A dietetic food enriched with selenium supplements

Abstract

The present application relates to L-selenium-methyl selenocysteine extraction technical field, disclose a kind of selenium-enriched supplement special dietary food, specifically: template molecule L-selenium-methyl selenocysteine carboxyl and the pyridyl of functional monomer acryloyloxy bipyridyl amine monomer is self-assembled into polymeric molecule by ionic interaction, by free radical polymerization method, preparation magnetic L-selenium-methyl selenocysteine extractant;Magnetic L-selenium-methyl selenocysteine extractant is added to selenium-containing amino acid free type selenium-enriched rice protein solution, oscillation extraction, magnetic separation magnetic L-selenium-methyl selenocysteine extractant, using mixed solvent oscillation precipitation, obtain L-selenium-methyl selenocysteine.The present application provides a kind of extraction L-selenium-methyl selenocysteine in selenium-enriched rice protein, L-selenium-methyl selenocysteine prepared as selenium supplement, supplemented with other nutrient substance preparation selenium-enriched supplement special dietary food.

Description

Technical Field

[0001] This invention relates to the field of L-seleno-methylselenocysteine ​​extraction technology, specifically to a special dietary food rich in selenium supplements. Background Technology

[0002] Selenium is an essential trace element for the human body. Selenium deficiency can lead to diseases such as Keshan disease, Kashin-Beck disease, heart disease, hypothyroidism, and weakened immune function. Lowering blood selenium levels can increase the risk of cancer, and selenium plays a very clear role in preventing tumors and in radiotherapy and chemotherapy. L-Seleno-methylselenocysteine ​​is a natural selenium-containing amino acid, a methylated derivative of selenocysteine, the 21st essential amino acid for the human body. It is an important precursor of methylselenocyanide and has functions such as cancer prevention and treatment, anti-oxidation, anti-aging, treatment of cardiovascular and cerebrovascular diseases, and detoxification of heavy metal poisoning.

[0003] Compared with existing selenium supplements such as sodium selenite, L-seleno-methylselenocysteine ​​has advantages such as low toxicity, good selenium supplementation effect, and strong anti-cancer bioactivity. It was approved by the Ministry of Health as a new type of nutritional fortifier in 2009 and has been included in the national food safety standard for the use of food nutritional fortifiers. It can be widely used in food and health care products. Summary of the Invention

[0004] This invention provides a method for extracting L-seleno-methylselenocysteine ​​from selenium-enriched rice protein, using L-seleno-methylselenocysteine ​​as a selenium supplement, and supplementing it with other nutrients to prepare a special dietary food rich in selenium supplements.

[0005] A method for extracting L-seleno-methylselenocysteine ​​from selenium-enriched rice protein includes the following steps:

[0006] Step 1, Preparation of magnetic L-seleno-methylselenocysteine ​​extractant: The carboxyl group of the template molecule L-seleno-methylselenocysteine ​​and the pyridyl group of the functional monomer acryloyloxybispyridylamine monomer self-assemble into a polymer molecule through ionic interaction, and the magnetic L-seleno-methylselenocysteine ​​extractant is prepared by free radical polymerization.

[0007] Step 2, Extraction of L-seleno-methylselenocysteine ​​from selenium-enriched rice protein: Add magnetic L-seleno-methylselenocysteine ​​extractant to the free selenium-enriched rice protein solution containing selenium amino acids, place it in a constant temperature shaker, shake and extract, magnetically separate the magnetic L-seleno-methylselenocysteine ​​extractant, and precipitate it by shaking with a mixed solvent to obtain L-seleno-methylselenocysteine.

[0008] Preferably, the method for preparing the acryloyloxybispyridylamine monomer is as follows:

[0009] In step S1, under the co-catalysis of potassium carbonate and potassium iodide, the chlorine functional group of 4-chloromethylpyridine undergoes a nucleophilic substitution reaction with the amino functional group of ethanolamine to generate a bispyridylethanolamine monomer.

