CN114259061B

CN114259061B - Lactation promoting product composition containing codonopsis pilosula oligopeptide and preparation method and application thereof

Info

Publication number: CN114259061B
Application number: CN202111547456.6A
Authority: CN
Inventors: 梁瑛杰; 包卫洋; 张延胜; 王祖哲; 赵林英
Original assignee: Dalian Deep Blue Peptide Technology Research And Development Co ltd; Jilin Guorui Pharmaceutical Co ltd
Current assignee: Dalian Deep Blue Peptide Technology Research And Development Co ltd; Jilin Guorui Pharmaceutical Co ltd
Priority date: 2021-12-16
Filing date: 2021-12-16
Publication date: 2023-11-07
Anticipated expiration: 2041-12-16
Also published as: CN114259061A

Abstract

A lactation promoting product composition containing codonopsis pilosula oligopeptide, a preparation method and application thereof belong to the technical field of processing of functional foods. A lactation promoting product composition containing radix Codonopsis Lanceolatae oligopeptide comprises radix Codonopsis Lanceolatae oligopeptide, ginseng radix oligopeptide, longan extract, fructus Lycii extract, fructus Jujubae extract, fructus Myrtilli extract, purple sweet potato extract, radix astragali extract, radix Platycodi extract, fructus Citri extract and Bulbus Allii Macrostemi extract as raw materials, has good effects of dispersing stagnated liver qi, promoting qi circulation, replenishing blood, invigorating spleen, and removing dampness, and is suitable for postpartum symptoms such as lack of milk, hypogalactia or hypogalactia due to qi and blood deficiency, stagnation of liver qi or phlegm dampness, etc. The application can be used for preparing liquid beverage with pleasant taste and convenient eating, and meets the requirements of female friends with postpartum hypogalactia.

Description

Lactation promoting product composition containing codonopsis pilosula oligopeptide and preparation method and application thereof

Technical Field

The application belongs to the technical field of processing of functional foods, and relates to a lactation promoting product composition containing codonopsis pilosula oligopeptide.

Background

Breast milk is the best natural food for infants, the national United states of health organization is based on foundation for children and China advocates that infants should be fed purely by breast milk within 4-6 months after birth, and nutrient substances contained in the breast milk are very important for the growth and development of infants, and are the best way for the infants to strengthen the digestive system and obtain natural immunity, and meanwhile, breast feeding has important significance for the recovery and health of puerperal bodies.

In recent years, with the increase of advanced puerpera, the number of the puerperal hypogalactia patients tends to rise year by year due to the increase of the number of caesarean products, and a plurality of factors such as large working pressure of females, unreasonable living diet and the like. The postpartum hypogalactia is characterized by that the female postpartum has no milk or insufficient milk in the lactation period, and the need of the infant cannot be met, also called as postpartum galactostasis or postpartum galactostasis, which is a common postpartum disease.

At present, western medicine has no effective treatment method aiming at the disease, so that the treatment is carried out by adopting a traditional Chinese medicine method. The products for improving the postpartum hypogalactia problem in the market are mostly external massage instruments or traditional Chinese medicines of lactation promotion type for soup cooking and eating, and the effect is poor.

Disclosure of Invention

The application aims to overcome the defects of the existing solution, and provides a lactation promoting product composition containing codonopsis pilosula oligopeptide, which is suitable for postpartum symptoms such as no milk, insufficient milk or milk deficiency and the like caused by qi and blood deficiency, liver depression qi stagnation or phlegm dampness obstruction and the like, and is safe and reliable and has no side effect.

The technical scheme of the application is as follows: a lactation promoting composition contains radix Codonopsis Lanceolatae oligopeptide, ginseng radix oligopeptide, arillus longan extract, fructus Lycii extract, fructus Jujubae extract, fructus Myrtilli extract, purple sweet potato extract, radix astragali extract, radix Platycodi extract, fructus Citri extract and Bulbus Allii Macrostemi extract.

Further, the composition comprises the following components in parts by weight: 50-150 parts of codonopsis pilosula oligopeptide, 5-50 parts of ginseng oligopeptide, 10-80 parts of longan extract, 10-80 parts of medlar extract, 10-80 parts of jujube extract, 10-80 parts of blueberry extract, 10-80 parts of purple sweet potato extract, 10-90 parts of astragalus extract, 5-60 parts of platycodon root extract, 15-50 parts of citron extract and 15-50 parts of allium macrostemon extract.

Further, the Codonopsis lanceolata oligopeptide is prepared from Codonopsis lanceolata in Changbai mountain area.

