JPH0687751B2 - Method for collecting lysolecithin containing high concentration of lysophosphatidylcholine - Google Patents

Description

TECHNICAL FIELD The present invention relates to a method for collecting necessary components from lysolecithin as a raw material, and in particular, it contains lysophosphatidylcholine (LPC) at a high concentration among various components. The present invention relates to a method for collecting lysolecithin.

(Background Art) Lysolecithin (lysophospholipid) has a hydroxyl group in the molecule, so it has a greater hydrophilicity than lecithin (phospholipid) and has the characteristic of forming an emulsion stable against changes in pH and temperature. Among them, lysophosphatidylcholine (LPC) behaves like a nonionic surfactant, and is said to maintain surface activity even in a solution with high salt concentration. Therefore, LPC is expected to be widely applied in various fields such as food, cosmetics, and pharmaceuticals.

By the way, commercially available lysolecithin, animal and vegetable lecithin such as egg yolk lecithin and soybean lecithin, by acting an enzyme, by hydrolysis of the lecithin, it is produced, although the composition is slightly different depending on the raw material, Generally, in addition to the lysophosphatidylcholine (LPC),
It contains lysophosphatidylethanolamine (LPE), lysophosphatidic acid (LPA), and various unreacted unreacted phospholipids, and in many cases, neutral fats and oils derived from the raw material lecithin. So LPC
The concentration is not so high.

Thus, in order to produce lysolecithin containing a higher concentration of LPC, a method is known in which neutral oils and fats are removed using a solvent such as acetone and then alcohol fractionation is performed.
However, this method is troublesome to operate, and when lysolecithin obtained from plant lecithin such as soybean lecithin is used as a raw material, since the raw material contains a large amount of alcohol-soluble components other than LPC. However, even if the alcohol fractionation operation is repeated, it is very difficult to raise the LPC concentration to 70% or more.

The chromatographic method using silica gel, etc.
Can be fractionated, but it has poor productivity, is expensive, and is not an industrial method.

(Problem to be solved) The present invention has been made against the background of such circumstances, and the problem to be solved is that, from a raw material lysolecithin (lysophospholipid mixture), lysophosphatidylcholine ( LPC) is efficiently extracted, and lysolecithin containing a high concentration of LPC is advantageously collected.

(Solution) As a result of intensive studies conducted by the present inventors in order to solve the above problems, various lysolins were obtained by performing an extraction operation of defatted lysolecithin with a predetermined mixed solvent of carbon dioxide and alcohol. Based on this finding, we found that among the unreacted phospholipids that remained unreacted with lipids, lysophosphatidylcholine (LPC) was easily extracted, and unreacted phospholipids were difficult to extract. Thus, the present invention has been completed.

That is, the present invention, after defatting lysolecithin obtained by acting an enzyme on lecithin, using a mixed solvent consisting of carbon dioxide in a liquid state or supercritical state and an alcohol having 1 to 4 carbon atoms as an extraction solvent, In the mixed solvent, an extraction operation of the defatted lysolecithin is performed, and the mixed solvent is removed from the extract, thereby collecting lysolecithin containing a high concentration of lysophosphatidylcholine. Is.

(Specific configuration) By the way, in the present invention, the lysolecithin to be subjected to the extraction operation is obtained by enzymatically hydrolyzing lecithin of animal or plant origin, for example, egg yolk, bovine brain, pig brain, etc. The obtained animal-derived lecithin, soybean,
It can be obtained by allowing a given enzyme to act on plant-derived lecithin obtained from plant seeds such as corn and rapeseed. Although phospholipase A ₂ is usually used as the enzyme, a commercially available preparation can be used for this. In addition, lecithin and lysolecithin can be used because various types of lecithin and lysolecithin have been commercialized and are commercially available.

Then, when the raw material lysolecithin obtained by such an enzyme treatment contains neutral fats and oils, the fats and oils are removed in advance prior to the LPC extraction operation. As a method for removing oil and fat, various conventionally known methods can be used. For example, the oil and fat can be extracted and separated by acetone extraction or supercritical carbon dioxide extraction.

Further, in particular, when the raw material lysolecithin is derived from vegetable lecithin, in the extraction operation of LPC, in order to facilitate contact between the raw material lysolecithin and the extraction solvent, the lysolecithin is a powder insoluble in the extraction solvent. It is desirable to premix with the granules. This is because, in general, the extraction solvent permeates the solid or liquid lysolecithin phase very poorly, and the dissolution rate of the lysolecithin phase in the extraction solvent is also slow. This is because not only is it time consuming and the extraction solvent is used more than necessary, but there is a risk of unstable extraction with poor reproducibility due to the difference in permeation of the extraction solvent. Therefore, the raw material lysolecithin,
If the powder and granules are mixed in advance prior to the extraction operation, the contact area of the lysolecithin with the extraction solvent can be greatly increased and the extraction can be facilitated. Thus, the flow of the solution can be made uniform.

