A formulation of a selenium protein peptide of cotoneaster horizontalis and albumin peptide and a preparation method thereof

Abstract

The application discloses a kind of pectinatocarpus selenium protein peptide and albumin peptide formula and preparation method, the core active ingredient of the formula is the selenium protein peptide of pectinatocarpus source and egg white albumin peptide, wherein the number average molecular weight of selenium protein peptide is 461Da, total selenium is 412.67mg / kg and selenocysteine and pectinatocarpus plant selenium peptide ratio is more than 90%, and the biological potency of albumin peptide is 94, and the molecular weight is 200-800Da;The application promotes active transport in intestinal tract by albumin peptide as carrier, and is prepared by combining dry granulation and tabletting process, the selenium absorption rate of the application is greatly improved, realizes the synergistic effect of hypoglycemic, hypolipidemic and antioxidant, and provides a new scheme for selenium supplement functional food.

Description

Technical Field

[0001] This invention relates to the field of functional dietary food technology, and in particular to a formulation and preparation method of violet leaf broken rice selenoprotein peptide and albumin peptide. Background Technology

[0002] In the current functional food sector, selenium supplements are mainly divided into three categories: inorganic selenium (such as sodium selenite), organic selenium (such as selenium yeast and selenoproteins), and plant-derived selenium-enriched extracts. Among these, the super-selenomeric plant Cardamine violifolia, due to its ability to naturally accumulate high levels of selenium, has been developed into a raw material for selenoproteins and selenium peptides. Meanwhile, albumin peptides, as small molecule peptides derived from egg white, are widely used in nutritional supplements to promote the intestinal absorption of minerals such as calcium, iron, and zinc due to their high similarity to the amino acid composition of human serum proteins and a bioavailability as high as 94.

[0003] Existing selenium supplements, primarily composed of selenoproteins or selenopeptides, have relatively large molecular weights (greater than 1000 Da), resulting in limited passive absorption efficiency in the intestines. Furthermore, the lack of specific carriers for selenium transport within the body leads to low bioavailability. While albumin peptides are known to promote mineral absorption, current technologies lack research on targeted formulations combining albumin peptides with specific molecular weight ranges and selenium forms from *Violet spp.* selenoprotein peptides. Moreover, the current technology offers no insights into whether albumin peptides can serve as active carriers for selenoamino acids, or whether there is a synergistic effect between the two in selenium supplementation. Additionally, most existing selenium supplements exist as single selenium sources, lacking complex formulation designs that leverage carrier peptides to enhance the active intestinal transport efficiency of selenium, thus limiting the effectiveness of selenium supplementation.

[0004] Therefore, in response to the problems mentioned above, this invention proposes a formulation and preparation method of violet leaf broken rice selenoprotein peptide and albumin peptide. Summary of the Invention

[0005] To overcome the problems of low selenium absorption efficiency, lack of specific transport carriers, and unclear synergistic mechanism between selenoprotein peptides and carrier peptides in existing selenium supplements, this invention proposes a combined formulation with specific molecular weight selenoprotein peptides and egg albumin peptides derived from Corydalis yanhusuo. By using albumin peptides as transport carriers, the active transport of selenoprotein peptides and their chelated selenium in the intestine is promoted, thereby achieving the technical effect of improving selenium absorption rate and synergistic effect of antioxidant effect.

[0006] The technical solution of this invention is: a formulation of violet leaf broken rice selenoprotein peptide and albumin peptide:

[0007] The core active ingredients of the formula include selenoprotein peptides derived from corydalis leaf and albumin peptides derived from egg white;

[0008] The violet leaf crushed rice selenoprotein peptide is a mixture of peptides with a molecular weight of less than 1000 Da obtained by extraction, enzymatic hydrolysis and ultrafiltration fractionation of the super-selenophytic plant violet leaf crushed rice. Its number average molecular weight is 461 Da and the total selenium content is 412.67 mg / kg. Its selenium form is mainly selenocysteine ​​and violet leaf crushed rice plant selenopeptide (SeP, 100% organic selenium, mainly selenocysteine).

[0009] The albumin peptide is an ovalbumin peptide with a biological titer of 94 and contains all 20 amino acids.

[0010] The composition uses albumin peptides as a carrier to promote the active transport of selenoprotein peptides and their chelated selenium in the intestine, thereby achieving the unexpected technical effect of improving selenium absorption and synergistic antioxidant effect.

[0011] Preferably, the formula comprises the following components in parts by weight: 25-40 parts sorbitol, 10-20 parts Poria cocos powder, 10-20 parts albumin peptide, 3-6 parts bitter melon peptide powder, 3-5 parts Polygonatum sibiricum extract, 3-5 parts selenium-enriched corydalis leaf and rice bran protein peptide powder, 2-4 parts kudzu root extract, 2-4 parts green tea extract, 2-4 parts selenium-enriched Eucommia ulmoides leaf instant tea, 2-4 parts hydroxypropyl methylcellulose, and 0.8-1.6 parts magnesium stearate.

[0012] Preferably, the selenium content provided by the violet leaf broken rice selenoprotein peptide is 52-70 μg, the peptide mass provided by the albumin peptide is 80-100 mg, and the albumin peptide is a specific enzymatic hydrolysis product derived from egg white, with a molecular weight distribution range of 200-800 Da.

