Acrylamide generation inhibitor and method for inhibiting generation of acrylamide

Abstract

The invention provides an acrylamide generation inhibitor and a method for inhibiting acrylamide generation. The acrylamide inhibitor is syringic acid, vanillic acid or sinapic acid. The syringic acid and the vanillic acid can passivate polyphenol oxidase in the food processing process and prevent polyphenol oxidase from oxidizing polyphenol in food into quinone, and the sinapic acid can directly inhibit quinone generated by oxidizing polyphenol, so that the content of Maillard reaction substrates is reduced, and the yield of the Maillard reaction products is improved. The structure of quinone is extremely similar to that of an intermediate product dicarbonyl compound in a path for producing acrylamide through Maillard reaction, so that quinone and asparagine can be subjected to Maillard reaction under a high-temperature condition to generate acrylamide. Therefore, the Maillard reaction is prevented by passivating the activity of polyphenol oxidase or inhibiting the generation of quinone oxidized by polyphenol, the generation of 2A cancerogen acrylamide easily generated in the high-temperature food processing process is further inhibited, the purpose of healthy diet is achieved, and a new thought is provided for controlling the generation of acrylamide in the food processing process.

Description

Technical Field

[0001] The invention belongs to the technical field of food production safety, and in particular relates to an acrylamide generation inhibitor and a method for inhibiting acrylamide generation. Background Art

[0002] Acrylamide (AA), with the chemical formula CH2CHCONH2, is an unsaturated amide and a colorless and odorless crystal at room temperature. Acrylamide can be absorbed by the human body through various pathways, such as intact skin, mucous membranes, and lungs. The fastest absorption is through the digestive tract, and acrylamide is widely distributed in various tissues in the body. Acrylamide has strong neurotoxicity and reproductive toxicity. In 2002, the International Agency for Research on Cancer (IARC) identified acrylamide as a Class 2A carcinogen. At present, it is generally recognized that the main formation pathway of acrylamide is the Maillard reaction between reducing sugars and asparagine, which leads to the formation of acrylamide. In the process of forming acrylamide through the Maillard reaction, the amino group of asparagine and the carboxyl group of reducing sugar combine to form a Schiff base, which can form acrylamide through the Strecker pathway and the N-glycoside pathway. In the Strecker pathway, asparagine can be further oxidized to form Strecker aldehyde after decarboxylation, and then reduced to form Strecker alcohol, and dehydrated to form acrylamide. In addition, Schiff base rearranges to form Amadori compounds, which are cleaved through different pathways to form a variety of dicarbonyl compounds. Under the premise of the presence of these dicarbonyl compounds, asparagine is further decarboxylated and deaminated to form acrylamide through the Strecker degradation mechanism. In the presence of oxygen, polyphenols are easily oxidized to quinones by polyphenol oxidase (PPO). Quinones are organic compounds with a six-membered cyclic diketone structure (containing two carbonyl groups) with two double bonds, which are very similar to the dicarbonyl compounds that are intermediate products in the Maillard reaction to produce acrylamide. Studies have shown that polyphenol oxidase can oxidize polyphenols in fruits and vegetables into quinones, and quinones can react with asparagine to produce acrylamide under high temperature conditions. At present, the technology of inhibiting the formation of acrylamide has been widely studied. Some studies have inhibited the formation of acrylamide by adding polyphenol compounds. For example, patent CN201911360202.6 discloses a method for inhibiting the production of acrylamide in baked foods and cookies, and inhibits the production of acrylamide by adding black wolfberry anthocyanins, with an inhibition rate of 69.46%; for example, patent CN201911049742.2 provides a method for reducing the content of acrylamide in fried foods, by combining dregs extract with phlorizin to reduce the intermediate harmful substances in the Maillard reaction of fried foods, while inhibiting the formation of acrylamide; for example, patent CN201510915459.9 inhibits acrylamide by subjecting potatoes soaked in an electrolytic solution to ultra-high pressure treatment. The method used in the above patent requires the extraction of additives from other ingredients or the addition of additional processing steps, which undoubtedly increases the complexity of production operations and requires production personnel to have higher professional skills and fine operation capabilities to ensure the accuracy and efficiency of extraction and addition. At the same time, the cumbersome process may also have a certain impact on production costs and efficiency, requiring enterprises to comprehensively consider the balance between costs and benefits when implementing. Therefore, it is urgent to provide an acrylamide generation inhibitor and a method for inhibiting acrylamide generation, which can prevent the occurrence of the Maillard reaction, thereby effectively inhibiting the generation of acrylamide, which not only helps to improve the shelf life and quality stability of food, but also better protect the health of consumers, showing broad application prospects and important practical value. Summary of the invention

