CN110540557B - A kind of method for preparing glycidin using soybean navel powder as raw material - Google Patents

Abstract

The invention discloses a method for preparing glycidin using soybean navel powder as a raw material. The volume of the ethanol solution added is 500-600ml to obtain an extract, and the extract is evaporated to dryness to obtain a dry residue; after adding n-hexane to the dry residue for extraction, the n-hexane supernatant is discarded to obtain a residue; ethyl acetate is added to the residue Ester extraction, centrifugation to obtain the supernatant, and the supernatant was evaporated to dryness to obtain glycidin. When the soybean navel powder is extracted with a specific process, a single component of isoflavone-glyzein can be extracted, and after removing fat with n-hexane and removing impurities with ethyl acetate, the pure glycidin product can be obtained. The method is simple, fast and low in cost, and can be applied to industrial production.

Description

A kind of method for preparing glycidin using soybean navel powder as raw material

technical field

The present invention specifically relates to a method for preparing glycidin by using soybean navel powder as a raw material.

Background technique

The information disclosed in this Background section is only for enhancement of understanding of the general background of the invention and should not necessarily be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person of ordinary skill in the art.

Soybean is a traditional food in my country and contains a large number of physiologically active substances, including oligosaccharides, phospholipids, vitamins, isoflavones and soybean saponins. more and more attention from all over the world. The navel of soybeans, also known as the germ, is an important part of soybean seeds. Soybean germ contains 28% protein, 8.7% fat (the content of unsaturated fatty acids is as high as 80%, such as linoleic acid, linolenic acid and oleic acid, etc.), a large amount of vitamin E, 1.4-1.76% soybean isoflavones and about 0.4% sterols. The isoflavone content in soybean cotyledons is 0.15-0.32%, and that in seed coat is 0.01%-0.02%. Therefore, isoflavones in soybean are mainly concentrated in germ. So the germ can be used to prepare soybean isoflavones.

The inventors have found through research that, because each component in soybean isoflavones has a common basic structure, the structure and properties of the various components differ little, and it seems that they coexist and are difficult to separate. Moreover, there are many kinds of oils and fats in soybean navel, and it is not easy to separate from the weakly polar soybean isoflavones. The primary extraction methods reported in the literature mainly include alkali extraction, acid precipitation, organic solvent extraction and carbon dioxide supercritical extraction, and solvent extraction is the more commonly used method. The solvents used in the solvent extraction method include ethanol, methanol, acetone and weakly alkaline aqueous solution. The extract includes various components of isoflavone, and also contains impurities such as oil, protein, cellulose, monosaccharide and polysaccharide. In the prior art, when n-hexane or petroleum ether or 6# gasoline is used as a solvent to remove oil from soybean meal and soybean, no soybean isoflavones are detected in the extracted oil.

SUMMARY OF THE INVENTION

Aiming at the technical problems existing in the above-mentioned prior art, the purpose of the present invention is to provide a method for preparing glycidin with soybean navel powder as a raw material.

In order to solve the above technical problems, the technical scheme of the present invention is:

An object of the present invention is to provide a kind of method for preparing glycidin with soybean navel powder as raw material, comprising the following steps:

Extract 80-100 mesh bean navel powder in 55-65% ethanol for 45-60min, add 500-600ml of ethanol solution per 100g of bean navel powder to obtain an extract, and evaporate the extract to dryness to obtain dry residue ;

After adding n-hexane to the dry residue for extraction, discard the n-hexane supernatant to obtain the residue;

Ethyl acetate was added to the residue for extraction, and the supernatant was obtained by centrifugation, and the supernatant was evaporated to dryness to obtain glycidin.

The inventor unexpectedly found in the test that when using 55-65% (volume fraction) ethanol to extract soybean navel powder for 45-60min, a single component of isoflavone-glyzein can be extracted, if the concentration of ethanol deviates from this range. (less than or greater than this range) or the volume of ethanol deviates from this range (less than or greater than this range) or the extraction time deviates from this range (less than or greater than this range), the extracts are all complexes of isoflavones, not single Composition of isoflavones - glycidin.

Since there are still some components such as oil, salt, cysteine and polypeptide in the extract, after n-hexane extraction, the oil can be removed, and after selective extraction with ethyl acetate, the salt, half of the glycidin can be removed. Cystine and polypeptides and other components are obtained to obtain pure glycidin.

In some embodiments, 15-20 ml of n-hexane is added to the dry residue prepared from 100 g of soybean navel powder to remove grease.