Further, the codonopsis pilosula oligopeptide is prepared based on the following steps: after the codonopsis pilosula sample is coarsely crushed, adding water with the mass volume of 10-15 times to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass of 0.5-1.0% of the codonopsis pilosula sample, carrying out enzymolysis for 1-2 hours at 45-50 ℃, and controlling the pH value of enzyme reaction to be 5.0-6.0; then adding compound protease with 3-5% of the protein mass of codonopsis pilosula, carrying out enzymolysis for 3-4 hours at 50-60 ℃, controlling the pH value of the enzyme reaction to 8.5-9.5, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain the codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, then adopting a ceramic membrane with the molecular weight cut-off of 3000Da to carry out membrane separation, refining to desalt and remove heavy metals, and concentrating until the content of soluble solids accounts for 30-40% of the total mass of the concentrated solution; the mass ratio of the compound protease is as follows: papain neutral proteinase alkaline proteinase= (2-4): (1-2): (4-6).

Further, the codonopsis pilosula oligopeptide comprises a peptide fragment sequence NH2-Phe-Gly-Arg-Trp-Arg.

Further, the ginseng oligopeptide is prepared based on the following manner: after coarse crushing treatment of ginseng sample, adding 15-20 times of water to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass of 0.5-1.0% of ginseng, carrying out enzymolysis for 1-2 hours at 45-50 ℃, controlling the pH value of enzyme reaction to be 5.0-6.0, adding compound protease with the mass of 3-5% of ginseng protein, carrying out enzymolysis for 3-4 hours at 50-55 ℃, controlling the pH value of enzyme reaction to be 9.0-10.0, heating to 90 ℃ after enzymolysis is finished, inactivating enzyme for 10 minutes to obtain a codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, carrying out membrane separation by adopting a ceramic membrane with the molecular weight cutoff of 3000Da, refining to desalt heavy metals, and concentrating until the content of soluble solids accounts for 30-40% of the total mass of the concentrated solution; the mass ratio of the compound protease is as follows: alkaline protease bromelain flavourzyme= (4-6): (2-4).

Further, more than 90% of peptides in the ginseng oligopeptide have a molecular weight of less than 1000Da.

Further, longan, medlar, jujube, blueberry, purple sweet potato, astragalus, platycodon grandiflorum, citron and allium macrostemon are respectively screened, purified, crushed, soaked in water, boiled and filtered to obtain effective extract liquid, and the effective extract liquid is concentrated to obtain extract extractum to obtain corresponding extracts.

A preparation method of a lactation promoting product composition containing codonopsis pilosula oligopeptide is characterized in that a concentrated solution of the codonopsis pilosula oligopeptide and ginseng oligopeptide and an extract of longan, medlar, chinese date, blueberry, purple sweet potato, astragalus, platycodon grandiflorum, citron and allium macrostemon are prepared, and spray drying is carried out after the codonopsis pilosula oligopeptide and the ginseng oligopeptide are fully and uniformly mixed.

Use of a composition according to any one of the preceding claims in a lactation food or pharmaceutical product.

Human prolactin is a polypeptide containing 198 amino acid residues and is secreted by pituitary gland eosinophils, decidua of pregnancy, immune cells, etc. Prolactin plays an important physiological role in human body, and has important effects in regulating ovarian function and maintaining pregnancy besides promoting development and growth of mammary glands and stimulating and maintaining lactation. For example, prolactin can act directly on receptors on mammary gland lactation cell membranes, activate activities of membrane-bound enzymes and adenylate cyclase to play a role in lactation, and also stimulate follicular LH receptor production and the like.

The serum prolactin level of the parturient with hypogalactia is very low, and the active peptide fragment (NH 2-Phe-Gly-Arg-Trp-Arg) in the codonopsis pilosula oligopeptide can improve the serum prolactin level, improve the lactation yield and improve the hypogalactia symptoms.

The lack of lactation is the disease name of traditional Chinese medicine, the understanding of the traditional Chinese medicine on lack of lactation can be traced to the inertia, and the cause of postpartum hypogalactia is considered to be related to deficiency of qi and blood, liver depression and spleen deficiency, phlegm turbidity blockage and the like. Therefore, the Chinese medicinal preparation starts from the aspects of soothing liver, strengthening spleen, enriching blood, nourishing qi, reducing phlegm, removing stagnation and the like, and has good auxiliary effect on treating hypogalactia.

Radix Codonopsis Lanceolatae, which has white milk, is a perennial herb of Codonopsis genus of Campanulaceae, and is also called goat milk, four-leaf ginseng, mountain carrot, etc., which is described by "Sucus Toosendan, zhengzhang Dao" in Minghu Kagao, and "Sucus Tonic, zhejiang folk herbal medicine". The codonopsis pilosula oligopeptide is prepared by biological enzymolysis, and simultaneously, the components such as triterpenoid saponin, sterol, alkaloid, lignan and the like and the nutrient substances such as polysaccharide, dietary fiber and trace elements are reserved, so that the codonopsis pilosula oligopeptide has the effects of nourishing and tonifying, tonifying qi and promoting lactation.

Ginseng has slightly warm nature, sweet and slightly bitter taste, enters spleen, lung, heart and kidney channels, and has the effects of reinforcing primordial qi, promoting fluid production, nourishing blood, tonifying spleen and soothing nerves. The macromolecular protein is changed into micromolecular active peptide through biological enzymolysis technology, and meanwhile, the components such as ginseng saponin, ginseng polysaccharide and the like are reserved, so that the nutrition components are easier to absorb, and the functions of tonifying qi, nourishing blood and strengthening spleen are stronger.

