A sheep liver peptide composition and preparation method for helping to replenish qi and blood and alleviate dysmenorrhea

Abstract

The present application belongs to the technical field of fermentation engineering, and particularly relates to a sheep liver peptide composition and a preparation method thereof, which are helpful for tonifying qi and blood and relieving dysmenorrhea. The sheep liver peptide composition comprises the following components in mass fractions: sheep liver peptide powder 60-70 parts, red date powder 5-15 parts, ginger powder 3-8 parts, licorice powder 3-8 parts, rose powder 2-6 parts, inulin 5-10 parts, astragalus peptide 5-10 parts, polygonatum kingianum peptide 5-10 parts, pepper powder 1-3 parts, cassia powder 1-3 parts, turmeric powder 2-5 parts, vitamin C 0.5-2 parts, and vitamin E 0.5-2 parts. The sheep liver peptide powder prepared by the present application has high contents of protein, total amino acids, free amino acids and vitamin A. The sheep liver peptide composition prepared by compounding the sheep liver peptide powder with the other components has the effects of tonifying qi and blood and relieving dysmenorrhea.

Description

Technical Field

[0001] This invention belongs to the field of fermentation engineering technology, specifically relating to a sheep liver peptide composition and preparation method that helps replenish qi and blood and relieve menstrual cramps. Background Technology

[0002] In recent years, with the widespread application of fermentation engineering in the field of food bioprocessing, the use of microbial fermentation to transform animal protein has become an important means of improving nutritional quality and bioactivity. Fermentation engineering can alter the nutritional properties of food at the molecular level. Through fermentation, not only can large protein molecules be significantly degraded to generate more small peptides and free amino acids, but the nutritional composition of raw materials can also be changed. In addition, fermentation can effectively remove undesirable flavors and anti-nutritional factors from raw materials, improve the sensory quality of products, and transform animal protein raw materials from traditional crude processing to precision nutritional functional foods.

[0003] Menstrual blood deficiency and dysmenorrhea are common physiological discomforts in women, closely related to insufficient iron reserves, protein and essential amino acid deficiencies, and disordered free radical metabolism caused by menstrual blood loss. Lamb liver, a byproduct of lamb processing, is rich in crude protein, iron, vitamin A, and various bioactive substances. Traditional Chinese medicine considers it to have "liver-nourishing and blood-enriching" effects, making it a high-quality raw material for preparing products that replenish blood and qi and relieve menstrual pain. Modern research shows that the protein in lamb liver, after being broken down into small molecule peptides, is more easily absorbed and utilized by the body. Its amino acid composition (especially essential amino acids) can not only directly replenish nutritional losses during menstruation but also indirectly alleviate dysmenorrhea-related discomfort by regulating the balance of free radicals in the body through antioxidant effects. Therefore, lamb liver peptide powder has become a key raw material for developing related functional products. The protein, total amino acid, and essential amino acid content of lamb liver peptide powder are core indicators for measuring its nutritional efficacy: high protein content provides sufficient nutritional substrates, comprehensive total amino acids, and a high proportion of essential amino acids can directly meet the body's metabolic needs during menstruation. The synergistic effect of these three factors achieves the dual benefits of replenishing blood and qi and relieving menstrual pain.

[0004] The article "Preparation and Antioxidant Activity Study of Sheep Liver Protein Peptides" (Wen Fei) found that sheep liver hydrolysate was obtained through enzymatic hydrolysis, and its protein and amino acid composition was determined. However, the existing technology for preparing sheep liver peptide powder has significant technical defects: on the one hand, the degree of hydrolysis of sheep liver protein in the enzymatic hydrolysis process is insufficient, and some large molecular proteins are not effectively degraded, resulting in a low protein content in the sheep liver peptide powder; on the other hand, amino acid release is insufficient, and the proportion of essential amino acids and other functional amino acids is low, which cannot meet the nutritional balance requirements for replenishing qi and blood and relieving menstrual pain. In addition, the sheep liver peptide powder prepared by the existing technology does not take into account the vitamin A content. Vitamin A participates in hematopoietic iron metabolism and regulates the physiological function of the reproductive system, resulting in a disconnect between the product's nutritional indicators and functional requirements, making it difficult to achieve the synergistic effect of replenishing qi and blood and relieving menstrual pain.

