CN112226480A - Method for preparing hydrophilic phytosterol dibasic acid sugar ester in organic phase by holoenzyme method - Google Patents
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Abstract
The invention provides a method for preparing hydrophilic phytosterol dibasic acid sugar ester in an organic phase by a holoenzyme method. The preparation steps are as follows: s1: preparation of phytosterol dibasic acid vinyl ester: respectively weighing phytosterol, divinyl ester dibasic acid and lipase in a reaction container, adding a reaction solvent subjected to dehydration treatment in advance, and placing in a constant-temperature oscillator for oscillation reaction; s2: preparation of phytosterol dibasic acid sugar ester: filtering the reaction solution to remove enzyme, performing rotary evaporation to remove solvent, adding saccharide/sugar alcohol, protease and dehydrated solvent, and continuing to react to obtain hydrophilic plant sterol diacid sugar ester. The method disclosed by the invention has the advantages of no need of functional group protection, simplicity in operation, low energy consumption, high yield, stable quality and easiness in storage, and can effectively overcome the defects of high reaction temperature, high solvent toxicity, large catalyst using amount, complex post-treatment, low edible safety coefficient and the like in the chemical synthesis process.
Description
Technical Field
The invention relates to the technical field of chemical synthesis, in particular to a method for preparing hydrophilic phytosterol dibasic acid sugar ester in an organic phase by a holoenzyme method.
Background
The phytosterol is a compound which has a chemical structure similar to that of cholesterol, has a hydrophobic steroid skeleton, has a beta-hydroxyl group at the C-3 position and is provided with a side chain on a D ring. The common phytosterols mainly comprise four types of beta-sitosterol, stigmasterol, campesterol and campesterol. They are mainly found in plants and marine organisms and cannot be synthesized in the human body. In food processing, phytosterols are valuable by-products produced during the refining of fats and oils.
Since the first discovery that phytosterol has the function of reducing blood fat and cholesterol in 1950 s, researchers have conducted extensive research on phytosterol, and guidelines and consensus on phytosterol consumption have been continuously released. Generally, the recommended intake of phytosterols is 1.5-3 g/day, which can reduce the low density cholesterol level by 8-15%. Meanwhile, a large number of animal and clinical experiments are carried out, and the phytosterol is further proved to have other biological activities, such as anticancer, anti-inflammatory, antioxidant, anti-tumor and the like.
Because the unique chemical structure and crystal form of the phytosterol determine that the phytosterol has the characteristics of poor fat solubility, water insolubility, low bioavailability and the like, the practical application of the phytosterol is greatly limited. Many documents have demonstrated that phytosterols, either used alone in powder form or added directly to food substrates, produce an undesirable texture and may significantly affect cholesterol-lowering efficacy due to their poor solubility.
In the last decade there have been a number of literature and patent reports on the synthesis of phytosterol esters from phytosterols and fatty acids by esterification or transesterification reactions to improve their lipid solubility. However, free phytosterols or phytosterol fatty acid esters are primarily used in lipid-based food matrices, which is not desirable for a low-fat diet. Moreover, the solubility of phytosterols in lipids affects their rate of diffusion, thereby extending the time required to lower blood cholesterol levels. In addition, it has also been reported that bioactive substances can increase their rate of entry into tissues and increase bioavailability in vivo by improving water solubility. In recent years, embedding, micro-emulsifying and other methods are used successively to improve the dispersibility of phytosterol in a water phase, but the prepared water-soluble phytosterol micro-emulsion, nano-dispersion, nano-liposome and the like have the defects of poor stability, difficult long-term storage, easy deterioration and the like.
The promising research of synthesizing hydrophilic phytosterol derivatives through chemical modification is still just started, and the following patents mainly exist:
CN 106755252B discloses a method for synthesizing hydrophilic phytosterol/stanol derivative by taking an ionic liquid as a catalyst through a one-pot method. The reaction temperature of the method is 80-130 ℃, the reaction temperature is too high, the food safety of the ionic liquid is unknown, after the two-step reaction is finished, the high-purity product is obtained by repeatedly extracting for many times, and the post-treatment is complex.
