A meat processing method for reducing the content of bound heterocyclic amines

Abstract

The present invention belongs to the field of food processing technology, and specifically relates to a meat processing method for reducing the content of bound heterocyclic amines. The meat processing method comprises the following steps: providing fresh meat and amino acids, wherein the amino acids are a mixture of cysteine, histidine, and proline; processing the fresh meat into minced meat, adding the amino acids, stirring and pressing into shape to obtain meat patties; and baking the meat patties to obtain baked meat patties with a low content of bound heterocyclic amines. The present invention blocks common aldehyde intermediates by compounding amino acids, thereby significantly reducing the content of bound heterocyclic amines in the baked meat patties, making the baked meat patties safer and more reliable to eat, and can also be applied to factory production; the added cysteine, histidine, and proline are all naturally occurring nutrients in food raw materials, and have the advantages of safety, low cost, and high degree of standardization.

Description

Technical Field

[0001] The invention belongs to the technical field of food processing, and in particular relates to a meat processing method for reducing the content of bound heterocyclic amines. Background Art

[0002] Meat and meat products are an important part of our daily diet and a rich source of valuable nutrients and minerals. Traditionally grilled or fried meat products are highly popular among consumers. However, these high-temperature processed products contain high levels of carcinogenic and mutagenic heterocyclic amines. Epidemiological studies have shown that excessive intake of fried and grilled meats increases the risk of various cancers, including colon, breast, and pancreatic cancer. Based on their formation pathways and chemical structures, these heterocyclic amines can be divided into aminoimidazoazarenes (AIAs) and aminocarbolines (ACs). Among them, AIAs heterocyclic amines all contain imidazole rings and are produced at temperatures between 150 and 300°C. They mainly include quinolines (IQ, MeIQ, IQ[4,5-b], etc.), quinoxalines (IQx, MeIQx, 4,8-DiMeIQx, etc.) and pyridines (PhIP, DMIP, etc.); ACs heterocyclic amines are generally believed to be produced by thermal decomposition of amino acids or proteins at high temperatures above 300°C, mainly including α-carbolines (MeAαC and AαC), β-carbolines (Norharman and Harman), etc.

[0003] Heterocyclic amines can exist in both free and bound forms. Food ingredients like meat, which are prone to producing heterocyclic amines, often have a higher protein content (20%) than small molecules like glucose, creatine (indene), and free amino acids (0.1% to 0.5%). This generates bound heterocyclic amines, which bind to muscle protein and are found to contain levels significantly higher than those in the free form, reaching levels as high as μg / g in beef patties. These bound heterocyclic amines are then converted into free heterocyclic amines by digestive enzymes, thereby increasing human exposure to heterocyclic amines.

[0004] Therefore, methods to effectively control and reduce the content of bound heterocyclic amines in meat products have received widespread attention in the academic community and have become one of the hot topics in the field of food safety. Summary of the Invention

[0005] The present invention aims to solve the above problems and provides a meat processing method for reducing the content of bound heterocyclic amines. The common aldehyde intermediates are blocked by amino acid compounding, thereby significantly reducing the content of bound heterocyclic amines in roast beef patties.

[0006] According to the technical solution of the present invention, the meat processing method for reducing the content of bound heterocyclic amines comprises the following steps:

[0007] S1: providing fresh meat and amino acids, wherein the amino acids are a mixture of cysteine, histidine and proline;

[0008] S2: processing the fresh meat into minced meat, adding the amino acid, stirring evenly, and pressing into shape to obtain meat patties;

[0009] S3: baking the meat patty to obtain a baked meat patty with a low content of bound heterocyclic amines.

[0010] The present invention blocks the common aldehyde intermediates by combining amino acids, thereby significantly reducing the content of bound heterocyclic amines in roast beef patties. Among them, the inhibitory pathway of the cysteine-histidine-proline combination on bound heterocyclic amines is as follows: Figure 1 shown.

[0011] Furthermore, the fresh meat is fresh beef, specifically fresh beef tenderloin, etc.

[0012] Furthermore, among the amino acids, the mass ratio of cysteine, histidine and proline is 1-2:1-2:1-2; for example, it can be 1:1:1, 2:1:1, 1:2:1, 1:1:2, or a range formed by any two ratios.

