JPH067853B2 - Heat-sterilized container containing blood preservation solution and method for producing the same - Google Patents

Description

TECHNICAL FIELD The present invention relates to a heat-sterilized plastic container containing a blood preservation solution and a method for producing the same.

<Prior Art> In recent years, component transfusion has become mainstream, and, for example, in red blood cell transfusion, a concentrated red blood cell solution (hereinafter referred to as CRC) is generally used, and its expiration date is 21 days from the date of blood collection. In addition, other blood components such as platelets have their respective effective periods, and it is necessary to store them during that period.

In order to preserve these blood cell components in a better condition during the effective period, it is considered to further add a blood preservation solution to the preserved blood components.

This blood preservation solution is generally called an additive solution, and its components are effective for maintaining nutrients, monosaccharides such as glucose for adjusting osmotic pressure, sugar alcohols such as mannitol for adjusting osmotic pressure, and ATP amount in erythrocytes. Adenine, sodium chloride as an electrolyte, etc., and ascorbic acid-2 for maintaining the amount of 2,3-diphosphoglyceric acid (hereinafter referred to as 2,3-DPG) necessary for maintaining the oxygen transport function of red blood cells. -Sodium phosphate is said to be effective and its use in blood preservation solutions is being considered.

<Problems to be Solved by the Invention> In the CRC prepared by the current formulation method, plasma remains sufficiently, and the components in this plasma have a buffering action, so it is necessary to add a buffering action to the blood preservation solution to be added. There is no.

Therefore, the pH of the blood preservation solution should be such that the contents do not deteriorate.
It should have been set to.

However, in order to use blood more efficiently, the plasma yield in the CRC will be increased by reducing the plasma in the CRC as much as possible in the future, and instead, a preservative solution having a buffering action and a hematocrit adjustment function will be added. Storage of red blood cells is being considered.

That is, it becomes necessary to increase the amount of the blood preservation solution and to add a phosphate, citrate or the like to the blood preservation solution so as to have a buffering action.

However, when the pH is set to a predetermined value (between about pH 6 and 8) in this way, when glucose is added or ascorbic acid-2-phosphate sodium salt is added, the pH of the blood preservation solution is increased by glucose. Is 6 or more, and ascorbic acid-
The sodium 2-phosphate salt becomes a problem when it is sterilized in an autoclave at a pH of 8 or less, since it is deteriorated.

Therefore, an object of the present invention is to solve the above-mentioned problems of the prior art, and to provide a container containing a blood preservative solution that has been sterilized by heat and that does not substantially change the quality of the blood preservative solution even when heat sterilized, and a method for producing the same. There is to try to serve.

<Means for Solving the Problems> According to the first aspect of the present invention, a container and ascorbic acid-2-phosphate sodium salt in a state of pH 8.0 or less contained in the container, or pH 6. A blood preservation solution containing a monosaccharide and / or an oligosaccharide in a state of 0 or more is included, and the blood preservation solution is heat-sterilized in a substantially oxygen-blocking state. The absorbance at 410 nm measured by a photometer was 0.1 when water was used as a control.
There is provided a container containing a heat-sterilized blood preservation solution, which is characterized in that it is 05 or less.

According to the second aspect of the present invention, when manufacturing a container containing a blood preservative solution that has been sterilized by heating, the blood preservative solution is bubbled with an inert gas and packaged with a packaging material made of a gas impermeable material. Thus, there is provided a method for producing a container containing a heat-sterilized blood preservation solution, which is characterized by heat sterilization.

In the above invention, the blood preservation solution is ascorbic acid-2.
-Phosphate sodium salt, and mono- and / or oligosaccharides, the pH of which is preferably 6.0 to 8.0.

The heat sterilization is carried out in a substantially oxygen-blocking state, and the oxygen-blocking is carried out by dispensing a blood preservation solution into a container made of a gas impermeable material filled with an inert gas. After dispensing the blood preservation solution in a container, wrap it in a packaging material made of gas impermeable material filled with an inert gas, or replace the gas phase of the device used for heat sterilization with an inert gas. be able to.

The present invention will be described in more detail below.

