A method and system for grading numbness

Abstract

The present invention discloses a method for rapidly evaluating and grading numbing flavors, which is applicable to the field of food numbing flavor perception and discrimination. The method uses silica gel column chromatography to separate and purify key numbing flavor components, and analyzes and identifies the numbing flavor components through high-performance liquid chromatography. Simultaneously, the stimulation threshold, stimulation time, and salivation amount of characteristic numbing flavor substances are measured to determine the sensory properties of each numbing flavor substance. The content of numbing flavor substances in numbing flavor products is determined by high-performance liquid chromatography, and a mathematical model for evaluating numbing flavor levels is established based on the numbing flavor substance content, the Scoville index value of numbing flavor monomers, and the conversion coefficient. This method can provide new ideas for companies engaged in sesame oil production. The rapid, comprehensive, and accurate numbing flavor grade detection method can help companies better position their products.

Description

Technical Field

[0001] The present invention relates to the technical field of food numbing taste perception and discrimination, and more particularly to a numbing taste grading method and system. Background Art

[0002] Sichuan peppercorns, a type of Sichuan peppercorn, are widely popular for their unique numbing aroma. Sichuan peppercorn products primarily include Sichuan peppercorn oil. There's no standardized evaluation method for different types of Sichuan peppercorn oil on the market, and each method has its own advantages and disadvantages. Currently, the Scoville Index (GB / T38495-2020) is the primary method used to assess the numbing flavor. However, this method requires guidance from experienced analysts and is cumbersome. Therefore, developing a rapid and scientific method for evaluating the numbing flavor of Sichuan peppercorn oil has become a pressing challenge for the Sichuan peppercorn industry. Because numbing is a tactile sensation, its sensation is very similar to spiciness. It's well known that spiciness is caused by capsaicin in chili peppers activating the TRPV1 and TRPA1 transient receptor potentials (TRPs), producing a primarily burning sensation. There are two theories about the mechanism of numbing. One hypothesis is that when numbing substances are stimulated, TRPV1 and TRPA1 depolarize, leading to an influx of Ca2+ and an inward current, resulting in a numbing sensation. The other hypothesis is that numbing substances activate sensory neurons by inhibiting diaphanous potassium channels, causing the numbing sensation. For Sichuan pepper and its products, whether a rapid evaluation method can be established is the focus of this study.

[0003] Therefore, how to provide a comprehensive and accurate method and system for grading the numbness has become a technical problem that those skilled in the art urgently need to solve. Summary of the Invention

[0004] In view of this, the present invention provides a method and system for grading numbing taste, which can comprehensively and accurately grade numbing taste.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] In one aspect, the present invention discloses a method for extracting, separating, purifying and identifying numbing substances from Sichuan pepper, the specific steps of which are as follows:

[0007] Ultrasound-assisted extraction of fresh Sichuan peppercorns was performed using anhydrous ethanol, water, ethyl acetate, and petroleum ether. The extracts were then concentrated, organic solvents removed, and reconstituted with drinking water. Sensory evaluation experiments were conducted to determine which solvent produced the strongest numbing flavor in Sichuan peppercorns.

[0008] The extract was subjected to silica gel column chromatography using petroleum ether, ethyl acetate, anhydrous ethanol, and water as the separation solvents. The isolated fractions were then concentrated, the organic solvent removed, and reconstituted. The numbing intensity of the isolated fractions was evaluated using sensory evaluation.

[0009] HPLC analysis was performed to identify the components with a numbing odor, and the numbing compounds were identified using an external standard method. The HPLC spectra of the eluate with the highest numbing irritation intensity were analyzed, and the eluted compounds were collected and enriched, then prepared at a concentration of 1 mg / mL. The numbing irritation intensity was evaluated through sensory analysis, and the numbing compounds were finally identified using an external standard method. This process aimed to identify the primary numbing compounds in Sichuan pepper, providing evidence for the following experiments.

[0010] In another aspect, the present invention discloses a method for grading numbing taste, which comprises the following steps:

[0011] Determining the content of numbing substances in the target test object by high performance liquid chromatography, and simultaneously determining the Scoville index value of the target test object;

[0012] Conducting a correlation analysis between the numbing substances and the Scoville index value;

[0013] Sensory evaluation of numbing flavor monomers was conducted, including their stimulation threshold, stimulation time, and salivation amount. The numbing flavor activity value in Sichuan pepper oil was calculated based on the stimulation threshold and the corresponding content to verify the numbing flavor contribution rate of various numbing flavor substances.

[0014] To verify the numbing intensity and contribution of the individual numbing substances, we compared raw Sichuan pepper oil with compounds consisting of four numbing substances. We then prepared multiple compounds lacking these numbing substances and compared them with the original samples. Finally, we scored the compounds based on sensory properties, including numbness, tingling, burning, duration of stimulation, and bitterness. This was to validate the aforementioned numbing activity values.