[0013] Furthermore, in step S2, the amount of amino acids added is 0.04-0.06% of the mass of the meat patty.

[0014] Furthermore, in step S2, the fresh meat is processed into minced meat by a meat grinder.

[0015] Furthermore, in step S2, the meat patty has a thickness of 1.5-2.5 cm. Specifically, the minced meat after adding amino acids can be pressed into a meat patty with a thickness of 1.5-2.5 cm, a diameter of about 10 cm, and a weight of 70-90 g.

[0016] Furthermore, in step S3, baking is performed in an oven.

[0017] Furthermore, in step S3, the baking temperature is 220-230°C, preferably 225°C.

[0018] Furthermore, in step S3, the baking time is 16-24 minutes; wherein, the baking time on one side is 8-12 minutes.

[0019] The technical solution of the present invention has the following advantages over the prior art:

[0020] (1) The method of the present invention can significantly reduce the content of bound heterocyclic amines in roast beef patties, making cooked meat products safer to eat. The method is low-cost and suitable for large-scale factory production.

[0021] (2) The method of the present invention can reduce the content of bound heterocyclic amines while ensuring that the flavor of the grilled beef patties is not affected;

[0022] (3) The cysteine, histidine and proline added by the method of the present invention are all naturally occurring nutrients in food raw materials, and have the advantages of being safe, inexpensive and highly standardized, and are in line with the development trend of being natural, green and healthy. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The content of aldehyde intermediates in grilled beef patties after adding a cysteine-histidine-proline combination.

[0024] Figure 2 This is the inhibitory pathway of the cysteine-histidine-proline combination on bound heterocyclic amines. DETAILED DESCRIPTION

[0025] The present invention will be further described below with reference to the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it. However, the embodiments are not intended to limit the present invention.

[0026] Taking beef as an example, the present invention provides a beef processing method for reducing the content of bound heterocyclic amines, comprising the following steps:

[0027] S1: providing fresh beef and amino acids, wherein the amino acids are a mixture of cysteine, histidine, and proline in a mass ratio of 1-2:1-2:1-2;

[0028] S2: mincing fresh beef using a meat grinder, adding amino acids, and stirring to ensure uniform distribution and better function; pressing and molding the minced beef into patties having a thickness of 1.5-2.5 cm, a diameter of approximately 10 cm, and a weight of 70-90 g; wherein the amount of amino acids added is 0.04-0.06% of the weight of the patties;

[0029] S3: Put the prepared beef patties into an oven for baking at a temperature of 220-230° C. for 16-24 minutes, wherein the baking time on one side is 8-12 minutes.

[0030] Example 1

[0031] Fresh beef was minced using a meat grinder, and a cysteine-histidine-proline combination was added to the minced beef. Patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the added amount of the cysteine-histidine-proline combination accounted for 0.05% of the weight of the beef patties, and the corresponding mass ratios were 2:1:1. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0032] Example 2

[0033] Fresh beef was minced using a meat grinder, and a cysteine-histidine-proline combination was added to the minced beef. Patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the added amount of the cysteine-histidine-proline combination accounted for 0.05% of the weight of the beef patties, and the corresponding mass ratios were 1:2:1. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0034] Example 3

[0035] Fresh beef was minced using a meat grinder, and a cysteine-histidine-proline combination was added to the minced beef. Patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the added amount of the cysteine-histidine-proline combination accounted for 0.05% of the weight of the beef patties, and the corresponding mass ratios were 1:1:2. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0036] Comparative Example 1

[0037] Fresh beef was minced using a meat grinder, cysteine was added to the minced beef, and patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the amount of cysteine added accounted for 0.05% of the weight of the beef patties. The patties were then placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0038] Comparative Example 2

[0039] Fresh beef was minced using a meat grinder, histidine was added to the minced beef, and patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the amount of histidine added accounted for 0.05% of the weight of the beef patties. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0040] Comparative Example 3

[0041] Fresh beef was minced using a meat grinder, proline was added to the minced beef, and patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the amount of proline added accounted for 0.05% of the weight of the beef patties. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0042] Comparative Example 4

[0043] Fresh beef was minced using a meat grinder, and a histidine-proline combination was added to the minced beef. Patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the added amount of the histidine-proline combination accounted for 0.05% of the weight of the beef patties, and the corresponding mass ratio was 1:1. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0044] Comparative Example 5

