Gelsmium elegans extract and application of gelsmium elegans extract in traditional Chinese veterinary medicine oral liquid

Abstract

The invention relates to the technical field of veterinary drugs, and provides a gelsmium elegans extract and application thereof in traditional Chinese veterinary drug oral liquid, a preparation method of the gelsmium elegans extract comprises the following steps: S1, mixing gelsmium elegans with an extraction aid, performing extraction aid, and filtering to obtain filter residues; s2, adding the filter residues into a phosphoric acid aqueous solution, performing first extraction and filtration to obtain a first extracting solution, extracting the first extracting solution with an extracting agent, and concentrating an organic phase to obtain a concentrate; s3, adding an extracting agent into the concentrate, carrying out secondary extraction, filtering to obtain a second extracting solution, adjusting the pH value of the second extracting solution to 5.2-5.4, adsorbing with macroporous resin, eluting with ethanol, concentrating the eluent, and drying to obtain a gelsmium elegans extract; the extraction aid comprises n-pentane; the extraction agent comprises esters; the extracting agent comprises an organic acid aqueous solution. According to the technical scheme, the problems that in the prior art, the yield of the gelsmium elegans extract is low, and the content of medicinal components is low are solved.

Description

Technical Field

[0001] The invention relates to the technical field of veterinary medicines, and in particular to a Gelsemium elegans extract and application thereof in a traditional Chinese veterinary oral liquid. Background Art

[0002] Gelsemium elegans, also known as Gelsemium elegans and pig ginseng, is mainly distributed in Zhejiang, Fujian, Guangdong, Guangxi and other provinces or autonomous regions. It is an evergreen woody vine of the genus Gelsemium in the family Loganaceae ( Gelsemium elegans Benth). Gelsemium alkaloids are both the active and toxic components of Gelsemium, of which the most abundant are collonein and collonein A. The pharmacology of Gelsemium can be divided into three aspects, namely, enhancing immunity, anti-inflammatory and analgesic, and promoting growth. Gelsemium is not only non-toxic to pigs and sheep, but also can increase the gloss of their hair and increase their weight. At present, the extraction methods of Gelsemium extract mainly include water decoction, acid water extraction and alcohol extraction, but the yield is low, the content of active ingredients is low, and the efficacy is unclear. Summary of the invention

[0003] The invention provides a Gelsemium elegans extract and application thereof in a traditional Chinese veterinary oral liquid, which solves the problems of low yield and low content of medicinal components of the Gelsemium elegans extract in the related art.

[0004] The technical solution of the present invention is as follows: The present invention provides a method for preparing a Gelsemium elegans extract, which is characterized by comprising the following steps: S1, mixing Gelsemium elegans with a lifting aid, lifting aid, filtering, and obtaining a filter residue; S2, adding the filter residue into a phosphoric acid aqueous solution, performing a first extraction, filtering, obtaining a first extract, extracting the first extract with an extractant, concentrating the organic phase, and obtaining a concentrate; S3, adding an extractant to the concentrate, performing a second extraction, filtering, and obtaining a second extract, wherein the pH value of the second extract is adjusted to 5.2-5.4, adsorbed by a macroporous resin, eluted with ethanol, concentrated, and dried to obtain a Gelsemium elegans extract; The lifting agent includes n-pentane; The extractant includes esters; The extractant includes an aqueous solution of an organic acid.

[0005] As a further technical solution, the mass volume ratio of Gelsemium elegans to n-pentane in step S1 is 1 g:10~20 mL.

[0006] As a further technical solution, during the assisted lifting, the temperature is 20-30°C and the time is 2-4 hours.

[0007] As a further technical solution, the mass volume ratio of the filter residue to the phosphoric acid aqueous solution in step S2 is 1 g: 20-40 mL.

[0008] As a further technical solution, the mass concentration of the phosphoric acid aqueous solution in step S2 is 0.3%~0.7%.