If the amount of n-hexane is added too little, the grease cannot be completely removed, and if the amount of n-hexane is added too much, it is easy to cause waste of n-hexane.

Further, the extraction time after adding n-hexane is 10-20min.

Further, after the system extracted with n-hexane is centrifuged, the n-hexane supernatant is discarded, the centrifugal speed is 1000-1500 r/min, and the centrifugation time is 5-10 min. Centrifugal separation makes it easier to separate the n-hexane supernatant from the solids, avoiding the loss of solids when the supernatant is discarded.

In some embodiments, the time for ethyl acetate extraction is 10-20 min.

The second object of the present invention is to provide the second method for preparing glycylate with soybean navel powder as a raw material, comprising the steps:

Add n-hexane to the soybean navel powder, stir and extract to obtain an extract, and evaporate the extract to dryness to obtain soybean navel oil;

Add absolute ethanol to soybean navel oil, the volume ratio of absolute ethanol to soybean navel oil is 9-15:1, extract for 0.5-1 h, stand for stratification, take the upper layer solution, spin steam to obtain a paste ;

Absolute ethanol is added to the paste for extraction, the supernatant is taken, and the supernatant is evaporated to dryness to obtain glycidin.

Before using n-hexane to extract soybean navel powder, the inventors used n-hexane or petroleum ether or 6# gasoline to extract soybean meal, soybean and soybean oily foot in order to verify the prior art, and detected that there was no soybean isotope in the extract. The presence of flavonoids. The inventor tried to extract the bean navel with n-hexane, petroleum ether or 6# gasoline. The result unexpectedly found that when the extraction solvent was n-hexane, the extract contained a small amount of glycidin, but did not contain other soybean isoforms. flavonoids. It may be that the oil in bean navel promotes the dissolution of glycidin in n-hexane.

In order to further separate the oil and glycidin in the extract and prepare the pure glycidin product, the inventors carried out a series of experiments and found that when absolute ethanol was added to the prepared soybean navel oil, the volume of absolute ethanol was reduced. The volume ratio to soybean navel oil is 9-15:1, and the extraction is performed for 0.5-1 h, so that the glycidin in the soybean navel oil can be selectively extracted and further purified to obtain the pure glycidin product.

In some embodiments, n-hexane is used to repeatedly extract soybean navel powder 5-8 times, and the dosage ratio of n-hexane to soybean navel powder is: 1.5-3:1, V/M during each extraction.

Due to the low amount of glycidin in bean navel extracted by n-hexane in a single time, it is necessary to repeat the extraction of bean navel powder to extract the glycidin in soybean navel powder as completely as possible. When n-hexane and bean navel are extracted each time The dosage ratio of the powder is: 1.5-3:1. When V/M, the glycidin in the bean navel can be selectively extracted. The test found that the amount of n-hexane is too large or too small, which is not conducive to the selective extraction of glycidin. .

Further, the time for each n-hexane extraction of soybean navel powder is 1-1.5h, and the extraction temperature is 15-30°C.

In some embodiments, the volume ratio of soybean navel oil to absolute ethanol added thereto is 1:8-12.

In some embodiments, the mass ratio of the paste to absolute ethanol is 100 times.

The beneficial technical effects of the present invention are:

The inventor unexpectedly found that when the soybean navel powder is extracted with a specific process (ethanol extraction with a specific concentration, n-hexane degreasing, ethyl acetate removal; or n-hexane extraction with a specific volume ratio, absolute ethanol removal), when extracting, Single-component isoflavone-glyzein can be obtained by extracting, and after removing fat and impurities, pure glycidin can be obtained. The method is simple, fast and low in cost, and can be applied to industrial production.

Description of drawings

The accompanying drawings that constitute a part of the present application are used to provide further understanding of the present application, and the schematic embodiments and descriptions of the present application are used to explain the present application and do not constitute an improper limitation of the present application.

Figure 1 is a 3D scan of the thin layer separation of daidzein, genistein and daidzein;

FIG. 2 is a thin-layer developed scanning peak diagram of the product prepared in Example 1. FIG.

Figure 3 is a thin-layer developed scanning peak diagram of a glycitein standard solution.

Detailed ways

It should be noted that the following detailed description is exemplary and intended to provide further explanation of the application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

It should be noted that the terminology used herein is for the purpose of describing specific embodiments only, and is not intended to limit the exemplary embodiments according to the present application. As used herein, unless the context clearly dictates otherwise, the singular is intended to include the plural as well, furthermore, it is to be understood that when the terms "comprising" and/or "including" are used in this specification, it indicates that There are features, steps, operations, devices, components and/or combinations thereof.