Longan has warm nature, sweet taste, and heart and spleen meridian homing, and has the effects of tonifying heart and spleen, nourishing blood and tranquillizing.

The medlar has the effects of tonifying liver and kidney, replenishing vital essence and improving eyesight, and has the advantages of being mild in nature and sweet in taste, and entering liver and kidney channels.

The jujube has warm nature and sweet taste, enters spleen, stomach and heart meridian, can tonify middle-jiao and qi, nourish blood and tranquilize mind, and can be used for spleen deficiency.

Astragalus root has slightly warm nature, sweet taste, and can enter lung and spleen meridians, contains polysaccharides, saponins, flavonoids, amino acids and microelements, and has effects of invigorating qi, raising yang, nourishing blood and activating stagnancy.

Platycodon root has flat property, bitter and pungent taste, enters lung meridian, contains various chemical components such as saponin and the like, and has the effects of dispersing lung qi, relieving sore throat, eliminating phlegm and expelling pus.

Citron is warm in nature, pungent in flavor, bitter in taste, sour, and enters liver, spleen and lung meridians, and has the effects of soothing liver, regulating qi, and resolving phlegm.

The allium macrostemon is warm in nature, pungent and bitter in taste, enters heart, lung, stomach and large intestine meridians, contains various chemical substances with biological activity, such as saponins, alkaloids, volatile oils, polysaccharides and the like, wherein the steroid saponins are main active ingredients, and can activate yang, dissipate stagnation, promote qi circulation and remove stagnation.

The blueberry fruit contains rich nutritional ingredients, especially anthocyanin, and has good nutrition and health care effects, and also has the functions of preventing cerebral nerve aging, strengthening heart, resisting cancer, softening blood vessels, enhancing human immunity and the like.

Purple sweet potato contains abundant dietary fiber, vitamins, microelements, anthocyanin, glycoprotein, carotene, dehydroepiandrosterone and other functional factors, and has the functions of resisting oxidation, repairing cardiac muscle, relieving liver dysfunction, enhancing organism immunity and the like.

The application has the beneficial effects that:

the application provides a preparation method of a novel low polymer peptide from codonopsis pilosula, which is characterized in that the molecular weight of the obtained small molecular active peptide with more than 90 percent is less than 1000Da, and the peptide contains peptide segments (NH 2-Phe-Gly-Arg-Trp-Arg) with lactation promoting activity, the preparation process is simple, the low polymer peptide is easy to obtain, belongs to food-borne low polymer peptide, can improve serum prolactin level after being eaten for a long time, improves hypogalactia, and has wide application prospect.

In order to improve the availability of effective materials, the ginseng is prepared into ginseng oligopeptide by adopting a targeted enzymolysis technology, longan, medlar, chinese date, blueberry, purple sweet potato, astragalus, platycodon grandiflorum, citron and allium macrostemon are extracted and concentrated, various functional components are reserved, and the absorption and utilization rate is greatly improved.

Prolactin plays an important physiological role in human bodies, and women with hypogalactia usually show low serum prolactin level, and meanwhile, hypogalactia is related to deficiency of qi and blood, liver depression, spleen deficiency, phlegm turbidity blockage and the like. According to the application, the active peptide segment (NH 2-Phe-Gly-Arg-Trp-Arg) in the codonopsis pilosula oligopeptide is used for directly improving the serum prolactin level, and meanwhile, ginseng, longan, medlar, chinese date, blueberry, purple sweet potato, astragalus mongholicus, platycodon grandiflorum, citron and allium macrostemon are matched, so that the effects of soothing liver, strengthening spleen, enriching blood, nourishing qi, reducing phlegm and removing stagnation are achieved, milk formation is increased under the synergistic effect, milk channels are unblocked, and a very good effect is achieved on postpartum hypogalactia.

Drawings

FIG. 1 is a graph showing the effect of Codonopsis lanceolata on significantly increasing serum prolactin levels in milk deficient mice.

Detailed Description

Example 1

A method for preparing Codonopsis lanceolata oligopeptide with serum prolactin increasing effect in a lactation promoting product composition containing Codonopsis lanceolata oligopeptide comprises the following steps: coarsely pulverizing radix Codonopsis Lanceolatae (Codonopsis lanceolata) sample, adding 15 times of water to obtain homogenate, placing in enzymolysis tank, adding cellulase 1.0% of radix Codonopsis Lanceolatae (dry weight), performing enzymolysis at 50deg.C for 1 hr, and controlling pH of enzyme reaction at 5.5; then adding compound protease accounting for 4% of the mass of the codonopsis pilosula protein, wherein the mass ratio of the compound protease is as follows: papain neutral protease and alkaline protease=3:2:5, and performing enzymolysis for 4 hours at 55 ℃, controlling the pH value of the enzyme reaction to be 8.5, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain codonopsis pilosula enzymatic hydrolysate, centrifuging to remove impurities, performing membrane separation by adopting a ceramic membrane with the molecular weight cutoff of 3000Da, refining, desalting, removing heavy metals, and concentrating until the content of soluble solids accounts for 40% of the total mass of the concentrated solution.