[0013] In the Viola yedoensis selenoprotein peptide, the sum of selenocysteine ​​and Viola yedoensis plant selenopeptide (SeP, 100% organic selenium, mainly selenocysteine) accounts for more than 90% of its total selenium content.

[0014] The bitter melon peptide powder contains at least 0.3% 1-deoxynojirimycin, the polygonatum extract contains ≥20% polygonatum polysaccharide, and the selenium-enriched Eucommia ulmoides leaf instant tea contains no less than 2mg / kg of total selenium.

[0015] This invention proposes a method for preparing a formulation of violet leaf and rice porridge selenium protein peptide and albumin peptide, comprising the following steps:

[0016] S1. Sorbitol, Poria cocos powder, albumin peptide, bitter melon peptide powder, Polygonatum sibiricum extract, selenium-enriched Viola yedoensis and Coix lacryma-jobi protein peptide powder, Pueraria lobata extract, green tea extract, selenium-enriched Eucommia ulmoides leaf instant tea, and hydroxypropyl methylcellulose are dried separately at 60℃ for 4-6 hours to reduce the moisture content to ≤8%. Then, they are sieved through 80-mesh and 100-mesh double-stage sieves to remove coarse particles. Finally, the filtered materials are mixed according to the formula ratio and stirred in a horizontal mixer at 60-100 rpm for 10-15 minutes.

[0017] S2, the obtained mixture is fed into a roller press dry granulator, the roller pressure is set to 5-8MPa, the roller gap is controlled at 0.6-1.0mm, and it is pressed into a strip or sheet compacted material. Then the compacted material is fed into a high-speed rotary pulverizer for crushing, the speed is set to 800-1200rpm, the fragment particle size is controlled at 0.2-1.0mm, and finally the crushed material is sieved through a 0.8mm sieve to obtain uniform particles;

[0018] S3, add magnesium stearate (magnesium stearate addition amount is 0.5-1.0% of the total formula mass) to the granules obtained by dry granulation, and then mix in a horizontal mixer for 3-5 minutes, with the stirring speed controlled at 50-80 rpm;

[0019] S4. The granules are fed into a rotary tablet press and compressed into tablets. The mold diameter is 8-10mm. The weight of the tablet is controlled at 0.9-1.2g / tablet. The tableting pressure is set at 30-50kN. The hardness of the tablets after compression is 60-100N and the brittleness is less than 1.0%. The finished tablets are packaged in aluminum-plastic blister packs, with 10 tablets per blister pack and 0.5-1g of silica gel desiccant. After packaging, the tablets are stored in a dry environment with humidity ≤60% and temperature ≤25℃.

[0020] The beneficial effects of this invention are:

[0021] 1. This invention combines a specific molecular weight selenoprotein peptide derived from the super-selenomeric plant *Corydalis yanhusuo* with an egg albumin peptide. Using the albumin peptide as a carrier, the apparent intestinal permeability coefficient of the selenoprotein peptide is increased by 136.5% and the serum selenium concentration in rats is increased by 42.3% through a PepT1-mediated active transport pathway. This solves the technical problems of low passive absorption efficiency and lack of specific carriers in existing selenium supplements, achieving an unexpected and significant increase in selenium absorption rate.

[0022] 2. This invention achieves hypoglycemic and hypolipidemic functions in a diabetic mouse model through the synergistic effect of albumin peptides and selenoprotein peptides, while significantly increasing glutathione peroxidase activity and spleen and thymus indices. Its overall effect is significantly better than using selenoprotein peptides or inorganic selenium alone.

[0023] 3. This invention can significantly reduce the estimated glycemic index in noodle, steamed bun, and milk powder bases, providing a novel selenium-supplemented dietary product with high absorption rate and multiple functions for selenium-deficient individuals, patients with diabetes and hyperlipidemia, and those with cognitive decline and chronic low-grade inflammation. Attached Figure Description

[0024] Figure 1 The diagram illustrates the effect of NaOH concentration on the yield and selenium content of selenoproteins according to the present invention.

[0025] Figure 2 The diagram illustrates the effect of the liquid-to-solid ratio on the yield and selenium content of selenoproteins according to the present invention.

[0026] Figure 3 The diagram illustrates the effect of extraction time on selenoprotein yield and selenium content according to the present invention.

[0027] Figure 4 The diagram illustrates the effect of the number of extractions on the yield and selenium content of selenoproteins according to the present invention.

[0028] Figure 5 The diagram illustrates the effect of extraction temperature on the yield and selenium content of selenoproteins according to the present invention.

[0029] Figure 6 The diagram illustrates the effect of enzyme dosage on the degree of hydrolysis in the enzymatic preparation of the Viola yedoensis oligopeptide of the present invention.

[0030] Figure 7 The diagram illustrates the effect of substrate concentration on the degree of hydrolysis in the enzymatic preparation of the Viola yedoensis oligopeptide of the present invention.

[0031] Figure 8 The diagram illustrates the effect of pH on the degree of hydrolysis during the enzymatic hydrolysis preparation of the Viola yedoensis oligopeptide of the present invention.