[0015] A mathematical model for evaluating numbing flavor levels was established based on the content of numbing substances and the Scoville index values ​​of individual numbing substances. To optimize the model's quality, a simple linear regression fit was performed between the established model and the Scoville value of Sichuan peppercorn oil measured using the Scoville index method, resulting in a regression equation. Weighting coefficients were applied to the model to further enhance its accuracy.

[0016] Preferably, in the above-mentioned method for extracting, separating, purifying and identifying numbing substances from Sichuan peppercorns, the specific steps for extracting numbing substances from Sichuan peppercorns are as follows:

[0017] Remove the leaves and branches of freshly harvested peppercorns, retaining only the fruit. Soak 500g of peppercorn fruit in petroleum ether, ethyl acetate, anhydrous ethanol, and water, respectively, and extract using ultrasonic technology for 1 hour. Filter the extract and concentrate it on a rotary evaporator to 100mL for storage. Repeat this process three times. Prepare an extract of equal concentration (5mg / mL) and determine the extraction solvent with the strongest numbing flavor through sensory analysis.

[0018] Preferably, in the above-mentioned method for extracting, separating and purifying numbing substances from Sichuan peppercorns and identifying numbing substances, the specific operating steps for separating and purifying numbing substances from Sichuan peppercorns are as follows:

[0019] Take 200g of concentrated viscous extract and separate it by layer-by-layer chromatography on silica gel. The sampling method adopts wet loading, and the volume of the silica gel column used is 400mL. Petroleum ether, ethyl acetate, anhydrous ethanol and water are used for elution and separation respectively, and each solvent elutes three column volumes, collecting 200mL in each bottle, and a total of 24 bottles are collected. The 24 bottles of eluate are numbered A1-A6, B1-B6, C1-C6, D1-D6, and finally they are concentrated for use. Prepare 5mg / mL equal concentration eluate, and determine whether the 24 tubes of eluate contain hemp taste through sensory analysis.

[0020] Preferably, in the above-mentioned method for extracting, separating, purifying and identifying numbing substances from Sichuan peppercorns, the specific operating steps for identifying numbing substances from Sichuan peppercorns are as follows:

[0021] The eluate containing a numbing odor was analyzed by high-performance liquid chromatography, and the approximate numbing substance was determined by the external standard method. The liquid phase spectrum of the eluate with the highest numbing irritation intensity was analyzed, and the peak substances were collected and enriched, and prepared into a 1 mg / mL concentration. The numbing irritation intensity was evaluated by sensory analysis, and finally the numbing substance was determined by the external standard method. The HPLC conditions were a 30-minute gradient elution of 60%-80% acetonitrile-acetic acid water, followed by a 40-minute elution of 90% acetonitrile-acetic acid water, a flow rate of 1 mL / min, and a UV detection wavelength of 270 nm.

[0022] Preferably, in the above-mentioned method for extracting, separating, purifying and identifying numbing substances from Sichuan pepper, the specific steps of the sensory evaluation experiment are as follows:

[0023] Each sensory evaluation stage consisted of eight panelists, who were instructed to evaluate samples with a numbing taste. At each stage, the samples were subjected to a vacuum rotary evaporator or vacuum freeze concentrator to evaporate the organic solvent and ensure that the samples were safe and non-toxic. The samples were then dissolved in a small amount of food-grade ethanol and added to drinking water to prepare an aqueous solution of equal concentration for tasting. The numbing taste stimulation score was scored using a 9-point scale, with 0 representing no numbing taste, 5 representing moderate numbing taste, and 9 representing extremely strong numbing taste.

[0024] Preferably, in the above-mentioned numbing taste classification method, the specific steps of determining the numbing taste substance content in the target test object by high performance liquid chromatography are as follows:

[0025] Utilizing ethanol as an extractant to extract the target substance, and combining the supernatants to obtain an extract;

[0026] The extract is sequentially fixed to volume and filtered through a filter membrane to obtain a sample to be tested;

[0027] Set up chromatographic conditions and determine the content of numbing substances.

[0028] Preferably, in the above-mentioned method for grading the numbness, the Scoville index method is used to extract unsaturated fatty acid amide substances in the sample to be tested, and the extract is diluted into alcohol-water solutions of different concentrations in proportion. The maximum dilution factor of the sample to be tested at which the evaluation panel just identifies the numbness is determined by paired comparison test to obtain the Scoville index value.

[0029] Preferably, in the above-mentioned numbing taste classification method, the specific steps of performing correlation analysis between the numbing taste substances and the Scoville index value are as follows:

[0030] Pearson correlation analysis was performed on the measured numbing substance content of the pepper oil and the corresponding Scoville value of the pepper oil to obtain the correlation coefficient between the numbing substance content and the Scoville value.