Example 2

A method for preparing Codonopsis lanceolata oligopeptide with serum prolactin increasing effect in a lactation promoting product composition containing Codonopsis lanceolata oligopeptide comprises the following steps: after the codonopsis pilosula sample is coarsely crushed, adding 10 times of water with the mass volume to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase accounting for 0.5% of the codonopsis pilosula (based on dry weight), carrying out enzymolysis for 2 hours at 45 ℃, and controlling the pH value of enzyme reaction to be 5.0; then adding the compound protease with the mass of 5% of the codonopsis pilosula protein into the mixture, wherein the mass ratio of the compound protease is as follows: papain neutral protease and alkaline protease=3:1:4, carrying out enzymolysis for 3 hours at 50 ℃, controlling the pH value of the enzymatic reaction to 8.8, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain codonopsis pilosula enzymatic hydrolysate, centrifuging to remove impurities, carrying out membrane separation by adopting a ceramic membrane with the molecular weight cutoff of 3000Da, refining, desalting, removing heavy metals, and concentrating until the content of soluble solids accounts for 30% of the total mass of the concentrated solution.

Example 3

A method for preparing Codonopsis lanceolata oligopeptide with serum prolactin increasing effect in a lactation promoting product composition containing Codonopsis lanceolata oligopeptide comprises the following steps: after the codonopsis pilosula sample is coarsely crushed, adding water with the mass volume of 13 times to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass (calculated by dry weight) of 0.7% of the codonopsis pilosula sample, carrying out enzymolysis for 2 hours at 48 ℃, and controlling the pH value of enzyme reaction to be 6.0; then adding 3% of compound protease of codonopsis pilosula protein by mass, wherein the mass ratio of the compound protease is as follows: papain neutral protease and alkaline protease=2:2:4, and performing enzymolysis for 4 hours at 56 ℃, controlling the pH value of the enzyme reaction to be 9.0, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain codonopsis pilosula enzymatic hydrolysate, centrifuging to remove impurities, performing membrane separation by adopting a ceramic membrane with the molecular weight cutoff of 3000Da, refining, desalting, removing heavy metals, and concentrating until the content of soluble solids accounts for 35% of the total mass of the concentrated solution.

More than 90% of the Codonopsis lanceolata oligopeptides prepared in examples 1, 2 and 3 have a molecular weight of less than 1000Da and each contain a peptide fragment sequence of NH2-Phe-Gly-Arg-Trp-Arg.

Measurement of serum prolactin enhancement:

appropriate amounts of healthy SD rats (male: female=1:3, and female mice were not pregnant) were housed in cages, and after 15 days, male mice were removed and female mice confirmed to be pregnant were transferred to single cage for breeding until parity. Female mice with a parity interval of not more than 24h were selected and randomly divided into 5 groups (control group, model group, experimental group 1, 2, 3) of 10 animals each.

Control group: from the 2 nd day after delivery, the stomach was irrigated 1 time with physiological saline (10 mL/kg) for 7 days in the morning.

Model group: from day 2 after delivery, the stomach was lavage with 0.05mg/mL bromocriptine mesylate (10 mL/kg) 1 time in the morning for 7 days with normal saline (10 mL/kg).

Experimental groups 1, 2, 3: from day 2 after delivery, stomach was lavage with 0.05mg/mL bromocriptine mesylate (10 mL/kg) daily for 7 days at the same time as 1 time with the Codonopsis pilosula oligopeptide (10 mL/kg) prepared in examples 1, 2, 3, respectively.

After the end of the experiment, all the female mice were centrifuged for 5min at 4000r/min with about 2mL of abdominal aortic blood, serum was separated, and the Prolactin (PRL) content in the serum of each group of female mice was determined by enzyme-linked immunosorbent assay (ELISA).

The experimental results are shown in figure 1, and the results show that the prepared codonopsis pilosula can obviously improve the serum prolactin level of the milk-deficient mice.

Example 4

A lactation promoting product composition containing codonopsis pilosula oligopeptide comprises the following components in parts by weight: 100 parts of codonopsis pilosula oligopeptide, 30 parts of ginseng oligopeptide, 50 parts of longan extract, 50 parts of medlar extract, 60 parts of jujube extract, 40 parts of blueberry extract, 40 parts of purple sweet potato extract, 50 parts of astragalus extract, 35 parts of platycodon root extract, 20 parts of citron extract and 25 parts of allium macrostemon extract.

The preparation method comprises the following steps:

(1) Preparation of Codonopsis lanceolata oligopeptide: prepared according to the method of example 1.