The container 10 containing the blood preservation solution of the present invention has various containers 11a, 11b, 11c and 11 as illustrated in FIG.
d, each has a required mixed injection port 12, and each is connected by a tube 13. The container 11 a has a puncture needle 14 at the tip of the tube 13. The anticoagulant 15 is enclosed in the container 11a, and the blood preservation solution 16 is enclosed in the container 11d.

The blood collected in the container 11a is separated by centrifugation, and the platelet concentrate is fractionated in the container 11b and plasma is fractionated in the container 11c. The preservation solution in the container 11d is centrifuged and then dispensed into the container 11a.

10 containers 11a, 11b, 1 containing the blood preservation solution of the present invention
1c and 11d are formed of a plastic material having plasticity, for example, polyvinyl chloride, a cross-linked ethylene-vinyl acetate copolymer, high density polyethylene, linear low density polyethylene, etc., and are connected by a tube. These containers can be manufactured by stacking two sheets of the above plastic material and sealing the peripheral edges. Then, the blood preservation solution 16 is enclosed in a state in which the blood preservation solution 16 is not substantially denatured by heat sterilization. Furthermore, it is preferable that the blood preservation solution is deoxygenated by degassing or bubbling with an inert gas prior to heat sterilization.

The container 10 containing a blood preservation solution of the present invention can be packaged with a gas-impermeable packaging material 20 before or after heat sterilization described later. As the packaging material, an aluminum foil, a laminated film containing aluminum foil, or a laminated film having a polyvinyl alcohol film as an intermediate layer, a biaxially oriented polypropylene film as an outer layer, and a uniaxially oriented polyethylene film as an inner layer can be used. This packaging material can prevent the container from coming into contact with the outside air even after sterilization.

Therefore, in order to reduce the amount of air remaining in the packaging material as much as possible, it is possible to create a vacuum, enclose an oxygen scavenger, or fill an inert gas such as nitrogen gas.

Further, the container of the present invention may be composed of the above-mentioned gas impermeable material, filled with an inert gas together with the blood preservation solution, and subjected to high-pressure steam sterilization. At this time, the outer packaging material 20 is not always necessary.

The blood preservation solution in the present invention is a solution containing ascorbic acid-2-phosphate sodium salt, a monosaccharide such as glucose and / or an oligosaccharide such as maltose. The former is for preventing a decrease with time of 2,3-DPG which is responsible for oxygen transport in CRC. When it is sterilized by high-pressure steam at a pH of 8 or less, its decomposition, oxalic acid formation, and coloration occur. Therefore, heat sterilization should be carried out at a pH of 8 or higher, but this will reduce the preservation of blood, and when the blood preservation solution contains monosaccharides and / or oligosaccharides, they will decompose and become colored. I will end up.

Examples of monosaccharides include glucose, fructose, xylose and mannose, and examples of oligosaccharides include saccharose, palatinose and maltose.

In addition, it is preferable to put the oxygen absorber 31 in the packaging material 20 if necessary.

The osmotic pressure of the blood preservation solution after heat sterilization is 280-500mo.
smol, preferably 300 to 450 mosmol.
That is, if it is smaller than 280 mosmol, there is a risk of hemolysis.
This is because blood cells are not destroyed at 50 mosmol or more, but their functions may be deteriorated.

Also, the absorbance at 410 nm when water is used as a control is 0.05 or less, and if it is higher than this value, the degree of alteration affects the quality, which is not preferable.

The volume of the preservation solution contained in the container of the present invention is, for example, C
In the case of RC, the ratio is about 1 to 2 of the separated red blood cell fraction.

The present invention solves the above-mentioned various problems at once, and next, a method for producing a container containing a blood preservation solution without substantially degrading the blood preservation solution even by heat sterilization will be described.

First, oxygen is expelled from a blood preservation solution containing ascorbic acid-2-phosphate sodium salt, glucose and the like and adjusted to pH 8.0 or less.

As this means, the following examples are used alone or in combination.

Degassing: Reduced pressure or ultrasonic treatment under reduced pressure, stirring,
Boiling Replace with inert gas: bubbling or combination of ultrasonic treatment, stirring, boiling and decompression Chemical treatment: Addition of deoxidizer such as metal powder of iron or metal powder coated with metal halide Alternatively, the inert gas may be nitrogen, argon, helium, or the like, and nitrogen is particularly preferable.