By the way, as a specific example of such a granular material, one that is insoluble in the extraction solvent consisting of carbon dioxide and alcohol described later and is inactive to the raw material lysolecithin (that is, reacts with the raw material lysolecithin or has a strong adsorption action) There is no particular limitation, and starch powder, soybean protein, egg white, cellulosic powder, porous silica and the like can be used. In addition, the alcohol blended in the extraction solvent is 2
Dextrin powder can also be used if it does not contain more than 50% water.

Further, the mixing ratio of the granular material and the raw material lysolecithin may be appropriately determined to a ratio at which uniform mixing can be performed, but since it is not economical to use the granular material more than necessary, the raw material lysolecithin is used. 1 part by weight of
It is suitable to mix in the range of 0.2 to 4 parts by weight.

Furthermore, when the raw material lysolecithin is in the form of powder or liquid, it is easy to mix it uniformly with the granular material,
In the case of a solid or a liquid of extremely high viscosity, it is advisable to dissolve the raw material lysolecithin in a suitable solvent, mix it with the granular material, and then dry it. At that time, when the solvent used for dissolving the raw material lysolecithin is alcohol which is one component of the extraction solvent described later, the drying after mixing can be omitted.

In addition, in lysolecithin derived from egg yolk, the protein in egg yolk plays the role of the above-mentioned powder and granules, so that it is not necessary to add the powder or granules in particular. In short, it suffices to consider the addition of the powdery or granular material so that the extraction can be tried evenly according to the properties of the raw material.

The composition of the raw material lysolecithin thus prepared is
In addition to lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE) and lysophosphatidic acid (LPA), as well as unreacted phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol that remained unresolved. (PI) etc. are included. Then, in the present invention, such a raw material lysolecithin is subjected to an extraction operation using the following extraction solvent so that LPC can be efficiently extracted and the concentration thereof is remarkably increased.

That is, in the present invention, a mixed solvent of carbon dioxide in a liquid state or a supercritical state and an alcohol having 1 to 4 carbon atoms is used as an extraction solvent. Carbon dioxide among them is well known as an extraction solvent, but in the present invention, it also has the function of controlling the extraction ratio of alcohol-soluble lysolecithin and unreacted lecithin. The supercritical state of carbon dioxide is
Temperature is critical temperature: 31.1 ℃ or more, pressure is critical pressure: 7.38MPa
(75.2 kg / cm ² ) or more, and the liquid state is
A state under a temperature lower than the critical temperature.

On the other hand, alcohol works as a solvent for LPC, LPE, PC, PE, etc., and any of those having 1 to 4 carbon atoms can be used, and among them, one kind or two or more kinds are appropriately combined. Will be selected. If the carbon number is larger than 4, the solubility of lysolecithin will be low and, in effect, it will not be able to act as an extraction solvent. Also,
The water content of the alcohol is preferably about 10% by weight or less. If the water content exceeds that level, not only will the solubility of lysolecithin be reduced, but also LP
This is because there is a risk of adversely affecting the extraction of high-concentration LPC, such as reducing the sorting ability of C.

The mixing ratio of the alcohol and the carbon dioxide is appropriately determined according to the composition of the raw material lysolecithin and the like, but the blending amount of the alcohol is preferably about 4 to 20% by weight. If the amount of alcohol is less than this range, the solubility of LPC tends to be small and the yield tends to decrease.On the contrary, if the mixing ratio is larger than this, the action of carbon dioxide to control the solubility of lysolecithin This is because there is a concern that the LPC concentration will decrease and it will be difficult for the LPC concentration to increase.

By the way, FIG. 1 is a flow sheet showing an example of an extraction apparatus to which the method of the present invention is applied. As shown therein, carbon dioxide is supplied from the cylinder 2 while a predetermined amount which is an entrainer from the tank 4 is supplied. The alcohol is supplied, and they are mixed in a predetermined ratio on the flow path, whereby the extraction solvent is prepared. Then, the extraction solvent is supplied into the extractor 6 in which the raw material lysolecithin is contained and which is controlled at a predetermined temperature and a predetermined pressure, whereby LPC is extracted.

Then, the extract in which the soluble component in the raw material lysolecithin is dissolved is taken out of the extractor 6, and is first returned to normal pressure in the separators 8 and 10 to separate into carbon dioxide and alcohol solution. To be done. Then, the alcohol is removed by vacuum distillation by a conventional method to obtain the desired lysolecithin containing LPC at a high concentration. In addition, 3,5
Is a pump, 12 is a flow meter, and 14 is an integrating gas meter.

