METHOD FOR DETERMINING DEGREE OF DONENESS OF STURGEON MEAT BASED ON FLAVOR FINGERPRINTING AND PARTIAL LEAST SQUARES (PLS) REGRESSION

Abstract

A method for determining a degree of doneness of a sturgeon meat based on flavor fingerprinting and a partial least squares (PLS) regression is provided. The method determines the degree of doneness of a sturgeon meat sample and predicts a flavor characteristic based on gas chromatography-ion mobility spectrometry (GC-IMS) data and a linear regression equation of the sturgeon meat sample. The method includes conducting headspace (solid-phase) micro-extraction of sturgeon meat samples with different degrees of doneness, conducting a qualitative analysis by gas chromatography-mass spectrometry (GC-MS), analyzing fingerprinting by GC-IMS, establishing a regression prediction model by performing a regression on a myofibrillar protein extraction ratio and a number of different types of volatile flavor substances, comparing characteristic regions acquired by the fingerprinting, conducting a principal component analysis (PCA), and determining, by the prediction model, a doneness degree and a flavor characteristic of a target sturgeon meat sample effectively.

Claims

1. A method for determining a degree of doneness of a sturgeon meat based on flavor fingerprinting and a partial least squares (PLS) regression, comprising the following steps: (1) shaping the sturgeon meat to a fixed size and heating the sturgeon meat at different temperatures at different times to obtain a heated sturgeon meat; (2) pulping the heated sturgeon meat in step (1) to obtain a pulped sturgeon meat; pipetting 11-15 mL of the pulped sturgeon meat into a first 20 mL headspace vial; sealing the first headspace vial; equilibrating the first headspace vial at 50-52° C. for 25-30 min; inserting an extraction head into the first headspace vial to allow adsorption at a distance of 1-2 cm from a liquid surface for 25-30 min; inserting the extraction head into a gas chromatography (GC) injection port desorbing at 235-245° C. for 2 min; and conducting a gas chromatography-mass spectrometry (GC-MS) analysis; (3) loading 1-5 g of the heated sturgeon meat in step (1) into a second 20 mL headspace vial; sealing the second headspace vial; incubating the second headspace vial at 45-55° C. for 15-25 min; and taking, by a syringe at 80-90° C., 490-510 μL of headspace gas from the second headspace vial for a gas chromatography-ion mobility spectrometry (GC-IMS) analysis; (4) measuring a myofibrillar protein content of the heated sturgeon meat in step (1), and calculating a myofibrillar protein extraction ratio indicating a ratio of a myofibrillar protein content of the heated sturgeon meat to a myofibrillar protein content of the sturgeon meat before heating; (5) establishing a multivariate linear fit curve by the PLS regression by taking the myofibrillar protein extraction ratio as a response variable and an amount of volatile flavor substances as independent variables, directly comparing characteristic regions of a fingerprint by GC-IMS, and conducting a principal component analysis (PCA); and (6) determining a degree of doneness of a target sturgeon meat sample by acquiring GC-MS and GC-IMS data of the target sturgeon meat sample, analyzing through the multivariate linear fit curve established in step (5), and comparing characteristic regions of a fingerprint of the target sturgeon meat sample and conducting the PCA according to the step (5).

2. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein step (1) further comprises: shaping a raw sturgeon meat to 3 cm×3 cm×1 cm and vacuum-heating at 50° C., 70° C., and 100° C. each for 15 min and 30 min, respectively.

3. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein in the GC-MS analysis in step (2), a GC analysis is conducted by a chromatographic column comprising a splitless injection at an inlet temperature of 240° C. and a constant flow rate of 1 mL/min, and the GC analysis is subjected to a temperature programming: holding 35° C. for 3 min, raising the temperature to 220° C. at 3° C./min, and holding at 220° C. for 10 min; and an MS analysis is conducted by a 230° C. electron impact (EI) ion source and 150° C. quadrupoles with a mass range of 33-500 amu.

4. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein in the GC-IMS analysis in step (3), a GC analysis is conducted by programming a flow rate of high-purity nitrogen (99%), wherein the high-purity nitrogen serves as a carrier gas: holding an initial flow rate of 2 mL/min for 2 min, and raising the flow rate to 10 mL/min at 10 min, 100 mL/min at 20 min, and 150 mL/min at 30 min; and an IMS analysis is conducted by controlling a flow rate of a drift gas at 150 mL/min.

5. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein in step (4), the myofibrillar protein content is determined by using a Coomassie brilliant blue method; and the myofibrillar protein extraction ratio is calculated by $A\left(\%\right)=\frac{P}{C}\times 100,$ wherein A denotes the myofibrillar protein extraction ratio (%); P denotes the myofibrillar protein content of the heated sturgeon meat (mg/100 g); and C denotes the myofibrillar protein content of the sturgeon meat before heating (mg/100 g).

6. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein in step (5), the multivariate linear fit curve between the myofibrillar protein extraction ratio and a number of different types of volatile flavor substances is expressed by: Y=16.8553+0.0496382X.sub.1−0.0167546X.sub.2+0.0284132X.sub.3−0.0359706X.sub.4+0.0106525X.sub.5−0.0796625X.sub.6−0.0192646X.sub.7+0.0360119X.sub.8+0.0194102X.sub.9+0.0196761X.sub.10, wherein X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, X.sub.7, X.sub.8, X.sub.9, and X.sub.10 denote contents of 2-butanone monomer, ethyl acetate monomer, acetylacetone monomer, acetyl dimer, n-nonanal, octanoic acid, hexanal dimer, heptanal monomer, 1-hexanol monomer, and cyclohexanone, respectively.

7. The method for determining the degree of doneness of the sturgeon meat based on flavor fingerprinting and the PLS regression according to claim 1, wherein step (5) further comprises: conducting a comparative analysis of the fingerprint by GC-IMS to acquire characteristic regions of the volatile flavor substances, wherein the characteristic regions characterize a flavor characteristic of the sturgeon meat, and conducting a dynamic PCA by a Dynamic PCA plug-in to cluster volatile flavor substances and quickly determine a type of an unknown volatile flavor substance.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIGS. 1 to 6 show total ion chromatograms (TICs) of flavor components of modeling standard sturgeon meat samples with different degrees of doneness according to the examples of the present disclosure.

[0023] FIG. 7 shows myofibrillar protein contents of the modeling standard sturgeon meat samples with different degrees of doneness according to the examples of the present disclosure.

[0024] FIG. 8 shows a mean squared error of prediction (MSEP) and R.sup.2 as a function of a number of principal components according to an example of the present disclosure.

[0025] FIG. 9 shows a regression of a training set of principal components of different dimensions according to an example of the present disclosure.

[0026] FIG. 10 shows a score for a myofibrillar protein extraction ratio according to an example of the present disclosure.

[0027] FIG. 11 shows a validation regression of a prediction model according to an example of the present disclosure.

[0028] FIG. 12 shows a fingerprint of flavor components of modeling standard sturgeon meat samples with different degrees of doneness according to an example of the present disclosure.

[0029] FIG. 13 shows a principal component analysis (PCA) plot of the flavor components of the modeling standard sturgeon meat samples with different degrees of doneness according to the example of the present disclosure.

[0030] FIGS. 14A and 14B show a TIC of flavor components of a target sturgeon meat sample according to an example of the present disclosure.

[0031] FIGS. 15A and 15B show a fingerprint of the flavor components of the target sturgeon meat sample according to an example of the present disclosure.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0032] In order to make the objectives, technical solutions, and advantages of the present disclosure clearer, the implementations of the present disclosure will be further described in detail in conjunction with the drawings.

Example 1

[0033] The present disclosure provides a method for determining a degree of doneness of sturgeon meat based on flavor fingerprinting and a partial least squares (PLS) regression, which specifically includes the following steps:

(1) Sample Treatment

[0034] A frozen male Russian sturgeon was thawed for 0.5 h by flowing water, cut, and sampled along the spine. The cut sturgeon meat was shaped to 3 cm×3 cm×1 cm and washed through sterile water.

[0035] The temperatures of vacuum pans were respectively set at 50° C. (−880 bar), 70° C. (−700 bar), and 100° C. (1.01 bar), corresponding to a boiling point under vacuum. The sturgeon meat was completely immersed in water and heated for 15 min and 30 min, respectively. Then the sturgeon meat was removed from the vacuum pan and placed into a polyamide (PA)+cast polypropylene (CPP) bag for vacuum packaging. Raw meat samples were washed with sterile water and directly placed into PA+CPP bags for vacuum packaging, sequentially numbered as RAW, LTVH5015, LTVH5030, LTVH7015, LTVH7030, and TC10015.

(2) Gas Chromatography-Mass Spectrometry (GC-MS)

1) Extraction of Volatile Flavor Substances

[0036] 10 g of the sturgeon meat was taken and pulped with 20 mL of distilled water. 13 mL of sturgeon meat pulp was pipetted into a 20 mL headspace vial, and the headspace vial was sealed with a cap (with a silicone septum). The headspace vial was equilibrated at 50° C. for 30 min. An extraction head was inserted into the headspace vial to allow adsorption at a distance of 1 cm from a liquid surface for 30 min. Then the extraction head was inserted into a GC injection port, and the desorption was conducted at 240° C. for 2 min for a GC-MS analysis.