(2) The preparation method of the ginseng oligopeptide comprises the following steps: after coarse crushing treatment of ginseng samples, adding 15 times of water by mass volume to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase accounting for 0.8% of the mass (calculated by dry weight) of ginseng, carrying out enzymolysis for 1 hour at 48 ℃, controlling the pH value of enzyme reaction to be 5.5, and adding compound protease accounting for 3% of the mass of ginseng protein, wherein the mass ratio of the compound protease is as follows: alkaline proteinase, bromelain, flavourzyme=4:2:3, enzymatic hydrolysis is carried out for 4 hours at 52 ℃, the pH value of the enzymatic reaction is controlled at 9.0, the temperature is raised to 90 ℃ after the enzymatic hydrolysis is finished, enzyme is inactivated for 10 minutes, codonopsis pilosula enzymatic hydrolysate is obtained, impurities are centrifugally removed, then ceramic membrane with the interception molecular weight of 3000Da is adopted for membrane separation, and the membrane separation is carried out, the desalination and heavy metal removal are refined, and the concentration is carried out until the content of soluble solids accounts for 40% of the total mass of the concentrated solution.

(3) The longan, the medlar, the Chinese dates, the blueberries, the purple sweet potatoes, the astragalus, the platycodon grandiflorum, the citron and the allium macrostemon are respectively screened, purified, crushed, soaked in water, decocted and filtered to obtain effective extract liquid, and the effective extract liquid is concentrated to obtain extract.

(4) And (3) fully and uniformly mixing the concentrated solution and the extract of the codonopsis pilosula oligopeptide and the ginseng oligopeptide, and then performing spray drying to obtain powder.

Example 5

A lactation promoting product composition containing codonopsis pilosula oligopeptide comprises the following components in parts by weight: 120 parts of codonopsis pilosula oligopeptide, 40 parts of ginseng oligopeptide, 40 parts of longan extract, 60 parts of medlar extract, 50 parts of jujube extract, 35 parts of blueberry extract, 40 parts of purple sweet potato extract, 50 parts of astragalus extract, 35 parts of platycodon root extract, 30 parts of citron extract and 20 parts of allium macrostemon extract.

The preparation method comprises the following steps:

(1) Preparation of Codonopsis lanceolata oligopeptide: prepared according to the method of example 2.

(2) The preparation method of the ginseng oligopeptide comprises the following steps: after coarse crushing treatment of ginseng samples, adding water with the mass volume of 20 times to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass (calculated by dry weight) of 1.0% of ginseng, carrying out enzymolysis for 1 hour at 50 ℃, controlling the pH value of enzyme reaction to be 5.0, and adding compound protease with the mass of 3% of ginseng protein, wherein the mass ratio of the compound protease is as follows: alkaline proteinase, bromelain, flavourzyme=5:2:2, enzymatic hydrolysis is carried out for 4 hours at 50 ℃, the pH value of the enzymatic reaction is controlled at 9.5, the temperature is raised to 90 ℃ after the enzymatic hydrolysis is finished, enzyme is inactivated for 10 minutes, codonopsis pilosula enzymatic hydrolysate is obtained, impurities are removed by centrifugation, then ceramic membrane with the molecular weight cutoff of 3000Da is adopted for membrane separation, and the membrane separation is carried out for refining, desalting and heavy metal removal, and concentration is carried out until the content of soluble solids accounts for 30 percent of the total mass of the concentrated solution.

Example 6

A lactation promoting product composition containing codonopsis pilosula oligopeptide comprises the following components in parts by weight: 90 parts of codonopsis pilosula oligopeptide, 40 parts of ginseng oligopeptide, 40 parts of longan extract, 55 parts of medlar extract, 40 parts of jujube extract, 20 parts of blueberry extract, 20 parts of purple sweet potato extract, 46 parts of astragalus extract, 50 parts of platycodon root extract, 40 parts of citron extract and 40 parts of allium macrostemon extract.

The preparation method comprises the following steps:

(1) Preparation of Codonopsis lanceolata oligopeptide: prepared according to the method of example 3.

(2) The preparation method of the ginseng oligopeptide comprises the following steps: after coarse crushing treatment of ginseng samples, adding 17 times of water by mass volume to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase accounting for 1.0% of the mass (calculated by dry weight) of the ginseng, carrying out enzymolysis for 1 hour at 50 ℃, controlling the pH value of enzyme reaction to be 5.8, and adding compound protease accounting for 5% of the mass of ginseng protein, wherein the mass ratio of the compound protease is as follows: alkaline proteinase, bromelain, flavourzyme=6:3:2, enzymatic hydrolysis is carried out for 3 hours at 55 ℃, the pH value of the enzymatic reaction is controlled at 9.7, the temperature is raised to 90 ℃ after the enzymatic hydrolysis is finished, enzyme is inactivated for 10 minutes, codonopsis pilosula enzymatic hydrolysate is obtained, impurities are removed by centrifugation, then ceramic membrane with the interception molecular weight of 3000Da is adopted for membrane separation, and the membrane separation is carried out for refining, desalting and heavy metal removal, and concentration is carried out until the content of soluble solids accounts for 35 percent of the total mass of the concentrated solution.

