JPH01317502A - Method for removing pyrogen - Google Patents

Abstract

PURPOSE:To efficiently remove a pyrogenic material (pyrogen) by treating a liquid containing pyrogen with a cation exchanger and then by further treating the treated liquid with a pyrogen adsorption substance. CONSTITUTION:A pyrogen containing liquid 2 is treated with a cation exchanger 4 and then the treated liquid is further treated with a pyrogen adsorbing substance 5 (e.g., L-histidine holding agarose). Pyrogen is thus efficiently removed from the liquid containing it. This de-pyrogened water is usable for medical use such as dialysis water of artificial dialysis for a patient with renal insufficiency, supply water for filtration-type artificial kidney, preparation of dilute solution from highly concentrated instillation medicine at bed side, etc., or for washing medical tools.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a method for removing pyrodiene from a pyrodiene-containing liquid by using both a cation exchanger and a pyrodiene adsorbent.

In particular, the method of the present invention is useful for producing depyrodiene water and pyrogen-free pharmaceutical products.

The depyrogenized water produced by the method of the present invention is used for pharmaceutical purposes, such as dialysis water for artificial dialysis performed on patients with renal insufficiency, replacement fluid for artificial kidneys with high filtration rate, and dilution of highly concentrated drugs for bedside infusion. manufacturing liquids or cleaning medical equipment;
Furthermore, it can be used for pharmaceutical purposes, such as the production of pure water for injections and the cleaning of containers for injections.

Furthermore, the production of pyrogen-free pharmaceuticals refers to a method for removing pyrogenes from an aqueous solution containing pharmaceuticals to be administered into the blood, and the method of the present invention is also useful as a method for producing such pharmaceuticals. .

Aqueous solutions containing such pharmaceuticals include, for example, various pharmaceuticals to which the pyrogenic substance test method is applied (
(fructose injection, physiological saline, dextran 40 injection, glucose injection, Ringer's solution, sodium citrate injection for blood transfusion, etc.), and others administered into the blood.
The present invention can be applied to any pharmaceutical product that requires removal of pyrodiene during its manufacturing process.

[Conventional technology and its problems]

Pyrogenic substances (pyrogienes), particularly endotoxins, are derived from the cell walls of Gram-negative bacteria, and their constituent components are polysaccharides, whose molecular weight varies depending on the fungus from which they are derived, and whose subunits are several. However, because they exist in an aggregated state in an aqueous solution, they are said to have a size of several hundred thousand to several million, and when an ultra-trace amount enters the body, it causes a noticeable fever. It is known that it can be fatal in some cases.

Recently, it has also become known that lipid A (molecular weight approximately 2,000), which is a component of endotoxin, is also pyrogenic.

On the other hand, the Japanese Pharmacopoeia requires a pyrogenicity test using rabbits to detect endotoxin, but it is laborious and time-consuming. Simple detection reagents have become commercially available, and measurements can now be carried out in a short time.

In addition, a colorimetric assay method using a synthetic substrate that utilizes the reaction mechanism between endotoxin and horseshoe crab blood cell components has also been established. Furthermore, the detection limit of quantitative methods using synthetic substrates also increases, and n
There has been a shift from g/- units to pg/@Z units, and even ultra-trace amounts of endotoxins have come to be detected.

Conventionally, methods for removing pyrodiene include physical adsorption methods (activated carbon, ion exchange resin, etc.), chemical decomposition methods (acid, alkali treatment, etc.), and adsorbents that specifically adsorb pyrodiene. Methods are known. Among these methods, the method using an adsorbent that specifically adsorbs pyrodiene is less likely to cause the main drug to be decomposed or the recovery rate to be poor, and is easy to operate.

However, these adsorbents have the drawback that their adsorption performance may be reduced depending on the type of pyrodiene.

[Means for solving problems]

As a result of various studies on a method for removing pyrodiene from a pyrodiene-containing liquid that does not have the above-mentioned drawbacks, the present inventor found that
The inventors have discovered that a method for removing pyrodiene with excellent removal effects can be obtained by using a cation exchanger and a pyrodiene adsorbent in combination, leading to the present invention.

Examples of the pyrodiene-containing liquid that is the object of the present invention include the following.

