Fat-soluble rutin ester as well as preparation method and application thereof

Abstract

The invention provides fat-soluble rutin ester as well as a preparation method and application thereof, and belongs to the technical field of food. Comprising the following steps: S1, respectively weighing rutin, an acyl donor and a biocatalyst, putting the rutin, the acyl donor and the biocatalyst into a reaction flask, and adding a mixed solvent for reaction; s2, after the reaction in the step S1 is finished, filtering out the biocatalyst, and collecting clear liquid; and S3, removing the reaction solvent to obtain the fat-soluble rutin ester product. According to the method, edible vegetable oil which is in a liquid state at room temperature is used as an acyl donor, and lipase is used as a biocatalyst to synthesize the fat-soluble rutin ester. The biocatalyst used in the method is low in cost and high in catalytic efficiency, and the acyl donor used in the method is low in cost and high in safety. The synthesized fat-soluble rutin ester is good in safety and can be directly applied to grease and grease-rich systems without separation and purification. The technology has the advantages of simplicity, high efficiency and the like and is suitable for industrial production.

Description

Technical Field

[0001] The invention belongs to the technical field of food and medicine, and specifically relates to a fat-soluble rubutan ester and a preparation method and application thereof. Background Art

[0002] Oxidation is the main cause of quality deterioration of oils and oil-rich systems. Adding antioxidants can delay oxidative rancidity. At present, less than 15 antioxidants are allowed to be used in oils and oil-rich foods in my country's national food safety standard-Food Additives Use Standard GB 2760-2014. Most of these antioxidants are artificially synthesized, such as 2,6-di-tert-butyl-p-cresol (BHT), butylated hydroxyanisole (BHA), tert-butylhydroquinone (TBHQ), etc., and their safety is facing more and more doubts. Therefore, it is urgent to develop natural, efficient and safe antioxidants.

[0003] Rutin is a flavonoid compound found in plants such as buckwheat, rue, and sophora japonica. Studies have shown that rutin has excellent antioxidant activity and is an ideal natural antioxidant ingredient. However, rutin has poor fat solubility (oil solubility, hydrophobicity), which limits its application in oils and related systems. Therefore, how to improve the fat solubility of rutin and then exert its antioxidant activity has become a current problem.

[0004] CN 101704866A discloses a rutin fatty acid ester derivative, a preparation method and application thereof. The technology uses rutin and fatty acids as raw materials, and synthesizes rutin fatty acid esters through esterification reaction under the catalytic action of Novozym 435 lipase. Although the prior art can improve the fat solubility of rutin to a certain extent, it still has the following problems: first, the Novozym 435 lipase used is expensive and cannot be used for mass production; second, high-purity fatty acids are used as acyl donors, which is expensive; third, the synthesized rutin ester contains free fatty acids and cannot be directly added to oil and fat systems, and needs to go through a series of separation and purification links, which has the disadvantages of complicated steps. Therefore, it is necessary to do further research to solve the above technical problems. Summary of the invention

[0005] In view of the above technical problems, the present invention provides a fat-soluble rutin ester and a preparation method thereof, which improves the fat solubility of rutin and facilitates its application in oils and fat-rich food systems.

[0006] The present invention also provides an application of the fat-soluble rutin ester in oils and fats and oil-rich food systems.

[0007] The biocatalyst used in the invention has low cost and high catalytic efficiency, and the synthesized fat-soluble rubutan ester has high safety.

[0008] The modifier used in the present invention is edible vegetable oil, which has the advantages of low cost and high safety, can be directly used without separation and purification, and has the advantages of simplicity and high efficiency.

