Production method of rare ginsenoside and feed composition of rare ginsenoside

Abstract

The invention relates to the technical field of production of rare saponin nutritional eggs, in particular to a production method of ginseng rare saponin and a feed composition of the ginseng rare saponin. The production method of the rare ginseng saponin and the feed composition thereof are characterized in that black phoenix laying hens eat the feed composition of rare ginseng saponin rg-3 and rh-2 eggs which are uniformly stirred with water every day, the black phoenix laying hens are fed three times every day, the black phoenix laying hens are fed once in the morning, at noon and at night respectively, the total feeding amount of each laying hen every day is 120 + / -10 g, the illumination time every day is 18 h, and the feed composition of the rare ginseng saponin rg-3 and rh-2 eggs is obtained. The cage opening and stocking time is 8 hours every day. The method has the beneficial effects that an environment convenient for conversion or absorption of the rare ginsenoside rg-3 and rh-2 is provided for poultry bodies, so that the rare ginsenoside rg-3 and rh-2 are transferred into poultry eggs, the human body can conveniently absorb the rare ginsenoside rg-3 and rh-2, and the method is very beneficial to human health.

Description

Technical Field

[0001] The invention relates to the technical field of rare saponin nutritious egg production, and in particular to a production method of rare ginseng saponin and a feed composition thereof. Background Art

[0002] As protein energy synthesis promoting factors, ginseng rare saponins rg-3 and rh-2 act on the G2 phase of the cell reproductive cycle, express in multiple gene networks in the regulation, defense, repair, and metabolism systems, improve energy metabolism, promote nucleic acid and protein synthesis, and regulate the dynamic balance of protein synthesis and degradation (speed), remove harmful metabolites (garbage in the body), inhibit diseased cells, activate and repair damaged and decayed cells, thereby exerting the body's anti-fatigue, anti-aging, anti-lesion and anti-cancer effects, and deeply mobilize the body's vitality. Ginseng rare saponins rg-3 and rh-2 can inhibit tumor angiogenesis by inhibiting the formation of endothelial cells, the activity of VEGF and the expression of VEGF2, and effectively block the channels for tumors to absorb nutrients and spread and metastasize. The content of ginsenoside rg-3 in naturally grown ginseng is extremely low, and currently it is mainly achieved through chemical, enzymatic and microbial methods. The rich ginsenosides (Rb-1, Rb-2, Rc, Rd, Re, Rg-1, Rf, etc.) in ginseng are converted into rare ginsenosides with higher medicinal value and stronger intestinal absorption function. However, the price is too high for ordinary people to afford, so it cannot be promoted on a large scale.

[0003] Functional eggs are eggs that not only have the nutrition of ordinary eggs but also have other functions that are beneficial to human health or regulate human vital activities. By adjusting the trace elements, vitamins, unsaturated fatty acids and the components of different feed compositions, and then adding the amount and feeding time to the laying hen feed, eggs enriched with functional substances higher than normal values ​​can be obtained.

[0004] There is an urgent need in the market for rare ginsenosides or materials containing rare ginsenosides that are easily popularized and low in cost and that are easy for the human body to absorb and utilize. Summary of the invention

[0005] In order to achieve the above object, the present invention aims to provide a method for producing rare ginseng saponins and a feed composition thereof.

[0006] A method for producing rare ginsenosides of ginseng and a feed composition thereof, characterized in that: the feeding method for laying hens is to let the laying hens eat a feed composition containing ginseng extract residues that is evenly stirred with drinking water every day, feeding them three times a day, once in the morning, noon and evening, the total amount of feed for each laying hen per day is 120±10 grams, the daily lighting time is 18 hours, and the daily cage-opening and free-range time is 8 hours.

[0007] The laying hen is a black phoenix chicken.

[0008] The feeding time interval is 5h.

[0009] The illumination time is 5h in the morning, 5h in the afternoon and 8h in the evening.

[0010] The daily cage-opening and free-range feeding time is 5 hours in the morning and 3 hours in the afternoon.

