Method of removing protein compounds from aqueous solutions

FIELD: chemistry.

SUBSTANCE: invention relates to biochemistry and can be used for removing proteins and amino nitrogen from aqueous solutions. The method involves adsorption of proteins on a hydroaluminosilicate natural sorbent, which contains such minerals as clay, zeolite, feldspars, mica, calcite, and filtration. Adsorption takes place at pH ranging from 1 to 3 for 1 to 10 minutes.

EFFECT: invention allows for fast and quality deproteinisation of a solution.

3 tbl, 2 ex

 

The invention relates to the field of biochemistry and can be used to remove proteins and amino nitrogen from aqueous solutions.

Known methods for removal of proteins from aqueous solutions, based on the precipitation of proteins by heating or by treatment with acids (Workshop on biochemistry / edited Sieberana. - M, MSU. - 1979. - S). Disadvantages removal of proteins by treatment with acids include pollution protein-free filtrate aggressive against chemical substances. Boiling unsuitable for deproteinization dilute solutions.

The technical essence and the achieved positive effect closest to the offer are the following two ways to remove proteins from aqueous solutions.

The first involves the adsorption of proteins on the sorbent, which is used as the hydroxide of zinc, and the Department deproteinizing the solution by filtration (Guide to laboratory studies of biological chemistry, Ed. Thetav. - M.: Medicine, 1976. - S). This method is complicated and long, since before removing proteins need to get the sorbent by mixing zinc sulfate with alkali and subsequent boiling and filtering. Very deproteinization also includes a step of heat treatment. Deproteinisation solutions contaminated zinc ions.

The second method is selected for FR is the type, involves the use as a sorbent hydrophilic Aerosil a-3 00 and a-175 with a specific surface area 300 and

175 m2/g (the Description of the invention to the author's testimony (19) SU (11) 1122354 And / Neblock, Picsanimated, Mvoc and others, 1983). Aerosil deproteinized aqueous solutions almost instantly. This method is effectively used to remove proteins from solutions containing polysaccharides, chlorophos, heparin and organic acid at a pH of from 1 to 5.6. It includes the adsorption of proteins on the sorbent and the Department deproteinizing solution by filtration.

However, hydrophilic Aerosil manufactured abroad (Ukraine), which causes some difficulties when purchasing it.

The objective of the invention is the expansion of technical means for removing proteinaceous compounds from aqueous solutions.

This object is achieved in that according to the method of removing protein compounds from aqueous solutions, including the adsorption of proteins on the sorbent and the Department deproteinizing solution by filtration, as a sorbent use hydroelasticity drug "ECOS", and the adsorption spend at least one minute at pH 1-3.

Hydroelasticity drug LPCD "ECOS" - preparation of domestic production of mineral raw materials deposits in the Belgorod region. He designed the La prevention of digestive disorders and normalize the function of the intestines of animals due to the ability to bind and excrete heavy metals and radioactive isotopes, nitrates, nitrites, nitrates and pesticide residues, as well as toxins, pathogens, and is a powder in light gray with yellowish, greenish or brownish shades, without peculiar smell. The size of the particles in the groundmass ranges from 0.03 to 1000 μm. Specific surface area of the drug is 1.2-1.9 m2/g Sorbent contains the following elements in terms of oxides (in wt.%):

wt.%10-3wt.%
SiO250.0-51.2Ti80
Al2About313.8-15.7V6
CaO12.6-13.3Mn5
Fe2About34.27-4.47Cr4
MgO1.66-1.96Zn3
Tio20.92-0.92 Ni1
To2O0.84-0.84Co0.6
Na2O0.22-0.22Cu0.6
CO28,58-9,14Pb0.3
H2About4,76-4,82Mo0.1
wN/a
AgH/o
CdN/a

General specific radioactivity level 115,4±8,16 Bq/kg, which does not exceed MPC. In the preparation includes montmorillonite, kaolinite and clinoptilolite, calcite, opal, feldspar, white mica and glauconite. (The use of natural hydroaluminosilicates in animal husbandry and veterinary medicine. Method. recommendations / Any, Nasasira, Aeeseap etc. - Belgorod, (in Belgorod is Oh, SAA, 2003. - P.4-5).