[0010] In step S2, through a nucleophilic substitution reaction mechanism, under the catalysis of potassium carbonate, the primary hydroxyl functional group of the bispyridylethanolamine monomer undergoes an esterification reaction with the acyl chloride functional group of acryloyl chloride to generate the acryloyloxybispyridylamine monomer.

[0011] Preferably, the mixed solvent consists of 4 parts by volume of methanol and 1 part by volume of acetic acid.

[0012] L-Selenium-methylselenocysteine ​​obtained by extracting L-Selenium-enriched rice protein is used as a selenium supplement. It is then combined with protein, fat, dietary fiber, fruit powder, vitamins, and minerals to prepare a special dietary food rich in selenium supplements.

[0013] Compared with the prior art, the present invention has the following beneficial technical effects:

[0014] This invention synthesizes a novel functional monomer: acryloyloxybispyridylamine monomer; using L-seleno-methylselenocysteine ​​as a template molecule, the carboxyl group of L-seleno-methylselenocysteine ​​and the pyridyl group of acryloyloxybispyridylamine monomer self-assemble into a polymer molecule through ionic interaction; and prepares a magnetic L-seleno-methylselenocysteine ​​extractant by free radical polymerization. Experiments show that the magnetic L-seleno-methylselenocysteine ​​extractant can extract L-seleno-methylselenocysteine ​​from selenium-enriched rice protein. Attached Figure Description

[0015] Figure 1 The chemical structural formula of the bispyridylethanolamine monomer;

[0016] Figure 2 The chemical structural formula of the acryloyloxybispyridylamine monomer;

[0017] Figure 3 The chemical structural formula of L-seleno-methylselenocysteine ​​is shown.

[0018] Figure 4 The chemical structural formula is for the self-assembly of L-seleno-methylselenocysteine ​​and acryloyloxybispyridylamine monomers into a polymer. Detailed Implementation

[0019] Experimental Example 1:

[0020] The synthesis of the bispyridylethanolamine monomer follows a reaction mechanism: under the co-catalysis of potassium carbonate and potassium iodide, the chlorine functional group of 4-chloromethylpyridine undergoes a nucleophilic substitution reaction with the amino functional group of ethanolamine to generate the bispyridylethanolamine monomer, the chemical structure of which is shown below. Figure 1 As shown;

[0021] The specific steps for synthesizing the bispyridylethanolamine monomer are as follows: 4.92 g of 4-chloromethylpyridine hydrochloride and 20 mL of acetonitrile were added to a round-bottom flask and stirred to dissolve. Then, 5.5 g of anhydrous potassium carbonate and 0.16 g of potassium iodide were added. Nitrogen gas was purged for protection, and stirring was started. 0.6 mL of ethanolamine was added dropwise using a dropper. The mixture was refluxed for 8 h and filtered using a sintered glass funnel. The filtrate was then rotary evaporated to remove the acetonitrile solvent, yielding the bispyridylethanolamine monomer.

[0022] Experimental Example 2:

[0023] The synthesis of acryloyloxybispyridylamine monomers follows a nucleophilic substitution mechanism. Catalyzed by potassium carbonate, the primary hydroxyl functional group of the bispyridylethanolamine monomer undergoes an esterification reaction with the acyl chloride functional group of acryloyl chloride to generate the acryloyloxybispyridylamine monomer, whose chemical structure is shown below. Figure 2 As shown;

[0024] The specific steps for synthesizing acryloyloxybispyridylamine monomer are as follows: 1.94 g of bispyridylethanolamine monomer was dissolved in 20 mL of acetonitrile, and 5 g of anhydrous potassium carbonate was added. Under nitrogen protection, a mixture of 0.8 mL of acryloyl chloride and 10 mL of acetonitrile was added dropwise using a constant pressure dropping funnel while stirring in an ice bath. After the addition was complete, the reaction was stirred in an ice bath for 4 h. The product was filtered and rotary evaporated to obtain acryloyloxybispyridylamine monomer. Its chemical structure was characterized using CDCl3 as solvent. 1 The H NMR characterization results are as follows: 1 H NMR (400MHz, δ / ppm): 2.94-2.97 (t, 2H), 3.96 (s, 4H), 4.24-4.27 (t, 2H), 5.96-6.11 (m, 3H), 7.35-7.36 (m, 4H), 8.55-8.536 (m, 4H).