[0009] The preparation method of the present invention comprises the following steps: step S1, respectively weighing rutin, acyl donor and biocatalyst in a reaction bottle, adding a mixed solvent for reaction; step S2, after the reaction is finished, filtering out the biocatalyst, and collecting the clear liquid; step S3, removing the reaction solvent to obtain a fat-soluble rutin ester product; the acyl donor is an edible plant oil that is liquid at room temperature, including but not limited to soybean oil, sunflower oil, peanut oil, corn oil, rapeseed oil, olive oil, camellia oil, peony seed oil, and linseed oil. The method of the present invention has the advantages of simplicity, high efficiency, and low cost; the prepared fat-soluble rutin ester has high safety and can be directly applied to oils and oil-rich food systems without purification.

[0010] The present invention achieves the above technical objectives through the following technical means.

[0011] A method for preparing fat-soluble rubutan ester comprises the following steps:

[0012] Step S1, weighing rutin, acyl donor and biocatalyst respectively into a reaction bottle, adding a mixed solvent to react;

[0013] Step S2, after the reaction in step S1 is completed, the biocatalyst is filtered out and the clear liquid is collected;

[0014] Step S3, removing the reaction solvent to obtain a fat-soluble rubutan ester product.

[0015] In the step S1, the acyl donor is an edible vegetable oil that is liquid at room temperature, including but not limited to soybean oil, sunflower oil, peanut oil, corn oil, rapeseed oil, olive oil, camellia oil, peony seed oil or linseed oil. The main component of edible vegetable oil is triglyceride, and its content accounts for more than 99% of vegetable oil. Rutin and triglyceride are subjected to ester exchange reaction to generate rutin ester and diglyceride. The specific rutin ester and diglyceride (1,2-diaceryl or 1,3-diaceryl) generated depend on the type and distribution of fatty acids in vegetable oil (triglyceride). Diaceryl is a new food raw material allowed for use in my country, so after the reaction is completed, fat-soluble rutin ester can be obtained without any separation and purification steps, and the product can be directly applied to oils and oil-rich food systems.

[0016] In the above scheme, the biocatalyst is lipase Lipozyme RM IM, Candida rugosa, Lipozyme TL IM or Candida sp.99-125.

[0017] In the above scheme, the biological mixed solvent is isooctane / tert-butanol, n-hexane / tert-butanol, petroleum ether / tert-butanol, isooctane / tert-amyl alcohol, n-hexane / tert-amyl alcohol, petroleum ether / tert-amyl alcohol, isooctane / acetone, n-hexane / acetone or petroleum ether / acetone.

[0018] In the above scheme, the concentration of rutin is 10mmol / L to 120mmol / L.

[0019] In the above scheme, the concentration of the acyl donor is 20 mmol / L to 240 mmol / L.

[0020] In the above scheme, the amount of lipase added is 5g / L to 100g / L.

[0021] In the above scheme, the reaction temperature of step S1 is 35°C to 80°C, and the reaction time is 12h to 144h.

[0022] A fat-soluble rubutan ester obtained according to the preparation method of the fat-soluble rubutan ester.

[0023] An application of the fat-soluble rubutin ester is an application of the fat-soluble rubutin ester in oils and fat-rich foods.

[0024] In the above scheme, a fat-soluble rubutan ester product that can be directly used is obtained without any separation step.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] 1. The method of the present invention is simple and efficient; the biocatalyst used is low in cost and high in catalytic efficiency; the modifier edible plant oil used is widely available and low in cost.

[0027] 2. The fat-soluble rubutan ester obtained by the method of the present invention is highly safe and can be used directly without separation. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 Schematic diagram of the preparation of fat-soluble rubutyrate in the present invention;

[0029] Figure 2 Infrared spectrum of fat-soluble rutin ester;

[0030] Figure 3 Liquid chromatography-mass spectrometry of fat-soluble rubutyrate, where Figure 3 (A) is the total ion current diagram, Figure 3 (B) Mass spectrum of rutin oleate, Figure 3 (C) Mass spectrum of rutin linoleate. DETAILED DESCRIPTION

[0031] The preferred embodiments of the present invention are described below. It should be understood that the embodiments are for better explanation of the present invention and are not intended to limit the present invention. The experimental methods used in the following embodiments are conventional methods unless otherwise specified; the materials, reagents, etc. used in the following embodiments are all commercially available unless otherwise specified.