Hydroelasticity drug "ECOS" is not composed of chemicals that can adversely affect animals and the quality of products. The additive is non-toxic to animals, has no cumulative properties. Embryotoxicity, teratogenicity and irritating effect was experimentally established (Certificate of conformity. Mineral feed additive "ECOS" for livestock No. POCC.RU. PA from 12.03.01 year).

The technical result - the use of the proposed hydroalumination drug "ECOS" as sorbent protein compounds can deproteinizing solutions without preparation of the sorbent within 1 minute. Additional technical result - at the same time from the solution is partial removal of the amine nitrogen.

The proposed method is that for deproteinization aqueous solutions, including the adsorption of proteins on the sorbent and the Department deproteinizing solution by filtration, as a sorbent use hydroelasticity drug "ECOS", and adsorption are at least 1-10 minutes at pH 1-3.

Novelty and inventive step of the proposed method proves detected for the first time the ability of the drug to prevent digestive disorders and normalize the function of the gut of animals and in order to adsorb from aqueous solutions of proteins and amino nitrogen. The deproteinization should be carried out at pH 1-3, further increase in pH leads to a decrease of efficiency of sorption of proteins hydroalumination drug "ECOS". Aqueous solutions deproteinized hydroalumination drug "ECOS" instantly. Not established difference in deproteinised action hydroalumination drug "ECOS" in contact with the protein solution within 1-10 minutes For removal from aqueous solution 0,76 mg protein you need to make 70 mg hydroalumination drug "ECOS".

The advantage of this method is its simplicity when fast process deproteinization. In addition, the proposed drug is more available and relatively cheaper than the recommended Neblock et al. (1983) hydrophilic Aerosil (respectively 15 USD and 20 USD per 1 kg).

The method is illustrated by the following examples.

Example 1. In a test tube filled with 9.9 ml of isotonic sodium chloride solution (pH 5) and 0.1 ml of the blood serum of cattle; make 700 mg hydroalumination drug "ECOS". The contents of the tube mixed and filtered through filter paper of "blue ribbon" (for small and very small precipitation), pre-washed with two portions of isotonic sodium chloride solution (pH 5). The lack of protein in the filtrate set a break from sulfosalts the gross acid (FCS). To 2 ml of the filtrate add 4 drops of a solution SCQ.

The transparency of the mixture indicates the absence of protein in the filtrate. All the protein in 0.1 ml serum (7,6 mg) binds 700 mg of the drug "ECOS". In table. 1 shows studies deproteinization aqueous solutions containing different amounts of serum, by introducing different amounts of hydroalumination drug "ECOS". In the described experience of 700 mg of the drug can bind the whole protein in 0.1 ml of serum, i.e. to 7.6 mg, representing 1.09% of the mass of the sample of the drug introduced into the solution.

Table 1
Minimum deproteinizing dose hydroalumination drug "EKOS
Isotonic solution ml9,5the 9.79,89,910,0
Serum, ml0,50,3 0,20,1-
The concentration of protein,3,802,281,520,760,0
mg/ml
Test with FCS*
20+++++++-
Dose "Akasa",30+++++++-
mg/ml40+++++++-
solution50++ ++++±-
serum60+++++±-
70++++±--
*SCQ - sulfosalicylic acid
"-"- negative reaction
"±" - opalescence
"+" - opacity
"++" is the appearance of flakes in the filtrate

In table. 2 shows data about the objectivity of the binding protein in 1%aqueous solution of serum at different pH values. The decrease in pH from 5 to 3 and 1 led to a decrease in the amount of sorbent required to fully deproteinization solution. At pH 3 the test with FCS was negative even when making 300-400 mg hydroalumination drug "ECOS" in 10 ml serum (7,6 mg protein). At pH 1 DOS is enough quantities of the drug are 200-300 mg of drug in 10 ml of the protein-containing solution.