[0032] Figure 9 The diagram illustrates the effect of enzymatic hydrolysis temperature on the degree of hydrolysis in the enzymatic preparation of the Viola yedoensis oligopeptide of the present invention.

[0033] Figure 10 The diagram illustrates the effect of selenoprotein in lowering serum total cholesterol (TC) of the present invention.

[0034] Figure 11 The diagram shown illustrates the effect of the selenoprotein in lowering serum triglycerides (TG) of the present invention.

[0035] Figure 12 The diagram illustrates the effect of enzymatic hydrolysis temperature on the degree of hydrolysis in the enzymatic preparation of the Viola yedoensis oligopeptide of the present invention.

[0036] Figure 13 The diagram illustrates the effect of the selenoprotein of the present invention on serum glutathione peroxidase (GSH-Px) activity.

[0037] Figure 14 The diagram illustrates the effect of the selenoprotein of the present invention on the spleen index.

[0038] Figure 15 The diagram illustrates the effect of the selenoprotein of the present invention on serum insulin (INS) levels.

[0039] Figure 16 The diagram illustrates the effect of the selenoprotein of the present invention on the thymus index. Detailed Implementation

[0040] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are some embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0041] This invention provides Embodiment 1:

[0042] This embodiment provides a method for extracting, enzymatically hydrolyzing, and purifying selenoprotein peptides from the hyperselenophoretic plant Corydalis yanhusuo. These selenoprotein peptides, as one of the core active ingredients in the formulation of this invention, have specific molecular weight distribution, specific selenium forms, and high selenium content.

[0043] (I) Pretreatment and nutritional analysis of Viola yedoensis raw material

[0044] The whole plant of *Vigna oleifera*, an artificially cultivated plant rich in selenium, was harvested during the bolting stage. After washing and drying, it was blanched at 105℃ for 5 minutes, then dried with hot air at 80℃, pulverized, and sieved to obtain *Vigna oleifera* vegetable powder. Analysis showed that the vegetable powder contained 22.29% protein, 1.13g / 100g amino acids, 2.69μg / g vitamin C, and 2.83μg / g vitamin B1.

[0045] To clarify the nutritional value of *Corydalis yanhusuo*, this invention conducted a comprehensive nutritional component analysis, the results of which are shown in Table 1:

[0046] Table 1. Vitamin content in Viola yedoensis and Citrus aurantium

[0047] Vitamin Name Content (mg / kg) Vitamin A 10.1 Vitamin C 0.39 Vitamin D 1.7 Vitamin E 45.2 Vitamin B1 2.76 Vitamin B2 17.08 Vitamin B6 1.24 Vitamin B3 4.24 Vitamin B5 0.78

[0048] In Viola yedoensis, fat-soluble vitamins account for 68.27% of the total vitamin content, while water-soluble vitamins account for 31.73%. Vitamin E has the highest content, accounting for 54.14% of the total vitamin content, reaching 45.2 mg / kg, followed by vitamin B2, accounting for 20.46% of the total vitamin content, at 17.08 mg / kg. It also contains vitamin A 10.1 mg / kg, vitamin C 0.39 mg / kg, vitamin D 1.7 mg / kg, vitamin B1 2.76 mg / kg, vitamin B6 1.24 mg / kg, vitamin B3 4.24 mg / kg, and vitamin B5 0.78 mg / kg.

[0049] Table 2. Amino acid content in Viola yedoensis and Citrus aurantium

[0050] Amino acid name content(%) Amino acid name content(%) Aspartic acid 2.03 Cystine 0.039 glutamic acid 3.3 Valine 1.06 Serine 0.97 Methionine 0.11 Histidine 0.44 Phenylalanine 1.29 glycine 1.01 Isoleucine 0.81 threonine 1.02 Leucine 1.82 Arginine 0.11 Lysine 1.29 alanine 1.42 proline 0.81 Tyrosine 0.95 Total amino acid 18.48

[0051] As shown in Table 2, the seven essential amino acids required by the human body account for 40.04% of the total amino acids in *Corydalis yanhusuo*, while non-essential amino acids account for 59.96%; the eight essential amino acids required by infants account for 42.42%. The ratio of essential to non-essential amino acids is 0.67, which conforms to the ideal protein pattern recommended by FAO / WHO. The total amino acid content is 18.48%, with glutamic acid having the highest content (3.3%), followed by aspartic acid (2.03%), leucine (1.82%), alanine (1.42%), lysine (1.29%), phenylalanine (1.29%), etc.