CN 109053843B discloses a phytosterol polybasic acid inositol ester and a preparation method thereof, wherein the method comprises the following three steps: firstly, reacting phytosterol with succinic anhydride to synthesize phytosterol succinic acid monoester; secondly, preparing phytosterol vinyl succinate from the phytosterol succinate monoester and vinyl acetate; and thirdly, synthesizing phytosterol inositol succinate from the phytosterol ethylene succinate and inositol. Wherein, the reaction in the third step is catalyzed by biological enzyme, and the rest two steps are catalyzed by chemical enzyme. Meanwhile, the application range of the method is smaller and the public acceptance is poorer compared with that of the method which adopts a plurality of functional saccharides as hydrophilic agents.
CN 103965278A discloses a preparation method of phytosterol organic diacid sugar ester. The invention adopts the technical scheme that dicyclohexylcarbodiimide, 4-dimethylaminopyridine and p-toluenesulfonic acid composite catalyst are used for catalyzing the esterification of phytosterol organic diacid monoester and functional sugar. The first step of the reaction of the patent is carried out in toluene for 6 hours at the temperature of 110 ℃, and the selected reaction solvent has high toxicity and overhigh reaction temperature. In the second step, a plurality of chemical catalysts are compounded to synthesize the phytosterol organic diacid sugar ester, and the catalysts are not easy to remove, complex in post-treatment and poor in safety.
Disclosure of Invention
The technical problem to be solved is as follows: the invention utilizes the saccharides/sugar alcohols as the hydrophilic agent and the dibasic acid divinyl ester as the linking agent, and synthesizes the hydrophilic phytosterol derivative through chemical modification, thereby solving the problem of physically improving the water solubility of the phytosterol. Meanwhile, the invention also provides a method for synthesizing the hydrophilic phytosterol dibasic acid sugar ester by one-pot two-step holoenzyme catalysis transesterification in the organic phase, which does not need the protection of functional groups, has simple operation, low energy consumption, high yield, stable quality and easy storage, and can effectively overcome the defects of high reaction temperature, high solvent toxicity, large catalyst consumption, complex post-treatment, low edible safety coefficient and the like in the prior chemical synthesis process.
The technical scheme is as follows: a method for preparing hydrophilic phytosterol dibasic acid sugar ester in organic phase by holoenzyme method comprises the following steps:
s1: preparation of phytosterol dibasic acid vinyl ester: respectively weighing phytosterol, divinyl ester dibasic acid and lipase in a reaction container, adding a reaction solvent subjected to dehydration treatment in advance, placing the mixture in a constant-temperature oscillator, and starting to react for 10-48 h at the temperature of 35-60 ℃ and the oscillation speed of 200-300 rpm;
s2: preparation of phytosterol dibasic acid sugar ester: filtering the reaction liquid to remove enzyme, performing rotary evaporation to remove the solvent, adding saccharides/sugar alcohols, protease and the solvent subjected to dehydration treatment in advance, and continuing to react for 24-120 h at the temperature of 35-60 ℃ to synthesize the hydrophilic phytosterol diacid sugar ester.
Further, the phytosterol is one or a mixture of more than one of beta-sitosterol, stigmasterol, campesterol and brassicasterol in any proportion.
Further, the dibasic acid divinyl ester is one or more of divinyl succinate, divinyl adipate, divinyl suberate and divinyl sebacate.
Further, the lipase is Novozyme immobilized lipase Novozyme 435, immobilized lipase Lipozyme RM, Lipozyme TL IM or Candida Rugosa lipase Candida Rugosa.
Further, the reaction solvent subjected to dehydration in advance in the step S1 is cyclohexane, n-hexane, n-heptane, n-octane, isooctane, or petroleum ether.
Further, the sugar/sugar alcohol is one or more of sorbitol, mannitol, xylitol, maltitol, isomaltitol, lactitol, galactitol, erythritol, glucose, fructose, mannose, sucrose, sorbose, xylose, lactose, maltose, and raffinose.
Further, the protease is a neutral protease, an alkaline protease or a bacillus protease.
Further, the solvent previously dehydrated in step S2 is pyridine, pyridine + acetone, N-dimethylformamide, or dimethyl sulfoxide.
Furthermore, in the step S1, the molar weight of the dibasic acid divinyl ester is 1-5 times of that of the phytosterol, and the dosage of the lipase is 10-50 mg/ml.
Furthermore, in the step S2, the molar weight of the phytosterol dibasic acid vinyl ester and the saccharides/sugar alcohols is 1: 1-1: 5, and the dosage of the protease is 10 mg/ml-50 mg/ml.