[0003] Technical problems to be solved: In view of the above technical problems, the purpose of the present invention is to provide an inhibitor of acrylamide production and a method for inhibiting acrylamide production, wherein the acrylamide inhibitor is syringic acid, vanillic acid or sinapinic acid. Syringic acid and vanillic acid can passivate polyphenol oxidase in the food processing process and prevent polyphenol oxidase from oxidizing polyphenols in food into quinones, while sinapinic acid can directly inhibit quinones generated by oxidation of polyphenols, and reduce the content of Maillard reaction substrates, because quinones are very similar in structure to dicarbonyl compounds, an intermediate product in the Maillard reaction to produce acrylamide, so quinones can react with asparagine to produce acrylamide under high temperature conditions. Therefore, by passivating the activity of polyphenol oxidase or inhibiting the generation of quinones oxidized by polyphenols, the occurrence of Maillard reaction is prevented, and the generation of acrylamide, a class 2A carcinogen, which is easily produced during high-temperature food processing, is further inhibited, so as to achieve the purpose of healthy diet and provide a new idea for controlling the generation of acrylamide in food processing.

[0004] Technical solution: An acrylamide generation inhibitor, wherein the acrylamide inhibitor is syringic acid, vanillic acid or sinapinic acid. A method for inhibiting the generation of acrylamide comprises the following steps: S1. Adding water to the acrylamide inhibitor to prepare an inhibitor solution; S2. After the pre-treatment, the food is placed in the inhibitor solution and soaked for 10-30 minutes to obtain the treated food; S3. The processed food materials are processed to obtain processed food. Furthermore, the molar concentration of the inhibitor solution in step S1 is 0.5-3 mmol / L. Furthermore, the food materials in step S2 are organic substances containing amino groups, sugars capable of generating acrylamide, substances that are susceptible to Maillard reaction to generate acrylamide, and raw materials containing polyphenols that are susceptible to being oxidized by polyphenol oxidase. Furthermore, the processing in step S3 includes a heating step. Furthermore, the heating step includes roasting, frying, baking, microwave heating, and pressurized heating. Beneficial effects: 1. The addition of syringic acid or vanillic acid in this patent can interact with the active center of polyphenol oxidase, causing the structure of polyphenol oxidase to rearrange, thereby inhibiting the activity of polyphenol oxidase. At the same time, syringic acid and vanillic acid can interact with the copper ions in the active center of polyphenol oxidase, causing the dissociation, distortion or formation of a stable complex of the copper ions, thereby passivating the catalytic activity of polyphenol oxidase, thereby ensuring the original flavor of the food while inhibiting the formation of acrylamide. 2. The addition of sinapinic acid in the present invention can effectively inhibit the oxidation of polyphenols in food to generate quinone substances, significantly reduce the content of Maillard reaction substrates in food, further prevent the occurrence of Maillard reaction, thereby effectively inhibiting the formation of acrylamide, ensuring the good quality of food ingredients, and helping to improve the quality stability of food. 3. The present invention proves that by inactivating the activity of polyphenol oxidase or directly inhibiting the quinone generated by the oxidation of polyphenols, the content of Maillard reaction substrate is reduced, and the formation of acrylamide in the food processing process is inhibited. The acrylamide inhibition rate is 34.18-54.07%, which is highly safe and has a good inhibition effect. It not only broadens the method ideas for inhibiting the formation of acrylamide, but also provides a technical reference for subsequent research on inhibiting the formation of acrylamide, laying a good foundation for better ensuring food quality and food safety. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 It is the action mechanism diagram of Example 1, Example 2 and Example 3; Figure 2 is the polyphenol oxidase activity inhibition rate of Example 1, Example 2 and Comparative Example 1; Figure 3 is the acrylamide content in the fried potatoes of Example 1, Example 2 and Comparative Example 1; Figure 4 It is the acrylamide inhibition rate of Example 1, Example 2 and Comparative Example 1. DETAILED DESCRIPTION The present invention will be further described below in conjunction with embodiments, which are explanations of the present invention and are not limited to the following embodiments: Example 1 A method for inhibiting the generation of acrylamide in potato processing comprises the following steps: S1. Preparation of syringic acid solution: weigh 10.19 mg of syringic acid powder, add 28 mL of ethanol to dissolve it, then add 28 mL of buffer, mix well to completely dissolve the polyphenols, and sonicate for 5 minutes to prepare a 1 mmol / L syringic acid solution; S2. Potato pretreatment: potatoes were cut in half in water from which dissolved oxygen had been removed by nitrogen blowing, and cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a die cutter exposed to air. After oxidation on a heating pad at 35° C. for 5 min, 25.0 g of potato slices were soaked in 100 mL of a 1 mmol / L syringic acid solution for 10 min to prepare pretreated potato slices. S3. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pre-treated potato slices into the electric fryer and fry at 180° C. for 3 min. Example 2 A method for inhibiting the generation of acrylamide in potato processing comprises the following steps: S1. Preparation of vanillic acid solution: Weigh 11.01 mg of vanillic acid powder, add 28 mL of ethanol to dissolve it, then add 28 mL of buffer, mix well to completely dissolve the polyphenols, and sonicate for 5 minutes to prepare a 1 mmol / L vanillic acid solution; S2. Potato pretreatment: potatoes were cut in half in water from which dissolved oxygen had been removed by nitrogen blowing, and cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a die cutter exposed to air. After oxidation on a heating pad at 35° C. for 5 min, 25.0 g of potato slices were soaked in 100 mL of a 1 mmol / L vanillic acid solution for 10 min to prepare pretreated potato slices. S3. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pre-treated potato slices into the electric fryer and fry at 180° C. for 3 min. Comparative Example 1 The difference between this comparative example and Example 1 and Example 2 is that the samples were not immersed in a syringic acid or vanillic acid solution, as follows: A method for producing acrylamide comprises the following steps: S1. Potato pretreatment: potatoes were cut in half in water from which dissolved oxygen had been removed by nitrogen blowing, exposed to air, cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a die cutter, and oxidized on a heating pad at 35° C. for 5 min to obtain pretreated potato slices; S2. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pretreated potato slices into the electric fryer and fry at 180° C. for 3 min. Performance Testing Example 1, Example 2 and Comparative Example 1 were placed in a ziplock bag, pre-frozen in a -80°C refrigerator for 5 h, and then freeze-dried in a vacuum freeze dryer for 48 h. After freeze-dried into powder, they were frozen and stored in a -20°C refrigerator for later use. (1) Inhibition rate of polyphenol oxidase activity Prepare extraction buffer: weigh 340 mg polyethylene glycol 6000 (PEG6000), 4 g polyvinyl pyrrolidone (PVPP), and 1 ml polyethylene glycol octyl phenyl ether (TritonX-100), dissolve them in 100 mmol / L, pH 6.8 phosphate buffer solution, dilute to 100 mL, and prepare an extraction buffer containing 1 mmol / L PEG, 4% PVPP and 1% TritonX-100. Preparation of catechol solution: Weigh 55 mg of catechol, dissolve it in 100 mmol / L, pH 6.8 phosphate buffer solution, and dilute to 10 ml to prepare a 50 mmol / L catechol solution. Determination of polyphenol oxidase activity: Take 5.0g of Example 1, Example 2 and Comparative Example 1 respectively, wipe off the surface liquid and place in a mortar, add 5.0ml enzyme extraction buffer, grind into a homogenate under ice bath conditions, place the homogenate in a centrifuge tube and centrifuge at 4°C, 10000r for 30min, collect the supernatant as the crude enzyme extract, and store at 4°C for standby use. Then take a 10mL test tube, add 4.0ml, 50mmol / L, pH6.8 phosphate buffer (take 2.0ml, 100mmol / L phosphate buffer, add 2ml distilled water) and 1.0ml, 50mmol / L catechol solution, and finally add 100μL enzyme extract, and start timing immediately. Add 200μL of the reaction mixture to a transparent 96-well plate and place it in an ELISA instrument for detection, use distilled water as a reference, and start recording from the reaction time of 15s. The absorbance of the reaction system at a wavelength of 420 nm was taken as the initial value, and then recorded every 1 minute. The measurement was continued to obtain data of at least 6 points, and repeated three times. (2) Determination of acrylamide content Sample pretreatment: Weigh 1.000g sample into a 50mL centrifuge tube, add 10mL of n-hexane to deesterify, shake for 1min, centrifuge at 4℃, 10000rpm for 15min, and remove the supernatant. Repeat the above degreasing process twice. Add 20μL of 200mg / L of 13 C3-acrylamide internal standard solution, then add 10mL of pure water, 10mL of acetonitrile, 1.0g of sodium chloride, 4.0g of anhydrous magnesium sulfate, shake and mix, and ultrasonicate for 15min. After the ultrasonication, centrifuge at 4°C and 10000rpm for 15min, take all the supernatant and blow dry with nitrogen, then re-dissolve with 1mL of ultrapure water. Then activate the Cleanert-PEPSPE column (60mg / 3mL) with 3mL of methanol and 3mL of ultrapure water in turn, pass the re-dissolved solution through the SPE column, and collect the effluent. Add 