More specifically, in the above extraction operation, generally, the lower the temperature or the higher the pressure, the greater the solubility of the raw material lysolecithin in the extraction solvent. On the other hand, when unreacted phosphatidylcholine (PC) remains in the raw material lysolecithin, PC is extracted together with LPC. In that case, it is better to set the extraction temperature slightly higher. Incidentally, more specific temperature conditions and pressure conditions, the LPC concentration and yield of the target, depending on the composition of the raw material lysolecithin, etc., the optimum conditions will be appropriately selected, generally, the temperature is 70 Above ℃, there is a problem that the thermal change of lysolecithin becomes significant, excessive low temperature extraction is not economical, and when the pressure exceeds 45 MPa, there is a problem in the design and maintenance of the equipment. As the carbon, when using a supercritical state, temperature: 31 ~ 65 ℃, pressure: preferably in the range of 8 ~ 45 MPa,
More preferably, the temperature is 40 to 60 ° C. and the pressure is 10 to 30 MPa. When using liquid carbon dioxide,
It is preferable that the temperature is -20 to 30 ° C and the pressure is 5 to 8 MPa.

In this way, the defatted raw material lysolecithin,
When applying an extraction solvent consisting of carbon dioxide and alcohol, among various lysolecithin and unreacted lecithin, LPC is most efficiently extracted, on the contrary, unreacted lecithin is difficult to extract, PI among them is It is hardly extracted. Therefore, most of PI remains as a residual component by passing through such extraction operation, and also in the extract.
Since a large amount of lysolecithin other than LPC and unreacted lecithin are also effectively suppressed, the LPC concentration of the extracted lysolecithin can be significantly increased.

The specific extraction time is set appropriately according to the scale of the extraction device, the feed rate of the mixed solvent of carbon dioxide and alcohol, other extraction conditions, the composition of the raw material lysolecithin, etc. If the time becomes too long, alcohol-soluble components other than LPC will gradually be mixed in, so it is generally desirable to collect the extraction category at a relatively early stage after the start of extraction.

(Examples) Hereinafter, several examples of the present invention will be shown to clarify the present invention more specifically, but the present invention does not impose any restrictions due to the description of such examples. Needless to say, it is not something to receive.

In addition to the following embodiments, the present invention further includes various changes and modifications based on the knowledge of those skilled in the art, in addition to the above specific description, without departing from the spirit of the present invention. It should be understood that improvements and the like can be added.

Example 1 A commercially available lysolecithin (Kyowa Hakko Kogyo Co., Ltd., Elmizer A) was used to remove the oil and fat using acetone according to a conventional method, and the solvent was removed under reduced pressure to obtain a powdered delipidated lysolecithin. And this defatted lysolecithin: 30 g, about 60
After dissolving in hexane (ml) and mixing with 60 g of dextrin powder (Matsutani Chemical Co., Pineflow), the solvent was removed under vacuum at 40 ° C. to prepare a powder mixture of lysolecithin and dextrin.

Then, this powder mixture was put into a 500 ml extraction container, and supercritical carbon dioxide and ethanol (containing 2% by weight of water).
A mixed solvent of and (ethanol ratio: 16% by weight) was continuously injected, and LPC was extracted for 8 hours. The extraction conditions are
The temperature was 40 ° C., the pressure was 30 MPa, and the average flow rate of the solvent was 200 N / h. The amount of lysolecithin obtained by such extraction is
The amount was 2.7 g, and the yield was 9.0%.

Then, the composition of the lysolecithin collected in this way was determined by the “standard oil and fat analysis method” (edited by Japan Oil Chemistry Association, 2.2.8.4a).
-86) "Phospholipid Phospholipid composition" was analyzed by a thin layer chromatographic method, and the results of main phospholipids are shown in Table 1 below. The composition of delipidated lysolecithin (control) used as an extraction raw material was also analyzed and shown in the same table.

As is clear from the results, the LPC concentration was increased from 30.1% (before extraction) to 64.8% (after extraction). Also,
Unreacted PE and PC were not extracted.

Example 2 Soybean lecithin fortified with phosphatidylcholine content (PC35, manufactured by Tsuru-Lecithin Industry Co., Ltd.): To 100 parts by weight,
Water: 50 parts by weight was added, and the mixture was stirred to form a uniform mixed solution. Then, add 0.1% phospholipase A ₂ preparation (N
OVO, lecitase 10L) was added, and the enzyme reaction was carried out at 50-60 ° C for 48 hours while adjusting the pH to 5.5-6.5 with 1N sodium hydroxide solution. Then, the obtained enzyme reaction product was vacuum dried at 50 to 60 ° C. to remove water, followed by washing with acetone and vacuum drying to remove fats and oils to obtain delipidated lysolecithin.