[0052] Table 3. Mineral content in Viola yedoensis and Citrus aurantium

[0053] Mineral Name Content (mg / kg) potassium 78.6 calcium 7.8 magnesium 8.22 phosphorus 4.88 sulfur 15.44 chlorine 2.00 iron 0.6 copper 5.52 Zinc 306 cobalt 1.65 manganese 114 chromium 4.82 molybdenum 0.46 boron 15.5

[0054] As shown in Table 3, *Cardamine hirsuta* is rich in the six essential macroelements (calcium, phosphorus, magnesium, potassium, sulfur, and chlorine), accounting for 20.62% of the total mineral content; the seven essential trace elements (iron, zinc, copper, cobalt, molybdenum, selenium, and chromium) account for 56.55% of the total content; and the potentially essential elements manganese and boron account for 22.83%. Specifically, the potassium content is 78.6 mg / kg, calcium 7.8 mg / kg, magnesium 8.22 mg / kg, phosphorus 4.88 mg / kg, sulfur 15.44 mg / kg, and chlorine 2.00 mg / kg; the trace elements include iron 0.6 mg / kg, copper 5.52 mg / kg, zinc 306 mg / kg, cobalt 1.65 mg / kg, manganese 114 mg / kg, chromium 4.82 mg / kg, molybdenum 0.46 mg / kg, and boron 15.5 mg / kg. It is worth noting that the zinc and manganese content is extremely high, at 306 mg / kg and 114 mg / kg respectively. These trace elements have a synergistic antioxidant effect with selenium.

[0055] Table 4. Fatty acid content in Viola yedoensis and Citrus aurantium

[0056] Fatty acid name content(%) Myristic acid 1.38 Palmitic acid 21.61 Hexadecanetrienoic acid 4.9 stearic acid 3.92 Oleic acid 2.4 Linoleic acid 18.39 Alpha-linolenic acid 30.22 Arachidonic acid 0.49 Erucic acid 9.11 Lignin acid 6.26 Nervonic acid 1.31

[0057] As shown in Table 4, polyunsaturated fatty acids accounted for 53.51% of *Cardamine hirsuta*, monounsaturated fatty acids accounted for 12.83%, and saturated fatty acids accounted for 33.66%. Among these, linolenic acid had the highest content at 30.22%, followed by linoleic acid at 18.39% and palmitic acid at 21.61%. It also contains 1.31% nervonic acid (cis-15-tetracosenoic acid), which plays an important role in repairing and unblocking damaged brain neural pathways, improving nerve cell activity, regulating blood sugar and lipids, enhancing immunity, and inhibiting cancer cell growth. The abundant polyunsaturated fatty acids provide a lipid synergistic environment for the function of selenoprotein peptides.

[0058] This invention uses high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) to determine the selenium speciation in different parts and at different growth stages of *Corydalis yanhusuo*. The results are shown in Table 5.

[0059] Table 5. Selenium content in different parts and at different growth stages of Viola yedoensis.

[0060] name Secys2 (mg / kg) SeP (mg / kg) Semet (mg / kg) Three selenoamino acids and (mg / kg) Total selenium (mg / kg) Selenized amino acids as a percentage of total selenium (%) The whole plant did not flower 1991.27 144.35 0.00 2135.62 4127.04 51.75 whole plant with stem 764.65 0.00 0.00 764.65 2765.04 27.65 whole plant producing leaves 2012.56 111.77 0.00 2124.32 3675.87 57.79 The whole plant blooms 1865.67 119.18 0.00 1984.85 2452.82 57.49 Small Leaf 2493.35 440.50 8.10 2941.95 5682.98 51.77 Large Leaf 1455.60 68.07 19.55 1543.21 4033.15 38.26 flower 2185.11 417.81 63.26 2666.17 8936.17 29.84 bud 3547.65 762.24 62.06 4371.95 7215.16 60.59 whole plant 2188.84 138.65 0.00 2327.49 4306.93 54.04

[0061] Before the plant bolts, the content of selenocysteine ​​(Secys2) was 1991.27 mg / kg, the content of plant selenophoretic peptide (SeP, 100% organic selenium, mainly selenocysteine) was 144.35 mg / kg, and selenomethionine (Semet) was not detected. The total content of the three selenoamino acids was 2135.62 mg / kg, and the total selenium was 4127.04 mg / kg, with selenoamino acids accounting for 51.75% of the total selenium. During the leafing stage, the content of Secys2 was 2012.56 mg / kg, SeP was 111.77 mg / kg, and the total content was 2124.32 mg / kg, with the total selenium being 3675.87 mg / kg, accounting for 57.79%. In the leaflets, Secys2 was as high as 2493.35 mg / kg, SeP was 440.50 mg / kg, Semet was 8.10 mg / kg, the total of the three selenoamino acids was 2941.95 mg / kg, and the total selenium was 5682.98 mg / kg, accounting for 51.77%. The flowers had the highest total selenium content, reaching 8936.17 mg / kg, but the selenoamino acids accounted for only 29.84%. In the buds, Secys2 was 3547.65 mg / kg, SeP was 762.24 mg / kg, Semet was 62.06 mg / kg, the total was 4371.95 mg / kg, and the total selenium was 7215.16 mg / kg, accounting for 60.59%, the highest proportion among all parts.

[0062] In this embodiment, the extraction of selenium protein from *Violet spp.* is described, specifically:

[0063] The above-mentioned violet leaf and water chestnut vegetable powder was used to extract selenoproteins using the alkali dissolution and acid precipitation method. Single-factor experiments were conducted to optimize the extraction conditions, investigating the effects of NaOH concentration, liquid-to-solid ratio, extraction temperature, extraction time, and extraction cycles on the yield and selenium content of selenoproteins.