1mL of pure water to rinse the SPE column, combine 2mL of the extract, shake and mix, pass through a 0.22μm polyethersulfone water filter, and inject into a brown sample bottle for liquid quality testing. Acrylamide was determined by ultra performance liquid chromatography-tandem mass spectrometry. The ultra-high performance liquid chromatography conditions were as follows: an ACQUITY UPLC HST3 column (2.1×100 mm, 1.8 μm, Waters, USA) was selected, the column temperature was 40° C., and the mobile phase and elution conditions were as shown in Table 1. The mass spectrometry conditions were as follows: capillary voltage 3KV; cone gas flow rate 150L / Hr; desolvation gas temperature 350℃; desolvation gas flow rate 650L / Hr. Acrylamide was quantified using an ESI electrospray ion source and multiple reaction monitoring (MRM) mode, with an ionization mode of ESI+, a quantitative ion pair of 72 / 55m / z, a cone voltage of 20V, and a collision voltage of 11V. 13 The ionization mode of C3-acrylamide was ESI+, the quantitative ion pair was 75 / 58 m / z, the cone voltage was 20 V, and the collision voltage was 11 V. To determine the accuracy and precision of this extraction method, three samples were randomly selected, and an acrylamide standard solution (2500 μg / L) with an intermediate concentration of the standard curve was added, and then the sample pretreatment was performed according to this extraction method. The accuracy of the method was expressed by the spiked recovery rate, and the coefficient of variation CV was expressed by the precision of the method. The average spiked recovery rate of this method was 100.4%, and the CV value was 0.046. The samples for detecting acrylamide and PPO enzyme activity in Example 1, Example 2 and Comparative Example 1 all came from the same potato, that is, a parallel data all came from the same potato. Table 1 ULTRA PERFORMANCE LIQUID CHROMATOGRAPHIC ELECTROPHOTOME Table 2 PPO enzyme activity inhibition rate, acrylamide content and inhibition rate of Example 1, Example 2 and Comparative Example 1 index Comparative Example 1 Example 1 Example 2 PPO enzyme activity inhibition rate (%) - 42.63 72.48 Acrylamide content (μmol / g) 0.0336748 0.0211502 0.0205821 Acrylamide inhibition rate (%) - 34.18 41.30 Note: - indicates no inhibitory effect. From Table 2, Figure 2 , Figure 3 and Figure 4 It can be seen that the PPO enzyme activity inhibition rate, acrylamide content and inhibition rate of Example 1 and Example 2 are better than those of Comparative Example 1. The inhibition rates of acrylamide by inactivating polyphenol oxidase activity are 34.18% and 41.30%, respectively. The mechanism of action is as follows: Figure 1 As shown, it is shown that both syringic acid and vanillic acid can interact with polyphenol oxidase, causing its active site to lose catalytic activity, thereby inactivating the activity of polyphenol oxidase, and preventing it from participating in the Maillard reaction, inhibiting the generation of Maillard reaction intermediates, and further inhibiting the generation of acrylamide. The present invention can significantly inhibit the generation of acrylamide by Maillard reaction of organic substances with amino groups contained in or caused to be contained in the food when the edible material that may generate acrylamide by heat treatment is subjected to heat treatment, and can be applied to the manufacturing process of various compositions such as food, cosmetics, pharmaceuticals or their intermediate products, and can manufacture a composition with low possibility of causing carcinogenesis and neurological disorders and high safety. Example 3 A method for inhibiting the generation of acrylamide in potato processing comprises the following steps: S1. Preparation of sinapinic acid solution: weigh 12.54 mg sinapinic acid powder, add 28 mL ethanol to dissolve it, then add 28 mL buffer, mix well to completely dissolve the polyphenols, sonicate for 5 min, and prepare a 1 mmol / L sinapinic acid solution; S2. Potato pretreatment: potatoes were cut in half in water from which dissolved oxygen had been removed by nitrogen blowing, and cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a die cutter exposed to air. After oxidation on a heating pad at 35° C. for 5 min, 50.0 g of potato slices were soaked in 100 mL of 1 mmol / L sinapinic acid solution for 10 min to prepare pretreated potato slices. S3. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pre-treated potato slices into the electric fryer and fry at 180° C. for 3 min. Comparative Example 2 The difference between this comparative example and Example 3 is that the soaking in the sinapinic acid solution is replaced by a blanching treatment. A method for inhibiting the generation of acrylamide in potato processing comprises the following steps: S1. Potato pretreatment: Potatoes were cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a die cutter in water. 50.0 g of potato slices were blanched at 95°C for 30 seconds. The surface water of the blanched potato slices was drained and oxidized on a heating pad at 35°C for 5 minutes. S2. