CN113545420A - Preparation and application of codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material - Google Patents
Preparation and application of codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material Download PDFInfo
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- CN113545420A CN113545420A CN202110855976.7A CN202110855976A CN113545420A CN 113545420 A CN113545420 A CN 113545420A CN 202110855976 A CN202110855976 A CN 202110855976A CN 113545420 A CN113545420 A CN 113545420A
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- attapulgite
- codonopsis pilosula
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/28—Silicates, e.g. perlites, zeolites or bentonites
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Abstract
The invention provides a preparation method of a codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material, which comprises the steps of firstly performing crude extraction on codonopsis pilosula by adopting a conventional graded alcohol extraction and water precipitation mode, and then performing graded precipitation on codonopsis pilosula polysaccharide by using an ethanol solution to obtain codonopsis pilosula polysaccharide; dispersing the purified attapulgite in distilled water to obtain an attapulgite dispersion liquid, dissolving the codonopsis pilosula polysaccharide in distilled water, mixing the obtained solution with the attapulgite dispersion liquid, stirring the obtained solution at normal temperature for 1-5 hours, and adding an ethanol solution to obtain the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material. The invention adopts a gel synthesis technology to prepare the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material, improves the antibacterial adsorption performance of attapulgite, has antibacterial stability and biocompatibility, has strong antibacterial effect on escherichia coli and staphylococcus aureus, and can be used as a natural antibacterial feed additive.
Description
Technical Field
The invention belongs to the technical field of composite materials and plant source feed additives, and relates to a preparation method of a codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material.
Background
In recent years, research on plant-derived antibacterial agents has been greatly advanced at home and abroad. At present, about 2000 plants can be used for extracting and preparing the antibacterial agent. The substances with plant-derived antibacterial activity mainly comprise terpenes, alkaloids, flavonoids, phenols, quinones, aldehydes, alcohols, phenylpropanoids, spices, lignans, steroids, organic acids, essential oils and the like.
The codonopsis pilosula has the main effects of tonifying middle-jiao and Qi, strengthening spleen and benefiting lung. It can be used for treating spleen and lung weakness, short breath, palpitation, internal heat, and diabetes. Modern pharmacological experiments show that the codonopsis pilosula polysaccharide is one of the main components of codonopsis pilosula, and has various pharmacological activities of removing free radicals, regulating the immunity of the organism, resisting inflammation, gastric ulcer, tumor and the like.
The gelation property is an important aspect of the biological function of polysaccharide macromolecules, and in various processing industries, the gelation property not only can obviously influence the texture and the sensory characteristics of a final product, but also can improve the instability, the biological activity and the bioavailability of functional components of the product.
The attapulgite is a natural green nano antibacterial material, and has a special pore structure, stability and biocompatibility. In 2019, the rural agricultural department clearly provides that from 7 and 1 month in 2020, feed production enterprises stop producing commercial feeds containing growth-promoting drug feed additives (except traditional Chinese medicines). The traditional Chinese medicine replaces the antibacterial era and provides opportunities and challenges for the development of the traditional Chinese medicine in the antibacterial field. The gel prepared by compounding the codonopsis pilosula polysaccharide and the attapulgite is used as an additive auxiliary material to feed matrixes, and is also an antibacterial feed with a very promising prospect.
Disclosure of Invention
The invention aims to provide a preparation method of a codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material, which adopts a gel synthesis technology, not only fully applies the advantages of easy degradation and environmental protection of natural product codonopsis pilosula polysaccharide, but also fully utilizes the adsorbability and biocompatibility of attapulgite to complement the advantages of the codonopsis pilosula polysaccharide and the attapulgite, completes the preparation of the codonopsis pilosula polysaccharide @ attapulgite gel material, and provides auxiliary materials for the development of natural antibacterial feed additives.
The invention relates to a preparation method of a codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material, which comprises the following steps:
(1) extracting radix Codonopsis with conventional fractional alcohol and water precipitation method, removing glycoprotein with trichloroacetic acid precipitation method, dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing, and vacuum freeze drying to obtain radix Codonopsis total polysaccharide; dissolving the total polysaccharide of the codonopsis pilosula in distilled water, carrying out fractional precipitation by using 30-80% ethanol solution, and carrying out vacuum freeze drying to obtain codonopsis pilosula polysaccharide;
(2) dispersing the purified attapulgite in distilled water to obtain an attapulgite dispersion liquid, dissolving the codonopsis pilosula polysaccharide in the distilled water, mixing the obtained solution with the attapulgite dispersion liquid, stirring for 1-5 hours at normal temperature, and adding a 20-80% ethanol solution to obtain the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material. Wherein the mass ratio of the purified attapulgite to the codonopsis pilosula polysaccharide is 1: 10-1: 15.
The purification steps of the attapulgite are as follows: taking attapulgite clay which is crushed and sieved by a sieve of 100-400 meshes, adding water for soaking to prepare turbid liquid, dispersing and pulping at a high speed, standing for settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying for later use.
