阅读说明：本技术 一种醇胺脂肪酸离子液体及其制备方法和应用 (Alcohol amine fatty acid ionic liquid and preparation method and application thereof ) 是由 张嘉恒 刘凌雯 费玉清 李志超 黎穗芝 陈正件 张基亮 余明远 于 2020-05-13 设计创作，主要内容包括：本发明属于天然产物提取技术领域,公开了一种醇胺脂肪酸离子液体,其结构通式如式(Ⅰ)：<Image he="303" wi="689" file="DDA0002489629290000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>其中X<Sup>n-</Sup>为脂肪酸脱去质子以后形成的阴离子,n＝1-3；R<Sub>1</Sub>、R<Sub>2</Sub>、R<Sub>3</Sub>独立选自氢原子、烷基或ω-羟基烷基,且R<Sub>1</Sub>、R<Sub>2</Sub>、R<Sub>3</Sub>中至少有一个为ω-羟基烷基；烷基选自甲基、乙基、丙基或丁基中的任意一种；ω-羟基烷基选自羟甲基、羟乙基或羟丙基中的任意一种。将该醇胺脂肪酸离子液体用于萃取细菌溶胞物,特别是乳酸菌溶胞物,不仅萃取效率高,而且无毒性。(The invention belongs to the technical field of natural product extraction, and discloses an alcohol amine fatty acid ionic liquid, which has a structural general formula as shown in formula (I): wherein X n‑ Is an anion formed after deprotonation of a fatty acid, n-1-3; r 1 、R 2 、R 3 Independently selected from hydrogen atom, alkyl or omega-hydroxyalkyl, and R 1 、R 2 、R 3 At least one of which is omega-hydroxyalkyl; the alkyl is selected from any one of methyl, ethyl, propyl or butyl; the omega-hydroxyalkyl is selected from any one of hydroxymethyl, hydroxyethyl or hydroxypropyl. The alcohol amine fatty acid ionic liquid is used for extracting bacteria lysate, particularly lactobacillus lysate, and has high extraction efficiency and no toxicity.)

1. An alcohol amine fatty acid ionic liquid is characterized in that the structural general formula of the alcohol amine fatty acid ionic liquid is as shown in formula (I):

wherein X^n-Is an anion formed after deprotonation of a fatty acid, n-1-3; r₁、R₂、R₃Independently selected from hydrogen atom, alkyl or omega-hydroxyalkyl, and R₁、R₂、R₃At least one of which is omega-hydroxyalkyl; the alkyl is selected from any one of methyl, ethyl, propyl or butyl; the omega-hydroxyalkyl is selected from any one of hydroxymethyl, hydroxyethyl or hydroxypropyl.

2. The alcohol amine fatty acid ionic liquid according to claim 1, wherein the carbon number of the fatty acid is any positive integer between 3 and 10.

3. A method for preparing the alcohol amine fatty acid ionic liquid as claimed in claim 1 or 2, which is characterized by comprising the following steps:

and (2) stirring and mixing alcohol amine and a solvent, then adding fatty acid, stirring and reacting at 25-60 ℃, and removing the solvent to obtain the alcohol amine fatty acid ionic liquid.

4. The method according to claim 3, wherein the alcohol amine is at least one selected from the group consisting of triethanolamine, diethanolamine, and ethanolamine.

5. The method according to claim 3, wherein the molar ratio of the alcohol amine to the fatty acid is 1 (0.3-1.2).

6. The method according to claim 3, wherein the stirring reaction is carried out for 1 to 5 hours.

7. Use of the alcohol amine fatty acid ionic liquid of claim 1 or 2 for extracting a bacterial lysate.

8. A method of extracting a lactic acid bacteria lysate, comprising the steps of:

inoculating lactobacillus seed liquid into a lactobacillus culture medium for culture, centrifuging, taking thalli, suspending the thalli by using the alcohol amine fatty acid ionic liquid as claimed in claim 1 or 2, carrying out ultrasonic crushing, centrifuging and filtering, and taking filtrate to obtain lactobacillus lysate.

9. The extraction method according to claim 8, wherein the mass ratio of the cell to the alcohol amine fatty acid ionic liquid is (0.05-0.5): 1.

10. The extraction process according to claim 8, characterized in that the ultrasonication time is 20 to 60 minutes.

Technical Field

The invention belongs to the technical field of natural product extraction, and particularly relates to an alcohol amine fatty acid ionic liquid, and a preparation method and application thereof.