[0031] Preferably, in the above-mentioned numbing taste classification method, the specific steps of obtaining the stimulation threshold, stimulation time and salivation amount of the numbing taste monomer substance are as follows:

[0032] After the hemp-flavored monomer substances were dissolved in a small amount of food-grade ethanol, drinking water was used as the solvent to prepare hemp-flavored aqueous solutions with different concentrations;

[0033] To evaluate the stimulation threshold of a single substance of numbing taste, start tasting from a high-concentration sample and continue tasting until the numbing taste is no longer felt. This is the stimulation threshold of the single substance of numbing taste.

[0034] For the evaluation of the stimulation time and saliva production of numbing monomer substances, different gradients are recorded; the sampling method is to accurately weigh the sample solution with a pipette and place it on the evaluator's tongue, and it is required to ensure that the sample solution is placed on the tongue for a fixed length of time. When evaluating the stimulation time, the evaluator is allowed to swallow, and when evaluating the saliva production, the evaluator is required not to swallow until the numbing taste disappears.

[0035] Preferably, in the above-mentioned numbing taste classification method, the numbing taste activity value is determined by the ratio of the content of a certain numbing taste substance to its stimulation threshold.

[0036] Preferably, in the above-mentioned method for grading the numbing taste, the mathematical model expression for evaluating the numbing taste level is established as follows:

[0037] X=a×(W1×b1+W2×b2+W3×b3+W4×b4+……+W n ×b n )×q;

[0038] W1, W2, W3 and W4 are the contents of numbing substances respectively, b1, b2, b3 and b4 are the corresponding Scoville index values ​​respectively, n represents the nth numbing substance, a represents the conversion coefficient; q represents the weight coefficient, and is greater than 1.

[0039] In another aspect, the present invention discloses a numbing taste grading system, which utilizes the above-mentioned numbing taste grading method, comprising:

[0040] The numbing substance extraction module uses ultrasound to assist in extracting numbing substances from Sichuan peppercorns;

[0041] The hemp flavor substance separation and purification module uses silica gel column chromatography to separate and purify the hemp flavor extract;

[0042] The numbing substance identification module uses high-performance liquid chromatography to determine the fingerprint of the separated components and uses the external standard method to perform qualitative analysis and identify the numbing substances;

[0043] The sensory evaluation module uses sensory evaluation experiments to evaluate the numbing intensity of substances during the extraction, separation, purification and identification processes;

[0044] The first determination module uses high performance liquid chromatography to determine the content of numbing substances in the target test object;

[0045] A second determination module is used to determine the Scoville index value of the target detection object;

[0046] A correlation analysis module, performing a correlation analysis on the numbing substance and the Scoville index value;

[0047] The numbing activity value calculation module obtains the stimulation threshold, stimulation time and salivation amount of the numbing monomer substance, and calculates the numbing activity value based on the stimulation threshold and content;

[0048] The recombinant validation module compares the original sesame oil with a mixture of four numbing monomers to verify the numbing intensity and contribution of the individual numbing compounds. Subsequently, multiple compound samples lacking these numbing compounds are prepared and compared with the original samples. Finally, scores are assigned based on sensory properties including numbness, tingling, burning, duration of stimulation, and bitterness. This aims to validate the aforementioned numbing activity value results.

[0049] The linear regression calculation module performs a simple linear regression fit on the established model and the Scoville value measured by the Scoville index method to optimize the quality of the model, and derives the regression equation. The weight coefficient is applied to the model to further improve the accuracy of the model.

[0050] It is important to understand that since the regression equation for the determined numbing monomers is obtained by ignoring substances of low importance, the regression equation needs to be adjusted to compensate for the ignored numbing monomers to improve accuracy.

[0051] The output module establishes a mathematical model for evaluating the numbing taste level based on the numbing taste substance content and the Scoville index value of the numbing taste monomer substance.

[0052] As can be seen from the above technical solutions, compared with the prior art, the present invention provides a method and system for grading sesame flavor. This method can quickly evaluate the sesame flavor level using an established sesame flavor evaluation formula. Compared to directly using the Scoville index method to evaluate the sesame flavor level, this method has certain improvements in detection speed and scientificity. Currently, high-performance liquid chromatography is widely used in various companies. When determining the sesame flavor level, it is only necessary to determine the content of the sesame flavor monomer substance using high-performance liquid chromatography, and then apply the measured content to the formula to obtain the SPU value. This reduces the error of the SPU value mainly due to manual evaluation, and improves the speed of detection. This method can provide new ideas for companies engaged in sesame oil production. The rapid sesame flavor level detection method can help companies better identify their products. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0054] Figure 1 is a flow chart of the method of the present invention;

