A reagentless method of separating a mixture of tyrosine and tryptophan

 

(57) Abstract:

The invention relates to methods for selection of individual amino acids (tyrosine and tryptophan) from their mixtures and can be used in chemical, food, microbiological industry and agriculture. The purpose of the invention is the separation of components of a mixture without the use of auxiliary reagents. It is achieved by passing hydrochloric acid solution of the amino acids at pH of 1.0 through a strong base anion exchange resin AV-17-2P in C1 form, followed by washing with water.

The invention relates to a method of allocating individual aromatic and heterocyclic amino acids from mixtures thereof and can be used in chemical, microbiological, food industry, agriculture.

Known methods reagentless division and separation of individual amino acids by methods of ion-exchange chromatography. The closest solution to this invention the method of separation of L-serine and glycine chromatographic method [BP 0213736, C 07 C 99/12, 1987]. In addition, similar to the solution of the present invention methods of separation of amino acids [A. S. USSR N 644782, C 07 C 99/12, 1979, A. S. USSR N 979991, G 01 N 31/08, 1981], the basis of Katie a large number of auxiliary reagents and flow in the system of the ion exchanger-sorbent various non-interactions, what complicates the process of excretion of amino acids.

The purpose of the invention is to simplify the process of separating a mixture of amino acids and reducing the number of auxiliary reagents used during the separation.

This goal is achieved by the use of the method inclusional chromatography, which is based on non-absorption of amino acids by ion exchange, by passing a mixture of cations of tyrosine and tryptophan through a strong basic anion exchange resin AV-17-2P in the Cl-form, followed by washing with water. The preferred use of this method due to the absence of ion-exchange interaction of adsorption with the sorbent, and thus eliminates the use of electrolyte solutions for regeneration of the sorbent.

The proposed method allows to obtain at the output of aqueous solutions of individual amino acids that are free of mineral ions.

Example. The division performed on a column with a strong basic anion exchange resin AB-17-2P in the Cl-form mass 3,0000 g; height of the layer of sorbent - 12 see Through the ion exchanger at a rate of 2 ml/min miss a 1 liter hydrochloric acid solution of a mixture of tyrosine and tryptophan containing 1,81 and 2.11 g of amino acids, respectively (pH=1, T=20oC). This cm control over the content of amino acids in solutions was carried out spectrophotometer Specord M40 half pans with 1=1 cm at 275 nm for tyrosine and 279 nm for tryptophan with the influence of amino acids on the analytical signal each other. At the outlet from the column of get a mixture of the same volume, containing 1.27 g of tyrosine and 0.27 g of tryptophan, i.e., sorbed 30% tyrosine and 87% of tryptophan. The regeneration is carried out with water at the same temperature (20oC). As a result of regeneration in 1 liter of eluate passes all previously sorbed tyrosine (0.54 g) and tryptophan in the number of 0.95, In the following 3 liters of the eluate passes all remaining column tryptophan (0,89 g). Thus have the solution of pure tryptophan, and the column goes into the initial state: the anionite AB-17-2P in the Cl-form, saturated with water. Then through the column again pass the mixture of amino acids obtained earlier at the outlet of the column (1 liter, containing 1.27 g of tyrosine and 0.27 g of tryptophan). As a result, the rest of the sorbed tryptophan (0.27 g) and 0.36 g of tyrosine and get a solution of pure tyrosine containing 0.9 g of this amino acid. After this, the process of regeneration of the column with water again. Thus, the allocation of tyrosine occurs at the stage of sorption of a mixture of amino acids, and the allocation of tryptophan - elution of the column with water, the resin enters the initial state. Control over the purity of amino acids was carried out by spectrophotometric and TLC, and the presence of a mineral is to 91.1%, and tryptophan - 97,2%.

The method of separating a mixture of tyrosine and tryptophan, including the transmission of hydrochloric acid solution of amino acid (pH 1,0) through a bed of anion exchange resin AV-17-2P with C1 to form, at the output of the columns have the solution tyrosine, and subsequent elution of the column with water to obtain a solution of tryptophan.

 

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