JP2016180702A - Method and kit for preparing endotoxin standard solution - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an endotoxin standard solution preparation method that enables preparation through a simple procedure, and an endotoxin standard solution preparation kit for the same.SOLUTION: The above mentioned problem is cleared by providing an endotoxin standard solution preparation method that involves: a preparation step for preparing three or more airtight containers, each having a predetermined amount of injection water provided therein; and a dilution step for diluting an endotoxin standard stock solution by injecting the same into one of the airtight containers, further diluting the diluted endotoxin standard stock solution by injecting the same into another airtight container, and repeating the above procedure of diluting the endotoxin standard stock solution until the last airtight container is used. Such a method allows for preparing three or more endotoxin standard solutions having different endotoxin concentrations.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a method for preparing an endotoxin standard solution for preparing a calibration curve in an endotoxin test method and an endotoxin standard solution preparation kit used therefor.

  Endotoxin is a substance present on the cell wall of Gram-negative bacteria and has various biological activities such as pyrogenicity. Therefore, when pharmaceuticals and medical devices are contaminated with endotoxin, even a very small amount can cause serious results, and the amount of endotoxin contamination must be strictly controlled.

  The endotoxin test method includes a qualitative test and a quantitative test. As a qualitative test, a gelation method and a turbidimetric method and a colorimetric method are generally known as quantitative tests. In the endotoxin test method, as a preliminary test, an endotoxin standard solution having at least three concentrations is prepared and a calibration curve is prepared.

  When preparing an endotoxin standard solution, first prepare an endotoxin standard stock solution by dissolving the endotoxin standard product with water for injection or endotoxin test water, and then dispense the endotoxin standard stock solution and water for injection into a test tube. Dilute. At this time, it is essential that the endotoxin standard solution is not mixed with endotoxin from the external environment, and preparation of the endotoxin standard solution requires a certain level of skill and requires complicated procedures and time. Therefore, it is required to simplify the preparation process of the endotoxin standard solution.

Seikagaku Biobusiness Co., Ltd. “Standard operation method of endotoxin test method in the 16th revision Japanese Pharmacopoeia”, August 1, 2011

  The present invention has been made in view of the above circumstances, and has as its main object to provide a method for preparing an endotoxin standard solution that can be prepared by a simple process and a kit for preparing an endotoxin standard solution used therefor. .

  In order to achieve the above object, the present invention provides a preparation step of preparing three or more airtight containers containing a predetermined amount of water for injection, and an endotoxin standard stock solution is injected into one of the above airtight containers for dilution. And diluting the endotoxin standard stock solution into another airtight container and further diluting the endotoxin standard stock solution by the number of the airtight containers, and having three or more kinds of endotoxin concentrations different from each other. Provided is a method for preparing an endotoxin standard solution, characterized by preparing an endotoxin standard solution.

  According to the present invention, since a predetermined amount of water for injection is previously stored in the airtight container, when diluting the endotoxin standard stock solution, it is only necessary to inject the endotoxin standard stock solution or the diluted endotoxin standard stock solution into the airtight container. It is possible to prepare an endotoxin standard solution by a simple process. Moreover, since the process can be simplified, contamination of endotoxin from the external environment can be suppressed.

  The present invention also provides a kit for preparing an endotoxin standard solution, which contains a predetermined amount of water for injection and has three or more airtight containers that can be closed again.

  According to the present invention, since a predetermined amount of water for injection is previously stored in an airtight container, when the endotoxin standard stock solution is diluted, it is only necessary to inject the endotoxin standard stock solution or the diluted endotoxin standard stock solution into the airtight container. . Therefore, by using the endotoxin standard solution preparation kit of the present invention, it is possible to prepare an endotoxin standard solution in a simple process. In addition, since the airtight container can be reclosed, when the endotoxin standard solution is prepared using the endotoxin standard solution preparation kit of the present invention, it is possible to prevent the endotoxin standard solution from being mixed with endotoxin in the external environment. it can.

  Moreover, in this invention, it is preferable that the said airtight container has a cover part. This is because the hermetic container can be easily closed again.

  Further, in the present invention, the airtight container has a main body portion, a head portion, the main body portion and the head portion, and a cutting portion that can be cut, and the head portion is the lid portion. Also good. Such an airtight container can be opened by twisting, pulling, or pressing the head portion toward the main body portion and separating it by the cutting portion, and the head portion in which a sterilized state is ensured can be used as the lid portion.

  In the above case, it is preferable that the head is connected to the main body via a hinge. This is because it is possible to prevent the head that becomes the lid from falling during the work.

  Further, in the present invention, the airtight container is disposed at the main body, the opening of the main body, the inner lid having the opening valve, the knob, the inner lid, and the knob can be connected and cut. And the inner lid may be the lid portion. In such an airtight container, the knob portion can be twisted, pulled, or pressed toward the main body portion and separated by a cutting portion, and then opened and closed by an opening valve. Therefore, working efficiency can be improved and contamination of endotoxin from the external environment can be further suppressed.

  In the present invention, the airtight container is preferably made of plastic. This is because a disposable plastic airtight container is convenient.

  The present invention has an effect that an endotoxin standard solution can be prepared by a simple process.

It is process drawing which shows an example of the preparation method of the endotoxin standard solution of this invention. It is a schematic diagram which shows an example of the kit for endotoxin standard solution preparation of this invention. It is a schematic diagram which shows an example of the airtight container in the kit for endotoxin standard solution preparation of this invention. It is a schematic diagram which shows the other example of the airtight container in the kit for endotoxin standard solution preparation of this invention. It is a schematic diagram which shows the other example of the airtight container in the kit for endotoxin standard solution preparation of this invention. It is a schematic diagram which shows the other example of the airtight container in the kit for endotoxin standard solution preparation of this invention. It is a schematic sectional drawing which shows the other example of the airtight container in the kit for endotoxin standard solution preparation of this invention.