[0032] A method for preparing fat-soluble rubutan ester comprises the following steps:

[0033] Step S1, weighing rutin, acyl donor and biocatalyst respectively into a reaction bottle, adding a mixed solvent to react;

[0034] Step S2: After the reaction is completed, the biocatalyst is filtered out and the clear liquid is collected;

[0035] Step S3, removing the reaction solvent to obtain a fat-soluble rubutan ester product;

[0036] The acyl donor is an edible vegetable oil that is liquid at room temperature, including but not limited to soybean oil, sunflower oil, peanut oil, corn oil, rapeseed oil, olive oil, camellia oil, peony seed oil, and linseed oil.

[0037] The biocatalyst is lipase Lipozyme RM IM, Candida rugosa, Lipozyme TL IM or Candida sp. 99-125.

[0038] The biological mixed solvent is isooctane / tert-butanol, n-hexane / tert-butanol, petroleum ether / tert-butanol, isooctane / tert-amyl alcohol, n-hexane / tert-amyl alcohol, petroleum ether / tert-amyl alcohol, isooctane / acetone, n-hexane / acetone or petroleum ether / acetone.

[0039] The rutin concentration is 10 mmol / L to 120 mmol / L, the acyl donor concentration is 20 mmol / L to 240 mmol / L, the lipase addition amount is 5 g / L to 100 g / L, the reaction temperature is 30° C. to 70° C., and the reaction time is 12 h to 144 h.

[0040] Without any separation process, the fat-soluble rubutan ester product that can be directly used is obtained.

[0041] Thin layer chromatography: the developing solvent is petroleum ether:ethyl acetate:methanol (1:3:1, v / v / v), and the color developer is iodine vapor.

[0042] Liquid chromatography analysis: Take 200 μL of the reaction solution, dissolve and dilute it with 1 mL of anhydrous ethanol, centrifuge at 3000 rpm for 2 min, take the lower clear liquid, and filter it with a 0.45 μm organic microporous filter membrane. Use LC-20AD high performance liquid chromatography to analyze the sample, and the column model is Symmetry C18 Column (5μm, 4.6mm×250mm, Waters), column oven 30℃, methanol:formic acid (1000:1, v / v) as mobile phase, flow rate 0.8mL / min. ZAM 4000 evaporative light scattering detector, nitrogen as carrier gas, pressure 0.3Mpa, drift tube temperature 70℃. The conversion rate was calculated by normalization method, specifically according to formula 1:

[0043] Conversion rate (%) = rubutyrate peak area / (rubutyrate peak area + rutin peak area) × 100 (Formula 1)

[0044] Liquid chromatography-mass spectrometry analysis: LXQ LC / MS liquid chromatography ion trap mass spectrometry was used to determine the relative molecular weight of the reaction sample. The chromatographic column was Symmetry C 18 Column (5 μm, 4.6 mm × 150 mm, Waters), mobile phase methanol: formic acid (1000: 1, v / v), column temperature 25 ° C, flow rate 0.8 mL / min. Mass scan range 100-1500 amu, evaporator temperature 350 ° C, sheath gas flow rate 25 arb, auxiliary gas flow rate 5 arb, discharge current 4.0 μ A, capillary temperature 275 ° C, capillary voltage 30 V, lens voltage 120 V.

[0045] Fourier transform infrared spectroscopy analysis: The total reflection attenuation method was used on a Nicolet iS10 FT-IR spectrometer at 4000-400 cm -1 Internal scanning 32 times, resolution 4cm -1 .

[0046] The present invention uses edible vegetable oil that is liquid at room temperature as an acyl donor and the above-mentioned lipase as a biocatalyst to synthesize fat-soluble rubutin ester. The biocatalyst used has low cost and high catalytic efficiency, and the acyl donor used has low cost and high safety. The synthesized fat-soluble rubutin ester has good safety and can be directly applied to oils and oil-rich systems without separation and purification. The present invention has the advantages of simplicity, high efficiency, etc. and is suitable for industrial production.