Background

Lactic acid bacteria are a general name for a group of bacteria capable of producing a large amount of lactic acid by utilizing fermentable carbohydrates, are widely distributed in the nature, and have abundant species diversity. The lactobacillus lysate contains thalli cell lysate, cytoplasm components, cell wall components and various organelle fragments, and various active components contained in the lactobacillus lysate comprise peptidoglycan, teichoic acid, protein, phospholipid, sterol, various enzymes, nucleotide, extracellular polysaccharide, vitamin, mineral substances, organic substances (alcohol, phenol and unsaturated fatty acid), amino acid, polypeptide and the like, so that the lactobacillus lysate can provide rich nutrition for skin, can capture free radicals and inhibit peroxidation of lipid, achieves a good anti-aging effect, can increase the metabolism speed of epithelial cells, promotes skin renewal and inhibits the activity of melanin, and can be used as a natural whitening agent.

However, in the prior art, the bacterial lysate (i.e. bacterial lysate) is mainly extracted by water and Phosphate Buffered Saline (PBS), but the yield of the effective components is low, other organic solvents such as ethanol, acetone, methanol and the like have high toxicity, and the subsequent separation process is easy to cause reagent residue. Therefore, the development of new green, high efficiency lysate effective component extraction material is very important.

Therefore, it is necessary to provide a novel solvent for extracting bacterial lysate, which can extract effective components in high yield and is non-toxic.

Disclosure of Invention

The present invention is directed to solving at least one of the problems of the prior art described above. Therefore, the invention provides the alcohol amine fatty acid ionic liquid, the preparation process of the alcohol amine fatty acid ionic liquid is simple, particularly the temperature required by the preparation is low, the time is short, and the extraction of bacterial lysate (namely bacterial lysate), particularly lactobacillus lysate by using the alcohol amine fatty acid ionic liquid has high extraction efficiency and no toxicity.

An alcohol amine fatty acid ionic liquid has a structural general formula shown in formula (I):

wherein X^n-Is an anion formed after deprotonation of a fatty acid, n-1-3; r₁、R₂、R₃Independently selected from hydrogen atom, alkyl or omega-hydroxyalkyl, and R₁、R₂、R₃At least one of which is omega-hydroxyalkyl; the above-mentionedThe alkyl is selected from any one of methyl, ethyl, propyl or butyl; the omega-hydroxyalkyl is selected from any one of hydroxymethyl, hydroxyethyl or hydroxypropyl.

Preferably, the carbon number of the fatty acid is any positive integer between 3 and 10; more preferably, the carbon number of the fatty acid is any positive integer between 3 and 8.

Preferably, the fatty acid is selected from at least one of propionic acid, butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, malonic acid, succinic acid, glutaric acid, adipic acid, suberic acid, pimelic acid, citric acid, maleic acid, or fumaric acid.

A preparation method of alcohol amine fatty acid ionic liquid comprises the following steps:

and (2) stirring and mixing alcohol amine and a solvent, then adding fatty acid, stirring and reacting at 25-60 ℃, and removing the solvent to obtain the alcohol amine fatty acid ionic liquid.

Preferably, the alcohol amine is selected from at least one of triethanolamine, diethanolamine or ethanolamine; further preferably, the alcohol amine is triethanolamine.

Preferably, the solvent is selected from at least one of water, ethanol, methanol, acetone, or isopropanol.

Preferably, the volume ratio of the alcohol amine to the solvent is 1 (1-8); further preferably, the volume ratio of the alcohol amine to the solvent is 1 (1-5); still more preferably, the volume ratio of the alcohol amine to the solvent is 1 (1-3).

Preferably, the molar ratio of the alcohol amine to the fatty acid is 1 (0.3-1.2); more preferably, the molar ratio of the alcohol amine to the fatty acid is 1 (0.5-1).

Preferably, the fatty acid is firstly mixed with the solvent to prepare a fatty acid solution, and then the fatty acid solution is dropwise added into a mixture formed by stirring and mixing the alcohol amine and the solvent.

Preferably, the temperature of the stirring reaction is 25-60 ℃; further preferably, the temperature of the stirring reaction is 25 to 40 ℃.

Preferably, the stirring reaction time is 1-5 hours; further preferably, the stirring time is 2 to 4 hours.

Preferably, the solvent removal is under negative pressure; the negative pressure is 0.04-0.09Mpa less than the atmospheric pressure; more preferably, the negative pressure is 0.07-0.08MPa less than the atmospheric pressure.

The application of the alcohol amine fatty acid ionic liquid in extracting bacterial lysate.

Preferably, the bacterial lysate is a lactic acid bacteria lysate.

A method of extracting a lactic acid bacteria lysate comprising the steps of:

inoculating the lactic acid bacteria seed liquid into a lactic acid bacteria culture medium for culture, centrifuging, taking thalli, suspending the thalli by using the alcohol amine fatty acid ionic liquid, carrying out ultrasonic crushing, centrifuging and filtering, and taking filtrate to obtain the lactic acid bacteria lysate.

