JPS61221298A - Purification of monoglyceride - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] The present invention relates to a novel method for purifying monoglycerides.

More specifically, by treating oils and fats containing monoglycerides, for example, oils and fats containing triglycerides, diglycerides, monoglycerides, free fatty acids, etc. obtained by enzymatic decomposition, transesterification, synthetic methods, etc., with an ion exchange resin. This is a method for purifying monoglycerides, which is characterized by separating monoglycerides with good yield and purity.

Conventionally, molecular distillation has been commonly used as a method for purifying monoglycerides, but there are problems as listed below.

(1) High vacuum is required and complicated and expensive equipment is required.

(2) The distillation fraction of monoglyceride cannot be clearly distinguished from other components, and when attempting to obtain highly pure monoglyceride, the yield is extremely reduced.

(3) Since the treatment is performed at high temperatures, the β-
The monoglyceride at the α-position undergoes a rearrangement reaction during the operation and changes to the monoglyceride at the α-position.

Furthermore, monoglycerides with a high degree of unsaturation undergo isomerization, polymerization, etc. at high temperatures, making their application difficult.

In addition, in the laboratory, column separation methods using adsorbents such as silica dal are known, but they have problems such as contamination deterioration and difficulty in regeneration, and have not been industrialized. Rather than using various methods, monoglyceride can be easily separated and recovered from a mixture with high purity and high yield by utilizing the interaction between the caldexyl group of the resin and the two alcohol groups of monoglyceride in a non-aqueous system. It has been found that the β-position monoglyceride does not undergo a rearrangement reaction in this treatment. Furthermore, this purification method does not require any special regeneration of the resin, only conditioning is required, and the operation is simpler than that of general ion exchange resin treatment. Furthermore, the present invention was completed based on the discovery that the chromatographic treatment method does not require any conditioning and can be fully applied to industrialization.

The monoglyceride-containing fats and oils used in the present invention are synthesized from glycerin using catalysts, enzymes, and microorganisms, including enzymatic hydrolysis of triglycerides and diglycerides.
Examples include fats and oils containing monoglycerides obtained by transesterification.

In addition, the ion exchange resins that can be used in the present invention include porous type and highly porous type weakly acidic Cal? Acid-type cation exchange resins are preferred, but are not limited thereto.

And as a specific example, "Diaion" WK-1
0, WK-11, WK-20 (manufactured by Mitsubishi Chemical Industries, Ltd.), [
Amberlight J IRC-50, IRC-75, IR
C-84 (manufactured by Rhom & Haas Co-),
11)owex, CCR-24 (The Dow・C
(manufactured by hemi cal Co.), rDuolit
e CC-3j (Diamond-8hamroc
(manufactured by K Chemical Co.).

Examples of the non-polar solvent used as an eluent include aliphatic hydrocarbons, aromatic hydrocarbons, and their non-rhodenated products, but from the viewpoint of boiling point and safety, C5 to C8 aliphatic hydrocarbons, such as pentane, are preferred. , hexane, hezotane, octane and the like are preferred. Further, as the polar solvent used as the eluent, water, lower alcohol, t[#'nol, ethanol, f-t4nol, etc. are preferable. First, as one method of the present invention, a solution containing a monoglyceride is brought into contact with an ion exchange resin in a batch or column operation, a non-polar solvent is used as an eluent, and compounds other than monoglycerides, such as free fatty acids, Elutes diglycerides, triglycerides, etc. The monoglyceride is then eluted using a polar solvent. The polar solvent eluate thus obtained can be further processed by conventional means such as concentration and freeze-drying to isolate and collect the desired monoglyceride.

Regarding the reuse of the resin, as long as the monoglyceride-containing oil and fat does not contain proteins or basic substances, there is no need to regenerate it, and it is only necessary to condition it using the nonpolar solvent used.

Furthermore, if it contains proteins or basic substances, it can be used by removing it by pretreatment or by regenerating it by ordinary hydrochloric acid aqueous solution treatment.