2) GC-MS Analysis Conditions

[0037] The GC analysis was conducted by a chromatographic column, which featured splitless injection, a constant flow mode, an inlet temperature of 240° C., and a flow rate of 1 mL.Math.min−1, and is subjected to temperature programming: hold 35° C. for 3 min, raise the temperature to 220° C. by 3° C..Math.min−1, and hold for 10 min. The MS analysis was conducted by a 230° C. electron impact (EI) ion source and 150° C. quadrupoles with a mass range of 33-500 amu.

3) Identification of Volatile Flavor Substances

[0038] The detected compounds were processed by MassHunter, and unknown substances were matched by NIST14 database. Only substances with forward and backward matching degrees greater than 750 were reported. Key flavor compounds were identified by a relative odor activity value (ROAV). That is, a component with the largest contribution to the flavor of the sample was defined as ROAV.sub.stan=100, and other volatile components are calculated as follows:

[00002]${\mathrm{ROAV}}_i\approx \frac{C_{\mathrm{ri}}}{C_{\mathrm{rstan}}}\times \frac{T_{\mathrm{stan}}}{T_i}\times 100$ [0039] C.sub.ri: relative percentage of the volatile component; [0040] C.sub.rstan: relative percentage of the component with the largest contribution to the overall flavor of the sample; [0041] T.sub.i: sensory threshold of the volatile component; [0042] T.sub.stan: sensory threshold of the component with the largest contribution to the overall flavor of the sample.

(3) Determination of Myofibrillar Protein Content

[0043] 15.00 g of the sample was weighed and added into 110 mL of buffer A (20 mmol/L phosphate buffer, including 100 mmol/L NaCl and 1 mmol/L EDTA, pH=7.0). The sample and the buffer were mixed well, homogenized at 15,000 r/min for 60 s, and centrifuged at 4° C. at 8,000 r/min for 10 min. Then the supernatant was removed. 50 mL of buffer A was added to a precipitate, and homogenization was conducted for 60 s. Centrifugation was conducted again under the same conditions to yield a precipitate. This operation was repeated twice. 30 mL of buffer B (25 mmol/L phosphate buffer, including 0.6 mol/L NaCl, pH=7.0) was added to the final precipitate, and homogenization was conducted. Then the mixture was placed in a 4° C. refrigerator overnight to dissolve. Centrifugation was conducted at 4° C. at 10,000 r/min for 10 min, and an insoluble fraction was removed to acquire a supernatant, namely a myofibrillar protein solution. Using bovine serum albumin as a standard curve, the concentration of the solution was determined by a Coomassie brilliant blue method.

(4) Gas Chromatography-Ion Mobility Spectrometry (GC-IMS)

[0044] 2 g of the mixed sample was weighed, placed into a 20 mL headspace vial, and incubated at 50° C. for 20 min. 500 μL of the sample was injected via an 85° C. syringe. Analytical detection was conducted by a GC-IMS flavor analyzer, and each sample was replicated 3 times. The GC was conducted in the conditions shown in the table below:

TABLE-US-00001 TABLE 1 GC conditions of Example 1 Time E1 E2 R 00:00,000 150 mL/min 2 ml/min Rec 02:00,000 150 mL/min 2 ml/min — 10:00,000 150 mL/min 10 ml/min — 20:00,000 150 mL/min 100 ml/min — 30:00,000 150 mL/min 150 ml/min Stop

[0045] GC-IMS data processing and analysis: The analysis software includes Laboratory Analytical Viewer (LAV), three plug-ins, and GC×IMS Library Search, which can analyze the samples from different angles. LAV is configured to view the analytical spectrum, where each point in the graph represents a volatile organic compound, and can be quantitatively analyzed by establishing a standard curve. The Reporter plug-in is configured to directly compare spectral differences between the samples. The Gallery Plot plug-in (Fingerprint Comparison) is configured to visually and quantitatively compare differences in the volatile organic compounds between different samples. The Dynamic PCA plug-in is configured to cluster substances and quickly determine the type of unknown substances. GC×IMS Library Search is the built-in NIST database and IMS database of the application software, which can conduct a qualitative analysis of substances.