[0047] Example 1

[0048] 10mmol / L rutin, 20mmol / L linseed oil (Golden Dragon Fish Oil Co., Ltd.) and 5g / L Lipozyme RM IM lipase were added to the reaction bottle in sequence, and 20mL petroleum ether:acetone (1:1, v / v) was added, and the mixture was placed in a 35°C constant temperature air bath oscillator for reaction for 72h. After the reaction was completed, the lipase in the reaction bottle was filtered out, the supernatant was collected, and the solvent was removed by rotary evaporation to obtain the product.

[0049] The structural characterization data of the synthesized product are as follows: Thin layer chromatography: the migration value of rutin is 0-0.2, and the migration value of fat-soluble rutin ester is 0.2-0.68. Figure 2 This is the infrared spectrum of the fat-soluble rutin ester prepared. As can be seen from the figure, 3251cm -1 The absorption peak is the absorption signal of unreacted -OH, 2924 cm -1 The absorption peak is -CH3 stretching vibration absorption, 2853cm -1 The absorption peak is the stretching vibration absorption of -CH2, 1443cm -1 The bending vibration absorption of -CH3 is 1356cm -1 is the bending vibration absorption of -CH2, 1715cm -1 The stretching vibration absorption signal of carbonyl C=O in the ester bond is 1602 cm -1 and 1651cm -1 The stretching vibration absorption signals of different C=C are at 1165cm -1 and 1198cm -1 The signals at are the stretching vibration absorptions of different CO. Mass spectrum: The molecular weight of rutin linolenate is 870. In the ES-mass spectrum, m / z 869 is the [MH] signal of rutin linolenate. Based on the above analysis, it is shown that rutin linolenate was successfully synthesized.

[0050] According to the analysis of high performance liquid chromatography, the conversion rate of rubutyrate is 85.2%, indicating that fat-soluble rubutyrate can be efficiently synthesized.

[0051] Example 2

[0052] 120mmol / L rutin, 240mmol / L olive oil (Olivilan) and 100g / L Lipozyme TL IM lipase were added to the reaction bottle in sequence, and 20mL isooctane: tert-butyl alcohol (1:1, v / v) was added, and the mixture was placed in a constant temperature air bath oscillator at 80 degrees to react for 12 hours. After the reaction was completed, the lipase in the reaction bottle was filtered out, the supernatant was collected, and the solvent was removed by rotary evaporation to obtain the product.

[0053] The structural characterization data of the synthesized product are as follows: Thin layer chromatography: the migration value of rutin is 0-0.23, and the migration value of fat-soluble rutin ester is 0.3-0.81. Infrared spectrum: 3250cm -1 The absorption peak is the absorption signal of unreacted -OH, 2931cm -1 The absorption peak is -CH3 stretching vibration absorption, 2846cm -1 The absorption peak is the stretching vibration absorption of -CH2, 1445cm -1 The bending vibration absorption of -CH3 is 1358cm -1is the bending vibration absorption of -CH2, 1724cm -1 The stretching vibration absorption signal of carbonyl C=O in the ester bond is 1601cm -1 and 1675cm -1 The stretching vibration absorption signals of different C=C are at 1151cm -1 and 1190cm -1 The signals at are the stretching vibration absorptions of different CO. Mass spectrometry: The molecular weights of rutin oleate and rutin linoleate are 874 and 872, respectively. Figure 3 is a liquid chromatography-mass spectrometry image of the prepared fat-soluble rutin ester, wherein Figure 3 (A) is the total ion current diagram, Figure 3 (B) Mass spectrum of rutin oleate, Figure 3 (C) Mass spectrum of rutin linoleate. In the ES-mass spectrum, m / z 873 and m / z 871 are the [MH] signals of rutin oleate and rutin linoleate, respectively. Based on the above analysis, it is shown that rutin linoleate and rutin oleate were successfully synthesized.