[0005] Most sheep liver peptide powder products currently on the market are positioned as ordinary protein supplements, without being specifically formulated for the unique physiological needs of women during menstruation, such as blood and qi deficiency and dysmenorrhea. More importantly, these products generally do not quantitatively test the core active ingredients related to replenishing blood and qi and relieving dysmenorrhea during the production and quality inspection stages. They do not measure the proportion of protein and amino acids in sheep liver peptides, nor do they specify the precise range of vitamin A content. As a result, the products lack support for their efficacy in "replenishing blood and qi" and "relieving dysmenorrhea", and the effects are difficult to accurately predict. Summary of the Invention

[0006] The purpose of this invention is to provide a sheep liver peptide composition and preparation method that helps to replenish qi and blood and relieve menstrual cramps, which can simultaneously maintain a high content of protein, amino acids, essential amino acids and vitamin A in the sheep liver peptide composition.

[0007] To achieve the above objectives, the present invention provides the following technical solution: A sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps comprises the following components in parts by weight: 60-70 parts sheep liver peptide powder, 5-15 parts jujube powder, 3-8 parts ginger powder, 3-8 parts licorice powder, 2-6 parts rose pollen, 5-10 parts inulin, 5-10 parts astragalus peptide, 5-10 parts polygonatum peptide, 1-3 parts Sichuan pepper powder, 1-3 parts cinnamon powder, 2-5 parts turmeric powder, 0.5-2 parts vitamin C, and 0.5-2 parts vitamin E.

[0008] The formula of this invention is based on the dual functions of replenishing qi and blood, warming the meridians and dispelling cold, regulating qi and relieving pain, and synergistic nutrition. Each component performs its specific function and works synergistically to enhance the effect. The specific principles are as follows: Sheep liver peptide powder can directly replenish the body with key nutrients for hematopoiesis; jujube powder is rich in polysaccharides, iron, and vitamin C, which not only helps replenish hematopoietic raw materials but also enhances the spleen and stomach's digestive function and promotes the absorption of sheep liver peptides; astragalus peptides are easily absorbed and can improve the body's qi and blood circulation efficiency; polygonatum peptides nourish yin and blood, strengthen the spleen and moisten the lungs; vitamin C effectively promotes the conversion and absorption of plant-based iron in sheep liver peptides and jujube powder; inulin, as a prebiotic, regulates the balance of intestinal flora, improves the intestinal absorption environment, and indirectly improves the absorption rate of peptides and nutrients, providing a guarantee for the generation of qi and blood. Ginger powder is warm in nature and pungent in taste, which can dilate blood vessels, improve pelvic blood circulation, and relieve dysmenorrhea and lower abdominal pain caused by cold stagnation and blood stasis. Sichuan pepper powder and cinnamon powder are both warm ingredients, which work synergistically with ginger powder to enhance the effect of dispelling cold; rose pollen has the effects of soothing the liver and regulating qi, promoting blood circulation and relieving pain. Turmeric powder contains curcumin, which has anti-inflammatory and antioxidant properties, and works synergistically with warming ingredients to reduce the severity of menstrual cramps. Licorice powder contains glycyrrhizin, which has anti-inflammatory and pain-relieving effects, and can neutralize the spiciness of ginger and Sichuan peppercorns, while enhancing the overall analgesic effect of the formula. Vitamin E has antioxidant properties, improves blood circulation, and further enhances the analgesic effect.

[0009] The preparation method of the sheep liver peptide powder includes the following steps: (1) Prepare lamb liver puree using fresh lamb liver; (2) Add water to sheep liver paste to obtain fermented material, add plant lactobacillus and bifidobacterium bifidum, carry out the first anaerobic fermentation, sterilize, and obtain the first fermentation product; (3) Add Lactobacillus paracasei to the first fermentation product, carry out a second anaerobic fermentation, sterilize, centrifuge, take the supernatant to obtain the fermentation broth; (4) The fermentation liquid was concentrated under reduced pressure, dried, and pulverized to obtain sheep liver peptide powder.