[0064] Please see Figure 1-5 The optimal process parameters were determined as follows: NaOH solution concentration 0.1 mol / L, liquid-to-solid ratio 50:1 mL / g, extraction temperature 60℃, extraction time 3 hours, and extraction times 2. Under these conditions, after centrifugation, the pH of the supernatant was adjusted to the isoelectric point (approximately 4.5), allowed to settle, and the precipitate was collected by centrifugation and freeze-dried to obtain crude selenoprotein. Further purification was performed using membrane separation technology (ultrafiltration membrane with a molecular weight cutoff of 100 kD) to obtain pure selenoprotein from *Corydalis yanhusuo*.

[0065] The selenoprotein was found to have a purity of 40.50%, a total selenium content of 2009.128 ± 26.193 μg / g, and a molecular weight concentrated around 100 kDa. Selenium speciation analysis showed that its characteristic form was selenocysteine, indicating that selenoamino acids did not undergo significant degradation during extraction. This selenoprotein can be further used for enzymatic hydrolysis to prepare selenoprotein peptides.

[0066] Please see Figure 6-9 In this embodiment, the enzymatic hydrolysis preparation of *Violet Leaf Broken Rice* oligopeptides (selenoprotein peptides) is described, specifically:

[0067] Using the selenoprotein from *Violet spp.* as a substrate, enzymatic hydrolysis was performed using alkaline protease (pH 8.0, 50℃). Single-factor experiments were conducted to investigate the effects of enzyme dosage, substrate concentration, hydrolysis temperature, and pH on the degree of hydrolysis. With increasing enzyme dosage, the degree of hydrolysis initially increased rapidly and then leveled off, with the optimal enzyme dosage being 6.0% (based on substrate protein). The degree of hydrolysis peaked at a substrate concentration of 3.0%. The highest degree of hydrolysis was achieved at a hydrolysis temperature of 60℃, and pH 9.5 was the optimal pH. Under optimal conditions, after hydrolysis, the hydrolysate was heated to inactivate the enzyme (95℃, 10 min), centrifuged, and the supernatant was collected. Fractions with molecular weights less than 1 kDa were collected by ultrafiltration and freeze-dried to obtain *Violet spp.* oligopeptides (selenoprotein peptides, hereinafter referred to as SeCPPs).

[0068] Testing revealed that the oligopeptide contained 5.25% moisture, 49.01% protein, 412.67 mg / kg total selenium, 44.2% acid-soluble protein (90% of total protein), and 1.33% free amino acids, resulting in a calculated peptide content of 42.87%. Its number-average molecular weight was 461 Da, indicating it is primarily composed of small molecules with a molecular weight distribution range less than 1000 Da, consistent with the characteristics of small-molecule bioactive peptides. Selenium speciation analysis showed that selenium mainly existed as selenocysteine ​​and *Violet spp.* selenium peptide (SeP, 100% organic selenium, primarily selenocysteine), with the two combined accounting for over 90% of its total selenium content.

[0069] This invention provides Embodiment 2:

[0070] The albumin peptide provided in this embodiment is an egg white albumin peptide, derived from fresh egg white, and cannot be replaced by other animal blood products. Its preparation method includes:

[0071] Fresh eggs were collected, egg whites were separated, and the pH was adjusted to 7.0-8.0. A complex protease (including trypsin and neutral protease) was added, and the enzymatic hydrolysis was performed at 50-55℃ for 3-5 hours. After hydrolysis, the enzyme was inactivated, and the mixture was separated by ultrafiltration (molecular weight cutoff 1000 Da). The permeate was collected, concentrated under vacuum, and spray-dried to obtain albumin peptide powder. Analysis showed that this albumin peptide contained all 20 amino acids, had a bioavailability of 94, a molecular weight distribution range of 200-800 Da, and a peptide content ≥85%. Its amino acid composition is highly similar to human serum albumin, making it one of the highest quality animal proteins currently recognized and a potential natural substitute for human serum albumin. Albumin peptides possess various physiological functions, including easy digestibility and absorption, low antigenicity, promotion of mineral absorption, and regulation of intestinal flora.

[0072] This invention provides embodiment 3:

[0073] This embodiment provides a composition formulation containing selenoprotein peptides and albumin peptides from *Corydalis yanhusuo*, with the core active ingredients being the selenoprotein peptides prepared in Example 1 and the albumin peptides prepared in Example 2.

[0074] The formula comprises the following components in parts by weight: 25-40 parts sorbitol, 10-20 parts Poria cocos powder, 10-20 parts albumin peptide, 3-6 parts bitter melon peptide powder, 3-5 parts Polygonatum sibiricum extract, 3-5 parts selenium-enriched corydalis leaf and rice bran protein peptide powder, 2-4 parts kudzu root extract, 2-4 parts green tea extract, 2-4 parts selenium-enriched Eucommia ulmoides leaf instant tea, 2-4 parts hydroxypropyl methylcellulose, and 0.8-1.6 parts magnesium stearate.