Example 1

Take 100 g of 80-100 mesh soybean navel powder, add 550 mL of 60% ethanol, stir and extract at 50°C for 1 h, and rotate 400 mL of the filtrate until the dry matter is 2 g. Add 20 mL of n-hexane to the dry matter, shake and extract for 10 min, centrifuge at 1000-1500 r/min for 5 min, discard the supernatant, and obtain the residue. Add 20 mL of ethyl acetate to the residue, shake and extract for 15 min, and centrifuge at 1000-1500 r/min for 10 min to obtain a supernatant, and remove the ethyl acetate by rotary evaporation to obtain 10.7 mg of glycidin, the purity of which is 88%.

Example 2

Take 100 g of 80-100 mesh bean navel powder, add 600 mL of 55% ethanol, stir and extract at 60°C for 1 h, and rotate 450 mL of the filtrate until the dry matter is 1.5 g. Add 15 mL of n-hexane to the dry matter, extract by shaking for 20 min, centrifuge at 1000-1500 r/min for 10 min, discard the supernatant, and obtain the residue. Add 15 mL of ethyl acetate to the residue, shake and extract for 20 min, and centrifuge at 1000-1500 r/min for 5 min to obtain a supernatant, and remove the ethyl acetate by rotary evaporation to obtain 11 mg of glycitein, the purity of which is 90%.

Example 3

Take 100 g of 80-100 mesh bean navel powder, add 500 mL of 65% ethanol, stir and extract at 55° C. for 45 minutes, and rotate 350 mL of the filtrate until the dry matter is 2 g. Add 20 mL of n-hexane to the dry matter, shake and extract for 15 min, centrifuge at 1000-1500 r/min for 8 min, discard the supernatant, and obtain the residue. Add 20 mL of ethyl acetate to the residue, shake and extract for 20 min, centrifuge at 1000-1500 r/min for 8 min to obtain a supernatant, remove the ethyl acetate by rotary evaporation to obtain 10.4 mg of glycidin, with a purity of 88%.

Comparative Example 1

The difference from Example 1 is that the particle size of the bean navel powder is 60-75 mesh, and the others are the same as in Example 1. The mass of the prepared glycidin was 7.6 mg.

Comparative Example 2

The difference from Example 1 is that the particle size of bean navel powder is 110-120 mesh, and the others are the same as Example 1. There are many fatty acids in the product, and impurities such as leucine, isoleucine, glycine, valine and polypeptides are increased, and the purity of glycitein is 63%.

Comparative Example 3

The difference from Example 1 is that the concentration of ethanol is 50%, and the others are the same as in Example 1. The extracted product contains daidzein.

Comparative Example 4

The difference from Example 1 is that the concentration of ethanol is 70%, and the others are the same as in Example 1. The product extracted at this concentration has many impurities such as triglycerides, and the purity of glycyxin is 65%.

Comparative Example 5

The difference from Example 1 is that the volume of ethanol added is 450 ml, and the others are the same as in Example 1. There are many impurities in amino acids and polypeptides in glycidin products, and the purity of glycidin is 54%.

Comparative Example 6

The difference from Example 1 is that the volume of ethanol added is 600 ml, and the others are the same as in Example 1. There are many impurities in oil, small peptides and fatty acids, and the purity of glycidin is 56%.

Example 4

Take 110 g of ground (80 mesh) soybean navel powder, add 300 mL of n-hexane, stir and extract at room temperature for 1 hour, pour out 150 mL of supernatant after standing still, add 150 mL of n-hexane and stir and extract at room temperature for 1 hour, pour out the supernatant after standing still 150mL. Repeat this 3 times for a total of 5 times. The supernatant was combined and the n-hexane was removed by rotary evaporation to obtain 218 mL of soybean navel oil.

Add 200 mL of absolute ethanol to 18 mL of soybean navel oil, stir and extract for 1 hour, pour out the ethanol extract (other oils and fats are below) after static, and remove the ethanol by rotary evaporation to obtain a small amount of paste (about <0.1 g).

Add 10 mL of absolute ethanol to the paste, shake and extract for 12 min, pour out the ethanol supernatant (discard the oil attached to the bottom of the bottle), and rotate it to dryness. After testing, glycidin is 19.88 mg (can be used as soybean isoflavones). Standard for detection), the purity of glycidin was 90.4%.