[0010] Existing technologies using enzymes or microorganisms to process sheep liver typically employ mixed fermentation, resulting in sheep liver powder with low protein and amino acid content. This leads to less than ideal effects of sheep liver peptide powder in replenishing qi and blood and alleviating menstrual cramps. This invention utilizes a gradient fermentation method with three commercially available bacteria: *Lactobacillus plantarum*, *Bifidobacterium bifidum*, and *Lactobacillus paracasei*. This bacterial combination and gradient fermentation method exhibit stronger compatibility with sheep liver, ensuring that the resulting sheep liver peptide powder maintains high protein, total amino acid, and essential amino acid content simultaneously, providing a core nutritional foundation for replenishing qi and blood and alleviating menstrual cramps.

[0011] Furthermore, the conditions for the first anaerobic fermentation are: fermentation temperature of 35-37℃ and fermentation time of 14-16h.

[0012] Furthermore, the dosage of *Lactobacillus plantarum* is (1-3) × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is (4-6) × 10 9 CFU / mL fermentation material.

[0013] Furthermore, the conditions for the second anaerobic fermentation are: fermentation temperature of 35-37℃ and fermentation time of 12-14h.

[0014] Furthermore, the dosage of Lactobacillus paracasei is (3-5) × 10⁻⁶. 9 CFU / mL fermentation material.

[0015] To improve vitamin A content, this invention, through research, discovered that using *Lactobacillus plantarum*, *Bifidobacterium bifidum*, and *Lactobacillus paracasei* yields better synergistic effects. Combined with a two-step gradient anaerobic fermentation process, using specific amounts of strains and fermentation times, this maximizes the retention of vitamin A and ensures efficient nutrient conversion. Analysis shows that the combination of *Lactobacillus plantarum*, *Bifidobacterium bifidum*, and *Lactobacillus paracasei* in gradient fermentation forms an optimal microbial community interaction, thereby efficiently converting nutrients. The synergistic combination of these three strains ensures high protein, high total amino acid, and essential amino acid content while providing a foundation for stable vitamin A retention. Combined with the two-step gradient anaerobic fermentation process, the synergistic effect of the strains is further enhanced, minimizing vitamin A loss and achieving efficient nutrient conversion and enrichment.

[0016] Further, the fermentation broth was concentrated under reduced pressure to a sheep liver peptide paste with a relative density of 1.13-1.16.

[0017] Furthermore, the total amino acid content in the sheep liver peptide composition is 69.8-70.2 wt%, and the content of essential amino acids is 28.0-28.5 wt%.

[0018] Furthermore, the vitamin A content in the sheep liver peptide composition is 8591-8643 μg / 100g. The protein content in the sheep liver peptide composition is 84.1-84.8 wt%.

[0019] The present invention also provides a method for preparing a sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps, comprising the following steps: mixing all components evenly to obtain a sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps.

[0020] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows: 1. This invention selects three strains, namely Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei, for gradient fermentation. These three strains work synergistically to increase the protein content, total amino acid content, essential amino acid content, and vitamin A content of sheep liver peptide powder, providing a core nutritional basis for replenishing qi and blood and relieving menstrual pain.

[0021] 2. This invention employs a two-step gradient anaerobic fermentation process and uses specific strains and fermentation conditions to efficiently convert the protein content, total amino acids, essential amino acids, and vitamin A content of sheep liver peptide powder.

[0022] 3. The sheep liver peptide powder prepared by this invention maintains a high level of vitamin A. Vitamin A plays a synergistic and auxiliary role in replenishing qi and blood and relieving dysmenorrhea by participating in hematopoietic iron metabolism and regulating the physiological function of the reproductive system. When used in combination with other ingredients, it can significantly enhance the efficacy.

[0023] 4. This invention breaks through the limitations of existing single-product lamb liver products by compounding lamb liver powder with multiple raw materials, achieving the dual goals of supplementing nutrition and improving blood stasis. It solves the problems of single nutrition and poor synergy of traditional products. Moreover, the water-soluble formula is more easily absorbed by the human body and is suitable for consumption during menstruation. Attached Figure Description

[0024] Figure 1 This is a schematic diagram of fresh sheep liver from Example 1.

[0025] Figure 2 This is a schematic diagram of the fermentation liquid after fermentation of sheep liver in Example 1.