The synthesis reaction equation of the phytosterol dibasic acid sugar ester is as follows:
the first step is as follows:
the second step is that:
wherein n =2, 4, 6 or 8;
wherein, R1 =
；
Wherein R2 is a glycosyl/glycosyl group.
Has the advantages that:
1. the invention uses the saccharides/sugar alcohols as a hydrophilic agent and the dibasic acid divinyl ester as a linking agent, synthesizes the phytosterol dibasic acid sugar ester through chemical modification, improves the hydrophilicity of the phytosterol to a certain extent, and enlarges the application range of the phytosterol in the field of food.
2. The invention adopts a two-step enzyme method to react under mild reaction conditions, fundamentally overcomes the defects of overhigh reaction temperature, large catalyst consumption, strong toxicity, complex post-treatment operation steps and the like in the traditional synthetic process of the hydrophilic phytosterol derivative, and lays a foundation for realizing green industrialized production in the later period.
3. The invention selects various sugar alcohols/saccharides as the hydrophilic agent to carry out hydrophilic modification on the phytosterol, and has wide coverage, high product safety and good public acceptance.
4. The direct glycosylation reaction of phytosterol and saccharide to prepare phytosterol glycoside has the following disadvantages: (1) the reaction conversion rate is low; (2) most of the used catalysts are expensive and have strong toxicity, so that the synthesis cost is increased and the ecology is deteriorated; (3) the corresponding glycosyltransferase is not easy to match, and the enzyme method cannot be adopted to synthesize the target product mildly and greenly. The invention selects two-step enzyme to catalyze the ester exchange to synthesize the hydrophilic phytosterol dibasic acid sugar ester, takes dibasic acid divinyl ester as a linking agent, has mild reaction conditions, simple operation and high yield, and avoids the defect of direct glycosylation.
5. The invention takes dibasic acid divinyl ester as a linking agent, synthesizes phytosterol dibasic acid sugar ester by an ester exchange method, generates acetaldehyde after reaction, releases the acetaldehyde in a gas form, promotes the normal phase of reaction balance, obtains higher conversion rate, and avoids that the byproduct water generated by the traditional esterification reaction obstructs the smooth reaction.
6. The enzyme has high specificity to the substrate, the two-step synthesis method of the invention sequentially selects the commercially available immobilized or solid lipase and protease as biocatalysts to carry out catalytic ester exchange, good conversion rates are respectively obtained, and the separation and recovery are easy after the reaction.
Drawings
FIG. 1 is a high performance liquid chromatogram of beta-sitosterol ethylene adipate.
FIG. 2 is a high performance liquid chromatogram of campesterol sucrose adipate.
FIG. 3 is a mass spectrum of beta-sitosterol ethylene adipate.
FIG. 4 is a mass spectrum of campesterol sucrose adipate.
FIG. 5 is a mass spectrum of beta-sitosterol succinate sorbitol ester.
FIG. 6 is an NMR spectrum of beta-sitosterol ethylene adipate and beta-sitosterol.
Detailed Description
The invention provides a method for preparing hydrophilic phytosterol dibasic acid sugar ester in an organic phase by a holoenzyme method, and in order to make the purpose, the technical scheme and the effect of the invention clearer and more definite, the invention is further explained in detail by matching with the embodiment. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The conversion rate and purity of the two-step reaction of phytosterol to phytosterol dibasic acid vinyl ester and phytosterol dibasic acid vinyl ester to hydrophilic phytosterol dibasic acid sugar ester were determined by High Performance Liquid Chromatography (HPLC) -Evaporative Light Scattering Detector (ELSD). The HPLC-ELSD analysis system comprises an Agilent 1260 high performance liquid chromatograph, an evaporative light scattering detector and data processing software. The samples were eluted isocratically at 35 ℃ by using ZORBAX 300SB-C18 (5 μm, 4.6X 250mm, Agilent). Methanol/formic acid (v/v, 1000/1) was used as the mobile phase, in a sample volume of 10. mu.L, at a flow rate of 1 mL/min. The ELSD detector was controlled at 60 ℃. High-purity nitrogen was used as carrier gas at a pressure of 0.5 bar.