Compared with the method for simply using attapulgite, the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material provided by the invention has the advantages that the antibacterial effect of escherichia coli and staphylococcus aureus is obviously enhanced, the minimum antibacterial concentration is lower than that of the attapulgite, the MIC =1.5mg/mL can be achieved, and the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material can be used as a feed additive after being freeze-dried.
In conclusion, the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material is prepared by adopting a gel synthesis technology, the antibacterial adsorption performance of attapulgite is improved, the material has antibacterial stability and biocompatibility, has a strong antibacterial effect on escherichia coli and staphylococcus aureus, and can be used as a natural antibacterial feed additive.
Detailed Description
The preparation and antibacterial performance of the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material are further explained by the following specific embodiments.
Example 1
(1) Crude extraction is carried out on the codonopsis pilosula by adopting a conventional grading alcohol extraction and water precipitation mode to obtain crude polysaccharide, and glycoprotein in the crude polysaccharide is removed by a trichloroacetic acid precipitation method. Dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing for 2d, removing residual ethanol and TCA small molecular substance (3500 Da), and vacuum freeze drying the retained solution to obtain radix Codonopsis total polysaccharide. Dissolving radix Codonopsis total polysaccharide in distilled water, performing fractional precipitation according to different final concentrations of ethanol, precipitating with 30% ethanol, vacuum freeze drying the precipitate to obtain radix Codonopsis polysaccharide, and freeze storing.
(2) Soaking crushed attapulgite clay which is sieved by a 200-mesh sieve in water to prepare turbid liquid, dispersing and pulping at a high speed, standing and settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying to obtain the purified attapulgite. Dissolving 0.1g of purified attapulgite in 10 mL of distilled water to obtain attapulgite dispersion, dissolving 1.0 g of prepared codonopsis pilosula polysaccharide in 10 mL of distilled water, mixing the prepared codonopsis pilosula polysaccharide with the attapulgite dispersion, stirring at normal temperature for 1 hour, adding 20% ethanol solution, and reaching the critical gel concentration to form codonopsis pilosula polysaccharide @ attapulgite gel. The gel was freeze dried and dissolved and the inhibitory experiment showed a MIC of 1.5 mg/mL.
Example 2
(1) Crude extraction is carried out on the codonopsis pilosula by adopting a conventional grading alcohol extraction and water precipitation mode to obtain crude polysaccharide, and glycoprotein in the crude polysaccharide is removed by a trichloroacetic acid precipitation method. Dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing for 2d, removing residual ethanol and TCA small molecular substance (3500 Da), and vacuum freeze drying the retained solution to obtain radix Codonopsis total polysaccharide. Dissolving radix Codonopsis total polysaccharide in distilled water, performing fractional precipitation according to different final concentrations of ethanol, precipitating with 40% ethanol, vacuum freeze drying the precipitate to obtain radix Codonopsis polysaccharide, and freezing and storing.
(2) Soaking crushed attapulgite clay which is sieved by a 200-mesh sieve in water to prepare turbid liquid, dispersing and pulping at a high speed, standing and settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying to obtain the purified attapulgite. Dissolving 0.1g of purified attapulgite in 10 mL of distilled water to obtain attapulgite dispersion, dissolving 1.2 g of prepared codonopsis pilosula polysaccharide in 10 mL of distilled water, mixing with the attapulgite dispersion, stirring at normal temperature for 2 hours, adding 50% ethanol solution, and obtaining codonopsis pilosula polysaccharide @ attapulgite gel after reaching the critical gel concentration. The gel was freeze dried and dissolved and the inhibitory experiment showed a MIC of 1.5 mg/mL.
Example 3
(1) Crude extraction is carried out on the codonopsis pilosula by adopting a conventional grading alcohol extraction and water precipitation mode to obtain crude polysaccharide, and glycoprotein in the crude polysaccharide is removed by a trichloroacetic acid precipitation method. Dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing for 2d, removing residual ethanol and TCA small molecular substance (3500 Da), and vacuum freeze drying the retained solution to obtain radix Codonopsis total polysaccharide. Dissolving radix Codonopsis total polysaccharide in distilled water, performing fractional precipitation according to different final concentrations of ethanol, precipitating with 60% ethanol, vacuum freeze drying the precipitate to obtain radix Codonopsis polysaccharide, and freeze storing.
(2) Soaking crushed attapulgite clay which is sieved by a 200-mesh sieve in water to prepare turbid liquid, dispersing and pulping at a high speed, standing and settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying to obtain the purified attapulgite. Dissolving 0.1g of purified attapulgite in 10 mL of distilled water to obtain attapulgite dispersion, dissolving 1.2 g of prepared codonopsis pilosula polysaccharide in 10 mL of distilled water, mixing with the attapulgite dispersion, stirring at normal temperature for 4 hours, adding 60% ethanol solution, and obtaining codonopsis pilosula polysaccharide @ attapulgite gel after reaching the critical gel concentration. The gel was freeze dried and dissolved and the inhibitory experiment showed a MIC of 1.5 mg/mL.