Example 7

A lactation promoting product composition containing radix codonopsis pilosulae oligopeptide and a preparation method thereof are characterized in that the composition comprises radix codonopsis pilosulae oligopeptide, ginseng oligopeptide, longan extract, medlar extract, jujube extract, blueberry extract, purple sweet potato extract, astragalus extract, balloonflower extract, citron extract and allium macrostemon extract.

A lactation promoting product composition containing codonopsis pilosula oligopeptide is characterized by comprising the following components in parts by weight: 50-150 parts of codonopsis pilosula oligopeptide, 5-50 parts of ginseng oligopeptide, 10-80 parts of longan extract, 10-80 parts of medlar extract, 10-80 parts of jujube extract, 10-80 parts of blueberry extract, 10-80 parts of purple sweet potato extract, 10-90 parts of astragalus extract, 5-60 parts of platycodon root extract, 15-50 parts of citron extract and 15-50 parts of allium macrostemon extract.

A lactation promoting product composition containing radix codonopsis pilosulae oligopeptide is characterized in that the raw material source of the radix codonopsis pilosulae oligopeptide is radix codonopsis pilosulae in Changbai mountain area.

The composition can be prepared into powder, granule, tablet, capsule, pill, oral solution, etc., preferably into powder or granule.

The application also relates to a preparation method of the composition.

(1) The preparation method of the codonopsis pilosula oligopeptide comprises the following steps: after the codonopsis pilosula sample is coarsely crushed, adding water with the mass volume of 10-15 times to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass (calculated by dry weight) of 0.5-1.0% of the codonopsis pilosula sample, carrying out enzymolysis for 1-2 hours at 45-50 ℃, and controlling the pH value of the enzymatic reaction to be 5.0-6.0; then adding compound protease with 3-5% of the codonopsis pilosula protein mass, carrying out enzymolysis for 3-4 hours at 50-60 ℃, controlling the pH value of the enzyme reaction to 8.5-9.5, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain the codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, then adopting a ceramic membrane with the molecular weight cut-off of 3000Da to carry out membrane separation, refining, desalting, removing heavy metals, and concentrating until the content of soluble solids accounts for 30-40% of the total mass of the concentrated solution. The mass ratio of the compound protease is as follows: papain neutral proteinase alkaline proteinase= (2-4): (1-2): (4-6).

(2) The preparation method of the ginseng oligopeptide comprises the following steps: after coarse crushing treatment of ginseng sample, adding 15-20 times of water to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass (calculated by dry weight) of 0.5-1.0% of ginseng, carrying out enzymolysis for 1-2 hours at 45-50 ℃, controlling the pH value of enzyme reaction to be 5.0-6.0, adding compound protease with the mass of 3-5% of ginseng protein, carrying out enzymolysis for 3-4 hours at 50-55 ℃, controlling the pH value of enzyme reaction to be 9.0-10.0, heating to 90 ℃ after enzymolysis is finished for inactivating enzyme for 10 minutes, obtaining a codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, adopting a ceramic membrane with the cut-off molecular weight of 3000Da to carry out membrane separation, refining to desalt heavy metals, and concentrating until the content of soluble solid accounts for 30-40% of the total mass of the concentrated liquid. The mass ratio of the compound protease is as follows: alkaline protease bromelain flavourzyme= (4-6): (2-4).

(4) And (3) fully and uniformly mixing the concentrated solution and the extract of the codonopsis pilosula oligopeptide and the ginseng oligopeptide, and then carrying out spray drying.

Wherein, the molecular weight of more than 90 percent of the codonopsis pilosula oligopeptide in the step (1) is less than 1000Da, and the codonopsis pilosula oligopeptide further refers to that the codonopsis pilosula oligopeptide contains peptide segment sequences NH2-Phe-Gly-Arg-Trp-Arg, so that the serum prolactin level can be improved.

In the step (2), more than 90% of the peptides of the ginseng oligopeptide have molecular weight smaller than 1000Da.

Comparative example 1

A lactation promoting product composition containing codonopsis pilosula oligopeptide comprises the following components in parts by weight: 100 parts of codonopsis pilosula oligopeptide, 30 parts of ginseng oligopeptide, 50 parts of longan extract, 50 parts of medlar extract, 60 parts of jujube extract, 40 parts of blueberry extract, 40 parts of purple sweet potato extract, 50 parts of astragalus extract and 35 parts of platycodon root extract.

The preparation method comprises the following steps:

(2) Preparation of Codonopsis lanceolata oligopeptide: prepared according to the method of example 4.

(2) The preparation method of the ginseng oligopeptide comprises the following steps: prepared according to the method of example 4.

(3) The longan, the medlar, the jujube, the blueberry, the purple sweet potato, the astragalus and the platycodon root are respectively screened, purified, crushed, soaked in water, decocted and filtered to obtain effective extract liquid, and the effective extract liquid is concentrated to obtain extract.