(5) Establishment of a Regression Prediction Equation for Flavor Components of Modeling Standard Sturgeon Meat

[0046] FIGS. 1 to 6 show total ion chromatograms (TICs) of modeling standard sturgeon meat samples with different degrees of doneness. The myofibrillar protein content was determined by the Coomassie brilliant blue method, and the changes in the myofibrillar protein extraction ratio of the modeling standard sturgeon meat samples with different degrees of doneness are shown in FIG. 7. Partial least squares (PLS) regression analysis was conducted by using RAW, LTVH50-15, LTVH50-30, LTVH70-15, LTVH70-30 and TC100-15 as training sets and LTVH60-15 and LTVH60-30 as validation sets. As shown in FIG. 8, when the number of principal components was 2, the mean squared error of prediction (MSEP) decreased significantly and then remained basically unchanged, and the R2 increased significantly and then remained basically unchanged. Therefore, the number of principal components for dimensionality reduction was determined to be 2. The model training results are shown in FIG. 9. The prediction accuracy of the PLS model using the first two components was almost the same as that of the full model, and the first two components were sufficient to represent the overall characteristics of the dataset. According to the score plot (FIG. 10), the myofibrillar protein extraction ratio had an increasing trend along the first and second principal components. It was further tested whether the model had desired predictive performance through the validation set. As shown in FIG. 11, for the test set data, the prediction effect of the constructed PLS regression model was still desired. After analyzing the contribution of all independent variables, 10 factors with the highest weight ratio were screened out, and their regression coefficients were calculated. Thus, the regression curve between the myofibrillar protein extraction ratio and a number of different types of volatile flavor substances was expressed by: Y=16.8553+0.0496382X.sub.1−0.0167546X.sub.2+0.0284132X.sub.3−0.0359706X.sub.4+0.0106525X.sub.5−0.0796625X.sub.6−0.0192646X.sub.7+0.0360119X.sub.8+0.0194102X.sub.9+0.0196761X.sub.10, where X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, X.sub.7, X.sub.8, X.sub.9, and X.sub.10 denoted the contents of 2-butanone monomer, ethyl acetate monomer, acetylacetone monomer, acetylacetone dimer, n-nonanal, octanoic acid, hexamethylene dimer, heptanal monomer, 1-hexanol monomer, and cyclohexanone, respectively. Through this expression, the myofibrillar protein extraction ratio could be inferred from the content of each flavor component to determine the degree of doneness of the sturgeon meat.

(6) Fingerprinting on Flavor Components of Modeling Standard Sturgeon Meat

[0047] According to FIG. 12, the flavor substances of the sturgeon meat gradually changed with the increase of the degree of doneness. In general, the contents of the volatile flavor substances in the sturgeon meat gradually increased and then decreased. Region 2 was a relatively concentrated region of aldehydes. In this region, nonanal, benzaldehyde, and heptanal did not change much in each treatment group, while octanal, hexanal, and pentanal first increased and then decreased. Regions 1 and 4 were concentrated by 2-pentanone, 3-octanol, and 1-hexanol, showing a trend of first increasing and then decreasing. In a left-to-right direction, Region 3 was occupied from left to right by acetone, 2-butanone, 3-methylbutyraldehyde, and acetoin, which all increased gradually. Therefore, the doneness of the sturgeon meat could be determined based on the contents of the volatile flavor substances in Region 3.

(7) PCA on Flavor Components of Modeling Standard Sturgeon Meat

[0048] According to FIG. 13, the contribution rates of the first principal component axis and the second principal component axis were 35% and 27%, respectively, and the total contribution rate was 62%. This indicates that the principal components could better reflect most of the characteristic information of the volatile flavor substances in the sturgeon meat with different degrees of doneness. The differences between the sturgeon meat samples in different treatment groups are clearly demonstrated in the figure. The sturgeon meat samples close to each other indicated a small difference in the components, and those far apart indicated an obvious difference in the components. With the increase in the degree of doneness, the sturgeon samples showed a regular distribution from the bottom left to the upper right, indicating that the PCA could well distinguish the sturgeon meat with different degrees of doneness.

(8) Determination and Analysis of Samples to be Tested

[0049] In this example, the sturgeon meat treated at 60° C. for 15 min was used as the test object, and the GC-MS test method is the same as the above method and will not be repeated herein.

[0050] The TIC of the sample to be tested is shown in FIG. 14A. According to the prediction of the regression curve between the myofibrillar protein extraction ratio and the number of different types of volatile flavor substances, the types and contents of the volatile flavor substances in LTVH60-15 are shown in Table 2. The myofibrillar protein content calculated by the prediction model was 19.47655%, and the actual myofibrillar protein content was 19.47652% with a relative deviation of prediction being 0.00020%. This shows that the regression curve could well predict the myofibrillar protein extraction ratio of the sample to determine the degree of doneness.