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pretreated potato slices into the electric fryer and fry at 180° C. for 3 min. Comparative Example 3 The difference between this comparative example and Example 3 is that the immersion in the sinapinic acid solution is replaced by an oxygen isolation treatment. A method for inhibiting the generation of acrylamide in potato processing comprises the following steps: S1. Potato pretreatment: Potatoes were cut into slices with a thickness of 2 mm and a diameter of 25 mm using a slicer and a cutting tool in water, and 50.0 g of potato slices were soaked in water at room temperature for 5 min after nitrogen blowing to remove dissolved oxygen; S2. Potato processing: Pour 3.6 L of soybean oil into an electric fryer and heat it to 180° C. Place 20.0 g of pretreated potato slices into the electric fryer and fry at 180° C. for 3 min. Performance Testing (1) Chlorogenquinone content Example 3 and Comparative Examples 1-3 were placed in ziplock bags, pre-frozen in a -80°C refrigerator for 5 h, and then freeze-dried in a vacuum freeze dryer for 48 h. After freeze-dried into powder, they were frozen and stored in a -20°C refrigerator for later use. The content of chloroquinone-benzenesulfinic acid (chloroquinone BSA) in Example 3 and Comparative Examples 1-3 was determined, and the determination method was as follows: 10 g of potato slices from each group were soaked in 20 mL of 14 mmol / L benzenesulfinic acid solution (BSA) for 3 min, and then immediately pre-frozen at -80 ° C for 5 h, and then freeze-dried in a vacuum freeze dryer for 48 h. After freeze-dried into powder, they were stored in a -20 ° C refrigerator to be tested for chloroquine BSA. The measured chloroquine BSA content represents the chloroquine content. Table 3 Chlorquinone content, acrylamide content and inhibition rate of Example 3 and Comparative Examples 1-3 index Comparative Example 1 Example 3 Comparative Example 2 Comparative Example 3 Chlorogen quinone content (μmol / g) 0.460470 0.181252 0.195825 0.21663 Acrylamide content (μmol / g) 0.0336748 0.0134636 0.0205821 0.0211502 Acrylamide inhibition rate (%) - 54.07％ 41.29％ 34.18％ Note: - indicates no inhibitory effect. As shown in Table 3, the chloroquinone content, acrylamide content and inhibition rate of Example 3 are better than those of Comparative Examples 1-3. The inhibition rate of acrylamide by sinapinic acid through reducing the chloroquinone content is 54.07%. The mechanism of action is as follows: Figure 1As shown, it means that sinapinic acid can inhibit the formation of chloroquinone which is oxidized from chlorogenic acid in food, reduce the content of Maillard reaction substrate during the processing of food, thereby inhibiting the progress of Maillard reaction and further inhibiting the formation of acrylamide, ensuring the high quality of food. It is also simple to operate, highly safe and has strong industrial applicability. The above is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any technician familiar with the art can make many possible changes and modifications to the technical solution of the present invention by using the above disclosed methods and technical contents without departing from the spirit and technical solution of the present invention, or modify it into an equivalent embodiment of equivalent changes. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still falls within the scope of protection of the technical solution of the present invention.

Claims

1. An acrylamide production inhibitor, characterized in that The acrylamide inhibitor is syringic acid, vanillic acid or sinapinic acid.

2. A method for inhibiting the formation of acrylamide, characterized in that: The following steps are involved: S1. Adding water to the acrylamide inhibitor to prepare an inhibitor solution; S2. After the pre-treatment, the food is placed in the inhibitor solution and soaked for 10-30 minutes to obtain the treated food; S3. The processed food materials are processed to obtain processed food.

3. A method for inhibiting the formation of acrylamide according to claim 2, characterized in that: The molar concentration of the inhibitor solution in step S1 is 0.5-3 mmol / L.

4. The method for inhibiting the formation of acrylamide according to claim 2, characterized in that: The food materials in step S2 are organic substances containing amino groups, sugars capable of generating acrylamide, substances that are susceptible to Maillard reaction to generate acrylamide, and raw materials containing polyphenols that are susceptible to being oxidized by polyphenol oxidase.

5. The method for inhibiting the formation of acrylamide according to claim 2, characterized in that: The processing in step S3 includes a heating step.

6. The method for inhibiting the formation of acrylamide according to claim 5, characterized in that: The heating step includes roasting, frying, baking, microwave heating, and pressurized heating.

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