[0055] Figure 2 (a) is a flow chart of the extraction, separation, purification and identification of the numbing substance of Sichuan pepper according to the present invention, wherein Figure 2(b)-Figure 2(e) They are the sensory evaluation test results of the extraction, separation and purification of numbing substances and substance identification process and the qualitative results of HPLC external standard method;

[0056] Figure 3 This is a schematic diagram of the numbing activity values ​​of S1-S14 of the present invention;

[0057] Figure 4 This is a correlation analysis diagram of the content of the numbing monomer substance and the SPU value of the present invention;

[0058] Figure 5(a)-Figure 5(b) These are line graphs showing the relationship between the content of different numbing monomer substances, stimulation time, and body fluid production in the present invention;

[0059] Figure 6 The radar chart shows the sensory evaluation results of the original sample and the compound samples of the four numbing substances of the present invention;

[0060] Figure 7(a)-Figure 7(e)Schematic diagrams of sensory evaluation results of the original sample and various compound samples of the present invention;

[0061] Figure 8 The linear regression scatter plot of Scoville value of Sichuan pepper oil measured by the mathematical model formula and Scoville index method established in the present invention, as well as the linear regression equation and R 2 value;

[0062] Figure 9 This is a schematic diagram comparing the Scoville index method used in the present invention and this method. DETAILED DESCRIPTION

[0063] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0064] The embodiment of the present invention discloses a method for grading numbing taste, and the specific steps are as follows:

[0065] S101 high performance liquid chromatography is used to determine the content of numbing substances in the target test object and the Scoville index value of the target test object;

[0066] S102 performs correlation analysis on the numbing substances and the Scoville index value;

[0067] S103 obtains the stimulation threshold, stimulation time, and salivation amount of the numbing taste monomer substance, and calculates the numbing taste activity value based on the stimulation threshold and content;

[0068] S104 performs pairwise comparisons between the target test object and complexes composed of different numbing flavor monomers, uses stimulation time and body fluid production to determine significant differences, performs feature correlation and linear regression analysis on complexes that do not show significant differences with the original target test object, determines the complex that is most similar to the original target test object, and extracts its correlation coefficient as a conversion coefficient for the numbing flavor evaluation formula;

[0069] S105 establishes a mathematical model for evaluating the numbing taste level based on the numbing taste substance content, the Scoville index value of the numbing taste monomer substance and the conversion coefficient.

[0070] Taking Sichuan pepper as an example, the following numbing monomer substances (including hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-ε-sanshool and hydroxy-γ-sanshool)

[0071] Remove the leaves and branches of freshly harvested peppercorns, retaining only the fruit. Soak 500g of peppercorn fruit in petroleum ether, ethyl acetate, anhydrous ethanol, and water, respectively, and extract using ultrasonic technology for 1 hour. Filter the extract and concentrate it on a rotary evaporator to 100mL for storage. Repeat this process three times. Prepare an extract of equal concentration (5mg / mL) and determine the extraction solvent with the strongest numbing flavor through sensory analysis.

[0072] Take 200g of concentrated viscous extract and separate it by layer-by-layer chromatography on silica gel. The sampling method adopts wet loading, and the volume of the silica gel column used is 400mL. Petroleum ether, ethyl acetate, anhydrous ethanol and water are used for elution and separation respectively, and each solvent elutes three column volumes, collecting 200mL in each bottle, and a total of 24 bottles are collected. The 24 bottles of eluate are numbered A1-A6, B1-B6, C1-C6, D1-D6, and finally they are concentrated for use. Prepare 5mg / mL equal concentration eluate, and determine whether the 24 tubes of eluate contain hemp taste through sensory analysis.

[0073] HPLC analysis was performed on the eluate containing a numbing odor, and the probable numbing substances were determined using the external standard method. The liquid phase spectrum of the eluate with the highest numbing irritation intensity was analyzed, and the peak substances were collected and enriched, and prepared into a 1 mg / mL concentration. The numbing irritation intensity was evaluated by sensory analysis, and finally the numbing substances were determined using the external standard method. The HPLC conditions were a 30-minute gradient elution of 60%-80% acetonitrile-acetic acid water, followed by a 40-minute elution of 90% acetonitrile-acetic acid water, a flow rate of 1 mL / min, and a UV detection wavelength of 270 nm.

[0074] Sensory evaluation. Each stage of sensory evaluation is conducted by 8 sensory evaluators, who are told to evaluate samples related to the numbing taste. At each stage, the samples will use a vacuum rotary evaporator or a vacuum freeze concentrator to evaporate the organic solvent and ensure that the samples are safe and non-toxic. They are then dissolved in a small amount of food-grade ethanol and added to drinking water to prepare an aqueous solution of equal concentration for tasting. The numbing taste stimulation score is scored using a 9-point scale, with 0 points representing no numbing taste, 5 points representing moderate numbing taste, and 9 points representing extremely strong numbing taste.

