JPS5821618B2 - Method for isolating creatine from meat canned and steamed waste liquid - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for isolating creatine, an important amino acid, and creatinine.

Creatine and creatinine have the following chemical formulas I and ■, respectively.
It is an amino acid with

Creatinine is an anhydride formed by dehydration within the molecule of creatine, and these two substances can be artificially changed into each other.

Hereinafter, creatine and creatinine will be collectively referred to as creatines.

Creatine compounds exist mainly in the muscles of higher animals such as fish and higher, and their content is approximately 0.1 to 0.6% for creatine and 0.01 to 0.1% for creatinine. It is useful as a test reagent and a reagent for physiological medical research on myasthenia and creatine metabolism disorders.

Recently, it has become useful as a raw material for the production of creatine phosphate, which is an energy storage substance for muscle exercise, and as a flavor improver in food.6 However, there have been very few publications regarding the extraction, separation, or isolation of creatine. However, after precipitating and removing proteins using heavy metal salts or organic agents, creatine compounds are simply precipitated and collected using alcohol, picric acid, etc., which is not satisfactory.

In other words, large amounts of chemicals are used for protein precipitation, and the resulting creatine may contain residual heavy metal salts and organic agents.
It has the disadvantage that these must be purified and removed.

The present inventors have found that creatine, an important substance as mentioned above, is present at 0.18 to 0.43% in the waste liquid from meat cooking, and also histidine, an important amino acid. It was discovered that 0.01 to 0.50% of The liquid from which mold and oil have been removed is brought into contact with a strongly acidic cation exchange resin having a sulfonic acid group as an exchange group, the amino acids are adsorbed onto the resin, separated from the non-adsorbed liquid, and a dilute ammonia aqueous solution is extracted from the adsorption resin. The amino acids are eluted in a stepwise manner, and the eluate is sequentially brought into contact with a strongly basic anion exchange resin having a quaternary ammonium group as an exchange group, and amino acids such as creatine and histidine other than creatinine are adsorbed onto the resin. This is a method for isolating creatine and/or creatinine from a meat cooking waste liquid, which is characterized by separating and concentrating and separating creatine and/or creatinine from a non-adsorbed liquid.

In the present invention, the waste liquid from cooking meat refers to the cooking liquid or broth produced when processing meat, such as fish, minced meat, and other animal meat, from which insoluble solids and fats and oils have been removed. Are suitable.

In addition to creatine and stidine, these aqueous solutions contain other amino acids, soluble proteins, coloring components, fishy odor components, salts, and others, which cannot be separated using normal separation methods, and many of them are Due to its fishy taste and odor, it is either concentrated and used as feed or is disposed of as waste.

When these solutions are brought into contact with a strongly acidic cation exchange resin, creatine, histidine, other amino acids,
Most of the coloring components and raw odor components are adsorbed by the exchange resin.

At this time, the liquid after adsorption has an acidic pH of about 1 to 3 due to the exchange action of the strongly acidic cation exchange resin, but this pH
The adsorption limit can be determined by the increase in H.

The adsorption limit can also be determined by the Barrett reaction, which is a reaction specific to creatine, or the Jaffe reaction, which is a reaction specific to creatinine.

As mentioned above, after washing the resin with water, the substances adsorbed on the strongly acidic cation exchange resin are eluted with a dilute alkaline aqueous solution, preferably dilute aqueous ammonia. , and finally a small amount of fishy odor components are eluted.

Fishy odor components are strongly adsorbed by cation exchange resins, so they are only slightly eluted with dilute alkaline aqueous solutions.

The color of the eluate is initially cloudy while other amino acids are eluting, but it is almost colorless and transparent while creatine and creatinine are eluting, and colored components begin to elute from the time when histidine has finished eluting. Since it starts, it can be easily separated into three parts if necessary.

However, it is also possible to contact the next strongly basic anion exchange resin in the order of elution without separating these eluates.

The end point of this elution can be easily determined by Jaffe's color reaction with creatinine, an increase in pH, or the outflow of colored components.

When the eluate from the above-mentioned strongly acidic cation exchange resin is brought into contact with the strongly basic anion exchange resin, either as it is or only the portion rich in creatine and histidine is contacted with the strong basic anion exchange resin, creatine is not adsorbed at all, but histidine and other amino acids are absorbed. , colored components, etc. are adsorbed.