Example 4
(1) Crude extraction is carried out on the codonopsis pilosula by adopting a conventional grading alcohol extraction and water precipitation mode to obtain crude polysaccharide, and glycoprotein in the crude polysaccharide is removed by a trichloroacetic acid precipitation method. Dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing for 2d, removing residual ethanol and TCA small molecular substance (3500 Da), and vacuum freeze drying the retained solution to obtain radix Codonopsis total polysaccharide. Dissolving radix Codonopsis total polysaccharide in distilled water, performing fractional precipitation according to different final concentrations of ethanol, precipitating with 80% ethanol, vacuum freeze drying the precipitate to obtain radix Codonopsis polysaccharide, and freeze storing.
(2) Soaking crushed attapulgite clay which is sieved by a 200-mesh sieve in water to prepare turbid liquid, dispersing and pulping at a high speed, standing and settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying to obtain the purified attapulgite. Dissolving 0.1g of purified attapulgite in 10 mL of distilled water to obtain attapulgite dispersion, dissolving 1.5 g of prepared codonopsis pilosula polysaccharide in 10 mL of distilled water, mixing with the attapulgite dispersion, stirring at normal temperature for 4 hours, adding 80% ethanol solution, and obtaining codonopsis pilosula polysaccharide @ attapulgite gel after reaching the critical gel concentration. The gel was freeze dried and dissolved and the inhibitory experiment showed a MIC of 1.5 mg/mL.
Example 5
(1) Crude extraction is carried out on the codonopsis pilosula by adopting a conventional grading alcohol extraction and water precipitation mode to obtain crude polysaccharide, and glycoprotein in the crude polysaccharide is removed by a trichloroacetic acid precipitation method. Dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing for 2d, removing residual ethanol and TCA small molecular substance (3500 Da), and vacuum freeze drying the retained solution to obtain radix Codonopsis total polysaccharide. Dissolving radix Codonopsis total polysaccharide in distilled water, performing fractional precipitation according to different final concentrations of ethanol, precipitating with 80% ethanol, vacuum freeze drying the precipitate to obtain radix Codonopsis polysaccharide, and freeze storing.
(2) Soaking crushed attapulgite clay which is sieved by a 200-mesh sieve in water to prepare turbid liquid, dispersing and pulping at a high speed, standing and settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying to obtain the purified attapulgite. Dissolving 0.1g of purified attapulgite in 10 mL of distilled water to obtain attapulgite dispersion, dissolving 1.5 g of prepared codonopsis pilosula polysaccharide in 10 mL of distilled water, mixing with the attapulgite dispersion, stirring at normal temperature for 5 hours, adding 80% ethanol solution, and obtaining codonopsis pilosula polysaccharide @ attapulgite gel after reaching the critical gel concentration. The gel was freeze dried and dissolved and the inhibitory experiment showed a MIC of 1.5 mg/mL.
Claims (5)
1. A preparation method of codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material comprises the following steps:
(1) extracting radix Codonopsis with conventional fractional alcohol and water precipitation method, removing glycoprotein with trichloroacetic acid precipitation method, dissolving the obtained crude extract in distilled water, concentrating the supernatant, dialyzing, and vacuum freeze drying to obtain radix Codonopsis total polysaccharide; dissolving the total polysaccharide of the codonopsis pilosula in distilled water, carrying out fractional precipitation by using 30-80% ethanol solution, and carrying out vacuum freeze drying to obtain codonopsis pilosula polysaccharide;
(2) dispersing the purified attapulgite in distilled water to obtain an attapulgite dispersion liquid, dissolving the codonopsis pilosula polysaccharide in distilled water, mixing the obtained solution with the attapulgite dispersion liquid, stirring the obtained solution at normal temperature for 1-5 hours, and adding an ethanol solution to obtain the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material.
2. The preparation method of the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material according to claim 1, is characterized in that: in the step (2), the mass ratio of the purified attapulgite to the codonopsis pilosula polysaccharide is 1: 10-1: 15.
3. The preparation method of the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material according to claim 1, is characterized in that: in the step (2), the concentration of the ethanol solution is 20-80%.
4. The preparation method of the codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material according to claim 1, is characterized in that: in the step (2), the purification steps of the attapulgite are as follows: taking attapulgite clay which is crushed and sieved by a sieve of 100-400 meshes, adding water for soaking to prepare turbid liquid, dispersing and pulping at a high speed, standing for settling, centrifuging, removing supernatant and non-clay impurities at the lower layer, and drying for later use.
5. The codonopsis pilosula polysaccharide @ attapulgite gel antibacterial material prepared by the method of claim 1 is used for feed additives.
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Application publication date: 20211026 |