Preferably, the lactic acid bacteria include at least one of lactobacillus, streptococcus, leuconostoc, bifidobacterium or cucurbitaceae.

Preferably, the lactobacillus seed solution is a lactobacillus seed solution of 2 nd to 3 rd generation recovered from lactobacillus taken out under freezing (-80 to 0 ℃).

Preferably, the volume ratio of the lactobacillus seed liquid to the lactobacillus culture medium is (1-10): 100.

Preferably, the temperature of the culture is 25-40 ℃.

Preferably, the incubation time is more than 24 hours, for example 24-72 hours.

Preferably, the rotation speed of the centrifugation is 3000-10000 rpm.

Preferably, the time of centrifugation is 10 to 30 minutes.

Preferably, the temperature for suspending the bacteria by the alcohol amine fatty acid ionic liquid is 4-40 ℃, and the suspension time is 5-50 minutes.

Preferably, the mass ratio of the bacteria to the alcohol amine fatty acid ionic liquid is (0.05-0.5): 1; more preferably, the mass ratio of the bacteria to the alcohol amine fatty acid ionic liquid is (0.1-0.3): 1.

preferably, in the process of suspending the bacteria by the alcohol amine fatty acid ionic liquid, water is firstly mixed with the alcohol amine fatty acid ionic liquid, and then the bacteria are suspended, so that the better mass and heat transfer effects are achieved; further preferably, the weight percentage of the water in the mixture of the alcohol amine fatty acid ionic liquid and the water is 10-40%.

Preferably, the temperature of the ultrasonication is 4 to 40 ℃.

Preferably, the ultrasonic power of the ultrasonic disruption is 100-600 w.

Preferably, the ultrasonication time is 20 to 60 minutes.

Further preferably, the ultrasonication takes 8 to 12 seconds per ultrasonication, then the ultrasonication is stopped for 8 to 12 seconds, and then the ultrasonication is continued for 8 to 12 seconds, and 20 to 60 minutes in total.

Triethanolamine, diethanolamine, ethanolamine have weak alkalinity, and are often used as pH regulators or neutralizers for moisturizing, nonionic surfactants, emulsifiers or lubricants in skin care products. In addition to triethanolamine, fatty acids, lactic acid, potassium hydroxide, sodium hydroxide, and the like are also used to adjust the pH of the skin care product. Similar to other amine compounds, because the nitrogen atom of the triethanolamine has lone pair electrons and is alkaline, the triethanolamine can react with inorganic acid or organic acid to generate ionic liquid (salt) which is closer to the weak acid environment of skin, thereby achieving the effects of safer thickening, moistening and moisturizing. As a novel green solvent, the precursor raw materials (such as alcohol amine and fatty acid) of the alcohol amine fatty acid ionic liquid are easy to obtain, the synthesis cost is far lower than that of the ionic liquid synthesized by matching other alkaloid organic acids, the industrial production is easy, and the method has important application in natural product extraction.

Compared with the prior art, the invention has the following beneficial effects:

(1) the preparation process of the alcohol amine fatty acid ionic liquid is simple, particularly the preparation temperature is low, the preparation time is short, the activity of the alcohol amine fatty acid ionic liquid can be well maintained by preparing the alcohol amine fatty acid ionic liquid at the low temperature, and the efficiency of extracting bacterial lysate is improved.

(2) In the prior art, the yield of the bacterial lysate extracted by using water or PBS buffer solution is low, and the problems of residue and large pollution exist when the bacterial lysate is extracted by using the traditional toxic solvent. However, the alcohol amine fatty acid ionic liquid prepared by the invention is used for extracting the bacterial lysate, particularly extracting the lactobacillus lysate, and has high extraction efficiency and no toxicity.

(3) According to the invention, the efficiency of extracting the bacterial lysate can be synergistically improved by utilizing the alcohol amine fatty acid ionic liquid and ultrasonic crushing, so that the active ingredient content of the extracted bacterial lysate is high.

Drawings

FIG. 1 is an HPLC chromatogram of a Lactobacillus plantarum lysate prepared in example 6;

FIG. 2 is an HPLC chromatogram of a Lactobacillus plantarum lysate prepared in comparative example 1;

FIG. 3 is an HPLC chromatogram of the Lactobacillus plantarum lysate prepared in comparative example 2.

Detailed Description

In order to make the technical solutions of the present invention more apparent to those skilled in the art, the following examples are given for illustration. It should be noted that the following examples are not intended to limit the scope of the claimed invention.

The starting materials, reagents or apparatuses used in the following examples are conventionally commercially available or can be obtained by conventionally known methods, unless otherwise specified.