[0025] Example 1:

[0026] Preparation of 3-(methacryloyloxy)propyltrimethoxysilane (γ-MPS) modified Fe3O4 nanoparticles: Under argon protection and in a water bath at 75°C, 80 mL of 0.5 mol / L NaOH solution was added to the reactor. 120 mL of a mixed solution containing 1.12 g of ferrous sulfate heptahydrate and 2.16 g of ferric chloride hexahydrate (composed of 1 part ultrapure water and 1 part anhydrous ethanol) was added dropwise to the reactor. After the addition was complete, the reactor was kept at 80°C for 0.5 h. Then, 1 mL of 3-(methacryloyloxy)propyltrimethoxysilane was added, and the mixture was cooled to 50°C for aging for 0.5 h. After cooling, the mixture was magnetically separated, washed until neutral, washed with anhydrous ethanol, and vacuum dried at 40°C for 24 h to obtain γ-MPS modified Fe3O4 nanoparticles.

[0027] Example 2:

[0028] Preparation of magnetic L-seleno-methylselenocysteine ​​extractant: 12 mg of L-seleno-methylselenocysteine ​​(its chemical structural formula is shown below) Figure 3 As shown), 0.12 g of acryloyloxybispyridylamine monomer was added to 20 mL of acetonitrile and shaken at room temperature for 2 h. This allowed the carboxyl group of L-seleno-methylselenocysteine ​​to self-assemble with the pyridyl group of the acryloyloxybispyridylamine monomer through ionic interactions into a polymer molecule (its chemical structure is shown in Figure 1). Figure 4 As shown in the figure, 50 mg of γ-MPS modified Fe3O4 nanoparticles, 0.75 mL of ethylene glycol dimethacrylate and 15 mg of azobisisobutyronitrile were added. After purging with nitrogen for 10 min, the mixture was sealed and reacted at 60 °C with shaking for 24 h. The product was magnetically separated and washed successively with anhydrous ethanol and distilled water. It was then extracted for 24 h with an extraction solution composed of 4 parts by volume of methanol and 1 part by volume of acetic acid to elute L-seleno-methylselenocysteine ​​molecules. The product was then vacuum dried at 60 °C for 12 h to obtain the magnetic L-seleno-methylselenocysteine ​​extractant.

[0029] Example 3:

[0030] Preparation of selenium-enriched rice protein solution containing free selenium amino acids: 10g of selenium-enriched rice was crushed and ground, then passed through a 150-mesh sieve. Following the Osborne fractionation method, albumin was extracted with distilled water, globulin with 2% sodium chloride solution, prolamins with 70% ethanol solution, and glutenin with 0.5% potassium hydroxide solution. Each step was repeated twice. The supernatants from the two extractions were combined to obtain selenium-enriched rice protein. A 1mg / mL solution of selenium-enriched rice protein was prepared with distilled water, and trypsin (enzyme activity: pH 8, 37℃, 2.5×10⁻⁶) was added. 5 Enzymatic hydrolysis was performed using protein (U / g protein) to obtain hydrolysate I; then proteinase K (with enzyme activity: pH 7.5, 55℃, 3×10) was added. 5The protein was further hydrolyzed to obtain hydrolysate II; then streptomycin (enzyme activity: pH 7, 37℃, 4000 U / g protein) was added, and hydrolysis was continued to obtain hydrolysate III; trypsin, proteinase K, and streptomycin were inactivated, and the supernatant obtained by centrifugation was the selenium-enriched rice protein solution containing free selenium amino acids. The content of free L-seleno-methylselenocysteine ​​was detected, and the result was 55.2 μg / mL;

[0031] Among them, the content of free L-seleno-methylselenocysteine ​​in the selenium-enriched rice protein solution containing selenium amino acids was determined by high performance liquid chromatography (HPLC), and the chromatographic conditions were as follows:

[0032] The following chromatographic column was used: EC250 / 4NUCLEOSIL Chiral-1 (4 mm × 250 mm, 5 μm), mobile phase was 0.4 mmol / L CuSO4·5H2O aqueous solution, column temperature was 35 ℃, flow rate was 1.0 mL / min, injection volume was 20 μL, and detection wavelength was 240 nm.

