Method of chitosan production

FIELD: medicine; pharmacology.

SUBSTANCE: offered invention concerns medicine and pharmacology, exactly to method of Chitosan production. Method includes milling of natural chitin-containing raw material, alternating by turn three stages of raw material deproteinisation, and three stages of decalcification, deproteinisation stages are carried out by sodium hydroxide solution of concentration 3-5% produced by diaphragm electrolysis method within 3-4 hours at temperature 60-65°C; decalcification stages are performed by 3-5% hydrochloric acid solution at temperature 20-25°C within 1-3 hours; deacetylation is performed by 49-55% sodium hydroxide produced by diaphragm electrolysis method; deproteinisation and decalcification stages are performed in series connected six steel devices with internal glass-enamel coating, equipped with stirrers, thermopools, connections for reagents feeding and disposal, jackets for reactionary mass cooling and heating. Afterwards deacetylation follows with 49-55% sodium hydroxide produced by diaphragm electrolysis method. Process is conducted at temperature 95-105°C within 7-9 hours and followed by washing, spinning and drying of finished Chitosan.

EFFECT: production of high-clean Chitosan.

 

The present invention relates to pharmacology and allows you to get a natural compound chitosan is widely used in pharmacology.

A method of obtaining chitosan, including disposable deproteinization of 3.5%sodium hydroxide solution for 2 hours at t=65°, filtering the suspension, washing the solid phase, the demineralization (decalcomania) 3,65%HCl solution for 30 min at room temperature, filtration, washing, extraction with acetone, drying, bleaching 0,315%sodium hypochlorite solution, washing and drying the obtained chitin, the deacetylation of 50%sodium hydroxide solution at 100°With a duration of 30 min, rinsing and drying of the obtained chitosan. (Meyers S.P., No NICHOLAS, for K.S. Lee Isolation and characterization of chitin from crabfish shell waste J. Agric. Food Chem. 1989, 37, 575).

The disadvantages of this method are the large number of operations to be performed manually because the process issued on a periodic method; the inability to remove a large part of protein and calcium hydroxide due to too short time of the process deproteinization and decalcomania; very little time for the stage of deacetylation; excessive conduction phase extraction, besides fire and explosion and toxic reagent acetone; bleaching ready chitin with sodium hypochlorite, the act is wny chlorine which to some extent destroys the chitin molecule; the appearance in the wastewater chlorine after prolonged washing of chitin to the complete lack of chlorine in the product. The result is a chitosan with deacetylation not above 60-70%, with a high content of protein and minerals, and low viscosity.

The closest analogue of the proposed invention is a method of producing chitosan, including grinding of natural chitin-containing raw material to a particle size of 100-200 μm; extraction of the powder with liquid carbon dioxide within 1-3 hours; the first decalcomanie powder 5-10-fold excess of 0.8-2.5%hydrochloric acid at 35-50°C for 0.5-2 hours; washing the solid phase with water at 50-75°to a pH of 5.4, holding 1st deproteinization with 4-7-fold volume of water and pH of 9.0 to 10.5 by adding to the water of solid sodium hydroxide. In the resulting suspension type enzyme in an amount of 0.1 to 0.5% by weight of the dry powder; pour the liquid phase; washing the residue with water; hold the second decalcomanie 5-7-fold by weight of the dry powder amount of 0.8-2.5% hydrochloric acid at a temperature of 35-50°C for 0.5-2 hours; washed with the reaction mass; add 4-7-fold volume of water and adjusted its pH to 9.0 to 10.5 by adding solid sodium hydroxide and 0.1 to 0.5% by weight of the dry powder of the enzyme. The deproteinization are 2-4 hours at a temperature of 50-70°; merge match the initial solution, add 2-4 volume of dry weight of chitin 45-50% sodium hydroxide and lead the deacetylation at a temperature of 85-95°C for 2-4 hours; washed chitosan water at a temperature of 60-80°to pH 6.5 to 7.4; chitosan centrifuged; dried and crushed (U.S. Pat. Of the Russian Federation No. 2221811).