[0009] As a further technical solution, during the first extraction, the temperature is 80-90° C. and the time is 2-4 hours.

[0010] As a further technical solution, the extractant includes one of propyl acetate, n-butyl acetate, and n-butyl propionate.

[0011] As a further technical solution, the extractant is n-butyl propionate.

[0012] As a further technical solution, the volume ratio of the extract to the extractant is 1:3~5.

[0013] As a further technical solution, the extraction temperature is 20-30°C.

[0014] Butyl propionate is the best extraction agent, which further improves the selectivity of the Gelsemium elegans extract, and further improves the quality of the obtained Gelsemium elegans extract and the content of the active ingredients in the extract.

[0015] As a further technical solution, the organic acid aqueous solution includes one of an acetic acid aqueous solution and a citric acid aqueous solution; The mass concentration of the organic acid aqueous solution is 0.3% to 0.7%; The mass volume ratio of the concentrate to the extractant is 1g:50-70mL.

[0016] As a further technical solution, the extractant further comprises an α-cyclodextrin aqueous solution, and the volume ratio of the α-cyclodextrin aqueous solution to the organic acid aqueous solution is 1:3-5; The mass concentration of the α-cyclodextrin aqueous solution is 1% to 3%.

[0017] The α-cyclodextrin aqueous solution and the organic acid aqueous solution have a synergistic effect, which can further improve the solubility of the Gelsemium elegans extract in water and further improve the yield of the Gelsemium elegans extract.

[0018] As a further technical solution, during the second extraction, the temperature is 20-30°C and the time is 20-40 minutes.

[0019] During the ethanol elution in step S3, the volume fraction of ethanol is 60% to 80%, and the volume of ethanol is equal to 3 to 6 column volumes of the macroporous resin.

[0020] The invention also provides a Gelsemium elegans extract prepared by the preparation method.

[0021] The invention also proposes the application of the Gelsemium elegans extract in a Chinese veterinary oral liquid.

[0022] As a further technical solution, the Chinese veterinary oral liquid includes the following components in parts by weight: 100 parts of water, 6-8 parts of Gelsemium elegans extract, 0.1-0.5 parts of calcium propionate, 0.01-0.05 parts of polyethylene glycol 400, and 0.01-0.05 parts of Tween 80.

[0023] As a further technical solution, the pH value of the Chinese veterinary oral solution is 3.6~4.2.

[0024] The working principle and beneficial effects of the present invention are: In the present invention, the processing sequence of Gelsemium elegans is auxiliary extraction, phosphoric acid aqueous solution extraction, extraction, organic acid aqueous solution extraction, and macroporous resin purification. The auxiliary extraction is first performed to remove impurities in the Gelsemium elegans to prevent the impurities from affecting the subsequent steps, thereby improving the quality of the obtained Gelsemium elegans extract and the content of the active ingredients in the extract; using ester as an extractant improves the selectivity of the Gelsemium elegans extract, thereby improving the quality of the obtained Gelsemium elegans extract and the content of the active ingredients in the extract; using macroporous resin purification effectively removes residual ineffective ingredients such as binders, oligosaccharides, and small molecular peptides, and the content of active ingredients in the Gelsemium elegans extract is higher. DETAILED DESCRIPTION

[0025] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] In the following examples and comparative examples, the macroporous resin is D133 macroporous resin.

[0027] Example 1 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: S1. Mix 100 g of Gelsemium elegans with n-pentane in a mass volume ratio of 1 g:20 mL, extract at 30° C. for 2 h, filter, evaporate the solvent, and obtain a filter residue; S2, adding the filter residue to a phosphoric acid aqueous solution with a mass concentration of 0.5%, the mass volume ratio of the filter residue to the phosphoric acid aqueous solution is 1g:40mL, extracting at 90°C for 2h, filtering to obtain an extract, extracting the extract with propyl acetate at 30°C, the volume ratio of the extract to propyl acetate is 1:5, concentrating the organic phase to obtain a concentrate; S3. Add 0.7% acetic acid aqueous solution to the concentrate, the mass volume ratio of the concentrate to the acetic acid aqueous solution is 1g:70mL, extract at 30°C for 30min, filter to obtain an extract, adjust the pH of the extract to 5.2, adsorb on a macroporous resin, elute with 70% ethanol by volume (the volume of ethanol is equal to 6 column volumes of the macroporous resin), concentrate the eluate, and dry to obtain a Gelsemium elegans extract.