TABLE-US-00002 TABLE 2 Types and peak volumes of volatile flavor substances in the sample in Example 1 LTVH50-15 LTVH50-30 LTVH60-15 LTVH70-15 LTVH70-30 TC100-15 Acetone 1092.515 3166.211333 1520.753667 2301.943667 2713.151 3875.865667 2-butanone 833.609 1451.395 804.9213333 1040.750333 1099.156333 1282.438333 monomer Ethyl acetate 635.4893333 539.739 525.6893333 402.8853333 492.49 414.754 monomer Ethyl acetate 188.5713333 142.6596667 143.4473333 76.10566667 120.1766667 118.434 dimer 3-methylbutanal 555.6446667 558.217 265.655 262.0243333 388.5626667 711.0326667 monomer 3-methylbutanal 254.9526667 158.7236667 60.83766667 53.41633333 96.98366667 371.9613333 dimer Acetylacetone 1316.502667 534.3076667 977.4813333 1484.413333 1096.660667 1881.913667 monomer Acetyl dimer 315.3886667 258.7856667 228.4673333 441.6536667 331.826 721.338 Benzaldehyde 156.3466667 274.7236667 152.481 149.5026667 167.2806667 151.2143333 N-nonanal 364.0623333 315.044 297.1036667 371.7013333 323.7786667 301.0543333 Octanoic acid 172.8633333 396.942 222.5953333 372.3293333 350.9216667 285.3846667 Pentanal 430.192 1015.541 809.396 727.9783333 879.769 383.2993333 monomer Hexanal monomer 1462.463333 2796.367667 2614.102333 2628.669333 2710.688333 2080.02 Hexanal dimer 829.22 5501.522 4014.124667 3197.962667 3930.321333 1767.170667 Heptanal dimer 59.59 220.8683333 75.939 98.40366667 107.1266667 64.89066667 1-hexanol dimer 43.097 43.744 54.921 191.5953333 200.226 216.6623333 Heptanal 417.7313333 1076.712 616.307 862.984 874.0033333 655.828 monomer 1-hexanol 73.35133333 99.75533333 253.6173333 683.6673333 684.4076667 705.0966667 monomer 3-octanol 65.024 193.4856667 128.9926667 180.3263333 206.214 131.8523333 2-butanone dimer 64.77933333 1087.649 119.427 415.128 652.547 1108.438333 2-heptanone 46.48566667 89.10033333 63.10266667 87.34566667 78.67833333 74.84433333 Pentanal dimer 32.607 588.2113333 151.7076667 124.424 243.453 53.91366667 2-pentanone 63.09333333 35.60466667 55.17166667 79.578 59.499 77.68733333 monomer 2-pentanone 25.54 160.3286667 45.65866667 76.48933333 70.797 72.98033333 dimer Cyclohexanone 35.36466667 195.4366667 49.95233333 58.00633333 69.797 48.689 Predicted 29.99896 27.32501 19.47651 8.463198 4.850957 8.159824 myofibrillar protein extraction ratio (%) Actual 29.99899 27.32503 19.47655 8.463249 4.851009 8.159876 myofibrillar protein extraction ratio (%) Relative deviation 0.000122247 9.66431E−05 0.000199197 0.000606532 0.001063612 0.000639678 of prediction (%)

(9) Verification of Prediction Results

[0051] According to FIG. 15A, the fingerprints of the sturgeon meat treated at 60° C. for 15 min and other treatment groups were significantly different. In general, the types of the volatile flavor substances were significantly less than those of other treatment groups, and the amount of other flavor substances were lower except for the ethyl acetate in Region 2. This shows that GC-IMS could effectively distinguish the characteristic flavors of different samples to determine their degree of doneness.

Example 2

[0052] The present disclosure provides a method for determining a degree of doneness of sturgeon meat based on flavor fingerprinting and a partial least squares (PLS) regression, which specifically includes the following steps:

(1) Sample Treatment

[0053] A frozen male Russian sturgeon was thawed for 0.5 h by flowing water and was cut and sampled along the spine. The cut sturgeon meat was shaped to 3 cm×3 cm×1 cm and washed with sterile water.

[0054] The temperatures of vacuum pans were respectively set at 50° C. (−880 bar), 70° C. (−700 bar), 100° C. (1.01 bar), corresponding to a boiling point under vacuum. The sturgeon meat was completely immersed in water and was heated for 15 min and 30 min, respectively. Then the sturgeon meat was removed from the vacuum pan and placed into a polyamide (PA)+cast polypropylene (CPP) bag for vacuum packaging. Raw meat samples were washed with sterile water and directly placed into PA+CPP bags for vacuum packaging, sequentially numbered as RAW, LTVH5015, LTVH5030, LTVH7015, LTVH7030, and TC10015.