This method has the advantage that monoglycerides can be obtained with a high purity of 95% or higher and a yield of 95% or higher.Moreover, the monoglycerides obtained by enzyme reaction are normally extremely difficult to produce. A product that remains in the same position and has almost no dislocations can be obtained.

Another method is to contact an ion-exchange resin packed in a column with a solution containing a monoglyceride-containing oil and a developing solvent, and develop it chromatographically using a mixed solvent of a non-polar solvent and a polar solvent.

As the non-polar solvent and the polar solvent, the same solvents as those shown in the above method can be used, but the mixing ratio is 2 to 20 parts by volume of the polar solvent to 100 parts of the non-polar solvent.
is preferred.

When developed using this mixed solvent, triglycerides, diglycerides, and free fatty acids are preferentially eluted in a mixed state, and then only monoglycerides are eluted.

Depending on the conditions, some free fatty acids may be mixed into the monoglyceride elution section as tailings, but in that case, a highly purified monoglyceride section can be obtained by removing the initial stream of the monoglyceride elution section. The solution of the monoglyceride fraction thus obtained can then be treated by conventional means such as concentration and lyophilization to isolate and collect the desired monoglyceride.

This method does not require any special treatment when reusing and can be used for the next operation, and the mixed solvent recovered in the concentration process does not require any special purification such as rectification, and can be eluted as is. It has the advantage of being able to be used as a liquid, greatly simplifying the process, and achieving a yield of 80 g or more.

According to these two methods, by specifically adsorbing and separating monoglycerides from various monoglyceride-containing solutions onto an ion exchange resin without pretreatment, monoglycerides with a high recovery rate and a purity of 95% or higher can be obtained. The process can be significantly simplified compared to the known purification methods. In addition, it is possible to obtain monoglycerides of β-position fatty acids without transfer of fatty acid groups, which is considered to be extremely difficult industrially.Furthermore, since this method does not require high temperatures, it is possible to obtain unsaturated monoglycerides that are unstable to heat. Its usefulness is clear, as it can be applied to systems containing fatty acids.

It is known that monolyluine, which is a type of monoglyceride, exhibits the same effect on lowering and suppressing the increase in blood cholesterol at one-hundredth the amount of conventional substances. (JP-A No. 59-53430) Among the monolyruines, β-monolyluine has a better effect than α-monolyluine. To selectively produce β-monoglyceride, there is a method of hydrolyzing fatty acids at the α and d positions of triglyceride using an enzyme, but according to the purification method of the present invention, as a method for concentrating β-monoglyceride, heat is used. Since no rearrangement occurs, stable separation from other components can be achieved.

The present invention will be explained in detail below with reference to Examples.

Example 1 Lysosisdelemer lipase (
400 g of acetic acid buffer solution (pH 5, 6) in which 100,000 units (manufactured by Seikagaku Kogyo Co., Ltd.) were dissolved was added, and after shaking at 40°C for 2 hours, the mixture was extracted with ethyl ether to obtain an enzymatic hydrolysis reaction solution.

This includes 35.2% free fatty acids and 7.5% monoglycerides.
%, diglyceride 27.2%,) diglyceride 30.1
%, and the β rate of monoglyceride (β-monocryceride/monoglyceride) was 95.4%.

"Diaion" WK-1, a weakly acidic ion exchange resin
0 (manufactured by Mitsubishi Chemical Industries, Ltd.) 365m7! After thorough washing and dehydration with anhydrous ethanol, it was packed into a glass column with an inner diameter of 32 mm, and was further washed with 365 ml of absolute ethanol, followed by conditioning by passing 365 ml of n-hexane.

After conditioning, 36.59 g of the above-adjusted enzyme hydrolysis reaction solution was fed to the top of the column, and 6.5 g of n-hexane was added to the top of the column. 10100O at the speed of I UlA+! Then, the solvent was changed to absolute ethanol and the monoglyceride was eluted by flowing 400 rILl.