The blood preservation solution deoxygenated in this manner is heat-sterilized in a substantially oxygen-blocked state.

Note that this step may be omitted when the oxygen content during storage of serum contains only an amount that does not matter.

The plastic container 10 containing the blood preservation solution 16 is sterilized by, for example, a high-pressure steam sterilizer shown in FIG.
This sterilizer is an autoclave 21 that is sterilized.
Is equipped with.

One end of the autoclave 21 is connected by a line 25 via a valve 28 to a steam supply source such as a boiler 22. Further, a supply source 23 of a gas inert to the blood preservation solution is connected to the autoclave 21 via a line 27, a valve 30, an inert gas storage tank 24, a line 26 and a valve 29. The storage tank 24 does not have to be provided (in that case,
Line 26 and valve 29 are also unnecessary).

Upon sterilization, a plastic container containing a blood preservation solution is housed in a plurality of units in the autoclave 21 and is evacuated to substantially remove oxygen, and then steam is introduced from the boiler 22 into the autoclave 21 to saturate it. . Then, if necessary, an inert gas such as argon, helium, and / or nitrogen (which is preferable) once stored in the storage tank 24 from the inert gas supply source 23 is introduced and pressurized, and then sterilized. . The sterilization temperature is generally 100 ° C to 130 ° C, usually 115 to 126 ° C. When supplying the inert gas, the sterilization atmosphere pressure is absolute pressure, which is about 10 to 20 times the saturated steam pressure at the sterilization temperature.
It is advisable to pressurize with 0% (for example, about 0.3 to 0.8 kg / cm ² ) of extra inert gas. Generally, the pressure during sterilization is about 1.2 to 2.0 kg / cm ² as a gauge pressure. A suitable sterilization time is 10 to 40 minutes.

When heat sterilization such as the above-described high-pressure steam is performed, the heat sterilization is performed in a substantially oxygen-blocked state. There are various methods for realizing the oxygen-blocking state, and typical examples are shown below.

(1) A blood preservation solution that has been deoxygenated as described above,
Dispense into a container made of gas impermeable material filled with an inert gas (preferably nitrogen gas).

(2) The deoxygenated blood preservation solution is dispensed into a container made of a gas permeable material and then packaged in a packaging material made of a gas impermeable material filled with an inert gas (preferably nitrogen gas).

(3) The gas phase in the autoclave in the high-pressure steam sterilizer as illustrated in FIG. 4 is replaced with an inert gas (preferably nitrogen gas).

According to the method as illustrated above, the once deoxidized blood preservation solution can be heat-sterilized in a substantially oxygen-blocked state. As a result, even if the blood preservation solution is sterilized by heating, ascorbic acid-2-phosphate sodium salt, glucose, etc. in the blood preservation solution are not decomposed, and ascorbic acid-
No problems such as oxalic acid formation and coloration, which occur as a result of decomposition of sodium 2-phosphate salt, occur.

Furthermore, since ascorbic acid-2-phosphate sodium salt is not oxidized, it also has a function of retaining 2,3-DPG in CRC for a long period of time.

<Examples> Next, the present invention will be specifically described based on Examples.

(Example 1) In order to investigate the effect of oxygen on the decomposition of ascorbic acid-2-phosphate sodium salt and glucose in autoclaving, the blood preservative solution in Table 1 was prepared and ascorbic acid-2-phosphate sodium salt and Decomposition of glucose,
Measurements were made on the production of oxalic acid and the coloration of the blood preservation solution.

Three kinds of solutions A, B and C having the compositions shown in Table 1
Was bubbled with 150 ml / min of nitrogen gas for 15 minutes. 90 ml of the solution thus obtained was dispensed into a polyvinyl chloride bag of 150 ml, and each bag was packaged in an aluminum packaging material filled with 500 ml of nitrogen gas as shown in FIG. did.

After sterilization, the absorbance (410 nm) of the blood preservation solution was measured with a spectrophotometer (Shimadzu MPS-2000) using water as a control. Further, ascorbic acid-2-phosphate sodium salt, oxalic acid and glucose were measured by liquid chromatography under the conditions shown in Tables 2 and 3, respectively.