[0028] Example 2 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: S1. Mix 100 g of Gelsemium elegans with n-pentane in a mass volume ratio of 1 g:10 mL, extract at 20° C. for 4 h, filter, evaporate the solvent, and obtain a filter residue; S2, adding the filter residue to a phosphoric acid aqueous solution with a mass concentration of 0.3%, the mass volume ratio of the filter residue to the phosphoric acid aqueous solution is 1g:20mL, extracting at 80°C for 4h, filtering to obtain an extract, extracting the extract with propyl acetate at 20°C, the volume ratio of the extract to propyl acetate is 1:3, concentrating the organic phase to obtain a concentrate; S3. Add 0.3% citric acid aqueous solution to the concentrate, the mass volume ratio of the concentrate to the citric acid aqueous solution is 1g:50mL, extract at 20°C for 40min, filter to obtain an extract, adjust the pH of the extract to 5.4, adsorb on a macroporous resin, elute with 65% ethanol by volume (the volume of ethanol is equal to 3 column volumes of the macroporous resin), concentrate the eluate, and dry to obtain a Gelsemium elegans extract.

[0029] Example 3 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: S1. Mix 100 g of Gelsemium elegans with n-pentane in a mass volume ratio of 1 g:15 mL, extract at 25° C. for 3 h, filter, evaporate the solvent, and obtain a filter residue; S2, adding the filter residue to a phosphoric acid aqueous solution with a mass concentration of 0.7%, the mass volume ratio of the filter residue to the phosphoric acid aqueous solution is 1g:30mL, extracting at 90°C for 3h, filtering to obtain an extract, extracting the extract with propyl acetate at 25°C, the volume ratio of the extract to propyl acetate is 1:4, concentrating the organic phase to obtain a concentrate; S3. Add 0.5% citric acid aqueous solution to the concentrate, the mass volume ratio of the concentrate to the citric acid aqueous solution is 1g:60mL, extract at 25°C for 30min, filter to obtain an extract, adjust the pH of the extract to 5.4, adsorb on a macroporous resin, elute with 80% ethanol by volume (the volume of ethanol is equal to 3 column volumes of the macroporous resin), concentrate the eluate, and dry to obtain a Gelsemium elegans extract.

[0030] Example 4 The only difference between this embodiment and embodiment 3 is that the extractant in this embodiment is replaced by n-butyl acetate.

[0031] Example 5 The only difference between this embodiment and embodiment 3 is that the extractant in this embodiment is replaced by n-butyl propionate.

[0032] Example 6 The only difference between this embodiment and embodiment 5 is that, in step S3 of this embodiment, the 0.5% mass concentration citric acid aqueous solution is replaced by an equal volume of a mixed solution consisting of a 1% mass concentration α-cyclodextrin aqueous solution and a 0.5% mass concentration citric acid aqueous solution, and the volume ratio of the α-cyclodextrin aqueous solution to the phosphoric acid aqueous solution is 1:10.

[0033] Example 7 The only difference between this embodiment and embodiment 6 is that the volume ratio of the α-cyclodextrin aqueous solution to the citric acid aqueous solution in this embodiment is 1:1.

[0034] Example 8 The only difference between this embodiment and embodiment 6 is that the volume ratio of the α-cyclodextrin aqueous solution to the citric acid aqueous solution in this embodiment is 1:3.