(2) Gas Chromatography-Mass Spectrometry (GC-MS)

1) Extraction of Volatile Flavor Substances

[0055] 10 g of the sturgeon meat was weighed and pulped with 20 mL of distilled water. 13 mL of sturgeon meat pulp was pipetted into a 20 mL headspace vial, and the headspace vial was sealed with a cap (with a silicone septum). The headspace vial was equilibrated at 50° C. for 30 min. An extraction head was inserted into the headspace vial to allow adsorption at a distance of 1 cm from a liquid surface for 30 min. Then the extraction head was inserted into a GC injection port, and the desorption was conducted at 240° C. for 2 min for a GC-MS analysis.

2) GC-MS Analysis Conditions

[0056] The GC analysis was conducted by a chromatographic column, which featured splitless injection, a constant flow mode, an inlet temperature of 240° C., and a flow rate of 1 mL.Math.min.sup.−1, and is subjected to temperature programming: hold 35° C. for 3 min, raise the temperature to 220° C. by 3° C..Math.min.sup.−1, and hold for 10 min. The MS analysis was conducted by a 230° C. electron impact (EI) ion source and 150° C. quadrupoles with a mass range of 33-500 amu.

3) Identification of Volatile Flavor Substances

[0057] The detected compounds were processed by MassHunter, and unknown substances were matched by NIST14 database. Only substances with forward and backward matching degrees greater than 750 were reported. Key flavor compounds were identified by a relative odor activity value (ROAV). That is, a component with the largest contribution to the flavor of the sample was defined as ROAV.sub.stan=100, and other volatile components are calculated as follows:

[00003]${\mathrm{ROAV}}_i\approx \frac{C_{\mathrm{ri}}}{C_{\mathrm{rstan}}}\times \frac{T_{\mathrm{stan}}}{T_i}\times 100$ [0058] C.sub.ri: relative percentage of the volatile component; [0059] C.sub.rstan: relative percentage of the component with the largest contribution to the overall flavor of the sample; [0060] T.sub.i: sensory threshold of the volatile component; and [0061] T.sub.stan: sensory threshold of the component with the largest contribution to the overall flavor of the sample.

(3) Determination of Myofibrillar Protein Content

[0062] 15.00 g of the sample was weighed and added into 110 mL of buffer A (20 mmol/L phosphate buffer, including 100 mmol/L NaCl and 1 mmol/L EDTA, pH=7.0). The sample and the buffer were mixed well, homogenized at 15,000 r/min for 60 s, and centrifuged at 4° C. at 8,000 r/min for 10 min. Then the supernatant was removed. 50 mL of buffer A was added to a precipitate, and homogenization was conducted for 60 s. Centrifugation was conducted again under the same conditions to yield a precipitate. This operation was repeated twice. 30 mL of buffer B (25 mmol/L phosphate buffer, including 0.6 mol/L NaCl, pH=7.0) was added to the final precipitate, and homogenization was conducted. Then the mixture was placed in a 4° C. refrigerator overnight to dissolve. Centrifugation was conducted at 4° C. at 10,000 r/min for 10 min, and an insoluble fraction was removed to acquire a supernatant, namely a myofibrillar protein solution. Using bovine serum albumin as a standard curve, the concentration of the solution was determined by a Coomassie brilliant blue method.

(4) Gas Chromatography-Ion Mobility Spectrometry (GC-IMS)

[0063] 2 g of the mixed sample was weighed, placed into a 20 mL headspace vial, and incubated at 50° C. for 20 min. 500 μL of the sample was injected via an 85° C. syringe. Analytical detection was conducted by a GC-IMS flavor analyzer, and each sample was replicated 3 times. The GC was conducted in the conditions shown in the table below:

TABLE-US-00003 TABLE 3 GC conditions of Example 2 Time E1 E2 R 00:00,000 150 mL/min 2 ml/min Rec 02:00,000 150 mL/min 2 ml/min — 10:00,000 150 mL/min 10 ml/min — 20:00,000 150 mL/min 100 ml/min — 30:00,000 150 mL/min 150 ml/min Stop

[0064] GC-IMS data processing and analysis: The analysis software includes Laboratory Analytical Viewer (LAV), three plug-ins and GC×IMS Library Search, which can analyze the samples from different angles. LAV is configured to view the analytical spectrum, where each point in the graph represents a volatile organic compound, and can be quantitatively analyzed by establishing a standard curve. The Reporter plug-in is configured to directly compare spectral differences between the samples. The Gallery Plot plug-in (Fingerprint Comparison) is configured to visually and quantitatively compare differences in the volatile organic compounds between different samples. The Dynamic PCA plug-in is configured to cluster substances and quickly determine the type of unknown substances. GC×IMS Library Search is the built-in NIST database and IMS database of the application software, which can conduct a qualitative analysis of substances.