[0026] Figure 3 This is a schematic diagram of the sheep liver peptide ointment in Example 1.

[0027] Figure 4 This is a schematic diagram of the sheep liver peptide powder in Example 1.

[0028] Figure 5 This is a schematic diagram of the sheep liver peptide composition of Example 1.

[0029] Figure 6 This is a schematic diagram of the sheep liver peptide composition from Example 1 after being mixed with warm water. Detailed Implementation

[0030] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and 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.

[0031] The raw materials and strains used in the following embodiments of the present invention are all commercially available products: Bifidobacterium bifidum: strain number CICC10395, from China Industrial Microbiology Preservation and Management Center. Lactobacillus paracasei: strain number SHIMCC D24719, from Shanghai Microbiology Preservation Center; Lactobacillus plantarum: strain number CICC10345, from China Industrial Microbiology Preservation and Management Center.

[0032] Alkaline protease, model: Xiasheng FDG-2202, Xiasheng (Beijing) Biotechnology Development Co., Ltd. Enzyme activity: 200,000 U / g.

[0033] Trypsin, model: Xiasheng FDG-2281, Xiasheng (Beijing) Biotechnology Development Co., Ltd.

[0034] Neutral protease, model: Xiasheng FDG-2230, Xiasheng (Beijing) Biotechnology Development Co., Ltd.

[0035] Papain, model: Xiasheng FDG-2203, Xiasheng (Beijing) Biotechnology Development Co., Ltd.

[0036] Red date powder, ginger powder, licorice powder, rose powder, inulin, astragalus peptide, polygonatum peptide, Sichuan pepper powder, cinnamon powder, and turmeric powder are all commercially available products.

[0037] Example 1 Please see Figure 1-6 This embodiment provides a sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps, comprising the following components in parts by weight: 65 parts sheep liver peptide powder, 9 parts jujube powder, 5 parts ginger powder, 4 parts licorice powder, 3 parts rose pollen, 7 parts inulin, 8 parts astragalus peptide, 6 parts polygonatum peptide, 2 parts Sichuan pepper powder, 2 parts cinnamon powder, 4 parts turmeric powder, 1 part vitamin C, and 1.3 parts vitamin E.

[0038] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented feed. Add *Lactobacillus plantarum* and *Bifidobacterium bifidum* to the fermented feed at the same time. The amount of *Lactobacillus plantarum* is 2 × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is 5×10⁻⁶. 9 The CFU / mL fermentation material was mixed evenly and subjected to the first anaerobic fermentation at a temperature of 35℃ for 15 hours. After fermentation, the mixture was sterilized and cooled to room temperature to obtain the first fermentation product. (3) Add Lactobacillus paracasei to the first fermentation product. The amount of Lactobacillus paracasei is 4 × 10⁻⁶. 9 The CFU / mL fermentation material was further subjected to a second anaerobic fermentation at 35℃ for 13 hours. After fermentation, the mixture was sterilized, centrifuged at 4000 rpm for 10 minutes, and the supernatant was collected to obtain the fermentation broth. (4) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0039] The method for preparing the sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps includes the following steps: mixing all components evenly to obtain the sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps.

[0040] Example 2 This embodiment provides a sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps, comprising the following components in parts by weight: 60 parts sheep liver peptide powder, 5 parts jujube powder, 8 parts ginger powder, 3 parts licorice powder, 6 parts rose pollen, 5 parts inulin, 10 parts astragalus peptide, 5 parts polygonatum peptide, 3 parts Sichuan pepper powder, 1 part cinnamon powder, 5 parts turmeric powder, 0.5 parts vitamin C, and 2 parts vitamin E.

[0041] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented feed. Add *Lactobacillus plantarum* and *Bifidobacterium bifidum* to the fermented feed at the same time. The amount of *Lactobacillus plantarum* is 1×10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is 6×10⁻⁶. 9 CFU / mL of fermentation material was mixed evenly and subjected to the first anaerobic fermentation at a temperature of 35℃ for 14 hours. After fermentation, the mixture was sterilized and cooled to room temperature to obtain the first fermentation product. (3) Add Lactobacillus paracasei to the first fermentation product. The amount of Lactobacillus paracasei is 3 × 10⁻⁶. 9 The CFU / mL fermentation material was further subjected to a second anaerobic fermentation at 35℃ for 12 hours. After fermentation, the mixture was sterilized, centrifuged at 4000 rpm for 10 minutes, and the supernatant was collected to obtain the fermentation broth. (4) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0042] The method for preparing the sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps includes the following steps: mixing all components evenly to obtain the sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps.