Comparative example 2

A lactation promoting product composition containing codonopsis pilosula comprises the following components in parts by weight: 120 parts of codonopsis pilosula, 40 parts of ginseng oligopeptide, 40 parts of longan extract, 60 parts of medlar extract, 50 parts of jujube extract, 35 parts of blueberry extract, 40 parts of purple sweet potato extract, 50 parts of astragalus extract, 35 parts of platycodon root extract, 30 parts of citron extract and 20 parts of allium macrostemon extract.

The preparation method comprises the following steps:

(1) The preparation method of the ginseng oligopeptide comprises the following steps: prepared according to the method of example 5.

(2) Respectively screening radix Codonopsis Lanceolatae, longan, fructus Lycii, fructus Jujubae, fructus Myrtilli, purple sweet potato, radix astragali, radix Platycodi, fructus Citri, and Bulbus Allii Macrostemi, purifying, pulverizing, soaking in water, decocting, filtering to obtain effective extract liquid, and concentrating to obtain extract.

(3) And fully and uniformly mixing the prepared ginseng oligopeptide concentrated solution and the extract, and then performing spray drying to obtain powder.

Experiment verification

The products obtained in examples 4, 5 and 6 and comparative examples 1 and 2 of the present application were used as samples, and the lactation promoting effect was verified by animal experiments.

Healthy SD rats in the parturient period are selected as experimental objects and are bred until the young rats are born, and then the female rats and the young rats are bred in the same cage according to the nest, so that the quality of each rat and the total quality of the corresponding mice in the nest are ensured to have no statistical difference. Establishment of a rat model with hypogalactia: the master mice were lavaged with 0.05mg/mL bromocriptine mesylate (10 mL/kg) for 7 days at morning and 1 time per day for 1h for 14 days with the mice placed in the mouse binder for binding on day 2.

70 experimental rats were selected and randomly divided into 7 groups (control, model, experimental groups 1, 2, 3, 4, 5), 10 each. According to the requirements of Table 1, the groups of rats were subjected to a gastric lavage test once daily for 7 consecutive days at 10 am.

Table 1 gastric lavage protocol for groups of mice

Group of	Rat class	Gastric lavage sample	Daily dosage
				Control group	Female mouse in normal lactation period	Physiological saline	1000mg/kg
Model group	Milk deficiency model female mouse	Physiological saline	1000mg/kg
				Experiment group 1	Milk deficiency model female mouse	Example 4 product	1000mg/kg
Experiment group 2	Milk deficiency model female mouse	Example 5 product	1000mg/kg
				Experiment group 3	Milk deficiency model female mouse	Example 6 product	1000mg/kg
Experiment group 4	Milk deficiency model female mouse	Comparative example 1 product	1000mg/kg
				Test group 5	Milk deficiency model female mouse	Comparative example 2 product	1000mg/kg

The observations during the gastric lavage test were as follows:

1. state of the master mouse: mental, fur, activity level, fecal character, response sensitivity, food intake, water intake, etc.

2. Milk secretion per hour in the female mice: the method comprises the steps of (1) performing gastric lavage on a female mouse at 10 am every day, after 2 hours of gastric lavage, weighing the total mass m1 of each litter mouse, and isolating the female mouse from the whole litter mouse after weighing; after 4 hours of isolation, weighing the total mass m2 of each litter; after weighing, the female mice and the mice are kept in the same cage for 1 hour, and then the total mass m3 of the whole litter mice is weighed. Milk secretion per hour for the female mice = m3-m2+ (m 1-m 2)/4, milk secretion per hour for each group of female mice is calculated as an average.

The experimental results are as follows:

1. the status of the groups of mice is shown in Table 2.

TABLE 2 status of groups of mice

2. The results of the lactation amount of each group of female mice per hour are shown in table 3, each experimental group can increase the lactation amount of the female mice with hypogalactia, the lactation level of the female mice in normal lactation period is gradually approached along with the extension of treatment time, and the effects of the experimental groups 1, 2 and 3 are better than those of the experimental groups 4 and 5.

Table 3 average milk yield per hour (unit: g) of the groups of mice

Group of	Control group	Model group	Experiment group 1	Experiment group 2	Experiment group 3	Experiment group 4	Experiment group 5
								Day 1	3.37	2.15	2.38	2.41	2.35	2.24	2.16
Day 2	3.75	2.08	2.65	2.71	2.67	2.43	2.28
								Day 3	4.18	1.93	3.21	3.18	3.15	2.84	2.67
Day 4	4.88	1.95	3.84	3.97	3.86	3.42	3.23
								Day 5	5.38	1.75	4.66	4.74	4.62	4.11	3.87
Day 6	5.94	1.62	5.57	5.62	5.48	4.68	4.32
								Day 7	6.71	1.48	6.38	6.48	6.24	5.28	5.14

The experimental results show that the composition of the application has good lactation promoting effect, and the compositions of examples 4, 5 and 6 have better effect compared with the compositions of comparative examples 1 and 2, so that the raw materials of the composition of the application have synergistic promoting effect.