[0035] Example 9 The only difference between this embodiment and embodiment 6 is that the volume ratio of the α-cyclodextrin aqueous solution to the citric acid aqueous solution in this embodiment is 1:4.

[0036] Example 10 The only difference between this embodiment and embodiment 6 is that the volume ratio of the α-cyclodextrin aqueous solution to the citric acid aqueous solution in this embodiment is 1:5.

[0037] Embodiment 11 The only difference between this embodiment and embodiment 9 is that the mass concentration of the α-cyclodextrin aqueous solution in this embodiment is 3%.

[0038] Comparative Example 1 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: S1. Add 100 g of Gelsemium elegans to a 0.3% phosphoric acid aqueous solution, the mass volume ratio of Gelsemium elegans to the phosphoric acid aqueous solution is 1 g:20 mL, extract at 80° C. for 4 h, filter to obtain an extract, extract the extract with propyl acetate at 20° C., the volume ratio of the extract to n-butyl propionate is 1:3, concentrate the organic phase to obtain a concentrate; S2. Add 0.3% citric acid aqueous solution to the concentrate, the mass volume ratio of the concentrate to the citric acid aqueous solution is 1g:50mL, extract at 20°C for 40min, filter to obtain an extract, adjust the pH of the extract to 5.4, adsorb on a macroporous resin, elute with 65% ethanol by volume (the volume of ethanol is equal to 3 column volumes of the macroporous resin), concentrate the eluate, and dry to obtain a Gelsemium elegans extract.

[0039] Comparative Example 2 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: S1. Mix 100 g of Gelsemium elegans with n-pentane in a mass volume ratio of 1 g:10 mL, extract at 20° C. for 4 h, filter, evaporate the solvent, and obtain a filter residue; S2, adding the filter residue to a phosphoric acid aqueous solution with a mass concentration of 0.3%, the mass volume ratio of the filter residue to the phosphoric acid aqueous solution is 1g:20mL, extracting at 80°C for 4h, filtering to obtain an extract, and concentrating to obtain a concentrate; S3. Add 0.3% citric acid aqueous solution to the concentrate, the mass volume ratio of the concentrate to the citric acid aqueous solution is 1g:50mL, extract at 20°C for 4h, filter to obtain an extract, adjust the pH value of the extract to 5.4, adsorb on a macroporous resin, elute with 65% ethanol by volume (the volume of ethanol is equal to 3 column volumes of the macroporous resin), concentrate the eluate, and dry to obtain a Gelsemium elegans extract.

[0040] Comparative Example 3 A Gelsemium elegans extract and a preparation method thereof, comprising the following steps: adding 100 g of Gelsemium elegans to a phosphoric acid aqueous solution with a mass concentration of 0.3%, wherein the mass volume ratio of Gelsemium elegans to the phosphoric acid aqueous solution is 1 g:20 mL, extracting at 80° C. for 4 h, filtering to obtain an extract, concentrating, and drying to obtain the Gelsemium elegans extract.

[0041] Experimental Example 1 Using koumine as an indicator, a koumine content detection experiment was performed on the koumine extracts in Examples 1 to 11 and Comparative Examples 1 to 3.

[0042] Detection conditions: Instrument: Wufeng LC-100 liquid chromatography system, mobile phase: methanol-water-0.1wt% ammonia water (volume ratio: 50:50:0.05), flow rate: 1.0mL / min, wavelength: 254nm, column temperature: room temperature.

[0043] The Gelsemium elegans extracts obtained in Examples 1 to 11 and Comparative Examples 1 to 3 were tested, and the test results are shown in Table 1.