[0075] Figure 2(a) illustrates the extraction, separation, identification, and sensory evaluation process of the numbing flavor of Sichuan pepper. During the Sichuan pepper numbing flavor extraction process, four Sichuan pepper numbing flavor extracts were evaluated using sensory evaluation methods. The results are shown in Figure 2(b). Because sanshool is readily soluble in ethanol, the Sichuan pepper extracted with anhydrous ethanol exhibited the strongest numbing flavor, while petroleum ether and water extracts contained almost no numbing flavor. Therefore, during the Sichuan pepper numbing flavor separation process, the ethanol extract was used as the raw material for silica gel column chromatography experiments. After column chromatography separation, 24 tubes of eluate were processed and subjected to sensory evaluation at equal concentrations. The results are shown in Figure 2(c). Sensory evaluation revealed that the eluate extracted with ethyl acetate exhibited the highest numbing flavor, followed by anhydrous ethanol. The eluate extracted with petroleum ether and water contained almost no numbing flavor. The third tube of ethyl acetate had the highest numbing flavor.

[0076] Subsequently, HPLC analysis was performed on ethyl acetate tube No. 3. The HPLC analysis was repeated several times and the corresponding substance peak eluates were collected, and the collected eluates were numbered, as shown in Figure 2(e). The sensory evaluation results of the differently numbered substances are shown in Figure 2(d). The substances numbered 3, 4, 5, and 8 contain a numbing taste. Moreover, it was judged by the external standard method that substances numbered 3, 4, 5, and 8 contain hydroxy-ε-sanshool, hydroxy-α-sanshool, hydroxy-β-sanshool, and hydroxy-γ-sanshool, respectively. Therefore, it can be judged that these four numbing monomer substances are the main substances that present a numbing taste in Sichuan pepper.

[0077] In S101, the content of numbing substances in Sichuan pepper oil was determined by high performance liquid chromatography.

[0078] According to GH / T 1291-2020, after shaking the Sichuan pepper oil, accurately weigh 0.2 g (accurate to 0.0001 g) and place it in a stoppered flask. Add 50 mL of ethanol and perform ultrasonic extraction for 0.5 h. Cool to room temperature, transfer to a 100 mL container, make up to volume with ethanol, filter through a 0.22 μm filter membrane, and wait for testing.

[0079] Chromatographic conditions: Column: C18 reversed-phase column, 4.6 × 150 mm, 5.0 μm; Mobile phase: acetonitrile: 1% glacial acetic acid aqueous solution (40:60, v / v); Flow rate: 1 mL / min; Injection volume: 20 μL; Detection wavelength: 270 nm.

[0080] The contents of numbing substances in 14 kinds of pepper oil were measured by high performance liquid chromatography, and their concentrations were calculated by external standard method. Figure 3The results shown in Table 3 below indicate that hydroxy-α-sanshool is the main component in all 14 types of pepper oil (accounting for approximately 70%), followed by hydroxy-β-sanshool, hydroxy-ε-sanshool, and hydroxy-γ-sanshool. The total numbing substance content of the 14 pepper oils was calculated, ranging from 3.86±0.01 to 70.06±0.68 mg / g.

[0081] Table 3

[0082]

[0083]

[0084] In S101, the SPU value of pepper oil was determined by the Scoville index method.

[0085] According to GB / T 38495-2020, five sensory evaluators were recruited to conduct sensory evaluation and numbing taste grading of 14 commercially available peppercorn oils. Their SPU values ​​are shown in Table 1.

[0086] Table 1 SPU values ​​of various pepper oils

[0087]

[0088] In S102, the correlation analysis between the content of each numb smell monomer substance in the pepper oil and the SPU value of the pepper oil is carried out. Figure 4 The results showed that hydroxy-α-sanshool, hydroxy-β-sanshool and hydroxy-ε-sanshool were positively correlated with the SPU value of pepper oil, and hydroxy-α-sanshool was highly positively correlated with the SPU value, which means that an increase in the hydroxy-α-sanshool content will directly affect the SPU value of the pepper oil; in addition, hydroxy-γ-sanshool was negatively correlated with the SPU value, proving that an increase in the hydroxy-γ-sanshool content in pepper oil basically does not lead to an increase in SPU.

[0089] S103 obtains the stimulation threshold, stimulation time, and salivation amount of the numbing taste monomer substance, and calculates the numbing taste activity value based on the stimulation threshold and content;

[0090] Sample preparation: Four numbing monomers (hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-ε-sanshool, and hydroxy-γ-sanshool) were dissolved in a small amount of food-grade ethanol and prepared into aqueous solutions of numbing substances with concentrations of 6.25 μg / mL, 12.5 μg / mL, 25 μg / mL, 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL using drinking water as the solvent.