When this absolute ethanol fraction was concentrated at 30° C. under reduced pressure, a 2.8% viscous liquid was obtained, and analysis showed that it contained 95.6% monoglycerides and 4.4% free fatty acids. Moreover, the β rate of monoglyceride was 92.3. Figure 1 shows the changes in each component in the column effluent.

Example 2 Weakly acidic ion exchange resin [Diaion WK-10J36
After thorough washing and dehydration of 5 ml with absolute ethanol in a glass beaker, 365rn7! Washing was performed four times with n-hexane.

This washed resin was mixed with 100% of the enzyme hydrolysis reaction solution used in Example 1. n-hexane solution containing li' 10100
After adding O and stirring gently for 30 minutes at room temperature, the filtration solution was removed, and the resin was further washed with n-hexane 730116.

The washed resin was transferred to 365ml of absolute ethanol, left to stand at room temperature for 30 minutes, filtered through the mouth, and further washed with 365ml of absolute ethanol. When the ethanol solution obtained by filtration and washing was concentrated at 30°C under reduced pressure, 7.3 g of a viscous liquid was obtained, and analysis revealed that monoglyceride 9.
The free fatty acid content was 7.4% and the free fatty acid content was 2.6%. Moreover, the β rate of monoglyceride was 93.7%.

Example 3 Weakly acidic ion exchange resin [Diaion JWK-1036
After thoroughly washing and dehydrating 5rrLl with absolute ethanol, it was packed into a glass column with an inner diameter of 32mm, and then t=570tra.
Further, 365 ml of absolute ethanol was passed through for cleaning, and then a mixed solvent of n-hexane and ethanol = 95:5 (volume ratio) was used for conditioning.

Enzyme hydrolysis reaction solution 3 prepared in Example 1 was placed at the top of the column.
Feed 6.59 and n-hexane:ethanol = 95
A mixed solvent of :5 (volume ratio) was passed through the solution at a rate of 5.1 mV at room temperature, and the eluted fraction after 500 rnl was collected and concentrated at 30°C under reduced pressure, resulting in 2.5 g of a viscous liquid. When analyzed, the monoglyceride content was 98.5%.
, the free fatty acid was 1.5%, and the recovery rate of this monoglyceride was 90%, and the β rate was 91%.
．． It was 7%. The diagram shows the changes in each component in the column effluent.
Shown in 2.

Example 4 Weakly acidic ion exchange resin “Diaion JWK-1011
After thorough washing and dehydration of 50 m with absolute ethanol, it was packed into a glass column with an inner diameter of 32 cm (t = 1800 mm).
Further, 1 ton of absolute ethanol was passed through the solution for cleaning, and then a mixed solvent of n-hexane:ethanol=90:10 (volume ratio) was used for 19 passes for conditioning.

36.5 g of the safflower hydrolyzate prepared in Example 1 was fed to the top of the column, and n-hexane:ethanol = 9
5. Mixed solvent of 0:10 (volume ratio) at room temperature. l m/
When the liquid was passed through at a rate of A+, the outflow section for L 1290 rnlnl and later was condensed under reduced pressure at 30°C, resulting in a viscous liquid of 2.
．． 3 g was obtained and analyzed, and the monoglyceride content was 99.5%, the recovery rate was 85%, and the β rate was 90.8%.

Example 5 400 ml of acetic acid buffer (pH 5, 6) in which 100,000 units of Rhizopus deremer lipase (Seikagaku Kogyo Co., Ltd.) was dissolved was added to 100 ml of soybean oil, shaken at 40°C for 2 hours, and then extracted with ethyl ether. An enzymatic hydrolysis reaction solution was obtained.

This includes 42.3% free fatty acids and 9.3% monoglycerides.
3%, diglyceride 23.6%, triglyceride 24.
81 is included, and the β rate of monoglyceride is 97.1.
It was Chi.