[0011] The feed composition containing ginseng extract residue is characterized by comprising the following ingredients by weight: 40-50 parts of soybean meal, 29-45 parts of corn flour, 15-30 parts of broken rice, 10-15 parts of bran, 6-18 parts of ginseng extract residue, 0.3-1 parts of lactic acid bacteria fermentation powder, 0.2-0.8 parts of yeast fermentation powder, 3-7 parts of glucose, 0-5 parts of litsea cubeba essential oil inclusion compound, 0-5 parts of star anise essential oil inclusion compound, and 1-5 parts of fine sand.

[0012] The ginsenoside residue in the ginseng extraction residue contains 0.015-0.036 g / kg of ginsenosides, and includes ginsenoside Rb-11, ginsenoside Rb-2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg-11, ginsenoside Rf, carbohydrates, ginseng polysaccharides, cellulose, vitamins, amino acids, crude protein, crude fat and other organic substances, as well as trace elements and other components.

[0013] The ginseng extract residue is prepared by crushing ginseng into 10 meshes, extracting ginseng three times, each time for 3 hours, filtering, and drying the ginseng extract residue.

[0014] The particle size of the fine sand is 0.2-0.5 mm.

[0015] Furthermore, the feed composition containing ginseng extract residue can also be added with a litsea cubeba essential oil inclusion compound, which can inhibit bacteria and resist oxidation. The preparation method of the litsea cubeba essential oil inclusion compound is as follows: 98 parts of β-cyclodextrin are added with 60 parts of 80-85°C geothermal water to fully wet and stir evenly, and then 2 parts of litsea cubeba essential oil are slowly added while stirring. After the addition is completed, continue stirring for 1 hour, and vacuum dry at 50-60°C for 10 hours to obtain the litsea cubeba essential oil inclusion compound.

[0016] Furthermore, the feed composition containing ginseng extract residue can also be added with anise essential oil inclusion compound, which can inhibit bacteria and resist oxidation. The preparation method of the anise essential oil inclusion compound is as follows: 98 parts of β-cyclodextrin are added with 60 parts of 80-85°C geothermal water to fully wet and stir evenly, then 2 parts of anise essential oil are slowly added while stirring, and after the addition is completed, stirring is continued for 1 hour, and vacuum drying is carried out at 50-60°C for 10 hours to obtain the anise essential oil inclusion compound.

[0017] Furthermore, when feeding the feed composition containing ginseng extract residue, the composition should be stirred evenly with water, and the feeding state of the feed composition is sticky flocculent.

[0018] Furthermore, as a preferred option, the feed composition containing ginseng extract residue can be further crushed, stirred, granulated and pressed into rice-sized feed composition particles for easy feeding.

[0019] Compared with the prior art, the beneficial effect of the invention is that the feed composition for producing rare ginsenosides of ginseng can provide a convenient environment for the transformation or absorption of rare ginsenosides RG-3 and RH-2 in the poultry body through reasonable nutritional matching, improving the gastrointestinal environment of the poultry body and improving the immunity of the poultry, so that the rare ginsenosides RG-3 and RH-2 in the poultry body can be transferred to the poultry eggs. This method is easy to popularize and apply the rare ginsenosides RG-3 and RH-2 in different types of poultry. At the same time, due to the appearance of rare ginsenosides RG-3 and RH-2 in poultry eggs, it is convenient for people to obtain rare ginsenosides RG-3 and RH-2 at a low cost, and it is also convenient for the human body to absorb the rare ginsenosides RG-3 and RH-2, which is very beneficial to human health. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The chromatogram of ginsenoside rg-3 by LC-MS of egg protein containing ginsenoside rg-3 in Example 1 of the present invention is shown;

[0021] Figure 2 The chromatogram of ginsenoside rg-3 contained in egg yolk by LC-MS / MS of ginsenoside rg-3 in Example 1 of the present invention;

[0022] Figure 3 The chromatogram of ginsenoside rg-3 by LC-MS of egg protein containing ginsenoside rg-3 in Example 2 of the present invention is shown;