[0043] Comparative Example 1 The difference between this comparative example and Example 1 is that Lactobacillus plantarum, Bifidobacterium bifidum, and Lactobacillus paracasei were fermented together simultaneously.

[0044] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented feed. Add *Lactobacillus plantarum*, *Bifidobacterium bifidum*, and *Lactobacillus paracasei* to the fermented feed at the same time. The amount of *Lactobacillus plantarum* is 2 × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is 5×10⁻⁶. 9 The dosage of CFU / mL fermentation material and Lactobacillus paracasei is 4 × 10⁻⁶. 9 The fermentation material was mixed with CFU / mL and stirred evenly for anaerobic fermentation at 35℃ for 28 hours. After fermentation, the mixture was sterilized, centrifuged at 4000r / min for 10 minutes, and the supernatant was collected to obtain the fermentation broth. (3) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0045] Comparative Example 2 The difference between this comparative example and Example 1 is that the amount of *Lactobacillus plantarum* used is 5 × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is 2×10⁻⁶. 9 CFU / mL fermentation material.

[0046] Comparative Example 3 The difference between this comparative example and Example 1 is that the amount of Lactobacillus paracasei used is 1×10⁻⁶. 9 CFU / mL fermentation material.

[0047] Comparative Example 4 The difference between this comparative example and Example 1 is that the amount of *Lactobacillus paracasei* used is 7 × 10⁻⁶. 9 CFU / mL fermentation material.

[0048] Comparative Example 5 The difference between this comparative example and Example 1 is that the total fermentation time is the same, but the fermentation times for the two fermentations are different: First anaerobic fermentation: fermentation temperature 35℃, fermentation time 10h. Second anaerobic fermentation: fermentation temperature 35℃, fermentation time 18h.

[0049] Comparative Example 6 The difference between this comparative example and Example 1 is that the order of the bacterial strains used in the gradient fermentation is different.

[0050] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented feed. Add *Lactobacillus plantarum* and *Lactobacillus paracasei* to the fermented feed at the same time. The amount of *Lactobacillus paracasei* is 4 × 10⁻⁶. 9 The amount of CFU / mL fermentation material and Bacillus plantarum was 2×10⁻⁶. 9 CFU / mL of fermentation material; mix well and carry out the first anaerobic fermentation at 35℃ for 15 hours. After fermentation, sterilize and cool to room temperature to obtain the first fermentation product. (3) Add Bifidobacterium bifidum to the first fermentation product and continue the second anaerobic fermentation. The amount of Bifidobacterium bifidum used is 5 × 10⁻⁶. 9 The fermentation material was prepared at CFU / mL, fermented at 35℃, and fermented for 13 hours. After fermentation, the mixture was sterilized, centrifuged at 4000 r / min for 10 minutes, and the supernatant was collected to obtain the fermentation broth. (4) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0051] Comparative Example 7 The difference between this comparative example and Example 1 is that sheep liver peptide powder was prepared using alkaline protease according to the method in "Preparation and Antioxidant Activity Study of Sheep Liver Protein Polypeptides".

[0052] The preparation method of sheep liver peptide powder specifically includes the following steps: S1. After cleaning the fresh lamb liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained lamb liver is 1:5, so that the particle size is less than 100 mesh, and you get lamb liver paste. S2: Add water at twice the mass of sheep liver paste to obtain enzymatic hydrolysate; add enzyme preparation, which is alkaline protease, at a concentration of 0.8% of the mass of fresh sheep liver raw material; the enzymatic hydrolysis temperature is 51℃, pH 8.5, and the enzymatic hydrolysis time is 4.1h. S3. Enzyme inactivation treatment: After the enzymatic hydrolysis reaction is completed, the enzymatic hydrolysate is rapidly heated to 85°C and maintained for 12 minutes. S4. The enzymatic hydrolysate was concentrated at 50℃ and -0.09MPa under reduced pressure to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and 12Pa under vacuum for 20 hours and then pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0053] Comparative Example 8 The difference between this comparative example and comparative example 9 is that the enzyme preparation, including trypsin, neutral protease, and papain, has a mass ratio of 2:4:3. The amount of enzyme preparation used accounts for 1.0% of the mass of fresh sheep liver raw material; the enzymatic hydrolysis temperature is 50℃, pH 7.0, and the enzymatic hydrolysis time is 5h.