[0075] Among them, the selenium-enriched corydalis leaf and rice bran protein peptide powder is the selenoprotein peptide prepared in Example 1, which provides a selenium content of 52-70 μg (based on each unit of finished product), and the albumin peptide provides a peptide mass of 80-100 mg. The bitter melon peptide powder contains at least 0.3% 1-deoxynojirimycin, which has an auxiliary hypoglycemic effect; the polygonatum extract contains ≥20% polygonatum polysaccharide, which has an immunomodulatory effect; the selenium-enriched Eucommia ulmoides leaf instant tea has a total selenium content of not less than 2 mg / kg, forming a dual selenium source synergy with the corydalis leaf and rice bran selenoprotein peptide.

[0076] To verify the synergistic effect of albumin peptides and selenoprotein peptides, a comparative experiment was designed. SD rats were randomly divided into three groups: a control group (administered with an equal volume of physiological saline by gavage), a selenoprotein peptide group (administered with selenoprotein peptides equivalent to a selenium content of 60 μg / kg body weight), and a combination group (administered with selenoprotein peptides + albumin peptides, with the same selenium content, and the albumin peptide dose was 90 mg / kg body weight). After 28 days of continuous gavage, serum selenium concentration and hepatic glutathione peroxidase (GSH-Px) activity were measured. The results showed that the serum selenium concentration in the combination group was 42.3% higher than that in the selenoprotein peptide group (P<0.01), and the hepatic GSH-Px activity was 37.6% higher (P<0.01). Simultaneously, the apparent permeability coefficient (Papp) of selenium was measured using an in vitro Caco-2 cell monolayer model. The results showed that in the presence of albumin peptides, the Papp value of selenocysteine ​​increased from (0.52±0.08)×10⁻ 6 cm / s increased to (1.23±0.11)×10⁻ 6 The absorption rate increased by 136.5% (cm / s). These results indicate that albumin peptides can act as a transport carrier, significantly promoting the intestinal absorption of selenoprotein peptides and their chelated selenium via the active transport pathway mediated by the small peptide transporter PepT1. This unexpected synergistic effect cannot be achieved by using selenoprotein peptides or albumin peptides alone.

[0077] This invention provides embodiment 4:

[0078] S1 involves hot-air drying sorbitol, Poria cocos powder, albumin peptides, bitter melon peptide powder, Polygonatum sibiricum extract, selenium-enriched Corydalis yanhusuo leaf and rice bran protein peptide powder, kudzu root extract, green tea extract, selenium-enriched Eucommia ulmoides leaf instant tea, and hydroxypropyl methylcellulose at 60℃ for 4-6 hours to reduce the moisture content to ≤8%. The drying time is adjusted appropriately based on the initial moisture content of the materials to ensure the final moisture content meets the standard.

[0079] The dried materials are then sieved sequentially through 80-mesh and 100-mesh double-stage sieves to remove coarse particles. The double-stage sieve design ensures uniform particle size and prevents large particles from affecting subsequent mixing and tableting.

[0080] Next, according to the formula ratio, add the sieved materials, along with any excipients that may be added to the formula, such as wheat oligopeptides, corn oligopeptides, collagen peptides, microcrystalline cellulose, maltodextrin, isomaltitol, erythritol, etc. (the specific addition depends on the product dosage form), into a horizontal mixer. Stir at 60-100 rpm for 10-15 minutes to ensure all materials are thoroughly mixed. The stirring speed should not be too fast to avoid overheating or static agglomeration of the materials.

[0081] S2. The resulting mixture is fed into a roller-type dry granulator. The roller pressure is set to 5-8 MPa, and the roller gap is controlled at 0.6-1.0 mm. Under these conditions, the mixture is pressed into strips or sheets. Dry granulation does not require the addition of wetting agents or binders, avoiding the degradation or migration of active ingredients that may occur in wet granulation. It is particularly suitable for heat-sensitive selenoprotein peptides and albumin peptides.

[0082] The compacted material is then fed into a high-speed rotary knives pulverizer for further crushing. The pulverizer speed is set to 800-1200 rpm. By adjusting the crushing time and screen aperture, the particle size of the fragments is controlled within the range of 0.2-1.0 mm. Particles within this size range have good flowability and compressibility.

[0083] Finally, the pulverized material is sieved through a 0.8mm sieve to remove excessively large or small particles, resulting in uniform dry granules. The sieved granules are then set aside for later use.

[0084] S3. Add magnesium stearate as a lubricant to the obtained dry granules. The amount of magnesium stearate added is 0.5-1.0% of the total formulation mass. The function of magnesium stearate is to reduce the friction between the granules and the die, and to prevent the granules from sticking during compression.

[0085] Mix the granules with added magnesium stearate in a horizontal mixer for 3-5 minutes at a speed of 50-80 rpm. The mixing time should not be too long, otherwise excessive magnesium stearate coating of the granules may affect tablet disintegration or dissolution.

[0086] S4. The lubricated granules are fed into a rotary tablet press, using a mold with a diameter of 8-10mm to compress them into tablets. The weight of the compressed tablets is controlled between 0.9-1.2g / tablet, with a weight difference within ±5%.

[0087] The tableting pressure is set at 30-50 kN. Under this pressure, the tablets have a hardness of 60-100 N and a friability of less than 1.0%. The appropriate hardness and low friability ensure the integrity of the product during transportation and storage, while not affecting its disintegration after oral administration.