[0023] Figure 4 The figure is a chromatogram of ginseng rare saponin rg-3 obtained by LC-MS of egg yolk containing ginseng rare saponin rg-3 according to Example 2 of the present invention;

[0024] Figure 5 The chromatogram of ginsenoside rg-3 by LC-MS of egg protein containing ginsenoside rg-3 in Example 3 of the present invention is shown;

[0025] Figure 6 The chromatogram of ginsenoside rg-3 contained in egg yolk by LC-MS / MS of ginsenoside rg-3 in Example 3 of the present invention;

[0026] Figure 7The chromatogram of ginsenoside rg-3 by LC-MS of egg protein containing ginsenoside rg-3 in Example 4 of the present invention is shown;

[0027] Figure 8 This is a chromatogram of ginsenoside rg-3 in egg yolk containing ginsenoside rg-3 by LC-MS / MS analysis in Example 4 of the present invention;

[0028] Fig. 9 The chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg protein in Example 1 of the present invention is shown;

[0029] Fig.10 The chromatogram of ginsenoside rh-2 contained in egg yolk containing ginsenoside rh-2 by LC-MS / MS in Example 1 of the present invention;

[0030] Fig.11 The chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg protein in Example 2 of the present invention;

[0031] Fig.12 This is a chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg yolk in Example 2 of the present invention;

[0032] Fig.13 The chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg protein in Example 3 of the present invention is shown;

[0033] Fig.14 This is a chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg yolk in Example 3 of the present invention;

[0034] Fig.15 The chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg protein in Example 4 of the present invention is shown;

[0035] Fig.16 This is a chromatogram of ginsenoside rh-2 containing ginsenoside rh-2 by LC-MS of egg yolk in Example 4 of the present invention;

[0036] Fig.17 The chromatogram of ginseng rare saponin rg-3 obtained by HPLC-MS / MS analysis of the ginseng extract residue of the present invention in which no ginseng rare saponin rg-3 was detected;

[0037] Fig.18 The chromatogram of ginsenoside rh-2 of ginsenoside rh-2 by liquid chromatography-mass spectrometry was not detected in the ginseng extract residue of the present invention. DETAILED DESCRIPTION

[0038] In order to make the purpose, technical solution and beneficial effects of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the embodiments described in this specification are only for explaining the present invention, not for limiting the present invention.

[0039] Example 1

[0040] The production method of rare ginseng saponins and feed composition in this example are carried out as follows:

[0041] Every day, the black phoenix laying hens are fed with a feed composition of ginseng extract residue containing 40 parts of soybean meal, 30 parts of corn flour, 16 parts of broken rice, 1 part of bran, 6 parts of ginseng extract residue, 0.3 parts of lactic acid bacteria fermentation powder, 0.2 parts of yeast fermentation powder, 3 parts of glucose, 0.5 parts of Litsea cubeba essential oil inclusion compound, 1 part of star anise essential oil inclusion compound, and 2 parts of fine sand, which is evenly mixed with water. The feed is fed 3 times a day, once in the morning, noon and evening. The total feed amount for each laying hen is 120±10 grams per day. The daily lighting time is 5h in the morning, 5h in the afternoon and 8h in the evening. The daily cage-opening and free-range time is 5h in the morning and 3h in the afternoon.

[0042] After laying eggs in this case, the eggs containing the rare ginsenosides rg-3 and rh-2 were tested for rg-3 using the Shimadzu LC+ABSCI EX 4500 liquid chromatography-mass spectrometry system. The test data are shown in Table 1 below. The detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg protein and the detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg yolk are shown in the attached table. Figure 1 , Attachment Figure 2 , Attachment Fig. 9 , Attachment Fig.10 shown.