[0091] 18 sensory evaluators were recruited to evaluate the stimulation threshold, stimulation time, and saliva production of numbing monomers. For the stimulation threshold of numbing monomers, the tasting started from the high-concentration sample and continued until the numbing taste was no longer felt. This was the stimulation threshold of the numbing monomer. For the stimulation time and saliva production of numbing monomers, four gradients of 50 μg / mL, 100 μg / mL, 200 μg / mL, and 400 μg / mL were used to record the results. The sampling method was to accurately weigh 200 μL of sample solution with a pipette and place it on the tongue of the evaluator. The evaluator was required to keep the sample solution on the tongue for 30 seconds as much as possible. The evaluator was allowed to swallow when evaluating the stimulation time, but was required not to swallow until the numbing taste disappeared when evaluating the saliva production.

[0092] The stimulation thresholds of four numbing monomers were measured, and the concept of numbing activity value (PAV) was proposed. Its purpose is to verify the numbing contribution rate of a numbing monomer in the overall tasting pepper oil by the ratio of the content of a numbing monomer in the tasting pepper oil to its stimulation threshold. From the results, the present invention measured the stimulation thresholds of four numbing monomers, and the stimulation thresholds of the four numbing substances are shown in Table 4. Among them, the lowest average stimulation threshold is hydroxy-γ-sanshool, with a threshold of 3.33±1.21, corresponding to an SPU value of 30030; the highest average stimulation threshold is hydroxy-β-sanshool, with a threshold of 7.50±2.57, corresponding to an SPU value of 13333. Figure 3 The results of PAV are shown. The contribution rates are ranked as follows: hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-ε-sanshool, and hydroxy-γ-sanshool. The average contribution rates are 66.94%, 22.29%, 9.78%, and 0.99%, respectively.

[0093] Table 4 Stimulation thresholds of various numbing substances

[0094]

[0095]

[0096] Specifically, Figure 5(a)-Figure 5(b) The results showed that there were significant differences in the stimulation time between the four numbing flavor monomers. The overall stimulation time variation pattern showed that the higher the concentration, the longer the stimulation time. The stimulation time of the four numbing flavor substances was as follows: hydroxy-γ-sanshool, hydroxy-α-sanshool, hydroxy-ε-sanshool, and hydroxy-β-sanshool. Similarly, there were significant differences in saliva secretion (the amount of fluid produced) between the four numbing flavor monomers. The variation pattern of different concentrations and saliva secretion were consistent with the stimulation time. This proves that the numbing flavor characteristics of different numbing flavor monomers are significantly different at the same concentration.

[0097] To verify the intensity and contribution of the numbing properties of individual numbing substances, a comparison was made between original Sichuan peppercorn oil and compounds consisting of four numbing substances. Subsequently, multiple compounds lacking these numbing substances were prepared and compared with the original samples. Finally, scores were given based on sensory properties including numbness, tingling, burning, duration of stimulation, and bitterness.

[0098] Experimental method: One of the peppercorn oils was used as the original sample, and the same amount of numb monomer substances was added to form different groups. The composition is shown in Table 2. 18 sensory evaluators were recruited and a 9-point scoring method was used as the sensory evaluation table. The scoring rules were as follows (taking numbness as an example): 0 points for no numbness, 5 points for moderate numbness, and 9 points for very strong numbness. Other sensory characteristics followed the same rules.

[0099] Sample preparation: S3 pepper oil diluted 10 times was used as the control sample (the contents of hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-ε-sanshool and hydroxy-γ-sanshool were 3.5 mg / g, 0.82 mg / g, 0.38 mg / g and 0.02 mg / g, respectively). Eight different compounds with the same content of numbing monomers were composed for pairwise comparison.

[0100] Table 2 Composition of compound

[0101]

[0102] In order to verify the above-mentioned numbing activity value results, a compounding and recombining experiment was conducted to verify it. This method is commonly used in the field of food aroma analysis. The present invention draws on the recombining and recombining method in the field of food aroma analysis to compound and recombinant the four numbing substances in pepper oil and compare them at the same content. The results are as follows Figure 6 , a sensory evaluation of the original and compound samples by 18 sensory evaluators showed that the scores of the two groups of samples in terms of bitterness, stinging, burning, numbness, and stimulation duration were very similar. This means that the compounding of numbing substances can well restore the numbing taste of the samples.