(5) Establishment of Regression Prediction Equation for Flavor Components of Modeling Standard Sturgeon Meat

[0065] FIGS. 1 to 6 show the TICs of modeling standard sturgeon meat samples with different degrees of doneness. The myofibrillar protein content was determined by the Coomassie brilliant blue method, and the changes in the myofibrillar protein extraction ratio of the modeling standard sturgeon meat samples with different degrees of doneness are shown in FIG. 7. Partial least squares (PLS) regression analysis was conducted by using RAW, LTVH50-15, LTVH50-30, LTVH70-15, LTVH70-30 and TC100-15 as training sets and LTVH60-15 and LTVH60-30 as validation sets. As shown in FIG. 8, when the number of principal components was 2, the mean squared error of prediction (MSEP) decreased significantly and then remained basically unchanged, and the R2 increased significantly and then remained basically unchanged. Therefore, the number of principal components for dimensionality reduction was determined to be 2. The model training results are shown in FIG. 9. The prediction accuracy of the PLS model using the first two components was almost the same as that of the full model, and the first two components were sufficient to represent the overall characteristics of the dataset. According to the score plot (FIG. 10), the myofibrillar protein extraction ratio had an increasing trend along the first and second principal components. It was further tested whether the model had desired predictive performance through the validation set. As shown in FIG. 11, for the test set data, the prediction effect of the constructed PLS regression model was still desired. After analyzing the contribution of all independent variables, 10 factors with the highest weight ratio were screened out, and their regression coefficients were calculated. Thus, the regression curve between the myofibrillar protein extraction ratio and the number of different types of volatile flavor substances was expressed by: Y=16.8553+0.0496382X.sub.1−0.0167546X.sub.2+0.0284132X.sub.3−0.0359706X.sub.4+0.0106525X.sub.5−0.0796625X.sub.6−0.0192646X.sub.7+0.0360119X.sub.8+0.0194102X.sub.9+0.0196761X.sub.10, where X.sub.1, X.sub.2, X.sub.3, X.sub.4, X.sub.5, X.sub.6, X.sub.7, X.sub.8, X.sub.9, and X.sub.10 denoted the contents of 2-butanone monomer, ethyl acetate monomer, acetylacetone monomer, acetylacetone dimer, n-nonanal, octanoic acid, hexamethylene dimer, heptanal monomer, 1-hexanol monomer, and cyclohexanone, respectively. Through this expression, the myofibrillar protein extraction ratio could be inferred from the content of each flavor component to determine the degree of doneness of the sturgeon meat.

(6) Fingerprinting on Flavor Components of Modeling Standard Sturgeon Meat

[0066] According to FIG. 12, the flavor substances of the sturgeon meat gradually changed with the increase of the degree of doneness. In general, the contents of the volatile flavor substances in the sturgeon meat gradually increased and then decreased. Region 2 was a relatively concentrated region of aldehydes. In this region, nonanal, benzaldehyde, and heptanal did not change much in each treatment group, while octanal, hexanal, and pentanal first increased and then decreased. Regions 1 and 4 were occupied by 2-pentanone, 3-octanol, and 1-hexanol, showing a trend of increasing first and then decreasing. In a left-to-right direction, Region 3 was occupied from left to right by acetone, 2-butanone, 3-methylbutyraldehyde, and acetoin, which all increased gradually. Therefore, the doneness of the sturgeon meat could be determined based on the amount of the volatile flavor substances in Region 3.

(7) PCA on Flavor Components of Modeling Standard Sturgeon Meat

[0067] According to FIG. 13, the contribution rates of the first principal component axis and the second principal component axis were 35% and 27%, respectively, and the total contribution rate was 62%. This indicates that the principal components could better reflect most of the characteristic information of the volatile flavor substances in the sturgeon meat with different degrees of doneness. The differences between the sturgeon meat samples in different treatment groups are clearly demonstrated in the figure. The sturgeon meat samples that cluster close to each other indicated a small difference in the components, and those far apart indicated an obvious difference in the components. With the increase in the degree of doneness, the sturgeon samples showed a regular distribution from the bottom left to the upper right, indicating that the PCA could well distinguish the sturgeon meat with different degrees of doneness.