The intermediate and final product conversion rates were defined as follows:
conversion (%) = new substance peak area/(new substance peak area + initial substance peak area) × 100
The chemical structures of the phytosterol dibasic acid vinyl ester and the hydrophilic phytosterol dibasic acid sugar ester are identified by adopting a high performance liquid chromatography-mass spectrometry instrument (HPLC-MS) and nuclear magnetic resonance spectrum (NMR). HPLC-MS analysis adopts Agilent1200 and Agilent 6310, Ion Trap LC/MS high performance liquid chromatography mass spectrometer. The liquid phase conditions were isocratic elution of the samples at 35 ℃ by using ZORBAX 300SB-C18 (5 μm, 4.6X 250mm, Agilent). Methanol/formic acid (v/v, 1000/1) was used as the mobile phase, in a sample volume of 10. mu.L, at a flow rate of 1 mL/min. The DAD detection wavelength was controlled at 205 nm. The mass spectrum condition adopts an ESI source (+) mode, the spray voltage is 3 KV, and the temperature is as follows: at 350 ℃. NMR analysis the purified product was dissolved in deuterated chloroform using a Bruke 400 MHz NMR spectrometer with tetramethylsilane as internal standard.
Example 1
Preparation of beta-sitosterol ethylene adipate
The preparation method comprises the following steps: respectively adding 3g of beta-sitosterol, 3 ml of divinyl adipate, 0.5 g of Candida Rugosa lipase Candida Rugosa and 100 ml of dehydrated isooctane into a reaction bottle in sequence, placing the reaction bottle in a constant-temperature shaking reactor, starting the shaking rotating speed to be 250 rpm, adjusting the temperature to be 50 ℃, and reacting for 14 hours. The conversion rate of the beta-sitosterol ethylene adipate detected by HPLC-ELSD can reach 95.7 percent. FIG. 1 is a high performance liquid chromatogram of beta-sitosterol ethylene adipate. The reaction solution was filtered to remove the enzyme and the solvent was removed by rotary evaporation to obtain 3.7 g of beta-sitosterol ethylene adipate.
And (3) structural identification: HPLC-MS: the relative molecular mass of beta-sitosterol is 414, respectively. The relative molecular mass of the beta-sitosterol ethylene adipate is 568. Beta-sitosterol ethylene adipate in ES+Under ionization, [ M + Na ] of beta-sitosterol ethylene adipate] +Molecular ion peaks 591 (i.e., 568+ 23) and [ M-B ]] + Molecular ion peak 397. FIG. 3 is a mass spectrum of beta-sitosterol ethylene adipate. NMR: the NMR spectra of beta-sitosterol ethylene adipate and beta-sitosterol are shown in FIG. 6. Since the hydroxyl group at C-3 position of free beta-sitosterol is converted into ester group by ester exchange with divinyl adipate, the hydrogen at C-3 position is transferred from 3.5ppm to 4.6ppm, and the characteristic peaks of the hydrogen of the beta-sitosterol vinyl adipate olefin double bond appear at 4.8ppm and 4.56 ppm. Therefore, the intermediate product was confirmed to be vinyl β -sitosterol adipate based on HPLC-MS and NMR results.
Example 2
Preparation of campesterol sucrose adipate
The preparation method comprises the following steps: (1) respectively adding 3g of campesterol, 3 ml of divinyl adipate, 1 g of Novozyme 435 and 100 ml of dehydrated petroleum ether into a reaction bottle in sequence, placing the reaction bottle in a constant-temperature oscillation reactor, starting oscillation at the rotation speed of 250 rpm, adjusting the temperature to 55 ℃, and reacting for 48 hours. The conversion rate of the campesterol ethylene adipate can reach 81.7 percent through HPLC-ELSD detection. (2) After sampling, filtering the reaction liquid in the first step to remove enzyme, performing rotary evaporation to remove the solvent, adding 6g of hydrophilic modifier sucrose, 0.2 g of alkaline protease and 30 ml of pyridine, placing the reaction bottle in a constant-temperature oscillation reactor, starting oscillation at the rotation speed of 250 rpm, adjusting the temperature to 55 ℃, and reacting for 96 hours. Sampling and analyzing by HPLC-ELSD, and detecting that the conversion rate of the final product phytosterol sucrose adipate can reach 84.1%. FIG. 2 is a high performance liquid chromatogram of campesterol sucrose adipate. (3) After the heating was stopped, the solvent was spin-dried. Dissolving the crude product in a certain amount of dichloromethane and methanol, dissolving by ultrasonic, and filtering. And (3) washing the organic layer by using 0.1M sodium hydroxide solution, saturated salt solution and distilled water in batches sequentially, finally collecting the organic layer, and removing n-butanol by rotary evaporation to obtain the phytosterol sucrose adipate. The HPLC purity was 92.1% and the calculated yield was 89.4%. The obtained phytosterol sucrose adipate had a solubility of 0.51 g/L in water at 30 ℃.