[0103] In order to continue to verify the results of the numbing activity value, the addition of numbing substances was reduced one by one and combined into different compound sample groups. Figure 7(a)-Figure 7(e)The sensory evaluation results of the original samples and various recombinant samples are shown. There are significant differences in these recombinant samples, such as Groups 2, 3, 7, 8 and 9, which all lack two or more numbing substances. However, there were no significant differences in the sensory evaluation of bitterness, tingling, burning, numbness and stimulation time among Groups 4, 5 and 6. Among them, Group 4 lacked hydroxy-γ-sanshool, Group 6 lacked hydroxy-ε-sanshool, and Group 5 was a completely recombinant sample. The results demonstrated the importance of hydroxy-α-sanshool and hydroxy-β-sanshool to the overall numbing taste, and the lack of hydroxy-γ-sanshool and hydroxy-ε-sanshool did not have a significant effect on the overall numbing taste. Therefore, this also proves the accuracy of the calculation results of the numbing activity value in this study.

[0104] In S105, since the above experiment verified that the numbing substances in the pepper oil are mainly hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-γ-sanshool and hydroxy-ε-sanshool, theoretically, the sum of the products of the numbing substance content in the pepper oil and the Scoville value of the corresponding numbing substance is the Scoville value of the pepper oil. In order to enhance the accuracy of the formula, the Scoville values ​​of 14 pepper oils were measured using the formula and the Scoville index method and a simple linear regression analysis was performed. Figure 8 , and the fitted regression equation is obtained. The following formula is obtained to establish the numbing taste level formula:

[0105] X=[(W1×23980+W2×13333+W3×17150+W4×30030)×1.1837×10]

[0106] Where:

[0107] 23980 – Scoville index of hydroxy-α-sanshool;

[0108] 13333 – Scoville index of hydroxy-β-sanshool;

[0109] 17150 – Scoville index of hydroxy-ε-sanshool;

[0110] 30030 – Scoville index of hydroxy-γ-sanshool;

[0111] 1.1837 – weight coefficient;

[0112] 10——Conversion factor;

[0113] W1, W2, W3 and W4 are the contents (g / g) of hydroxy-α-sanshool, hydroxy-β-sanshool, hydroxy-ε-sanshool and hydroxy-γ-sanshool in the pepper oil samples, respectively.

[0114] What needs to be understood is that the weight coefficient is calculated by converting the four main numbing flavor monomers and the Scoville index mentioned above to obtain the numbing flavor level. At the same time, the numbing flavor level of the sesame oil is known and verified by using different sesame oils. Therefore, multiple sets of calculated values ​​and true values ​​are linearly regressed to obtain the correction coefficient, i.e. the weight coefficient.

[0115] The numbing SPU values ​​of 14 kinds of pepper oils were recalculated according to the numbing grade formula and compared with the SPU values ​​measured directly using the Scoville index method. The results are as follows: Figure 9 As shown, Method 1 uses the Scoville Index method, while Method 2 uses a formula-based calculation. Of the 14 peppercorn oils, three (*) showed significant differences, while the remaining 11 (ns) showed no significant differences. If the formula is accurate based on the absence of significant differences, the accuracy rate is 78.57%. However, using the numbing flavor grading formula is more objective and scientific than calculating the peppercorn oil SPU value.

[0116] Another embodiment of the present invention discloses a numbing taste grading system, characterized by utilizing the above-mentioned numbing taste grading method, including:

[0117] The numbing substance extraction module uses ultrasound to assist in extracting numbing substances from Sichuan peppercorns;

[0118] The hemp flavor substance separation and purification module uses silica gel column chromatography to separate and purify the hemp flavor extract;

[0119] The numbing substance identification module uses high-performance liquid chromatography to determine the fingerprint of the separated components and uses the external standard method to perform qualitative analysis and identify the numbing substances;

[0120] The sensory evaluation module uses sensory evaluation experiments to evaluate the numbing intensity of substances during the extraction, separation, purification and identification processes;

[0121] The first determination module uses high performance liquid chromatography to determine the content of numbing substances in the target test object;

[0122] A second determination module is used to determine the Scoville index value of the target detection object;

[0123] A correlation analysis module, performing a correlation analysis on the numbing substance and the Scoville index value;

[0124] The numbing activity value calculation module obtains the stimulation threshold, stimulation time and salivation amount of the numbing monomer substance, and calculates the numbing activity value based on the stimulation threshold and content;

[0125] The recombinant compound verification module compared the original Sichuan pepper oil with compounds composed of four numbing monomers to verify the numbing intensity and contribution of the individual numbing substances. Subsequently, multiple compound samples lacking numbing substances were prepared and compared with the original samples. Finally, scores were assigned based on sensory characteristics including numbness, tingling, burning, duration of stimulation, and bitterness. This was to verify the numbing activity value results mentioned above.

[0126] The linear regression calculation module, in order to optimize the quality of the model, performs a simple linear regression fit on the established model and the Scoville value of Sichuan pepper oil measured by the Scoville index method to derive the regression equation. The weight coefficient is applied to the model to further improve the accuracy of the model.

[0127] The output module establishes a mathematical model for evaluating the numbing taste level based on the numbing taste substance content and the Scoville index value of the numbing taste monomer substance.