[0045] Fresh beef was minced using a meat grinder, and a cysteine-proline combination was added to the minced beef. Patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the added amount of the cysteine-proline combination accounted for 0.05% of the weight of the beef patties, and the corresponding mass ratio was 1:1. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0046] Comparative Example 6

[0047] Fresh beef was minced using a meat grinder, cysteine was added to the minced beef, and patties with a diameter of 10 cm, a thickness of 2 cm, and a weight of approximately 80 g were formed using a patty maker, wherein the amount of cysteine added accounted for 0.5% of the weight of the beef patties. The patties were placed in an oven and baked at 225°C for 20 minutes, with each side baked for 10 minutes.

[0048] Result Analysis

[0049] The contents of aldehyde intermediates and bound heterocyclic amines (ng / g) were measured in the blank beef patties, the grilled beef patties from Examples 1-3, and Comparative Examples 1-6. The aldehyde intermediates included benzaldehyde, phenylacetaldehyde, methylglyoxal, and glyoxal; the bound heterocyclic amines included bound 2-amino-1-methyl-6-phenyl-imidazo[4,5-b]pyridine (PhIP), bound 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeAαC), and bound β-carboline heterocyclic amines (Harman and Norharman).

[0050] 1. The method for determining the content of bound heterocyclic amines is as follows:

[0051] A blank beef patty, the roasted beef patties from Examples 1-4, and Comparative Examples 1-6 were pulverized using a grinder. 4 g of beef patty powder was accurately weighed and placed in a homogenizer tube. 30 mL of 1 mol / L NaOH solution was added and homogenized using an AH-30 fully automatic homogenizer for 1 minute. The minced meat was transferred to a 200 mL beaker containing 14 g of diatomaceous earth, and 50 mL of ethyl acetate was added and stirred. Ultrasonic extraction was performed for 30 minutes, followed by vacuum filtration. The filtrate contained the free heterocyclic amine. The remaining solid residue was dried and transferred to a 48 mL pressure bottle. 40 mL of 6 mol / L hydrochloric acid was added, nitrogen was added for 1 minute, and acid hydrolysis was performed at 110°C for 24 hours. The acid hydrolysis mixture was vacuum filtered to obtain a bound heterocyclic amine extract, which was then diluted to 100 mL with ultrapure water. After mixing, 10 mL of the extract was collected for solid-phase extraction.

[0052] Solid-phase extraction: First, an MCX cartridge was activated with methanol (6 mL), ultrapure water (6 mL), and 0.1 mol / L hydrochloric acid (6 mL) according to the solid-phase extraction cartridge's operating instructions at a flow rate of 1.0 mL / min. Then, 10 mL of the hydrolyzate was loaded at a flow rate of 0.6 mL / min. The cartridge was then eluted with 0.1 mol / L HCl (6 mL) and methanol (6 mL) in sequence. Finally, the cartridge was eluted with a methanol-ammonia solution (95:5, v / v) at 0.6 mL / min to obtain 6 mL of eluent. The eluent was dried with nitrogen, reconstituted with 300 μL of methanol, and filtered through a 0.22 μm syringe filter prior to UPLC-MS / MS analysis.

[0053] Liquid phase conditions: Chromatographic column selected Waters Acquity UPLC BEH C 18 The column was 2.1 mm × 100 mm, 1.7 μm, and the column temperature was set at 45°C. The mobile phases were A (100% acetonitrile) and B (0.1% formic acid). The gradient elution conditions were: 0-2 min, 2% A; 2-12 min, 2%-20% A; 12-14 min, 20%-100% A; 14-17 min, 100%-2% A; 17-20 min, 2% A. The flow rate was 0.3 mL / min, and the injection volume was 2 μL. Mass spectrometry conditions were: electrospray ionization mode (ESI) positive. + ); detection mode: multiple reaction monitoring (MRM); ion source temperature, 100°C; desolvation gas (nitrogen) temperature, 400°C; capillary voltage, 3.50 kV; cone gas (nitrogen) flow rate, 50 L / h; desolvation gas flow rate, 700 L / h; collision cell pressure, 3.15×10 -3 mbar; collision gas (argon) flow rate, 0.15 mL / min.