Weakly acidic ion exchange resin “Diaion WK-10J36
After thoroughly washing and dehydrating 5ml with absolute ethanol,
Fill a glass column with 2 pus at t=570. ) Further, washing was carried out by passing 365 mJ of anhydrous ethanol, and then switching to a mixed solvent of n-hexane:ethanol = 95:5 (volume ratio), passing 365 mJ of liquid, was carried out for conditioning.

Add the above soybean oil enzymatic hydrolysis reaction solution 36.5 to the top of the column.
9 and n-hexane:ethanol = 95:5 (
When a mixed solvent (volume ratio) was flowed at a rate of 6.1 m44 at room temperature, the eluted fraction after 500 m1 was collected and concentrated at 30°C under reduced pressure, 3.4 g of a viscous liquid was obtained, which was analyzed. The monoglyceride content was 98.0% and the free fatty acid content was 2.0%.

In addition, the recovery rate of this monoglyceride was 91.0%,
The β rate was 94.3%.

Example 6 [Power of Diamond Ion JWK-10 used in Example 2,
Solution containing free fatty acids and monoglycerides in rum (fatty acid: monoglyceride = 80:20 (weight ratio)) 28.5.
9 was fed and then developed under the same conditions as in Example 2.

When the effluent fractions from 400 to 400 were collected and concentrated at 30°C under reduced pressure, a viscous liquid of 5.54% was obtained, which was analyzed and found to be 97.5% monoglyceride, 2.5% fatty acid, and the recovery rate of monoglyceride was 94%. In addition, in the outflow section after 500 d, yield was 4.40 g Monoglyceride content 99.8% Recovery rate was 77.2 q/y Example 7 rAmberlystJ, a weakly acidic ion exchange resin
A-21 (manufactured by Rohm and Haas Co)
After thoroughly washing and dehydrating 365 mJ with absolute ethanol,
It was packed into a glass column with an inner diameter of 32 mm, and washed with 365 d of absolute ethanol, and then switched to a mixed solvent of n-hexane:ethanol = 95:5 (volume ratio) 36
Conditioning was performed by passing 5 ml of liquid through the tube.

Enzyme hydrolysis reaction solution 3 prepared in Example 1 was placed at the top of the column.
Feed 6.5g and n-hexane:ethanol = 95
:5 (volume ratio) mixed solvent at room temperature 6, 11rL415
), and the elution fraction after 500 d was collected and concentrated at 30°C under reduced pressure. 2.7 g of viscous liquid was obtained, and analysis of this revealed that the monoglyceride content was 89% and the free fatty acid content was 11%. Met. Furthermore, the recovery rate of monoglyceride was 88%, and the β rate was 92.1%.

[Brief explanation of the drawing]

Figures 1 and 2 are Example 1 and Example 3, respectively.
The results are shown below. The horizontal axis in the figure is the total amount of eluent <,vt'>
The t-1 vertical axis is the weight of each component in 10Ornl (gAoo
ml). a is triglyceride, b is free fatty acid,
C represents diglyceride and d represents monoglyceride.

Claims (4)

[Claims] (1) A feature of the method is to contact an oil or fat containing a monoglyceride with an ion exchange resin to selectively adsorb the monoglyceride onto the ion exchange resin, and then to contact the ion exchange resin with an eluent to elute the monoglyceride. A method for purifying monoglycerides. (2) A monoglyceride purification method characterized by chromatographically treating a liquid containing monoglyceride-containing fat and oil and a developing solvent in an ion exchange resin packed in a column to separate monoglyceride and components other than monoglyceride. . (3) The oil and fat containing monoglyceride is an oil and fat obtained by hydrolyzing natural oil and fat with an enzyme, Claim No. 1
) and (2). (4) The purification method according to claims (1) and (2), wherein the ion exchange resin is a weakly acidic carboxylic acid type ion exchange resin.