[0128] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. Reference can be made to the common and similar parts between the various embodiments. For the devices disclosed in the embodiments, since they correspond to the methods disclosed in the embodiments, the description is relatively simple, and the relevant parts can be referred to the method description.

[0129] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for grading numbness, characterized in that: The specific steps are as follows: Determining the content of numbing substances in the target test object by high performance liquid chromatography, and simultaneously determining the Scoville index value of the target test object; Conducting a correlation analysis between the numbing substances and the Scoville index value; Obtaining the stimulation threshold, stimulation time and salivation amount of the numbing taste monomer substance, and calculating the numbing taste activity value based on the stimulation threshold and content; Paired comparisons were conducted between the target test object and compounds composed of different numbing monomers. Significant differences were determined using stimulation time and salivation volume. Correlation analysis and linear regression analysis were performed on compounds that did not show significant differences with the original target test object. The compound most similar to the original target test object was determined, and its correlation coefficient was extracted as the conversion coefficient for the numbing taste evaluation formula. A mathematical model for evaluating the numbing taste level was established based on the numbing taste substance content, Scoville index value of numbing taste monomer substances and conversion coefficient.

2. A method for grading numbness according to claim 1, characterized in that: The specific steps for determining the content of numbing substances in the target test object by high performance liquid chromatography are as follows: The target substance is extracted using ethanol as an extractant, and the supernatants are combined to obtain an extract; the extract is successively fixed to volume and filtered through a filter membrane to obtain a sample to be tested; Set up chromatographic conditions and determine the content of numbing substances.

3. The method for grading numbness according to claim 2, characterized in that: The Scoville index method is used to extract unsaturated fatty acid amides from the test samples, and the extract is diluted proportionally into alcohol-water solutions of different concentrations. A paired comparison test is used to determine the maximum dilution factor of the test sample at which the evaluation team just identifies a numb feeling, and the Scoville index value is obtained.

4. The method for grading numbness according to claim 1, characterized in that: The specific steps for performing correlation analysis on the numbing substances and the Scoville index value are as follows: Pearson correlation analysis was performed on the measured numbing substance contents and the corresponding Scoville values ​​to obtain the correlation coefficients between the numbing substance contents and the Scoville values.

5. The method for grading numbness according to claim 1, characterized in that: The specific steps for obtaining the stimulation threshold, stimulation time, and salivation amount of the numbing taste monomer are as follows: After the hemp-flavored monomer substances were dissolved in food-grade ethanol, drinking water was used as a solvent to prepare hemp-flavored aqueous solutions with different concentrations; To evaluate the stimulation threshold of a single substance of numbing taste, start tasting from a high-concentration sample and continue tasting until the numbing taste is no longer felt. This is the stimulation threshold of the single substance of numbing taste. For the evaluation of the stimulation time and saliva production of numbing monomer substances, different gradients are recorded; the sampling method is to accurately weigh the sample solution with a pipette and place it on the evaluator's tongue, and it is required to ensure that the sample solution is placed on the tongue for a fixed length of time. When evaluating the stimulation time, the evaluator is allowed to swallow, and when evaluating the saliva production, the evaluator is required not to swallow until the numbing taste disappears.

6. The method for grading numbness according to claim 1, characterized in that: The numbing activity value is determined by the ratio of the content of a certain numbing substance to its stimulation threshold.

7. The method for grading numbness according to claim 1, characterized in that: The mathematical model expression for evaluating the numbing taste level is established as follows: X=a×(W1×b1+W2×b2+W3×b3+W4×b4+……+W n ×b n )×q; W1, W2, W3 and W4 are the contents of numbing substances respectively, b1, b2, b3 and b4 are the corresponding Scoville index values ​​respectively, n represents the nth numbing substance, a represents the conversion coefficient; q represents the weight coefficient, and is greater than 1.

8. A system for grading numbing taste according to any one of claims 1 to 7, characterized in that: include: The first determination module uses high performance liquid chromatography to determine the content of numbing substances in the target test object; A second determination module is used to determine the Scoville index value of the target detection object; A correlation analysis module, performing a correlation analysis on the numbing substance and the Scoville index value; The numbing activity value calculation module obtains the stimulation threshold, stimulation time and salivation amount of the numbing monomer substance, and calculates the numbing activity value based on the stimulation threshold and content; The linear regression calculation module compares the target test object with compounds composed of different numbing monomers in pairs. For compounds that do not show significant differences with the original target test object, correlation and linear regression analysis of each feature are performed to determine the compound that is most similar to the original target test object. The correlation coefficient is extracted as the conversion coefficient for the numbing taste evaluation formula. The output module establishes a mathematical model for evaluating the numbing taste level based on the numbing taste substance content, the Scoville index value of the numbing taste monomer substance and the conversion coefficient.

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