The above description is only a few embodiments of the present application and is not intended to limit the present application, but various modifications and variations will be apparent to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the protection scope of the present application.

Claims

1. A lactation promoting product composition comprising codonopsis pilosula oligopeptide, which is characterized by comprising codonopsis pilosula oligopeptide, ginseng oligopeptide, longan extract, medlar extract, jujube extract, blueberry extract, purple sweet potato extract, astragalus extract, platycodon root extract, citron extract and allium macrostemon extract;

the codonopsis pilosula oligopeptide is prepared based on the following steps: coarse pulverizing radix Codonopsis Lanceolatae sample, adding 10-15 times of water to obtain homogenate, and placing into enzymeAdding cellulase accounting for 0.5-1.0% of the codonopsis pilosula in the quality of the codonopsis pilosula in a decomposing tank, carrying out enzymolysis for 1-2 hours at 45-50 ℃, and controlling the pH value of the enzyme reaction to be 5.0-6.0; then adding compound protease with 3-5% of the protein mass of codonopsis pilosula, carrying out enzymolysis for 3-4 hours at 50-60 ℃, controlling the pH value of the enzyme reaction to 8.5-9.5, heating to 90 ℃ after the enzymolysis is finished, inactivating enzyme for 10 minutes to obtain the codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, then adopting a ceramic membrane with the molecular weight cut-off of 3000Da to carry out membrane separation, refining to desalt and remove heavy metals, and concentrating until the content of soluble solids accounts for 30-40% of the total mass of the concentrated solution; the mass ratio of the compound protease is as follows: papain neutral proteinase alkaline proteinase= (2-4): (1-2): (4-6); the codonopsis pilosula oligopeptide comprises a peptide segment sequence NH ₂ -Phe-Gly-Arg-Trp-Arg, 90% or more of the oligopeptides of said codonopsis pilosula have a molecular weight of less than 1000Da;

the ginseng oligopeptide is prepared based on the following steps: after coarse crushing treatment of ginseng sample, adding 15-20 times of water to prepare homogenate, placing the homogenate into an enzymolysis tank, adding cellulase with the mass of 0.5-1.0% of ginseng, carrying out enzymolysis for 1-2 hours at 45-50 ℃, controlling the pH value of enzyme reaction to be 5.0-6.0, adding compound protease with the mass of 3-5% of ginseng protein, carrying out enzymolysis for 3-4 hours at 50-55 ℃, controlling the pH value of enzyme reaction to be 9.0-10.0, heating to 90 ℃ after enzymolysis is finished, inactivating enzyme for 10 minutes to obtain a codonopsis pilosula enzymolysis liquid, centrifuging to remove impurities, carrying out membrane separation by adopting a ceramic membrane with the molecular weight cutoff of 3000Da, refining to desalt heavy metals, and concentrating until the content of soluble solids accounts for 30-40% of the total mass of the concentrated solution; the mass ratio of the compound protease is as follows: alkaline protease bromelain flavourzyme= (4-6): (2-4);

the longan extract, the medlar extract, the jujube extract, the blueberry extract, the purple sweet potato extract, the astragalus extract, the platycodon grandiflorum extract, the citron extract and the allium macrostemon extract are prepared based on the following modes: the longan, the medlar, the jujube, the blueberry, the purple sweet potato, the astragalus, the platycodon grandiflorum, the citron and the allium macrostemon are respectively screened, purified, crushed, soaked in water, decocted and filtered to obtain effective extract liquid, and the effective extract liquid is concentrated to obtain extract to obtain corresponding extracts.

2. The lactation promoting product composition of claim 1, wherein the composition is formulated in parts by weight: 50-150 parts of codonopsis pilosula oligopeptide, 5-50 parts of ginseng oligopeptide, 10-80 parts of longan extract, 10-80 parts of medlar extract, 10-80 parts of jujube extract, 10-80 parts of blueberry extract, 10-80 parts of purple sweet potato extract, 10-90 parts of astragalus extract, 5-60 parts of platycodon root extract, 15-50 parts of citron extract and 15-50 parts of allium macrostemon extract.

3. A lactation product composition according to claim 1, characterised in that the source of the codonopsis pilosula oligopeptide is codonopsis pilosula in the Changbai mountain area.

4. The prolapsed product composition according to claim 1, wherein more than 90% of peptides in the ginseng oligopeptide have a molecular weight of less than 1000Da.

5. A method for preparing a lactation promoting composition containing codonopsis pilosula oligopeptide-containing product according to any one of claims 1-4, wherein the codonopsis pilosula oligopeptide-containing product, the concentrated solution of ginseng oligopeptide-containing product and the extracts of longan, medlar, jujube, blueberry, purple sweet potato, astragalus, balloonflower, citron and allium macrostemon are prepared, fully and uniformly mixed and then spray-dried.

6. Use of a lactation promoting product composition comprising codonopsis pilosula oligopeptide of any one of claims 1-4 for the preparation of a lactation promoting food or drug.