And (3) structural identification: HPLC-MS: the relative molecular mass of campesterol sucrose adipate was 852, respectively. [ M + Na ] of sucrose campesterol adipate ester under ES + ionization] +And [ M-B] +Molecular ion peaks, 875 (i.e., 852+23) and 383, respectively. FIG. 4 is a mass spectrum of campesterol sucrose adipate.
Example 3
Preparation of beta-sitosterol succinic acid sorbitol ester
The preparation method comprises the following steps: (1) respectively adding 3g of beta-sitosterol, 3 ml of divinyl succinate, 1 g of immobilized lipase Lipozyme RM and 80 ml of dehydrated n-heptane into a reaction bottle in sequence, placing the reaction bottle in a constant-temperature oscillation reactor, starting the oscillation at the rotation speed of 250 rpm, adjusting the temperature to 60 ℃, and reacting for 48 hours. The conversion rate of phytosterol ethylene adipate detected by HPLC-ELSD can reach 97.1%. (2) After sampling, filtering the reaction liquid in the first step to remove enzyme, performing rotary evaporation to remove the solvent, adding 3g of hydrophilic modifier sorbitol, 0.5 g of neutral protease and 20 ml of binary mixed solvent of pyridine and acetone, placing the reaction bottle in a constant-temperature shaking reactor, starting the shaking rotating speed to be 250 rpm, adjusting the temperature to be 40 ℃, and reacting for 120 hours. Sampling and analyzing by HPLC-ELSD, and detecting that the conversion rate of the final product beta-sitosterol succinic acid sorbitol ester can reach 90.6%. (3) After the heating was stopped, the solvent was spin-dried. Dissolving the crude product in n-butanol, ultrasonic dissolving, and filtering. And (3) washing the organic layer by using 0.1M sodium hydroxide solution, saturated salt solution and distilled water in batches in sequence respectively, finally collecting the organic layer, and performing rotary evaporation to remove n-butanol to obtain the beta-sitosterol succinic acid sorbitol ester. HPLC purity 94.5% calculated yield 92.9%. The solubility of the beta-sitosterol succinic acid sorbitol ester in water at 30 ℃ is 0.39 g/L.
And (3) structural identification: HPLC-MS: the relative molecular masses of the beta-sitosterol sorbitan succinate are 678 respectively. [ M + Na ] of beta-sitosterol succinate sorbitol ester under ES + ionization] +And [ M-B] +Molecular ion peaks, 701 (i.e., 678+23) and 397, respectively. FIG. 5 is a mass spectrum of beta-sitosterol succinate sorbitol ester.
Claims (10)
1. A method for preparing hydrophilic phytosterol dibasic acid sugar ester in an organic phase by a holoenzyme method is characterized by comprising the following preparation steps:
s1: preparation of phytosterol dibasic acid vinyl ester: respectively weighing phytosterol, divinyl ester dibasic acid and lipase in a reaction container, adding a reaction solvent subjected to dehydration treatment in advance, placing the mixture in a constant-temperature oscillator, and starting to react for 10-48 h at the temperature of 35-60 ℃ and the oscillation speed of 200-300 rpm;
s2: preparation of phytosterol dibasic acid sugar ester: filtering the reaction liquid to remove enzyme, performing rotary evaporation to remove the solvent, adding saccharides/sugar alcohols, protease and the solvent subjected to dehydration treatment in advance, and continuing to react for 24-120 h at the temperature of 35-60 ℃ to synthesize the hydrophilic phytosterol diacid sugar ester.
2. The method for preparing the hydrophilic phytosterol dibasic acid sugar ester in the organic phase by the holoenzyme method according to claim 1, wherein the phytosterol is one or a mixture of more than one of beta-sitosterol, stigmasterol, campesterol and brassicasterol in any proportion.
3. The method for preparing the hydrophilic phytosterol dibasic acid sugar ester in the organic phase by the holoenzyme method according to claim 1, wherein the dibasic acid divinyl ester is one or more of divinyl succinate, divinyl adipate, divinyl suberate and divinyl sebacate.