[0054] Comparative Example 9 The difference between this comparative example and Example 1 is that only one type of bacteria is used for fermentation.

[0055] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented material. Add *Lactobacillus plantarum* to the fermented material at a dosage of 11 × 10⁻⁶. 9 CFU / mL of fermentation material; anaerobic fermentation was carried out at a fermentation temperature of 35℃ for 28 hours. After fermentation, the material was sterilized and cooled to room temperature to obtain the fermentation product. (3) Centrifuge the fermentation product at 4000 r / min for 10 min, take the supernatant, and obtain the fermentation broth; (4) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0056] Comparative Example 10 The difference between this comparative example and Example 1 is that only Lactobacillus plantarum and Bifidobacterium bifidum were used for fermentation.

[0057] The preparation method of the sheep liver peptide powder includes the following steps: (1) After cleaning the fresh sheep liver, remove the fascia and blood vessels, soak it in water for 25 minutes, drain the water, add ice water and stir. The mass ratio of ice water to the drained sheep liver is 1:5, so that the particle size is less than 100 mesh, and sheep liver paste is obtained. (2) Add water to the sheep liver paste at twice its weight to obtain fermented feed. Add *Lactobacillus plantarum* and *Bifidobacterium bifidum* to the fermented feed at the same time. The amount of *Lactobacillus plantarum* is 4 × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is 7×10⁻⁶. 9The fermentation material was mixed with CFU / mL and stirred evenly. The first anaerobic fermentation was carried out at a fermentation temperature of 35℃ for 28 hours. After fermentation, the mixture was sterilized and cooled to room temperature to obtain the fermentation product. (3) Centrifuge the fermentation product at 4000 r / min for 10 min, take the supernatant, and obtain the fermentation broth; (4) The fermentation liquid was concentrated at 50℃ and -0.09MPa to a relative density of 1.13 (measured at 60℃) to obtain sheep liver peptide paste. The sheep liver peptide paste was freeze-dried at -40℃ and vacuum degree of 12Pa for 20h and pulverized to less than 100 mesh to obtain sheep liver peptide powder.

[0058] Performance testing The sheep liver peptide powders of Examples 1-2 and Comparative Examples 1-10 were subjected to performance tests.

[0059] (1) The protein content in sheep liver peptide powder was determined with reference to the National Food Safety Standard - Determination of Protein in Food (GB 5009.5-2025).

[0060] (2) The amino acid composition of sheep liver peptide powder was determined according to the "Preparation and Antioxidant Activity Study of Sheep Liver Protein Polypeptides" (Wen Fei). The specific steps were as follows: 20 mg of sheep liver peptide powder was accurately weighed and placed in a hydrolysis tube; 5 mL of 6 mol / L hydrochloric acid aqueous solution was added to the tube, the cap was tightened to seal it, and then it was placed in an oven at 110℃ for 24 h. After naturally cooling to room temperature, the hydrolysate was obtained; using distilled water as a solvent, the hydrolysate was completely transferred to one-third of the volumetric flask; then 4.8 mL of sodium hydroxide solution was added for neutralization, and the pH of the solution was adjusted to 2.5 with 6 mol / L hydrochloric acid. Then, the volume was adjusted to 100 mL with distilled water to the mark of the volumetric flask. After the hydrolysate was adjusted to volume, it was filtered through a 0.45 μm filter membrane, and 1 mL of filtrate was collected into a sample bottle. The sample was injected through a fully automated amino acid analyzer to determine the amino acid composition.

[0061] Total amino acids: Aspartic acid, threonine, serine, glutamic acid, glycine, alanine, cysteine, valine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, lysine, histidine, arginine, proline Essential amino acids: The essential amino acids are: threonine, valine, methionine, isoleucine, leucine, phenylalanine, lysine, and histidine.