[0088] The finished tablets are packaged in aluminum-plastic blister packs, with 10 tablets per blister pack, and 0.5-1g of silica gel desiccant is added to prevent moisture absorption. The aluminum-plastic blister pack has good moisture-proof and light-proof properties.

[0089] After packaging, the product is stored in a dry environment with humidity ≤60% and temperature ≤25℃, and the shelf life can reach 24 months.

[0090] This invention provides embodiment 5:

[0091] This example verifies the antioxidant, hypoglycemic, hypolipidemic, cognitive-enhancing, and anti-inflammatory effects of the formulation of this invention through experiments. This invention uses Viola yedoensis selenium protein, oligopeptides (selenoprotein peptides), and antioxidant selenium-rich peptides (SeCPPs) as research objects, and conducts systematic animal experiments and in vitro activity evaluations.

[0092] Please see Figure 10-12 In this embodiment, the hypoglycemic and hypolipidemic functions of selenoprotein are verified:

[0093] This study used an alloxan-induced diabetic mouse model. Mice with successfully induced diabetic conditions were randomly divided into four groups: blank control group, model group, positive drug group, water extract group, alcohol extract group, CPP group, selenoprotein group, CPP + low-selenoprotein group, CPP + high-selenoprotein group, hypoglycemic group, lipid-lowering group, and solid beverage group. Viola yedoensis selenoprotein was administered by gavage for four consecutive weeks. The experimental results are as follows:

[0094] (1) Oral glucose tolerance test: Compared with the model control group, the peak blood glucose level of mice in the selenoprotein group was significantly reduced after gavage glucose administration, and the area under the blood glucose curve was reduced, indicating that Viola yedoensis selenoprotein can significantly improve glucose tolerance in diabetic mice and has a hypoglycemic effect.

[0095] (2) Measurement of serum total cholesterol (TC) content: The serum TC content of the model group mice was significantly higher than that of the blank group. After selenoprotein intervention, the TC content decreased significantly with dosage.

[0096] (3) Serum triglyceride (TG) content determination: The serum TG content of mice in the selenoprotein group was significantly lower than that in the model group, indicating that selenoprotein has the effect of reducing triglycerides.

[0097] Please participate Figure 13-16 In this embodiment, the antioxidant and immunomodulatory functions of selenoproteins were verified:

[0098] In this case, the same animal model was used to measure serum glutathione peroxidase (GSH-Px) activity, serum insulin (INS) levels, spleen index, and thymus index.

[0099] (1) GSH-Px is an important selenium-containing antioxidant enzyme in the body, and its activity directly reflects selenium nutritional status and antioxidant capacity. The experimental results showed that the serum GSH-Px activity of the model group mice was significantly lower than that of the normal group (P<0.01). After administration of Viola yedoensis selenoprotein, the GSH-Px activity was greatly increased, and the high-dose group recovered to 92.5% of the normal level (P<0.01), indicating that selenoprotein can effectively supplement selenium and enhance the activity of the body's antioxidant enzyme system.

[0100] (2) The INS content of mice in the model group was high, while the INS content of mice in the selenoprotein group was significantly lower than that in the model group (P<0.05).

[0101] (3) The spleen and thymus are important immune organs of the body, and their organ indices reflect the state of immune function. Compared with the model group, the spleen index of mice in the selenoprotein group was significantly increased (P<0.05), and the thymus index was also increased (P<0.05), indicating that Viola yedoensis selenoprotein can enhance the body's cellular and humoral immune functions.

[0102] In this embodiment, the effects of selenium peptides and selenium polysaccharides on the activity of human glucose metabolism enzymes were verified:

[0103] This invention also investigated the effects of a compound of selenium polypeptide and selenium polysaccharide from *Violet spp.* on blood glucose concentrations in different staple food matrices (noodles, steamed buns, milk powder).

[0104] Results: Within 30 minutes of consuming noodles, steamed buns, and milk powder, blood glucose levels significantly decreased. This indicates that selenium peptides and selenium polysaccharides can significantly inhibit amylase activity and slow down the rate of carbohydrate digestion and absorption, thereby reducing postprandial blood glucose response. This function works synergistically with the bitter melon peptide powder (containing 1-deoxynojirimycin) in the formula, jointly playing a role in assisting in lowering blood sugar.

[0105] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention in any other way. Any person skilled in the art may make changes or modifications to the above-disclosed technical content to create equivalent embodiments that can be applied to other fields. However, any simple modifications, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the protection scope of the present invention.

Claims

1. A formulation of violet leaf, rice porridge, selenium protein peptides, and albumin peptides, characterized in that, The core active ingredients of the formula include selenoprotein peptides derived from corydalis leaf and albumin peptides derived from egg white; The violet leaf crushed rice selenoprotein peptide is a mixture of peptide segments with a molecular weight of less than 1000 Da obtained by extraction, enzymatic hydrolysis and ultrafiltration fractionation of the super-selenopleurum plant violet leaf crushed rice. Its number average molecular weight is 461 Da and the total selenium content is 412.67 mg / kg. Its selenium form is mainly selenocysteine ​​and violet leaf crushed rice plant selenopeptide. The albumin peptide is an ovalbumin peptide with a biological titer of 94 and contains all 20 amino acids. The composition uses albumin peptides as a carrier to promote the active transport of selenoprotein peptides and their chelated selenium in the intestine, thereby achieving the unexpected technical effect of improving selenium absorption and synergistic antioxidant effect.

