroma substances in passion fruit according to claim 1, wherein the identification method of the key aroma substances in step 1) is as follows: an aroma substance with OAV more than or equal to 1 is taken as a component contributing to the whole aroma, and the component contributes more to the whole aroma when the OAV value is larger, namely a key aroma substance.
3. The method for researching interaction between sweetener and key aldehyde aroma substances in passion fruit as claimed in claim 1, wherein in step 2), the concentration of sweetener B is first fixed, and the absorbance value A corresponding to the maximum ultraviolet absorption wavelength at the concentration is measured0(ii) a Mixing key aldehyde substances A with multiple concentration points with a sweetener B with a fixed concentration, and measuring light absorption values A corresponding to the maximum absorption wavelengths of the sweetener B at different temperatures of the mixed solution; and (3) solving the binding constants among molecules by adopting a Lineweave-Burk double reciprocal curve and solving the binding constants at different reaction temperatures:
in the formula, C represents the molar concentration of a small molecule compound,k is a binding constant, and a and b are constants respectively.
4. The method for studying interaction between a sweetener and key aldehyde aroma substances in passion fruit according to claim 1, wherein the thermodynamic parameters of interaction between the key aldehyde substances A and the sweetener B in the step 2) are obtained by Van't Hoff equation:
in the formula, K is a binding constant when the temperature is T, R is a gas constant, and Delta H and Delta S are enthalpy change and entropy change respectively; drawing InK to 1/T, and obtaining entropy change delta H and entropy change delta S by the slope and intercept of the straight line; then calculating the free energy change deltaG of the combination reaction by the following formula:
ΔG＝ΔH-T·ΔS。
5. the method for studying interaction between a sweetener and key aldehyde aroma substances in passion fruit according to claim 4, wherein the judgment method for the type of interaction force between the key aldehyde substances A and the sweetener B is as follows: when the delta S is more than 0 and the delta H is more than 0, hydrophobic acting force exists between the delta S and the delta H; when the delta S is less than 0 and the delta H is less than 0, an oxygen bond and van der Waals force exist between the two; when Δ S > 0 and Δ H < 0, there is an electrostatic interaction between the two.
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| CN114935618B (en) * | 2022-06-07 | 2023-08-04 | 天津市生态环境科学研究院(天津市环境规划院、天津市低碳发展研究中心) | Non-target screening-based precise identification method for odor-causing substances |
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