[0054] The test results of the embodiments and comparative examples are shown in Table 1.

[0055] Table 1

[0056]

[0057] Note: The content of bound heterocyclic amines under each condition was repeated three times, and the results are expressed as mean ± standard deviation. The values in brackets are the inhibition rates of bound heterocyclic amines after adding the cysteine-histidine-proline combination of the present invention to the blank roast beef patty, “-” indicates inhibition, and “+” indicates promotion.

[0058] As can be seen from the table, the inhibitory effects of the three amino acids in Comparative Examples 1 to 3 when used alone are general, and Comparative Example 3 even has a promoting effect on the generation of these types of bound heterocyclic amines. Similarly, after adding two compound amino acids, Comparative Example 4 even has a promoting effect on these types of heterocyclic amines, and only Comparative Example 5 has an improved inhibition rate compared to when the amino acids act alone, indicating that the type of compound amino acids has a significant effect on the inhibitory effect. It can be seen from Comparative Examples 1 and 6 that too much amino acid addition will also promote the generation of bound heterocyclic amines, and the addition amount of 0.04% to 0.06% in the present invention is more appropriate. The cysteine-histidine-proline combination has a significant inhibitory effect on bound heterocyclic amines. Its simultaneous inhibition of multiple bound heterocyclic amines lies in the following points: first, aminoimidazole nitrogen heteroaromatic heterocyclic amines can be generated through the free radical pathway and then combined with muscle protein to generate bound heterocyclic amines, while cysteine, histidine and proline all have free radical scavenging and antioxidant effects, which can scavenge free heterocyclic amines to prevent them from binding to muscle protein; second, cysteine, histidine and proline have active amino or sulfhydryl groups, which can inhibit the production of multiple bound heterocyclic amines by combining with aldehyde intermediates.

[0059] The cysteine-histidine-proline combination described in the present invention exhibits a stable inhibitory effect on bound heterocyclic amines, with a further improved inhibition rate compared to the effects of individual amino acids. This is primarily due to the strong scavenging ability of cysteine, histidine, and proline for specific carbonyl compounds. Therefore, the combination can simultaneously scavenge the aldehyde intermediates of heterocyclic amines, achieving a synergistic inhibitory effect on bound heterocyclic amines.

[0060] 2. The method for determining the content of aldehyde intermediates is as follows:

[0061] Determination of methylglyoxal and glyoxal: Weigh 200 mg of meat powder into a centrifuge tube, add 2 mL of ultrapure water, and ultrasonically extract for 10 minutes. Then centrifuge at 10,000 g for 10 minutes, take 800 μL of supernatant, add an equal volume of methanol, and mix on a vortex oscillator at room temperature for 1 minute to precipitate the protein. After centrifugation at 10,000 g for 10 minutes, take the supernatant and determine creatinine. The liquid chromatography and mass spectrometry conditions refer to the creatinine determination in 2.2.4. Take another 500 μL of supernatant, add 100 μL of 0.08 mmol / L 2,3-hexanedione internal standard, add 200 μL of 5 mmol / L o-phenylenediamine reagent, vortex for 30 seconds, mix thoroughly, and derivatize at 4°C in the dark for 12 hours. After centrifugation at 10,000 g for 10 minutes, take the supernatant and determine. Liquid chromatography conditions: Waters Acquity UPLC BEH C 18 Chromatographic column (50mm×2.1mm, 1.7μm), column temperature 35℃. Mobile phase A was acetonitrile, phase B was 0.1% formic acid, gradient elution conditions: 0-0.1min, 10% A; 0.1-1min, 10%-20% A; 1-6min, 20%-70% A; 6-7min, 70%-100% A; 7-7.1min, 100%-10% A; 7.1-10min, 10% A. Injection volume: 5μL, flow rate: 0.3mL / min. Mass spectrometry conditions: Electrospray ionization positive mode (ESI + ) and MRM multiple reaction monitoring mode, with a capillary voltage of 3.55 kV, an ion source temperature of 110°C, and a desolvation temperature of 400°C. Desolvation gas (nitrogen) flow rate: 600 L / h; cone gas (nitrogen) flow rate: 50 L / h; and collision gas (argon) flow rate: 0.15 mL / min.