Example 5

Take 110 g of ground (80 mesh) soybean navel powder, add 250 mL of n-hexane, stir and extract at room temperature for 1.5 hours, pour out 150 mL of supernatant after standing still, add 150 mL of n-hexane, stir and extract at room temperature for 1.5 hours, pour out after standing still Supernatant 150mL. Repeat this 5 times for a total of 7 times. The supernatant was combined and the n-hexane was removed by rotary evaporation to obtain 21 mL of soybean navel oil.

Add 250 mL of anhydrous ethanol to 21 mL of soybean navel oil, stir and extract for 45 minutes, pour out the ethanol extract (other oils and fats are below) after static, and remove the ethanol by rotary evaporation to obtain a small amount of paste (about <0.1 g).

Add 10 mL of absolute ethanol to the paste, shake and extract for 10 min, pour out the ethanol supernatant (discard the grease attached to the bottom of the bottle), and rotate it to dryness. After testing, the glycidin is 21 mg (which can be used as soybean isoflavone detection). Using standard substance), the purity of glycidin was 90.8%.

Example 6

Take 110 g of ground (80 mesh) soybean navel powder, add 200 mL of n-hexane, stir and extract at room temperature for 1.3 hours, pour out 150 mL of supernatant after standing still, add 150 mL of n-hexane, stir and extract at room temperature for 1.3 hours, pour out after standing still Supernatant 150mL. This is repeated 4 times for a total of 6 times. The supernatant was combined and the n-hexane was removed by rotary evaporation to obtain 20 mL of soybean navel oil.

Add 300 mL of absolute ethanol to 20 mL of soybean navel oil, stir and extract for 0.5 h, pour out the ethanol extract (other oils and fats are below) after static, and remove the ethanol by rotary evaporation to obtain a small amount of paste (about <0.1 g).

Add 10 mL of absolute ethanol to the paste, shake and extract for 15 min, pour out the ethanol supernatant (discard the grease attached to the bottom of the bottle), and rotate it to dryness. After testing, the glycidin is 19.5 mg (which can be used as soybean isoflavones). Standard for detection), the purity of glycidin was 90.4%.

Comparative Example 7

The difference from Example 1 is: using petroleum ether to replace n-hexane, other steps are the same as those in Example 1, and the product does not contain glycidin.

Comparative Example 8

The difference from Example 1 is that the volume of n-hexane added to the soybean navel powder is 330 ml, and other steps are the same as those in Example 1. The amount of solvent is large, and the product has many impurities, and the purity of glycidin is 67%.

Comparative Example 9

The difference from Example 1 is: the volume of adding absolute ethanol to soybean navel oil is 150 ml, and other steps are the same as in Example 1. The dissolving of soybean navel oil is not good, the liquid is sticky, and the extraction is not thorough.

The products prepared in Examples 1-3 and Comparative Examples 1-9 are detected, and the detection method is as follows:

1. Prepare 6 standard solutions of daidzein, daidzein, genistein, genistein, glycidin, and daidzein into 6 standard solutions of 1 mg/ml respectively.

2. The samples prepared in Examples 1-3 and Comparative Examples 1-9 were respectively dissolved in 10 ml of ethyl acetate as sample solutions.

3. Using activated silica gel GF254 high-efficiency thin-layer plate (HPTLC) 10cm*20cm, take 1μl, 2μl, 3μl, 4μl, and 5μl of each of the 6 standard solutions to make 13 standard curves, corresponding to 6 Standard curve equation.

4. Detection of soybean isoflavone glycosides (daidzein, genistein, daidzein): 5 μl of each of the three external standard solutions. 5 μl of the ethyl acetate-dissolved sample solutions of the products of Examples 1-6 and Comparative Examples 1-9 were taken. Spot on the GF254HPTLC board and let it dry naturally. Expand in a double-slot glass developing cylinder, add 20 ml of developing agent, formula of developing agent: dichloromethane: methanol: acetic acid = 10:2:0.1, put the HPTLC plate into the developing cylinder and cover it, and start to expand after 10 minutes of saturation. Distance: 13 cm. Take out after exhibition. After drying, observe under 365nm UV lamp.

5. Scanning conditions of TLC scanner III with WINcats1.4.1 software (CAMAG, Switzerland): scanning speed 80nm/s, resolution 200μm/step, irradiation lamp D2 lamp, wavelength 260nm, scanning width 6.00mm*0.90 mm. daidzein Rf=0.41, genistein Rf=0.52, daidzein Rf=0.50, as shown in Figure 1, it can be seen that these three substances can be completely separated.