[0044]

[0045] Comparison of Comparative Examples 1 to 3 and Example 2 shows that the production sequence of auxiliary extraction, phosphoric acid aqueous solution extraction, extraction, organic acid aqueous solution extraction, and resin purification can significantly improve the quality of the obtained Gelsemium extract and the content of the active ingredients in the extract. Comparison of Example 5 with Examples 3-4 shows that the use of n-butyl propionate as an extractant can further improve the quality of the obtained Gelsemium extract and the content of the active ingredients in the extract. Comparison of Examples 6-11 with Example 5 shows that the α-cyclodextrin aqueous solution can synergize with the phosphoric acid aqueous solution to further improve the quality of the obtained Gelsemium extract and the content of the active ingredients in the extract, and comparison of Examples 8-10 with Examples 6-7 shows that when the volume ratio of the α-cyclodextrin aqueous solution to the organic acid aqueous solution is 1:3~5, the quality of the obtained Gelsemium extract and the content of the active ingredients in the extract can be further improved.

[0046] Experimental Example 2 Ten 58-day-old piglets with similar weight and an average weight of 19.8 kg were selected as experimental subjects and randomly divided into an experimental group and a control group. The control group was fed normally for 7 days. The experimental group was fed with a Chinese veterinary oral liquid containing the Gelsemium elegans extract of Example 2 for 7 days (the weight ratio of the Chinese veterinary oral liquid to the feed was 1.5:100, and the Chinese veterinary oral liquid included the following components by weight: 100 parts of water, 6 parts of Gelsemium elegans extract, 0.1 parts of calcium propionate, 0.01 parts of polyethylene glycol 400, 0.01 parts of Tween 80, and the pH value of the Chinese veterinary oral liquid was 3.9). After 7 days, the average weight gain of the pigs in the experimental group was 3.3 kg / pig, and the feed-to-weight ratio was 1.91; the average weight gain of the pigs in the control group was 2.6 kg / pig, and the feed-to-weight ratio was 2.43. This shows that the Gelsemium elegans extract extracted by the present invention can significantly promote the growth of pigs.

[0047] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a Gelsemium elegans extract, characterized in that: The following steps are involved: S1, mixing Gelsemium elegans with a lifting aid, lifting aid, filtering, and obtaining a filter residue; S2, adding the filter residue into a phosphoric acid aqueous solution, performing a first extraction, filtering, obtaining a first extract, extracting the first extract with an extractant, concentrating the organic phase, and obtaining a concentrate; S3, adding an extractant to the concentrate, performing a second extraction, filtering, and obtaining a second extract, wherein the pH value of the second extract is adjusted to 5.2-5.4, adsorbed by a macroporous resin, eluted with ethanol, concentrated, and dried to obtain a Gelsemium elegans extract; The lifting agent includes n-pentane; The extractant includes esters; The extractant includes an aqueous solution of an organic acid.

2. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The mass volume ratio of Gelsemium elegans to n-pentane in step S1 is 1 g:10-20 mL.

3. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The mass volume ratio of the filter residue in step S2 to the phosphoric acid aqueous solution is 1 g: 20-40 mL.

4. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The mass concentration of the phosphoric acid aqueous solution in step S2 is 0.3% to 0.7%.

5. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The extractant in step S2 includes one of propyl acetate, n-butyl acetate and n-butyl propionate.

6. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The organic acid aqueous solution includes one of an acetic acid aqueous solution and a citric acid aqueous solution; The mass concentration of the organic acid aqueous solution is 0.3% to 0.7%; The mass volume ratio of the concentrate to the extractant is 1g:50-70mL.

7. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: The extractant also includes an α-cyclodextrin aqueous solution, and the volume ratio of the α-cyclodextrin aqueous solution to the organic acid aqueous solution is 1-3:5; The mass concentration of the α-cyclodextrin aqueous solution is 1% to 3%.

8. The method for preparing a Gelsemium elegans extract according to claim 1, characterized in that: During the ethanol elution in step S3, the volume fraction of ethanol is 60-80%, and the volume is equal to 3-6 column volumes of the macroporous resin.

9. A Gelsemium elegans extract, characterized in that Prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the Gelsemium elegans extract according to claim 9 in a Chinese veterinary oral liquid.

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