This is a highly unexpected result.

That is, creatine is a neutral amino acid, but it has a carboxyl group, so it should be adsorbed to an anion exchange resin, but in reality it is not adsorbed at all.

This is clear from the fact that creatine was not adsorbed at all when the reagent creatine aqueous solution was brought into contact with the anion exchange resin, and the presence of creatine compounds could not be confirmed from the eluate of the adsorbed anion exchange resin with dilute hydrochloric acid. be.

Since the non-adsorbed liquid after contact with the strongly basic anion exchange resin contains only creatine compounds, it is directly concentrated to dryness to obtain crystals of creatine compounds with a high purity of 80% or more.

The yield of this creatine compound is 6% of the amount contained in the stock solution.
The yield is as high as 0% or more.

Furthermore, if fractional crystallization is performed during the concentration process, creatine and creatinine can be further separated into 95% L, 1. It can be isolated with high purity.

In other words, creatinine has a higher solubility in water than creatine, so if it is cooled during concentration, creatine will precipitate (so if you separate it and continue to concentrate the mother liquor again, creatinine crystals will precipitate out). After fractionation, crystalline creatinine can be obtained.

- If the histidine adsorbed on the strongly basic anion exchange resin is eluted with a dilute acid, preferably dilute hydrochloric acid, it will be separated from the colored components and eluted.

If this eluate is directly concentrated to dryness, histidine containing no creatine compounds can be obtained with a purity of 80% or more, and the yield is also 70% or more.

The end point of this elution can be determined by the ninhydrin reaction and the diazo reaction.

The strongly acidic cation exchange resin and the strongly basic anion exchange resin used in the present invention each have a sulfonic acid group or a quaternary ammonium group as an exchange group, and can be regenerated after use by following a conventional method. Bye.

That is, in the case of cation exchange resin, tap water, 1-2N hydrochloric acid,
It is sufficient to wash in the order of ion-exchanged water, and in the case of 7 anion-exchange resin, after histidine elution, ion-exchanged water, 1-2N.
It may be washed in the order of sodium hydroxide and ion-exchanged water.

As mentioned above, the method of the present invention has the advantage that it is possible to separate creatine, which is a useful substance, in a higher yield and with higher purity than other contaminant components in an extremely simple manner from the waste liquid of meat boiling. This method eliminates the disadvantage of using large amounts of heavy metal salts and organic agents as in conventional methods, and is also economically effective.

Furthermore, according to the method of the present invention, it is possible to effectively utilize the steamed liquid or broth of fish, livestock, meat, and joints, which has traditionally been discarded due to its fishy taste and odor, which has little utility value, and is one of the causes of polluting rivers. It also has the effect of reducing pollution.

Example 1 Steaming waste liquid produced during the production of tuna stories (94% water, 0.2% creatine, 0.4% histidine, 1.0% fat)
8%, ash 0.58%, crude protein 3.7%) 3850 kg
(total BOD 130 kg), add 9 to 3500 of the liquid from which insoluble solids and oil have been removed, and add 9 to the strongly acidic cation exchange resin (
After passing through the Amberlite 200.45 ol) column at a flow rate of 200 kg/min, it was washed with 640 ml of ion-exchanged water.

The liquid that passed through the resin tower was combined with this washing water, neutralized, and concentrated to obtain a seasoning extract.

Next, elution was carried out with 4050 kg of 0.06N aqueous ammonia, and the eluate was sequentially mixed with a strong basic anion exchange resin (Amber
lite IRA-900, 240, g) Flow rate inside the column: 1
00 kg/min, and the passed liquid was concentrated to dryness and then ground to obtain 6.2 kg of powder.

This product contains creatine with a purity of 81% (creatine 21%).
%, creatinine 60%), and the pure yield (hereinafter simply referred to as yield) was 65%.

Next, add 0.0% of a strong basic anion exchange resin. IN-Hydrochloric acid 450
kg was eluted.

During elution, a black stripe (colored component) appears from approximately the center of the resin column, and elution can be continued depending on how the stripe falls.

Concentrate 450 kg of the eluate, dry it, and crush it to 13.2 kg.
g of histidine powder was obtained.