That is, pyrodiene-containing amino acids (e.g., histidine, alanine, proline), nucleobases (e.g.,
cytosine), antibiotics (e.g. penicillin), hormones (e.g. insulin), vitamins (e.g. flavin adenine dinucleotide), serum proteins (e.g. albumin), enzymes (e.g. urokinase, asparaginase, lysozyme), antibodies. (eg, immunoglupurin) and other physiologically active substance solutions. The present invention can also be used to produce pyrodiene-free water from pyrodiene-containing water.

The cation exchanger used in the present invention means one that has a negative charge and has the ability to capture cations (metal ions, basic substances, etc.). Specifically, a charged group is covalently bonded to a three-dimensional mesh structure made of a polymer base material. As the polymer base material, spherical gels such as synthetic polymer gels, polymer membranes such as cellulose membranes, etc. can be used. As the charged group, a strong acidic group (such as a sulfo group) or a weakly acidic group (such as a carboxyl group) is used. Further, as the form of the cation exchanger, membrane, sheet, spherical, etc. are used. Therefore, it is necessary to select and use an appropriate cation exchanger depending on the target sample.

The pyrodiene adsorbent used in the present invention may be any material as long as it can adsorb pyrodiene selectively and efficiently.
Preferably, acetic acid or an antibiotic immobilized thereon can be used, but a combination of a nitrogen-containing heterocyclic compound such as L-histidine and a water-insoluble carrier described in JP-A-57-183172, or JP-A-57-183172, It is also possible to use an adsorbent comprising hyaluronic acid and an anion resin described in Japanese Patent Publication No. 54-67024. The treatment conditions for the adsorbent are determined by considering the pyrodiene removal rate and sample recovery rate for each sample, and p
It is necessary to set H, ionic strength, etc. to optimal processing conditions.

Next, preferred embodiments for carrying out the present invention will be explained with reference to the drawings.

In the apparatus shown in FIG. 1, 1 is a container containing an equilibration or elution buffer, 2 is a container containing a sample, 3 is a pump, 4 is a processing container equipped with a cation exchange sheet layer inside, 5 is a column filled with a pyrodiene adsorbent, and 6 is a fraction collector for collecting the treated eluate. First, an equilibration buffer (phosphate buffer) is pumped into the apparatus from a container 1 using a pump 3 and passed through a cation exchanger 4 and a pyrodiene adsorbent 5.

Next, a sample is sent into the apparatus from the container 2, and the liquid eluted from the pyrodiene adsorbent packed column 5 is collected by a fraction collector 6.

After sending all the samples into the device, elution buffer (same liquid as the equilibration buffer) is sent from container 1 to elute all the samples remaining in the device. Processing is performed using the above method.

〔Example〕

Next, Examples and Comparative Examples of the present invention will be shown.

Experiments were conducted using the system shown in FIG. Using a cation exchanger (Zetapretub SP, base material: hydrophilic linear cellulose sheet, charge: sulfopropyl group: sold by Seikagaku Kogyo ■) in which a strongly acidic sulfopropyl group was introduced into a cellulose membrane, a pyrodiene concentration of 1200 ng/ mZ
of 5% human serum albumin (phosphate buffer, po=
s, s, ionic strength (μ)”=0.01, hereinafter IS
The 11sA solution after passing through a pyrodiene adsorbent made of agarose fixed with L-histidine was treated with a Limulus test [measured with a toxinometer ET-201, manufactured by Wako Pure Chemical Industries, Ltd.]. The pyrodiene concentration was measured. For piping, use silicone tubes.

Table 1 shows the results obtained and the results of treating a similar ISA solution using only the above pyrodiene adsorbent (comparative example). As shown in Table 1, the combined use of a cation exchanger and pyrodiene adsorbent (example) showed a removal rate of 99.98%, while the removal rate of pyrodiene adsorbent alone (comparative example) was 98.98%.
The removal rate was 8%. In addition, IS of Examples and Comparative Examples
The recovery rate of A (human serum albumin) was 90% each.
%, 89%.

Table 1

[Brief explanation of the drawing]

FIG. 1 is a diagram schematically illustrating the steps of a preferred embodiment of carrying out the method of the present invention. 2...Sample 3...Cation exchanger 4...Pyrodiene adsorbent

Claims (1)

[Claims] A method for removing pyrogen, which comprises treating a pyrogen-containing liquid with a cation exchanger, and then treating the treated liquid with a pyrogen adsorbent.