6. Detection of soybean isoflavone aglycone (daidzein, genistein, glycidin): 5 μl of each of the three standard solutions of external standard. 5 μl of the ethyl acetate-dissolved sample solutions of the products of Examples 1-6 and Comparative Examples 1-9 were taken. Spot on the GF254HPTLC board and let it dry naturally. Expand in a double-slot glass developing cylinder, add 20 ml of developing agent, formula of developing agent: chloroform: methanol: acetic acid = 9:1:0.15, put the HPTLC plate into the developing cylinder and cover it, and start to expand after 10 minutes of saturation. Distance: 10 cm, take it out after exhibition, and observe under 365nm UV lamp after drying.

7. Scanning conditions of TLC scanner III with WINcats1.4.1 software (CAMAG, Switzerland): scanning speed 80nm/s, resolution 200μm/step, irradiation lamp D2 lamp, wavelength 260nm, scanning width 6.00mm*0.90 mm, daidzein Rf=0.50, genistein Rf=0.67, glycidin Rf=0.62.

The sample liquids of Examples 1-6 and Comparative Examples 1-9 were tested, and the test results were as follows: the sample liquids of Examples 1-6 only contained glycidin and no other soybean isoflavones. As shown in Fig. 2 and Fig. 3, the product prepared in Example 1 and the glycidin standard product have the same peak position, which is the same substance, and there are no other impurity peaks in the scan in Fig. 2, indicating that the prepared soybean yellow Vegetarian products are purer.

The components and purity of the products prepared in Examples 1-6 and Comparative Examples 1-9 are shown in Table 1.

Table 1

The above descriptions are only preferred embodiments of the present application, and are not intended to limit the present application. For those skilled in the art, the present application may have various modifications and changes. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of this application shall be included within the protection scope of this application.

Claims (9)

1. A method for preparing glycitein by taking soybean navel powder as a raw material is characterized by comprising the following steps: the method comprises the following steps:

extracting 80-100 mesh bean navel powder in 55-65% ethanol for 45-60min, adding ethanol solution with volume of 500-600ml into per 100g bean navel powder to obtain extractive solution, and evaporating to dry to obtain dry residue;

adding n-hexane into the dry residue, extracting, and removing the n-hexane supernatant to obtain residue;

adding ethyl acetate into the residue, extracting, centrifuging to obtain supernatant, and evaporating the supernatant to dryness to obtain glycitein.

2. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 1, characterized in that: adding 15-20ml of n-hexane into dry residue per 100g of bean navel powder for removing oil and fat.

3. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 1, characterized in that: adding n-hexane, and extracting for 10-20 min.

4. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 1, characterized in that: and (3) centrifugally separating the system after n-hexane extraction, discarding the n-hexane supernatant, wherein the centrifugal rotation speed is 1000-1500r/min, and the centrifugal time is 5-10 min.

5. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 1, characterized in that: the extraction time of ethyl acetate is 10-20 min.

6. A method for preparing glycitein by taking soybean navel powder as a raw material is characterized by comprising the following steps: the method comprises the following steps:

adding n-hexane into the bean navel powder, stirring and extracting to obtain an extracting solution, and evaporating the extracting solution to dryness to obtain bean navel oil;

adding anhydrous ethanol into the Sophora umbilicalis oil at a volume ratio of the anhydrous ethanol to the Sophora umbilicalis oil of 9-15:1, extracting for 0.5-1h, standing for layering, collecting the upper layer solution, and steaming to obtain paste;

adding anhydrous ethanol into the paste for extraction, taking supernatant, and evaporating the supernatant to dryness to obtain glycitein;

and (3) repeatedly extracting the bean navel powder by using n-hexane for 5-8 times, wherein the dosage ratio of the n-hexane to the bean navel powder is as follows: 1.5-3: 1, V/M.

7. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 6, characterized in that: the extraction time of the soybean navel powder by n-hexane is 1-1.5h each time, and the extraction temperature is 15-30 ℃.

8. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 6, characterized in that: the volume ratio of the soybean navel oil to the absolute ethyl alcohol added into the soybean navel oil is 1: 8-12.

9. The method for preparing glycitein from soy bean paste powder as a raw material according to claim 6, characterized in that: the mass ratio of the paste to the absolute ethyl alcohol is 100 times.

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