4. The method for preparing hydrophilic phytosterol dibasic acid sugar esters in organic phase by holoenzymatic method according to claim 1, wherein the lipase is Novozyme 435, Lipozyme RM, Lipozyme TL IM or Candida Rugosa lipase.
5. The method for preparing hydrophilic phytosterol dibasic acid sugar ester in organic phase by holoenzyme method according to claim 1, wherein the reaction solvent subjected to dehydration treatment in advance in step S1 is cyclohexane, n-hexane, n-heptane, n-octane, isooctane or petroleum ether.
6. The method for preparing hydrophilic phytosterol dibasic acid sugar ester in organic phase by holoenzymatic method according to claim 1, wherein the sugar/sugar alcohol is one or more of sorbitol, mannitol, xylitol, maltitol, isomaltitol, lactitol, galactitol, erythritol and glucose, fructose, mannose, sucrose, sorbose, xylose, lactose, maltose and raffinose.
7. The method for preparing hydrophilic phytosterol dibasic acid sugar esters in organic phase by holoenzymatic method according to claim 1, wherein the protease is neutral protease, alkaline protease or Bacillus protease.
8. The method for preparing hydrophilic phytosterol dibasic acid sugar ester by holoenzymatic method in organic phase as claimed in claim 1, wherein the solvent dehydrated in step S2 is pyridine, pyridine + acetone, N-dimethylformamide or dimethyl sulfoxide.
9. The method for preparing the hydrophilic phytosterol dibasic acid sugar ester in the organic phase by the holoenzyme method according to claim 1, wherein the total enzyme method comprises the following steps: in the step S1, the molar weight of the dibasic acid divinyl ester is 1-5 times of that of the phytosterol, and the dosage of the lipase is 10-50 mg/ml.
10. The method for preparing the hydrophilic phytosterol dibasic acid sugar ester in the organic phase by the holoenzyme method according to claim 1, wherein the total enzyme method comprises the following steps: in the step S2, the molar weight of the phytosterol dibasic acid vinyl ester and the saccharides/sugar alcohols is 1: 1-1: 5, and the dosage of the protease is 10 mg/ml-50 mg/ml.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845473A (en) * | 2010-06-03 | 2010-09-29 | 江南大学 | Method for effectively synthesizing phytosterol ester |
| CN102936613A (en) * | 2012-11-20 | 2013-02-20 | 江南大学 | Enzymatic catalysis method for preparing phytosterol-beta-D-glucoside |
| CN103965278A (en) * | 2014-04-16 | 2014-08-06 | 江南大学 | Preparation method for water-soluble phytosterin organic dibasic acid sugar ester |
| CN106755252A (en) * | 2016-11-30 | 2017-05-31 | 江苏大学 | The method that one kettle way prepares hydrophilic plant sterol/stanol derivative |
| CN106755254A (en) * | 2016-12-29 | 2017-05-31 | 江南大学 | A kind of method of the Enzyme catalyzed synthesis lipoic acid sterol ester in organic phase |
-
2020
- 2020-09-14 CN CN202010961124.1A patent/CN112226480B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101845473A (en) * | 2010-06-03 | 2010-09-29 | 江南大学 | Method for effectively synthesizing phytosterol ester |
| CN102936613A (en) * | 2012-11-20 | 2013-02-20 | 江南大学 | Enzymatic catalysis method for preparing phytosterol-beta-D-glucoside |
| CN103965278A (en) * | 2014-04-16 | 2014-08-06 | 江南大学 | Preparation method for water-soluble phytosterin organic dibasic acid sugar ester |
| CN106755252A (en) * | 2016-11-30 | 2017-05-31 | 江苏大学 | The method that one kettle way prepares hydrophilic plant sterol/stanol derivative |
| CN106755254A (en) * | 2016-12-29 | 2017-05-31 | 江南大学 | A kind of method of the Enzyme catalyzed synthesis lipoic acid sterol ester in organic phase |
Non-Patent Citations (1)
| Title |
|---|
| YU-YUAN HU ET AL: "Enzymatic synthesis of hydrophilic phytosterol polyol esters and assessment of their bioaccessibility and uptake using an in vitro digestion /Caco-2 cell model", 《FOOD CHEMISTRY》 * |
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