[0062] (3) The vitamin A content in sheep liver peptide powder was determined according to GB5009.82-2016.

[0063] The test results are shown in Table 1.

[0064] Table 1 Test Results As shown in Table 1, Examples 1-2, through two-step segmented anaerobic fermentation, specific strain combinations, and precise dosage, achieved high levels of all four indicators: protein content, total amino acid content, essential amino acid content, and vitamin A content. It should be noted that the detection principles and calculation benchmarks for protein and total amino acid are different, and the substances represented by these two values ​​overlap. For details, please refer to the article "Preparation and Antioxidant Activity Study of Sheep Liver Protein Peptides".

[0065] In Comparative Example 1, a one-step fermentation with mixed bacteria was used, which led to competition for nutrients and mutual inhibition of metabolites among the strains, resulting in incomplete protein hydrolysis, a significant reduction in the total release of amino acids, and a simultaneous decrease in essential amino acids. At the same time, the vitamin A content decreased significantly. Analysis showed that a large number of free radicals were generated under these conditions, and vitamin A was oxidized and destroyed.

[0066] In Comparative Example 2, changes in the dosage of *Lactobacillus plantarum* and *Bifidobacterium bifidum* led to a significant decrease in the content of essential amino acids. Analysis suggests that the altered dosage in Comparative Example 2 resulted in insufficient cleavage of peptide bonds related to essential amino acids, leading to a significant reduction in the release of essential amino acids.

[0067] In Comparative Example 3, the dosage of Lactobacillus paracasei was too low, and the insufficient number of strains led to a significant decrease in peptide bond breaking efficiency and insufficient release of total amino acids.

[0068] In Comparative Example 4, the excessive amount of *Lactobacillus paracasei* resulted in a decrease in essential amino acids. Analysis suggests that the excessive amount of *Lactobacillus paracasei* led to over-proliferation of subsequent fermentation strains, producing large amounts of organic acids that disrupted the stability of essential amino acids. Simultaneously, the excessive metabolites inhibited the specificity of peptide bond breaking, resulting in insufficient release of essential amino acids.

[0069] In Comparative Example 5, the total amino acid content decreased significantly with the change in fermentation time. Analysis suggests that the first fermentation step did not fully break down large protein molecules, and even with extended fermentation time in the second step, only a small amount of peptides were broken down, resulting in a significant deficiency in the release of total amino acids.

[0070] The fermentation order of the strains in Comparative Example 6 was different. Since Lactobacillus paracasei acted on the macromolecular protein first, its peptidase had a weak ability to decompose the macromolecular protein. Subsequently, Bifidobacterium bifidum could not efficiently decompose the remaining macromolecular protein, resulting in incomplete peptide bond breaking and a significant reduction in the release of total amino acids.

[0071] Comparative Example 7 uses alkaline protease hydrolysis disclosed in the prior art. Alkaline protease only breaks specific peptide bonds and does not completely decompose the peptide segments containing essential amino acids. Furthermore, some essential amino acids are destroyed during the hydrolysis process, resulting in a decrease in essential amino acid content. Under these conditions, more free radicals are generated, which attack vitamin A, leading to a decrease in vitamin A content.

[0072] In Comparative Example 8, the use of a mixed enzyme hydrolysis significantly reduced the total amino acid content. Analysis suggests that this condition led to insufficient protein degradation efficiency; simultaneously, the mutual inhibition of multiple enzymes resulted in disordered peptide bond breakage, significantly reducing the release of total amino acids; and the disordered action of the mixed enzymes produced more pro-oxidative factors, leading to the destruction of vitamin A.

[0073] Comparative Examples 9 and 10 showed significantly worse fermentation results than Example 1, using only one or two strains.

[0074] In summary, this invention, through the synergistic effect of two-step segmented anaerobic fermentation, specific strain combination, and precise dosage, can successfully achieve high levels of protein, total amino acids, free amino acids, and vitamin A in sheep liver peptide powder, resulting in excellent nutritional value. Existing technologies cannot simultaneously achieve such high levels of multiple nutritional indicators.

