JPH0367069B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel method for producing halogenated hemin (hereinafter referred to as "hemin").

Hemin is useful as a raw material for producing protoporphyrin disodium, which has liver function-promoting and anti-inflammatory effects, and protoporphyrin cobalt, which has antitumor effects.

Hemin is a halide of heme, and heme, which is an iron complex of protoporphyrin, forms a molecular compound with the protein globin to constitute hemoglobin. Therefore, hemin is generally produced by separating it from hemoglobin in blood.

The current industrial method for producing hemin is
Glacial acetic acid method by Fischer et al. [Org.Synth. 21 , 53
(1941)], a method in which fresh blood is treated with large amounts of hot glacial acetic acid and salt.

However, this method has high production costs because it uses a large amount of acetic acid, and produces a large amount of waste liquid in which unnecessary protein parts are dissolved in acetic acid, which is difficult to process and recover. This method is not necessarily satisfactory as an industrial method, as it requires corrosion-resistant equipment for the recovery of blood, and it also requires fresh blood, and if old blood is used, the yield and purity are significantly reduced. Ta.

Therefore, the inventor of the present invention conducted intensive research to solve these drawbacks and found that lower alcohol containing a specific basic substance decomposes hemoglobin into heme and globin, dissolves heme, and solidifies the protein globin. It was discovered that the two can be easily separated because they are precipitated as a substance, and the present invention has been completed.

That is, the present invention involves extracting raw blood or blood meal with a lower alcohol containing a basic substance selected from ammonia and di-lower alkylamine, and then treating the obtained extract with sodium chloride or strontium chloride. The present invention provides a method for producing hemin characterized by the following.

The raw material blood in the present invention is human or bovine blood,
Blood from vertebrate animals such as pigs and sheep can be used. Also,
In the present invention, in addition to fresh blood, it is also possible to use blood powder obtained by drying this blood by spray drying or the like.

To carry out the method of the present invention, first, a lower alcohol containing a basic substance is added to live blood or blood powder, and the mixture is mixed and stirred.

As the basic substance, di-lower alkylamine such as diethylamine or diisopropylamine or ammonia is used, and as the lower alcohol, methanol, ethanol, propanol, etc. are used.
Of these, the combination of diethylamine and methanol is most preferred. Although the concentration of the basic substance varies depending on its type, it is generally suitable to be about 1.5 to 3%. Mixing and stirring is carried out at a temperature of room temperature to 50° C. for 1 minute to 1 hour. When doing this, the molecular bonds of hemoglobin are separated and decomposed into heme and globin, heme is dissolved in a solvent and extracted in the form of hematin or a basic substance coordinated, and globin is precipitated as a solid. The two can be easily separated.

Next, hemin can be obtained by reacting the extract thus obtained with sodium chloride or strontium chloride. The hemin thus obtained has extremely high purity.

As mentioned above, the present invention can completely extract the heme portion, resulting in a high yield, and the protein in the extraction residue can be used as a protein source in animal feed etc. just by drying.
In addition, since acetic acid is not used in the extraction operation, there is no problem of corrosion of the equipment, and since blood powder can be used, raw materials are easily available.

Next, an example will be given and explained.

Reference Example 1 Put 2 ml of methanol containing 2.5% diethylamine into a 5 ml centrifugal spittoon, add 50 µ of fresh human blood while stirring, and perform extraction with stirring for 1 minute. After extraction, set the spittoon in a centrifuge and centrifuge at 3000 rpm for 15 minutes. At this time, the protein portion precipitates as a white precipitate, and heme is extracted into the supernatant liquid in the form of diethylamine-coordinated heme. When 20μ of this supernatant was separated and quantified using high performance liquid chromatography, the amount of heme was calculated as hemin.
4 μg was detected. That is, by this extraction method, blood 1
8 mg of heme was obtained per ml.

Example 1 Put 60 methanol into a 100 molar extractor, add 1 kg of diethylamine, and stir well to mix.
To this is added 5 kg of blood powder made by spray-drying fresh bovine blood at an internal temperature of 65°C or less, and the mixture is heated and stirred at 45°C for 1 hour to extract. After cooling, the separated proteins are separated and the liquid is concentrated under reduced pressure. Add glacial acetic acid to the solution concentrated to approx.
10 and 200 g of strontium chloride are added and heated to distill off the remaining methanol. 100 more
The reaction was carried out at ~102°C for 1 hour, and after the reaction, the precipitated hemin was collected and washed with glacial acetic acid, water, and acetone sequentially to obtain 85 g of hemin.

When the obtained hemin was dissolved in 2.8% ammonia and determined by absorbance at 390 nm, its purity was 98.3%.

Example 2 Put 1200 g of methanol and 30 kg of diisoprolamine into a 2000 extractor and stir and mix well. To this is added 300 kg of blood powder obtained by spray-drying fresh bovine blood at an internal temperature of 65°C or less, and the mixture is heated and stirred at 45°C for 1 hour for extraction. After cooling, separate the separated proteins and concentrate the liquid under reduced pressure. The concentrated solution was further evaporated to dryness under reduced pressure to obtain 45 kg of hematin. This was added to 100ml of glacial acetic acid containing 2 kg of sodium chloride and boiled under reflux for 1 hour. Separate the hemin produced after cooling,
After washing with 5 parts of glacial acetic acid and 20 parts of water, the mixture was dried to obtain 4.8 kg of hemin.

When the obtained hemin was dissolved in 2.8% aqueous ammonia and determined by absorbance at 390 nm, its purity was 98.8%.

Example 3 60 methanol and 5 kg of spray-dried bovine red blood cells are added to a 100 mm extractor, and after stirring and mixing thoroughly, 1 kg of ammonia gas is introduced. After introducing gas, heat to 40°C for 30 minutes, and after cooling, separate the protein. After concentrating the solution (50) under reduced pressure to a concentration of about 5, add glacial acetic acid.
10, and then add 100g of sodium chloride to 22ml of water.
Add the solution dissolved in , heat and stir, and distill off the remaining methanol. The residue was heated under reflux at 100°C for 1 hour, and after cooling, the precipitated hemin was collected. This was washed with glacial acetic acid and water and dried to obtain 65 g of hemin. The purity of the hemin obtained was 97.5%.

Claims (1)

[Claims] 1. Hemin, which is characterized by extracting fresh blood or blood powder with a lower alcohol containing a basic substance selected from ammonia and di-lower alkylamine, and then treating the obtained extract with sodium chloride or strontium chloride. manufacturing method.