[0062] Benzaldehyde and phenylacetaldehyde were determined using HS-SPME-GC-MS. HS / SPME conditions: 2 g of meat powder was placed in a 20 mL headspace vial. 2 μL of 2-methyl-3-heptanone (118.2 μL / mL) internal standard solution was added and the vial was immediately sealed with a polytetrafluoroethylene / butyl membrane. After equilibration in a 50°C water bath for 7 min, extraction was performed using an SPME fiber extractor (50 / 30 μm DVB / CAR / PDMS) for 30 min. After injection, desorption was performed at 250°C for 7 min. High-purity helium was used as the carrier gas at a flow rate of 1 mL / min. Chromatographic conditions: An SH-WAX (30 m × 0.25 mm × 0.25 μm) capillary column was used. The column temperature program was: 40°C for 3 min, then increased to 100°C at 6°C / min, and finally increased to 250°C at 10°C / min and held for 5 min. The detector and injection port temperatures were both 250°C, and the sample was injected without split flow. Mass spectrometry conditions: EI electron source, electron energy 70 eV, mass spectrometry scanning range 35-450 m / z. The ion source and interface temperature were both 250°C. Qualitative method: The qualitative results were compared with the mass spectrum and the WILEY9 database, and C8-C 26 The retention indices were calculated using a mixed standard of n-alkanes and compared with the KI values in the NIST17 database to perform qualitative analysis of the compounds.

[0063] The results are as follows Figure 1 As shown in the figure, it can be seen that compared with the action of a single amino acid, the cysteine-histidine-proline combination of the present invention has improved the clearance rate of benzaldehyde, phenylacetaldehyde, methylglyoxal, and glyoxal, indicating that the amino acid combination is to efficiently react with these aldehydes simultaneously to achieve an improvement in the inhibitory effect on bound heterocyclic amines. The specific structure of the adduct generated by the reaction can be referred to Figure 2 .

[0064] 3. The method for determining volatile substances is as follows:

[0065] HS-SPME-GC-MS was used for determination under the same conditions as for the determination of benzaldehyde and phenylacetaldehyde. The results are shown in Table 2.

[0066] Table 2

[0067]

[0068] As can be seen, compared to the blank roast beef patties, the levels of volatile compounds such as 2-methylpyrazine and 2,5-dimethylpyrazine in Examples 1 and 2 decreased slightly, but the total amount remained almost unchanged. This demonstrates that the amino acid combination of the present invention reduces the content of bound heterocyclic amines without compromising flavor characteristics.

[0069] Obviously, the above embodiments are merely examples for clarity of explanation and are not intended to limit the implementation methods. Those skilled in the art will readily appreciate that other variations or modifications based on the above descriptions are possible. It is not necessary and impossible to enumerate all implementation methods here. Obvious variations or modifications arising therefrom remain within the scope of protection of the present invention.

Claims

1. A meat processing method for reducing the content of bound heterocyclic amines, characterized in that: The following steps are included: S1: providing fresh meat and amino acids, wherein the amino acids are a mixture of cysteine, histidine and proline; S2: processing the fresh meat into minced meat, adding the amino acid, stirring evenly, and pressing into shape to obtain meat patties; S3: baking the meat patty to obtain a baked meat patty with a low content of bound heterocyclic amines; Wherein, among the amino acids, the mass ratio of cysteine, histidine and proline is 1-2:1-2:1-2; In step S2, the amount of amino acids added is 0.04-0.06% of the mass of the meat patty.

2. The meat processing method according to claim 1, wherein The fresh meat is fresh beef.

3. The meat processing method according to claim 1, wherein In step S2, the amount of amino acids added is 0.05% of the mass of the meat patty.

4. The meat processing method according to claim 1, wherein In step S2, the fresh meat is processed into minced meat by a meat grinder.

5. The meat processing method according to claim 1, wherein In step S2, the thickness of the meat patty is 1.5-2.5 cm.

6. The meat processing method according to claim 1, wherein: In step S3, baking is performed in an oven.

7. The meat processing method according to claim 1, wherein In step S3, the baking temperature is 220-230°C.

8. The meat processing method according to claim 1 or 7, characterized in that: In step S3, the baking time is 16-24 minutes; wherein, the baking time on one side is 8-12 minutes.

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