As a control, a solution not subjected to nitrogen substitution treatment with a nitrogen gas bubble was dispensed into a polyvinyl chloride bag similar to the above, and subjected to high-pressure steam sterilization under the same conditions as above. In addition, pH
Was adjusted with 1N NaOH or HCl. The results are shown in Table 4.

As a result, by performing high-pressure steam sterilization under oxygen-blocking conditions, decomposition of APS and glucose, production of oxalic acid, and coloring of the solution can be suppressed even when the pH of the solution is 6.5 to 7.5.

(Example 2) In order to investigate the influence of oxygen on the decomposition of ascorbic acid-2-phosphate sodium salt during high-pressure steam sterilization, solutions A, B, and C in Table 5 were prepared, and ascorbic acid-2-phosphate sodium salt was prepared. The decomposition of the salt and the coloring of the storage solution were measured.

A solution having the composition shown in Table 5 was adjusted to 3 kinds of pH with 6N HCl, and an experiment was conducted using the same method as in Example 1.

The results are shown in Table 6.

(Example 3) In order to investigate the effect of oxygen on the decomposition of Maltose and Glucose during autoclaving, solutions A, B and C in Table 7 were used.
Was prepared, and the decomposition of Maltose and Glucose and the coloring of the stock solution were measured.

A solution having the composition shown in Table 7 was adjusted to 3 types of pH with 6N NaOH, and an experiment was conducted using the same method as in Example 1.

The quantitative determination of Maltose and Glucose was carried out by using liquid chromatography under the conditions shown in Table 3.

The results are shown in Table 8.

As a result, by performing high-pressure steam sterilization under oxygen blocking conditions, even if the pH of the solution was between 6.5 and 7.5, Maltose and Glu
The decomposition of cose and the coloring of the solution can be suppressed.

<Effects of the Invention> According to the present invention, even if a blood preservation solution containing ascorbic acid-2-phosphate sodium salt, glucose or the like is sterilized by heating, it is decomposed to produce harmful oxalic acid, and causes adverse effects such as coloring. No, the blood preservation solution is substantially unchanged.
A container containing such an unaltered blood preservation solution is very useful as a blood bag.

[Brief description of drawings]

FIG. 1 is a plan view showing an example of a container containing a blood preservation solution of the present invention. FIG. 2 and FIG. 3 are plan views showing the container containing the blood preservation solution of the present invention in a state of being wrapped with a packaging material. FIG. 4 is a diagram of a high-pressure steam sterilizer used in the present invention. DESCRIPTION OF SYMBOLS 10 ... Container containing blood preservation solution of the present invention, 11a to 11d ... Plastic container, 12 ... Mixed injection port, 13 ... Tube, 14 ... Puncture needle, 16 ... Blood preservation solution, 20 ... Packaging material, 21 ... Autoclave, 22 ... Boiler, 23 Inert gas supply source, 24 ... Storage tank, 25, 26, 27 ... Line, 28, 29, 30 ... Valve, 31 ... Deoxidizer

Claims (3)

[Claims] 1. A container and a pH of 8.0 contained in the container.
A blood preservation solution containing ascorbic acid-2-phosphate sodium salt in the following state or a monosaccharide and / or oligosaccharide in a state of pH 6.0 or more, wherein the blood preservation solution is in a substantially oxygen-blocking state It has been sterilized by heat, and the blood preservation solution was measured by a spectrophotometer after the heat sterilization.
A container containing a heat-sterilized blood preservation solution, wherein the absorbance at nm is 0.05 or less when water is used as a control. 2. The blood preservation solution is ascorbic acid-2-
The heat-sterilized container containing a blood preservation solution according to claim 1, which contains sodium phosphate and a monosaccharide and / or oligosaccharide and has a pH of 6.0 to 8.0. 3. A method of producing a container containing a blood preservative solution that has been heat-sterilized according to claim 1 or 2, wherein the blood preservative solution is bubbled with an inert gas to form a packaging material made of a gas impermeable material. A method for manufacturing a container containing a heat-sterilized blood preservation solution, which comprises packaging and heat-sterilizing.