[0054] According to the analysis of high performance liquid chromatography, the conversion rate of rubutyrate is 92.8%, indicating that fat-soluble rubutyrate can be efficiently synthesized.

[0055] Example 3

[0056] 60mmol / L rutin, 180mmol / L soybean oil (Fulinmen) and 50g / L Candida sp.99-125 lipase were added to the reaction bottle in sequence, and 20mL n-hexane / tert-amyl alcohol (1:1, v / v) was added, and the mixture was placed in a 50-degree constant temperature air bath oscillator for reaction for 144h. After the reaction was completed, the lipase in the reaction bottle was filtered out, the supernatant was collected, and the solvent was removed by rotary evaporation to obtain fat-soluble rutin ester.

[0057] The structural characterization data of the product are as follows: Thin layer chromatography: the migration value of rutin is 0.03-0.25, and the migration value of fat-soluble rutin ester is 0.25-0.65. Infrared spectrum: 3248cm -1 The absorption peak is the absorption signal of unreacted -OH, 2935cm -1 The absorption peak is -CH3 stretching vibration absorption, 2859cm -1 The absorption peak is the stretching vibration absorption of -CH2, 1450cm -1 The bending vibration absorption of -CH3 is 1361cm -1 is the bending vibration absorption of -CH2, 1719cm -1 The stretching vibration absorption signal of carbonyl C=O in the ester bond is 1608cm -1 and 1657cm -1The stretching vibration absorption signals of different C=C are at 1159cm -1 and 1187cm -1 The signals at are the stretching vibration absorptions of different CO. Mass spectrum: The molecular weight of rutin linoleate is 872. In the ES-mass spectrum, m / z 871 is the [MH] signal of rutin linoleate. Based on the above analysis, it is shown that rutin linoleate was successfully synthesized.

[0058] According to the analysis of high performance liquid chromatography, the conversion rate of rubutyrate is 95.1%, indicating that fat-soluble rubutyrate can be efficiently synthesized.

[0059] Although the present invention has been disclosed as above in the form of a preferred embodiment, it is not intended to limit the present invention. Anyone familiar with this technology can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the scope of protection of the present invention should be based on the definition of the claims.

Claims

1. A method for preparing a fat-soluble rubutyrate, characterized in that: The following steps are involved: Step S1, weighing rutin, an acyl donor and a biocatalyst respectively into a reaction bottle, adding a mixed solvent to react, wherein the acyl donor is an edible vegetable oil that is liquid at room temperature, including soybean oil, sunflower oil, peanut oil, corn oil, rapeseed oil, olive oil, camellia oil, peony seed oil or linseed oil; Step S2, after the reaction in step S1 is completed, the biocatalyst is filtered out and the clear liquid is collected; Step S3, removing the reaction solvent to obtain a fat-soluble rubutan ester product.

2. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The biocatalyst is lipase Lipozyme RM IM, Candida rugosa, Lipozyme TL IM or Candida sp. 99-125.

3. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The biological mixed solvent is isooctane / tert-butanol, n-hexane / tert-butanol, petroleum ether / tert-butanol, isooctane / tert-amyl alcohol, n-hexane / tert-amyl alcohol, petroleum ether / tert-amyl alcohol, isooctane / acetone, n-hexane / acetone or petroleum ether / acetone.

4. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The rutin concentration is 10 mmol / L to 120 mmol / L.

5. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The acyl donor concentration is 20 mmol / L to 240 mmol / L.

6. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The lipase addition amount is 5g / L to 100g / L.

7. The method for preparing fat-soluble rubutan ester according to claim 1, characterized in that: The reaction temperature of step S1 is 35°C to 80°C, and the reaction time is 12h to 144h.

8. A fat-soluble rubutan ester obtained according to the method for preparing the fat-soluble rubutan ester according to any one of claims 1 to 7.

9. A use of the fat-soluble rubutan ester according to claim 8, characterized in that: The fat-soluble rutin ester is used in oils and fat-rich foods.

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