[0033] Static extraction of L-seleno-methylselenocysteine ​​from selenium-enriched rice protein: 1g of magnetic L-seleno-methylselenocysteine ​​extractant was added to 10mL of the above-mentioned free selenium-containing amino acid-enriched selenium-enriched rice protein solution, and placed in a constant temperature shaker. The solution was shaken for 2h at 25℃ and 100r / min. The magnetic L-seleno-methylselenocysteine ​​extractant was magnetically separated, and the content of free L-seleno-methylselenocysteine ​​was determined. The result was that no L-seleno-methylselenocysteine ​​was detected.

[0034] In this process, L-selen-methylselenocysteine ​​extracted by the magnetic L-selen-methylselenocysteine ​​extractant is precipitated by shaking with a mixed solvent (composed of 4 parts by volume of methanol and 1 part by volume of acetic acid), and the solvent is removed to obtain L-selen-methylselenocysteine.

[0035] Example 4:

[0036] Special dietary foods rich in selenium supplements: These are prepared by mixing L-seleno-methylselenocysteine ​​as a selenium supplement with different proportions of nutrients such as protein, fat, dietary fiber, fruit powder, vitamins, and minerals.

Claims

1. A method for extracting L-seleno-methylselenocysteine ​​from selenium-enriched rice protein, characterized in that, Includes the following steps: Step 1: Preparation of magnetic L-seleno-methylselenocysteine ​​extractant: The carboxyl group of the template molecule L-seleno-methylselenocysteine ​​and the pyridyl group of the functional monomer acryloyloxybispyridylamine self-assemble into a polymer molecule through ionic interaction. Under the action of ethylene glycol dimethacrylate and azobisisobutyronitrile, the polymer molecule undergoes free radical polymerization on the surface of 3-(methacryloyloxy)propyltrimethoxysilane modified Fe3O4 nanoparticles, eluting off the L-seleno-methylselenocysteine ​​molecule to obtain the magnetic L-seleno-methylselenocysteine ​​extractant. The chemical structural formula of the acryloyloxybispyridylamine monomer is as follows: ； Step 2, Extraction of L-seleno-methylselenocysteine ​​from selenium-enriched rice protein: Add magnetic L-seleno-methylselenocysteine ​​extractant to the free selenium-enriched rice protein solution containing selenium amino acids, place it in a constant temperature shaker, shake and extract, magnetically separate the magnetic L-seleno-methylselenocysteine ​​extractant, and precipitate it by shaking with a mixed solvent to obtain L-seleno-methylselenocysteine.

2. The method for extracting L-seleno-methylselenocysteine ​​from selenium-enriched rice protein according to claim 1, characterized in that, The preparation method of the acryloyloxybispyridylamine monomer is as follows: In step S1, under the co-catalysis of potassium carbonate and potassium iodide, the chlorine functional group of 4-chloromethylpyridine undergoes a nucleophilic substitution reaction with the amino functional group of ethanolamine to generate a bispyridylethanolamine monomer. In step S2, through a nucleophilic substitution reaction mechanism, under the catalysis of potassium carbonate, the primary hydroxyl functional group of the bispyridylethanolamine monomer undergoes an esterification reaction with the acyl chloride functional group of acryloyl chloride to generate the acryloyloxybispyridylamine monomer.

3. The method for extracting L-seleno-methylselenocysteine ​​from selenium-enriched rice protein according to claim 1, characterized in that, The mixed solvent consists of 4 parts by volume of methanol and 1 part by volume of acetic acid.

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