[0043] Table 1. Detection data of ginsenosides RG-3 and RH-2 in eggs

[0044]

[0045]

[0046] Example 2

[0047] The production method of rare ginseng saponins and feed composition in this example are carried out as follows:

[0048] Every day, the black phoenix laying hens are fed with a feed composition of ginseng extract residue containing 45 parts of soybean meal, 35 parts of corn flour, 20 parts of broken rice, 12 parts of bran, 12 parts of ginseng extract residue, 0.5 parts of lactic acid bacteria fermentation powder, 0.4 parts of yeast fermentation powder, 5 parts of glucose, 2 parts of Litsea cubeba essential oil inclusion compound, 2 parts of star anise essential oil inclusion compound, and 3 parts of fine sand, which is evenly mixed with water. The feed is fed 3 times a day, once in the morning, noon and evening. The total feed amount for each laying hen is 120±10 grams per day. The daily lighting time is 5h in the morning, 5h in the afternoon and 8h in the evening. The daily cage-opening and free-range time is 5h in the morning and 3h in the afternoon.

[0049] After laying eggs in this example, the eggs containing the rare ginsenosides rg-3 and rh-2 in this example were tested for rg-3 using the liquid chromatography-mass spectrometry system Shimadzu LC+ABSCIEX 4500. The test data are shown in Table 2 below. The detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg protein and the detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg yolk are shown in the attached table. Figure 3 , Attachment Figure 4 , Attachment Fig.11 , Attachment Fig.12 shown.

[0050] Table 2. Detection data of ginsenosides RG-3 and RH-2 in eggs

[0051] Eggs in this case Protein (peak area) Egg yolk(peak area) Protein (content%) Egg yolk (content%) <![CDATA[2 # rg-3(S)]]> 189073.944 207022.448 0.000013654 0.000030232 <![CDATA[2 # rg-3(R)]]> 471795.881 2224354.186 0.000039805 0.000379493 <![CDATA[2 # rh-2(S)]]> 8226.819 4076.597 0.000051638 0.000047922 <![CDATA[2 # rh-2(R)]]> 7091.986 1326.788 0.000119035 0.000041707

[0052] Example 3

[0053] The production method of rare ginseng saponins and feed composition in this example are carried out as follows:

[0054] Every day, the black phoenix laying hens are fed with a feed composition of ginseng extract residue containing 47 parts of soybean meal, 39 parts of corn flour, 25 parts of broken rice, 13 parts of bran, 10 parts of ginseng extract residue, 0.6 parts of lactic acid bacteria fermentation powder, 0.3 parts of yeast fermentation powder, 6 parts of glucose, 3 parts of star anise essential oil inclusion compound, and 2 parts of fine sand, which is evenly mixed with water. The feed is fed 3 times a day, once in the morning, noon and evening. The total feed amount for each laying hen is 120±10 grams per day. The daily lighting time is 5h in the morning, 5h in the afternoon, and 8h in the evening. The daily cage-opening and free-range time is 5h in the morning and 3h in the afternoon.

[0055] After laying eggs in this example, the eggs containing the rare ginsenosides rg-3 and rh-2 in this example were tested for rg-3 using the liquid chromatography-mass spectrometry system Shimadzu LC+ABSCIEX 4500. The test data are shown in Table 3 below. The detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg protein and the detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg yolk are shown in the attached table. Figure 5 , Attachment Figure 6 , Attachment Fig.13 , Attachment Fig.14 shown.

[0056] Table 3. Detection data of ginsenosides RG-3 and RH-2 in eggs

[0057] Eggs in this case Protein (peak area) Egg yolk(peak area) Protein (content%) Egg yolk (content%) <![CDATA[3 # rg-3(S)]]> 141341.146 20277.989 0.000009909 0.000002843 <![CDATA[3 # rg-3(R)]]> 122921.951 10449.103 0.000010068 0.000001711 <![CDATA[3 # rh-2(S)]]> 28002.715 1724.238 0.000170514 0.000020994 <![CDATA[3 # rh-2(R)]]> 20095.338 738.653 0.000327209 0.000024050

[0058] Example 4

[0059] The production method of rare ginseng saponins and feed composition in this example are carried out as follows:

[0060] Every day, the black phoenix laying hens are fed with a feed composition of ginseng extract residue containing 50 parts of soybean meal, 45 parts of corn flour, 30 parts of broken rice, 15 parts of bran, 18 parts of ginseng extract residue, 1 part of lactic acid bacteria fermentation powder, 0.8 parts of yeast fermentation powder, 7 parts of glucose, 5 parts of Litsea cubeba essential oil inclusion compound, 4 parts of star anise essential oil inclusion compound, and 5 parts of fine sand, which is evenly mixed with water. The feed is fed 3 times a day, once in the morning, noon and evening. The total feed amount for each laying hen is 120±10 grams per day. The daily lighting time is 5h in the morning, 5h in the afternoon and 8h in the evening. The daily cage-opening and free-range time is 5h in the morning and 3h in the afternoon.

[0061] After laying eggs in this case, the eggs containing the rare ginsenosides rg-3 and rh-2 were tested for rg-3 using the Shimadzu LC+ABSCI EX 4500 liquid chromatography-mass spectrometry system. The test data are shown in Table 4 below. The detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg protein and the detection spectra of the rare ginsenosides rg-3 and rh-2 in the egg yolk are shown in the attached table. Figure 7 , Attachment Figure 8 , Attachment Fig.15 , Attachment Fig.16 shown.

[0062] Table 4. Detection data of ginsenosides RG-3 and RH-2 in eggs

[0063] Eggs in this case Protein (peak area) Egg yolk(peak area) Protein (content%) Egg yolk (content%) <![CDATA[4 # rg-3(S)]]> 323613.756 10633.893 0.000023176 0.000001536 <![CDATA[4 # rg-3(R)]]> 3287171.832 14331.342 0.000275034 0.000002419 <![CDATA[4 # rh-2(S)]]> 6028.178 1222.61 0.000037497 0.000015341 <![CDATA[4 # rh-2(R)]]> 4235.617 816.66 0.000070453 0.000027401

[0064] The detection method used for detecting the rare ginsenosides RG-3 and RH-2 in the ginseng eggs and ginseng extraction residues produced in the above Examples 1-4 is:

[0065] (1) Saponin extraction

[0066] Boil the eggs; weigh the egg yolk (5g) and egg white (10g) respectively; put them into a blender, add 50ml of 85% ethanol, and stir well. Add 50mL of 85% ethanol to clean the blender in batches, and ultrasonically treat the collected liquid for 30min; let it stand at room temperature. Shake well, filter with a 0.45μm organic filter membrane, take 2mL of the sample and blow dry with nitrogen, then re-dissolve with 2mL of 80% methanol, and centrifuge again (6K); take 1mL of the supernatant for use in the machine.

[0067] (2) Instrument parameters

[0068] The data acquisition instrument system consists of Shimadzu LC+ABSCI EX 4500 liquid chromatography-mass spectrometry system.

[0069] (3) Liquid chromatography parameters

[0070] ABSCIEX 4500 was used with an Agile ent C18 column (2.1x100mm 1.8-M i cron);

[0071] Detection of rare ginsenosides rg-3 and rh-2;

[0072] Mobile phase A was 10 mM ammonium acetate (pH adjusted to 5.0 with acetic acid), and mobile phase B was acetonitrile. Gradient elution was performed with an injection volume of 5 μL.

[0073] The gradient program was set as:

[0074]

[0075]

[0076] (4) Mass spectrometry conditions

[0077] The ABSCI EX 4500 mass spectrometry system was used to collect the primary and secondary spectra of the samples. The conditions were as follows: electrospray ionization source (ESI), negative ion scanning, multiple reaction monitoring (MRM), ion source temperature: 500°C, electrospray voltage: -4500V, curtain gas: 30.0psi, ion source gas 1: 50psi, ion source gas 2: 50psi.