The disadvantages of this method are:

a large number of operations (8) in the production of the product is performed manually because the process is designed as a periodic;

excessively high degree of grinding of raw materials, resulting in the loss of the processed material will be high and will increase as the distance from the raw protein (40%), caso3(30%) and associated with this additional size reduction of grains;

dangerous operation of extraction of the powder with liquid carbon dioxide, which remains liquid under pressure is higher than 60 kgf/cm2. The operation is possible only in the autoclave and becomes under the supervision of the state;

difficulties in the release of sediment deposited on the bottom of the industrial reactor sludge powder after removal of the mother solutions and probable failure of the gears when turning on the engines mixers;

the need for devices to ensure the safe release of pressure of CO2from the reactor;

too high temperature process decalcomania and

connected the traveler with this increased degradation of chitin molecules;

hydrogen chloride contained in the input to decalcomania hydrochloric acid, is not sufficient for complete removal of calcium carbonate;

washing the obtained calcined itinalaga complex (CHBC) of hydrogen chloride to a pH of 5.4 requires a large consumption of water and prolonged in time;

use potable water for flushing is associated with an increase of hardness salts (non-combustible residue) in the finished product;

the preparation of the alkaline solution to deproteinization directly in the reactor by adding a suspension of solid alkali - reception dangerous associated with spontaneous temperature rise and local overheating in the place hit the alkali in the water;

temperature, time and duration of deacetylation insufficient to achieve a high degree of deacetylation.

With the aim of obtaining high-quality chitosan suitable for use in medicine, as well as to avoid the above disadvantages of the known method is proposed the following method of producing chitosan carried out by a continuous method and includes three stages of deproteinization and three stages of decalcomania performed alternately, one after the deproteinization with calzinirovnie, simulating thus the countercurrent principle, in which the processed material in the ore processing is in contact with fresh reagents. Depending on the quality and composition of raw materials (crushed shell crab at the grist crab"), the number, order, and duration of stages of decalcomania and deproteinization can be changed. If the grains crab fraction minus 3 mm greater than 3%, reduce the number and timing of decalcomania and deproteinization. When the content in the grains of the crab caso3more than 40% or proteins over 45% reduce the number and duration of deproteinization or decalcomania respectively.

The result is a chitin white, almost not containing caso3and with very little residue protein. Selected chitin is subjected to reaction high deacetylation with sodium hydroxide at elevated temperatures. All reactions carried out in nitrogen atmosphere, in a chain of standard steel apparatus with high-quality glass-coated, with a capacity of 6.3-2.5 m3equipped with agitators , thermowells, fittings for insertion and removal of reactive components, jackets for heating and cooling the reaction mass, as well as in the apparatus of the steel 18CR10NITI, as when carrying out the deacetylation glass-coating reactor is destroyed by high temperatures and concentrations of alkali.

Download operation component is in, reagents, removal of the fallopian solutions and wash water are mechanized. They are due to the vacuum generated in each of the devices.

In contrast to known methods, in the proposed as an alkaline agent is used a liquid solution of sodium hydroxide obtained by the method of diaphragm electrolysis. Through the use of liquid caustic soda diaphragm was no need for hazardous and time-consuming work on the fragmentation and dissolution of solid alkali.

All devices before loading reagents and processed material are filled with nitrogen.

The essence of the method is as follows.

1st deproteinization. Reactor a-1 fill with nitrogen and load it with 800 kg of crushed shell crab. This also serves 4000 liters of a 3%aqueous NaOH solution, obtained by means of a diaphragm electrolysis, at 60-65°C. Maintain the slurry with stirring and t=60-65°3-4 hours. After 10-20 min of sludge liquor is removed in the collection. The remaining calcinated chainarray complex (CHBC) washed 2400 liters of water temperature 60-65°With, mix the pulp 30-40 minutes, allow to sit for 10-20 minutes and remove the wash water in the collection. The operation is carried out twice.

1-St decalcomania. Reactor a-2 is filled with nitrogen, serves the pulp from a-1 and stirred for 10 minutes Allow to settle for 10 minutes, removed from the reality in the Torah And-2 water and serves 4000 liters of a 5%HCl solution with t=10-20° C. Maintain the slurry with stirring and t=20-25°C 1-1,5 hours. After 10 min of sludge liquor is removed in the collection. The remaining CHBC washed 2400 liters of water temperature 60-65°With, mix the slurry for 30 minutes, allow to settle for 10 min and remove the wash water in the collection. The operation is carried out twice.

2nd deproteinization. Reactor A-3 is filled with nitrogen, serves the pulp from a-2 and stirred for 10 minutes Allow to settle for 10 minutes, removed from the reactor a-3 water and serves 4000 liters of 4%aqueous NaOH solution, obtained by means of a diaphragm electrolysis t=55-60°C. Maintain the slurry with stirring and t=55-60°4 hours. After 10 min of sludge liquor is removed in the collection. The remaining CHBC washed 2400 liters of water temperature 60-65°With, mix the slurry for 30 minutes, allow to settle for 10 min and remove the wash water in the collection. The operation is carried out twice.