[0075] Application examples A 32-year-old female employee of our company, due to long hours of desk work and irregular schedules, had been experiencing symptoms of blood and qi deficiency and dysmenorrhea during her menstrual period for the past year. Two days before her period, she experienced lower abdominal distension and cramping pain, requiring the use of heat compresses for relief, accompanied by a sallow complexion, weakness in the limbs, dizziness, and blurred vision. Following a dosage of 10g once daily, she took the sheep liver peptide composition from Example 1, dissolved it in 200mL of 40℃ warm water, and took it 30 minutes after breakfast for 30 consecutive days. After 30 days of continuous use, coinciding with her menstrual period, the severity of her lower abdominal distension and cramping pain significantly decreased, the duration of the pain shortened, and she no longer needed heat compresses for relief. Her complexion remained rosy, and the symptoms of blood and qi deficiency, including weakness in the limbs and dizziness, were significantly reduced.

[0076] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A sheep liver peptide composition that helps replenish qi and blood and alleviate menstrual cramps, characterized in that, The product comprises the following components in parts by weight: 60-70 parts sheep liver peptide powder, 5-15 parts jujube powder, 3-8 parts ginger powder, 3-8 parts licorice powder, 2-6 parts rose pollen, 5-10 parts inulin, 5-10 parts astragalus peptide, 5-10 parts polygonatum peptide, 1-3 parts Sichuan pepper powder, 1-3 parts cinnamon powder, 2-5 parts turmeric powder, 0.5-2 parts vitamin C, and 0.5-2 parts vitamin E. The sheep liver peptide powder is produced by first fermenting sheep liver with a mixture of Lactobacillus plantarum and Bifidobacterium bifidum, sterilizing it, and then continuing to ferment it with Lactobacillus paracasei.

2. The sheep liver peptide composition according to claim 1, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that, The preparation method of the sheep liver peptide powder includes the following steps: (1) Prepare lamb liver puree using fresh lamb liver; (2) Add water to sheep liver paste to obtain fermented material, add plant lactobacillus and bifidobacterium bifidum, carry out the first anaerobic fermentation, sterilize, and obtain the first fermentation product; (3) Add Lactobacillus paracasei to the first fermentation product, carry out a second anaerobic fermentation, sterilize, centrifuge, take the supernatant to obtain the fermentation broth; (4) The fermentation liquid was concentrated under reduced pressure, dried, and pulverized to obtain sheep liver peptide powder.

3. The sheep liver peptide composition according to claim 2, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The conditions for the first anaerobic fermentation are: fermentation temperature of 35-37℃ and fermentation time of 14-16h.

4. The sheep liver peptide composition according to claim 3, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The dosage of *Lactobacillus plantarum* was (1-3) × 10⁻⁶. 9 CFU / mL fermentation material; the dosage of Bifidobacterium bifidum is (4-6) × 10 9 CFU / mL fermentation material.

5. The sheep liver peptide composition according to claim 2, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The conditions for the second anaerobic fermentation are: fermentation temperature of 35-37℃ and fermentation time of 12-14h.

6. The sheep liver peptide composition according to claim 3, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The dosage of Lactobacillus paracasei is (3-5) × 10. 9 CFU / mL fermentation material.

7. The sheep liver peptide composition according to claim 2, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The fermentation broth was concentrated under reduced pressure to a relative density of 1.13-1.16 to obtain sheep liver peptide paste.

8. The sheep liver peptide composition according to claim 1, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The total amino acid content in the sheep liver peptide composition is 69.8-70.2 wt%, and the content of essential amino acids is 28.0-28.5 wt%.

9. The sheep liver peptide composition according to claim 1, which helps to replenish qi and blood and relieve menstrual cramps, is characterized in that... The protein content in the sheep liver peptide composition is 84.1-84.8 wt%, and the vitamin A content in the sheep liver peptide composition is 8591-8643 μg / 100g.

10. A method for preparing a sheep liver peptide composition according to any one of claims 1-9 that helps replenish qi and blood and relieve menstrual cramps, characterized in that, The process includes the following steps: mixing all components of the sheep liver peptide composition evenly to obtain a sheep liver peptide composition that helps replenish qi and blood and relieve menstrual cramps.

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