[0078] (5) Linear calculation

[0079] Saponin Linear range (ppb) Standard curve r2 Rare saponins 0.5-50 y=2.33649*105x+3.08712*104 0.99728

[0080] calculate

[0081]

[0082] R——the content of rare saponins in ginseng in ng / g;

[0083] A——Chromatogram peak area of ​​the substance being tested in the solution;

[0084] AS——mass chromatogram peak area of ​​the substance being tested in the standard solution;

[0085] CS——mass concentration value of the substance being measured in the standard solution, in ng / mL;

[0086] V——The value of the volume of the extract, in milliliters (mL);

[0087] W is the numerical value of the sample mass, in grams (g).

[0088] The specific implementation modes of the present invention described above should be understood as explaining the technical solutions of the present invention, and do not constitute limitations on the protection scope of the present invention.

Claims

1. A method for producing rare ginsenosides, characterized in that: The feeding method for laying hens is to let the laying hens eat a feed composition containing ginseng extract residue evenly mixed with water every day, including the following ingredients by weight: 40-50 parts of soybean meal, 29-45 parts of corn flour, 15-30 parts of broken rice, 10-15 parts of bran, 6-18 parts of ginseng extract residue, 0.3-1 parts of lactic acid bacteria fermentation powder, 0.2-0.8 parts of yeast fermentation powder, 3-7 parts of glucose, 0-5 parts of Litsea cubeba essential oil inclusion compound, 0-5 parts of star anise essential oil inclusion compound, and 1-5 parts of fine sand. The feed is fed three times a day, once in the morning, noon and evening, the total feed amount for each laying hen is 120±10 grams per day, the daily lighting time is 18 hours, and the daily cage-opening and free-range time is 8 hours.

2. The method for producing rare ginsenosides according to claim 1, characterized in that The feed composition containing ginseng extract residue comprises the following ingredients by weight: 40-50 parts of soybean meal, 29-45 parts of corn flour, 15-30 parts of broken rice, 10-15 parts of bran, 6-18 parts of ginseng extract residue, 0.3-1 parts of lactic acid bacteria fermentation powder, 0.2-0.8 parts of yeast fermentation powder, 3-7 parts of glucose, 0-5 parts of litsea cubeba essential oil inclusion compound, 0-5 parts of star anise essential oil inclusion compound and 1-5 parts of fine sand.

3. The method for producing rare ginsenosides according to claim 1, characterized in that The feeding time interval is 5h.

4. The method for producing rare ginsenosides according to claim 1, characterized in that The illumination time is 5h in the morning, 5h in the afternoon and 8h in the evening.

5. The method for producing rare ginsenosides according to claim 1, characterized in that The daily cage-opening and free-range feeding time is 5 hours in the morning and 3 hours in the afternoon.

6. The method for producing rare ginsenosides according to claim 1 or 2, characterized in that The ginsenoside residue in the ginseng extraction residue contains 0.015-0.036 g / kg of ginsenosides, and includes ginsenoside Rb-1, ginsenoside Rb-2, ginsenoside Rc, ginsenoside Rd, ginsenoside Re, ginsenoside Rg-1, ginsenoside Rf, carbohydrates, ginseng polysaccharides, cellulose, vitamins, amino acids, crude protein, crude fat and other organic substances, as well as trace elements and other components.

7. The method for producing rare ginsenosides according to claim 1 or 2, characterized in that The ginseng extract residue is prepared by crushing ginseng into 10 meshes, extracting ginseng three times, each time for 3 hours, filtering, and drying the ginseng extract residue.

8. The method for producing rare ginsenosides according to claim 1 or 2, characterized in that The particle size of the fine sand is 0.2-0.5 mm.

9. The method for producing rare ginsenosides according to claim 1 or 2, characterized in that When feeding the feed composition containing ginseng extract residue, the composition should be stirred evenly with water, and the feeding state of the feed composition is sticky flocculent.

10. The method for producing rare ginsenosides according to claim 1 or 2, characterized in that The feed composition containing ginseng extract residue can also be crushed, stirred, granulated and pressed into rice-sized feed composition particles.

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