(8) Determination and Analysis of Samples to be Tested

[0068] In this example, the sturgeon meat treated at 60° C. for 30 min was used as the test object, and the GC-MS test method is the same as the above method and will not be repeated herein.

[0069] The TIC of the sample to be tested is shown in FIG. 14B. A prediction is based on the regression curve between the myofibrillar protein extraction ratio and the number of different types of volatile flavor substances. The types and amount of the volatile flavor substances in LTVH60-30 are shown in Table 2. The myofibrillar protein content calculated by the prediction model was 17.17123%, and the actual myofibrillar protein content was 17.17127% with a relative deviation of prediction being 0.00026%. This shows that the regression curve could accurately predict the myofibrillar protein extraction ratio of the sample to determine the degree of doneness.

TABLE-US-00004 TABLE 4 Types and amounts of volatile flavor substances in the sample in Example 2 LTVH50-15 LTVH50-30 LTVH60-30 LTVH70-15 LTVH70-30 TC100-15 Acetone 1092.515 3166.211333 1351.545667 2301.943667 2713.151 3875.865667 2-butanone 833.609 1451.395 747.687 1040.750333 1099.156333 1282.438333 monomer Ethyl acetate 635.4893333 539.739 558.3936667 402.8853333 492.49 414.754 monomer Ethyl acetate 188.5713333 142.6596667 187.5213333 76.10566667 120.1766667 118.434 dimer 3-methylbutanal 555.6446667 558.217 275.336 262.0243333 388.5626667 711.0326667 monomer 3-methylbutanal 254.9526667 158.7236667 62.76 53.41633333 96.98366667 371.9613333 dimer Acetylacetone 1316.502667 534.3076667 928.03 1484.413333 1096.660667 1881.913667 monomer Acetyl dimer 315.3886667 258.7856667 207.9383333 441.6536667 331.826 721.338 Benzaldehyde 156.3466667 274.7236667 141.395 149.5026667 167.2806667 151.2143333 N-nonanal 364.0623333 315.044 364.6003333 371.7013333 323.7786667 301.0543333 Octanoic acid 172.8633333 396.942 314.9603333 372.3293333 350.9216667 285.3846667 Pentanal 430.192 1015.541 861.7086667 727.9783333 879.769 383.2993333 monomer Hexanal monomer 1462.463333 2796.367667 2736.581667 2628.669333 2710.688333 2080.02 Hexanal dimer 829.22 5501.522 4556.180333 3197.962667 3930.321333 1767.170667 Heptanal dimer 59.59 220.8683333 148.9803333 98.40366667 107.1266667 64.89066667 1-hexanol dimer 43.097 43.744 90.586 191.5953333 200.226 216.6623333 Heptanal 417.7313333 1076.712 892.508 862.984 874.0033333 655.828 monomer 1-hexanol 73.35133333 99.75533333 414.8763333 683.6673333 684.4076667 705.0966667 monomer 3-octanol 65.024 193.4856667 171.4526667 180.3263333 206.214 131.8523333 2-butanone dimer 64.77933333 1087.649 165.296 415.128 652.547 1108.438333 2-heptanone 46.48566667 89.10033333 78.43733333 87.34566667 78.67833333 74.84433333 Pentanal dimer 32.607 588.2113333 262.999 124.424 243.453 53.91366667 2-pentanone 63.09333333 35.60466667 62.51666667 79.578 59.499 77.68733333 monomer 2-pentanone 25.54 160.3286667 61.76633333 76.48933333 70.797 72.98033333 dimer Cyclohexanone 35.36466667 195.4366667 56.81233333 58.00633333 69.797 48.689 Predicted 29.99896 27.32501 17.17123 8.463198 4.850957 8.159824 myofibrillar protein extraction ratio (%) Actual 29.99899 27.32503 17.17127 8.463249 4.851009 8.159876 myofibrillar protein extraction ratio (%) Relative deviation 0.000122247 9.66431E−05 0.00026921 0.000606532 0.001063612 0.000639678 of prediction (%)

(9) Verification of Prediction Results

[0070] According to FIG. 15B, the fingerprints of the sturgeon meat treated at 60° C. for 30 min and other treatment groups were significantly different. In general, the types of the volatile flavor substances were closer to those of the TC treatment group, and the contents of the flavor substances were closer to those of the TC treatment group except for the 2-butanone in Region 3. This shows that GC-IMS could effectively distinguish the characteristic flavors of different samples to determine their degree of doneness.

[0071] The above embodiments are intended to explain the present disclosure, rather than to limit the present disclosure. Any modifications and changes made to the present disclosure within the spirit and the protection scope defined by the claims should all fall within the protection scope of the present disclosure.