This product contained no creatine, had a purity of 83%, and a yield of 71%.

The used ion exchange resin was regenerated by a conventional method.

At this time, the total BOD of the cation exchange resin recycled waste liquid was 0.6.
kg, and the total BOD of the anion exchange resin recycled waste liquid is 0.
．． It weighed 3 kg.

Even if both were combined, the total BOD was 0.9 kg, which was a 99% decrease compared to the total BOD of the undiluted solution, which was 130/9.

Example 2 The eluate containing only creatine compounds in Example 1 was concentrated to 1/100 and cooled to 10°C to obtain a purity of 95%.
Crystalline Creatine 1. Oky was filtered off.

On the other hand, the mother liquor was further concentrated to just before dryness, 1.0 kg of hot water was added, the mixture was thoroughly stirred, and the mixture was cooled to 10° C. to obtain 2.0 kg of crystalline creatinine with a purity of 95%.

Example 3 3,500 kg of the liquid from which insoluble solids and oil were removed from 3,800 kg (total BOD 76 kg) of the broth after blood removal (creatine 0.40%, histidine 0.02%) produced during the production of Whale Tales was used as an example. The mixture was treated in the same manner as in 1 to obtain 10.9 kg of creatine powder and 0.6 kg of histidine powder.

Creatine had a purity of 85% and a yield of 61%, and histidine had a purity of 81% and a yield of 70%.

The total BOD of recycled waste liquid Q from ion exchange resin is 0.7 k.
g, and the BOD reduction rate was 99%.

Example 4 The broth produced when making mackerel flakes (creatine 0.1
8%, histidine 0.5%) 4000 kg (total BOD4
After solid-liquid separation of 4 kg), 3900 kg of the liquid was used as a strong acidic cation exchange resin, Duolite C-25D 4001, and as a strong basic anion exchange resin, Duoli
te A-101D was treated using 3001.

The procedure of Example 1 was repeated except that 2,500 kg of 0.08N ammonia water and 900% of 0.2N hydrochloric acid were used as the eluent to obtain 5.3 kg of creatine powder and 17.3 kg of histidine powder.

The creatine powder had a purity of 84% and a yield of 62%, and the histidine powder had a purity of 83% and a yield of 72%.

In addition, the total BOD of recycled waste liquid from ion exchange resin is 0.4 kg.
The BOD reduction rate was 99%.

Example 5 Boiled soup produced during the production of Yakitori Manga (no creatine)
．． 41%, Histidine 0.01%) After solid-liquid separation of 9 into 4000, 3750 kg of the liquid was used as a strongly acidic cation exchange resin, Diaion PK212.5401, and as a strong basic anion exchange resin, Diaion PA418.270.
1.

2500 kg of 0.06 N-ammonia water as eluent
and 0. Example 1 using 540 kg of IN-hydrochloric acid
In the same manner as above, 11.2 kg of creatine powder and 0.35 kg of histidine powder were obtained.

The creatine powder had a purity of 88% and a yield of 60%, and the histidine powder had a purity of 80% and a yield of 70%.

Example 6 Boiled soup produced during the production of beef stories (creatine 0.4
After solid-liquid separation of 4,000 kg (3% histidine, 0.05% histidine), 3,800 kg of the liquid was treated with the same type and amount of ion exchange resin as used for the height measurement, and extraction was performed in the same manner to obtain 12.6 kg of creatine powder. 1.8 kg of histidine powder was obtained.

The creatine powder had a purity of 82% and a yield of 60%, and the histidine powder had a purity of 81% and a yield of 72%.

Claims (1)

[Claims] 1. A liquid containing creatine and/or creatinine and amino acids such as histidine from which insoluble solids and oil have been removed from a meat boiling waste liquid is brought into contact with a strongly acidic cation exchange resin having a sulfonic acid group as an exchange group. , the amino acids are adsorbed on the resin, separated from the non-adsorbed liquid, the amino acids are eluted from the adsorption resin with a dilute aqueous ammonia solution, and the eluate is sequentially used as an exchange group to
creatine and amino acids such as histidine other than creatinine are adsorbed and separated by the resin, and creatine and/or creatinine are concentrated and separated from the non-adsorbed liquid. A method for isolating creatine and/or creatinine from meat boiling waste liquid.