Table 2
The influence of reaction medium on the sorption activity of hydroalumination drug "ECOS" in 1%solution of serum
PHpH 1pH 3pH 5pH 7pH 9
Test with FCS*
Dose "Akasa", mg/ml serum concentration of protein 0,76 mg/ml)20-±+++++
30--++++
40--++++
50--± ++
60--±+++
70---±++

Increasing the pH to 7 (neutral environment) significantly increased the minimum required dose hydroalumination drug "ECOS", causing the deproteinization: only 700 mg it made in 10 ml serum, caused partial binding proteins. Alkaline pH solution (pH 9) does not contribute to the manifestation discussed sorption properties hydroalumination drug "ECOS" in relation to serum proteins.

Example 2. To confirm the ability hydroalumination drug "ECOS" free aqueous solutions not only from protein molecules tested its activity against amino acids and their residues appearing in the solution during the hydrolysis of proteins. As the object made of liquid nutrient medium for cultivation of microorganisms - broth pancreatic hydrolysate of sprat (120 mg% amine nitrogen). About the number of amino acids (and their residues) with the Dili based on the results of formolite reaction titration according to Sørensen (Handbook of veterinary technician / Fesenkov, Iaase, Retlock and others; Ed. by Wealthow. - M.: Kolos, 1981. - S-23).

To 10 ml of the investigated broth was added 5 ml of freshly prepared formolite mixture (50 hours of formalin and 1 tsp of phenolphthalein, brought to 0.2 N. the alkali solution until slightly pink staining). Was titrated (in triplicate) of 0.2 N. the solution of alkali (sodium hydroxide) to bright red, then added dropwise with 0.2 n hydrochloric acid to slightly pink color of the liquid. Having sodimie titration results were calculated content of amino nitrogen from the difference between the amounts of alkali and acid followed by titration of the sample, and assuming that 1 ml of the spent alkali equivalent to 2.8 g of amine nitrogen.

Reducing the number of amine nitrogen 34.5% was detected in the broth pancreatic hydrolysate sprat after treatment hydroalumination drug "ECOS" at a concentration of 100 mg/ml at pH 3 (table 3).

In all cases experience an increase in the concentration hydroalumination drug "ECOS" and the lowering of the pH (acidification of the environment) led to a decrease in the content of amino nitrogen in the medium due to its binding "Ecosim".

Table 3 Results of titration of the broth from pancreatic hydrolysis is and sprat
No.Containing% loss
experienceConditions of experimentsthe Aminamine
aqueous nitrogennitrogen
mg %
1.Pure broth (background control)119,700
50 (pH 7)103,9013,2
2.The broth after processing100(rn)101,5015,2
"Ecosim" doses (mg/ml):50 (pH 5)95,5020,2
100(pH 5)96,2519,6
50 (pH 3)83,3330,4
100(pH 3)78,4034,5

Despite the fact that the specific surface hydroalumination drug "ECOS" significantly less than that of the hydrophilic Aerosil a-175, a-300, it is able effectively to deproteinizing solutions even without phase of the sorbent and boiling the protein-containing solutions. The protein binding hydroalumination drug ECOS for 1 min and the number of stages can significantly accelerate the deproteinization.

In addition, hydroelasticity drug "ECOS" partially binds and amine nitrogen liquid nutrient media for cultivation of microorganisms (pH 3-7).

The insolubility and chemical indifference hydroalumination drug "ECOS" allow you to get deproteinisation aqueous solutions and culture media, partially deprived of amino nitrogen containing chemically aggressive compounds.

Method of removing proteins from aqueous rest the ditch, including the adsorption of proteins on the sorbent and the Department deproteinizing solution by filtration, characterized in that the sorbent use natural hydroecological containing montmorillonite, kaolinite and clinoptilolite, calcite, opal, feldspar, white mica and glauconite, and the adsorption is carried out for 1-10 min at pH 1-3.



 

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