Next, 20 g of this defatted lysolecithin was dissolved in about 40 ml of ethanol and mixed with 60 g of dextrin, and then the solvent was removed under vacuum to prepare a powder mixture of lysolecithin and dextrin.

Then, this powder mixture was put in a 500 ml extraction container, a mixed solvent of supercritical carbon dioxide and ethanol (ethanol ratio: 16% by weight) was continuously injected, and LPC was extracted for 4 hours. The extraction conditions were temperature: 40 ° C., pressure: 30 MPa, average solvent flow rate: 200 N / h. The amount of lysolecithin obtained by this extraction was 5.8 g, and the yield was 29%.

The composition of the lysolecithin thus collected and the defatted lysolecithin used as the extraction raw material was analyzed, and the results shown in Table 1 were obtained, and it was found that the LPC concentration was increased to 88.1%.

Example 3 The same delipidated lysolecithin as in Example 2 (20 g) was dissolved in about 40 ml of ethanol, and after adding and mixing perlite (manufactured by Toko Perlite Industrial Co., Ltd., Topcoperlite No. 34): 30 g,
The solvent was removed under vacuum to prepare a powder mixture of lysolecithin and perlite.

Next, this powder mixture was put into a 500 ml extraction container, and a mixed solvent of liquefied carbon dioxide and ethanol (ethanol ratio: 8% by weight) was continuously injected to extract LPC for 8 hours. The extraction conditions were temperature: 5 ° C., pressure: 7 MPa, solvent average flow rate: 400 N / h.

The yield of lysolecithin thus obtained was 20.0%. Further, when the composition of the collected lysolecithin was analyzed, the LPC concentration was as high as 89.3%, and when liquefied carbon dioxide was added to the extraction solvent, similar to the case of adding supercritical carbon dioxide, good results were obtained. Was obtained.

Example 4 Commercially available powdered egg yolk: A dispersion was prepared by adding 80 parts by weight of water to 100 parts by weight, and 0.1% of phospholipase A ₂ preparation (NOVO, lecitase 10L) was added to this dispersion. Plus 1
While adjusting the pH to 5.5-6.5 with N sodium hydroxide solution,
The enzymatic reaction was carried out at 0-60 ° C for 24 hours. Then, the obtained enzyme reaction product was vacuum dried at 50 to 60 ° C. to obtain a lyzed powder egg yolk. Then, this lyzed powder egg yolk: 100 g, 500
The mixture was placed in a ml extraction tube, and supercritical carbon dioxide at a temperature of 40 ° C. and a pressure of 30 MPa was used for extraction for 8 hours at an average flow rate of 400 N / h to remove oil and fat in the lysed powder egg yolk.

Then, with respect to this defatted lysed powder egg yolk, temperature: 40 ° C., pressure: 30 MPa, average flow rate of solvent: 400 N / h under extraction conditions, a mixed solvent of supercritical carbon dioxide and ethanol (ethanol ratio: 16% by weight) for extraction of LPC
I went on time.

The composition of the thus-collected lysolecithin and the defatted lysed powder egg yolk used as the extraction raw material was analyzed.
The results shown in the table were obtained, and it was found that the LPC concentration was increased to 91.2%.

Examples 5 to 8 The powder mixture of lysolecithin and dextrin prepared in Example 1 was extracted using each of the four types of extraction solvents prepared by changing the type of alcohol mixed with supercritical carbon dioxide.

When the composition of the lysolecithin obtained in each extraction was analyzed, the results shown in Table 1 were obtained, and it was found that the LPC concentration was effectively increased in each case. Moreover, PE and PI were not extracted in any of the solvents.

Therefore, it was confirmed that any of the alcohols having 1 to 4 carbon atoms can be used as the extraction solvent, and particularly methanol and ethanol are good solvents.

(Effects of the Invention) As is apparent from the above description, according to the method of the present invention,
LPC can be efficiently extracted by a simple operation and with a good yield from a raw material lysolecithin containing various lysolecithins and various unreacted lecithins that have not been lysed.

Therefore, the method of the present invention is capable of simplifying the process and achieving good productivity, and is suitable for industrial production of high-concentration LP.
It is extremely useful as a method for producing lysolecithin containing C.

[Brief description of drawings]

FIG. 1 is a flow sheet showing an example of an extraction device to which the method of the present invention is applied. 2: cylinder, 4: tank 6: extractor, 8, 10: separator

Claims (1)

[Claims] 1. After defatting lysolecithin obtained by reacting lecithin with an enzyme, a mixed solvent comprising carbon dioxide in a liquid state or a supercritical state and an alcohol having 1 to 4 carbon atoms is used as an extraction solvent, A method for collecting lysolecithin containing a high concentration of lysophosphatidylcholine, which comprises extracting the defatted lysolecithin with a mixed solvent and removing the mixed solvent from the extract.