2nd, decalcomania. Reactor a-4 are filled with nitrogen, serves the pulp from a-3 and stirred for 10 minutes Allow to settle for 10 minutes, removed from the reactor a-4 water and serves 2400 liters of 4%aqueous HCl solution with t=20-25°C. Maintain the slurry with stirring and t=20-25°2 hours. After 10 min of sludge liquor is removed in the collection. The remaining CHBC washed 2400 liters of water temperature 60-65°With, mix the slurry for 30 minutes, allow to settle for 10 min and remove the wash water in the collection. The operation is carried out twice.

3-the deproteinization. Reactor A-5 are filled with nitrogen, serves the pulp from the a-4 and stirred for 10 minutes Allow to settle for 10 minutes, removed from the reactor A-5 water and serves 2400 liters of a 5%NaOH solution, obtained by means of a diaphragm electrolysis t=50-55°C. Maintain the slurry with stirring and t=55-60°2 hours. After 10 min of sludge liquor is removed in the collection. The remaining calcinated chitin (LOC) is washed with 2000 liters of water temperature 60-65°With, mix the slurry for 30 minutes, allow to settle for 10 min and remove the wash water in the collection. The operation is carried out twice.

3-e decalcomanie. Reactor A-6 are filled with nitrogen, serves the pulp from A-5 and stirred for 20 minutes Allow to settle for 10 minutes, removed from the reactor A-6 water and serves 2400 liters of 3%HCl solution with t=20°C. Maintain the slurry with stirring and t=20°2 hours. After 10 min of sludge liquor is removed in the collection. The remaining chitin was washed with 1600 liters of water temperature 40-45°With, mix the slurry for 30 minutes, allow to settle for 10 min and remove the wash water in the collection. The operation was performed to obtain a pH of 6.0. Chitin is obtained in white, practically does not contain caso3and proteins.

The deacetylation. Reactor a-7 are filled with nitrogen, serves the pulp from the A-6 and stirred for 20 min, Allow to settle for 10 minutes, removed from the reactor a-7 water and serves 2700 liters 49-55%NaOH solution, policerelated diaphragm electrolysis. The process is conducted at t=95-105°C for 7-9 hours. After the reaction time the mother liquor is removed in a separate collection, and prepared chitosan washed.

Pohovka. Primary moisture from chitosan removed in a centrifuge at 1400 rpm for 25-40 minutes.

Drying of chitosan. Taken from centrifuge the crude chitosan contains about 60% water. It is dried in a vacuum drum dryer at a residual pressure of 90-120 mm RT. Art. and t=60-65°C.

In the proposed method of obtaining chitosan yield is 94-95%and its quality indicators expressed as follows:

- mass fraction of water,%, not more than6
- dynamic viscosity of 1% R-RA chitosan
1% solution10-1000
acetic acid JV, within
the degree of deacetylation %, not less than90
- mass fraction of insoluble residue
1% R-RA chitosan0.1
1% p-d acetic acid,%, not more than
- mass fraction of residue after
on ignition %, max0.1
- protein content,%, not more than 0.15
the content of heavy metals (Hg, As, Pb, Sb)
ppm, no more than3
- pH of water extract, within the6-7 .3
the microflora of Mafang, no more than1×10-3

Thanks to the gradual and layer-by-layer processing of raw materials can minimize the damage to the molecules of chitin, almost completely remove all impurities, as modeled here, the countercurrent principle, in which each of the apparatuses as the movement of the processed grain crab from device to device, she meets up with fresh reagents.

A method of producing chitosan, characterized in that it includes the grinding of natural chitin-containing raw material, conducting alternately interleaved three stages of deproteinization raw materials and three stages of decalcomania, stage deproteinization carried out with a solution of sodium hydroxide concentration of 3-5%, prepared by diaphragm electrolysis, for 3-4 hours at a temperature of 60-65°; stage decalcomania spend 3-5%solution of hydrochloric acid at a temperature of 20-25°C for 1-3 h; deacetylation 49-55%sodium hydroxide, obtained by the method of diaphragm electrolysis; stage deproteinization and decalcomania spend in th is therefore the United six steel apparatus with an internal glass-coated, with stirring, thermowells, fittings for insertion and removal of reagents, shirts for cooling and heating the reaction mass, followed by the deacetylation 49-55%sodium hydroxide solution, obtained by means of a diaphragm electrolysis, the process is conducted at a temperature of 95-105°C for 7-9 hours; washing, fugovka and drying of chitosan.



 

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