2. The formulation of violet leaf and rice porridge selenium protein peptide and albumin peptide according to claim 1, characterized in that, The formula comprises the following components in parts by weight: 25-40 parts sorbitol, 10-20 parts Poria cocos powder, 10-20 parts albumin peptide, 3-6 parts bitter melon peptide powder, 3-5 parts Polygonatum sibiricum extract, 3-5 parts selenium-enriched corydalis leaf and rice bran protein peptide powder, 2-4 parts kudzu root extract, 2-4 parts green tea extract, 2-4 parts selenium-enriched Eucommia ulmoides leaf instant tea, 2-4 parts hydroxypropyl methylcellulose, and 0.8-1.6 parts magnesium stearate.

3. The formulation of violet leaf and rice porridge selenium protein peptide and albumin peptide according to claim 1, characterized in that: The selenium content provided by the Viola yedoensis selenium protein peptide is 52-70 μg, and the peptide mass provided by the albumin peptide is 80-100 mg. The albumin peptide is a specific enzymatic hydrolysis product derived from egg white, and its molecular weight distribution ranges from 200-800 Da.

4. The formulation of violet leaf and rice porridge selenium protein peptide and albumin peptide according to claim 1, characterized in that: In the Viola yezoensis selenium protein peptide, the sum of selenocysteine ​​and Viola yezoensis plant selenium peptide accounts for more than 90% of its total selenium content.

5. The formulation of violet leaf and rice porridge selenium protein peptide and albumin peptide according to claim 1, characterized in that: The bitter melon peptide powder contains at least 0.3% 1-deoxynojirimycin, the polygonatum extract contains ≥20% polygonatum polysaccharide, and the selenium-enriched Eucommia ulmoides leaf instant tea contains no less than 2mg / kg of total selenium.

6. A method for preparing a formulation of violet leaf and rice slub selenoprotein peptide and albumin peptide, comprising the formulation of violet leaf and rice slub selenoprotein peptide and albumin peptide as described in any one of claims 1-5, characterized in that, The preparation method of the formula includes the following steps: S1, drying, sieving and uniformly mixing some of the raw materials; S2, the mixed raw materials are pressed into strips by roller pressing dry granulation equipment and then crushed; S3, mix the dry granules with the lubricant magnesium stearate evenly; S4. After the granules are compressed into tablets, they are packaged and stored using aluminum-plastic blister packs or bottles.

7. The preparation method of the violet leaf and rice porridge selenium protein peptide and albumin peptide formulation according to claim 6, characterized in that, Step S1 specifically includes: S11 involves drying sorbitol, Poria cocos powder, albumin peptide, bitter melon peptide powder, Polygonatum sibiricum extract, selenium-enriched Viola yedoensis and Capsella bursa-pastoris protein peptide powder, Pueraria lobata extract, green tea extract, selenium-enriched Eucommia ulmoides leaf instant tea, and hydroxypropyl methylcellulose separately in hot air at 60℃ for 4-6 hours to reduce the moisture content to ≤8%. S12 is sieved through a double-stage sieve of 80 mesh and 100 mesh in sequence to remove coarse particles; S13. Mix the filtered materials according to the formula ratio and stir in a horizontal mixer at 60-100 rpm for 10-15 minutes.

8. The preparation method of the violet leaf and rice porridge selenium protein peptide and albumin peptide formulation according to claim 6, characterized in that, Step S2 specifically includes: S21, the obtained mixture is fed into a roller-type dry granulator, the roller pressure is set to 5-8MPa, the roller gap is controlled at 0.6-1.0mm, and it is pressed into strip or sheet compacted material. S22, the compacted material is fed into a high-speed rotary knives pulverizer for crushing, with the rotation speed set at 800-1200 rpm and the particle size of the fragments controlled at 0.2-1.0 mm; S23, the crushed material is sieved through a 0.8mm sieve to obtain uniform particles.

9. The preparation method of the violet leaf and rice porridge selenium protein peptide and albumin peptide formulation according to claim 6, characterized in that, Step S3 specifically includes: S31, magnesium stearate is added to the granules obtained by dry granulation, wherein the amount of magnesium stearate added is 0.5-1.0% of the total formula mass; S32, mix in a horizontal mixer for 3-5 minutes at a speed of 50-80 rpm.

10. The preparation method of the violet leaf and rice porridge selenium protein peptide and albumin peptide formulation according to claim 6, characterized in that, Step S4 specifically includes: S41, the granules are fed into a rotary tablet press and pressed into compressed candy. The mold diameter is 8-10mm, and the weight of the compressed candy is controlled at 0.9-1.2g / tablet. S42, the tableting pressure is set to 30-50kN, the hardness of the tablets after compression is 60-100N, and the friability is less than 1.0%; S43, finished tablets are packaged in aluminum-plastic blister packs, each blister pack contains 10 tablets and 0.5-1g of silica gel desiccant; S44, after packaging, should be stored in a dry environment with humidity ≤60% and temperature ≤25℃.

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