  The endotoxin standard solution preparation method and endotoxin standard solution preparation kit of the present invention will be described in detail below.

A. Method for preparing endotoxin standard solution The method for preparing the endotoxin standard solution of the present invention comprises a preparation step of preparing three or more airtight containers containing a predetermined amount of water for injection, and an endotoxin standard stock solution is injected into one of the above airtight containers. And diluting the diluted endotoxin standard stock solution into another airtight container to further dilute, and repeatedly diluting the endotoxin standard stock solution by the number of the airtight containers. It is a method characterized by preparing three or more different endotoxin standard solutions.

A method for preparing the endotoxin standard solution of the present invention will be described with reference to the drawings.
1A to 1B are process diagrams showing an example of a method for preparing an endotoxin standard solution of the present invention. First, as shown in FIG. 1A, five airtight containers 1 containing 0.9 mL of water for injection 11 and two airtight containers 1 containing 0.6 mL of water for injection 11 are prepared. This hermetic container 1 is a plastic ampule, and has a main body 2, a head 3, a main body 2 and a head 3, and a cutting part 4 that can be cut. The head 3 is integrally formed. Moreover, the airtight container 1 can be separated by the cutting part 4 by twisting, pulling or pressing the head part 3 toward the main body part 2 as shown in FIG. And can be closed again by the head 3.
Next, as shown in FIG. 1 (b), the head 3 is twisted to open the airtight container 1, and 0.1 mL of 10,000 EU / mL endotoxin standard stock solution 12 is accommodated in 0.9 mL of water for injection 11. Injected into the airtight container 1 and diluted 10-fold, and then 0.1 mL of the 10-fold diluted endotoxin standard stock solution 12 was injected into the airtight container 1 containing another 0.9 mL of water 11 for injection. Dilute 10-fold and repeat 10-fold dilution 5 times to prepare a 0.1 EU / mL endotoxin standard solution. Next, 0.2 mL of a 0.1 EU / mL endotoxin standard solution is injected into the airtight container 1 containing 0.6 mL of water 11 for injection, diluted 4-fold, and 4-fold dilution is repeated twice. Thus, three kinds of endotoxin standard solutions (0.1, 0.025, 0.00625 EU / mL) for preparing a calibration curve are prepared. At this time, after the endotoxin standard stock solution 12 or the diluted endotoxin standard stock solution 12 is injected into the airtight container 1 in order to prevent contamination of endotoxin from the external environment, the airtight container 1 is immediately closed with the head 3.

  In the present invention, a predetermined amount of water for injection is previously stored in an airtight container, and the endotoxin standard stock solution can be diluted stepwise by injecting the endotoxin standard stock solution or the diluted endotoxin standard stock solution into the airtight container. Therefore, an endotoxin standard solution can be prepared by a simple process. Therefore, the preliminary test process in the endotoxin test method can be simplified. Moreover, since the process can be simplified, contamination of endotoxin from the external environment can be suppressed.

  Moreover, in this invention, it is possible to apply to the preliminary test of various endotoxin test methods by adjusting the quantity of the water for injection accommodated in an airtight container suitably.

  Hereinafter, each step in the method for preparing the endotoxin standard solution of the present invention will be described.

1. Preparation Step The preparation step in the present invention is a step of preparing three or more airtight containers in which a predetermined amount of water for injection is stored.

  Here, the “airtight container” is an airtight container specified by the Japanese Pharmacopoeia. Under normal handling, transportation or storage conditions, solid or liquid foreign substances do not enter, and the content of the medicinal product is lost. Or the container which can prevent evaporation. The airtight container is a container that is not contaminated with endotoxin.

As an airtight container, a sealed container can also be used.
Here, the “sealed container” is a sealed container defined by the Japanese Pharmacopoeia, and refers to a container into which gas does not enter during normal handling, transportation or storage.

  Since the airtight container is described in detail in “B. Endotoxin standard solution preparation kit” described later, description thereof is omitted here.

  Water for injection is water for injection specified by the Japanese Pharmacopoeia, which is sterilized after purifying normal water by distillation, ion exchange, reverse osmosis, ultrafiltration, or a combination thereof, and is used for endotoxin tests. It is suitable. The water for injection does not contain endotoxin exceeding the detection limit of the lysate reagent used for the endotoxin test, and any water suitable for performing the endotoxin test may be used. For example, Otsuka distilled water manufactured by Otsuka Pharmaceutical Co., Ltd. can be used.

  The amount of water for injection stored in the airtight container is appropriately selected according to the endotoxin test method and the type of lysate reagent. For example, a kinetic colorimetric method (microplate method) using Endspecy ES-50M manufactured by Seikagaku Corporation as the lysate reagent, and a kinetic colorimetric method using Endspecy ES-24S manufactured by Seikagaku Corporation as the lysate reagent. (Single vial method) and kinetic turbidimetric method (microplate method) using Pyrotel-T manufactured by Associates of Cape Cod as a lysate reagent, 5 airtight containers containing 0.9 mL of water for injection were used. Prepare two airtight containers containing one and 0.6 mL of water for injection. Further, for example, in the case of the kinetic colorimetric method (microplate method) using Pyrochrome manufactured by Associates of Cape Cod as a lysate reagent, six airtight containers containing 0.9 mL of water for injection and 0.5 mL Prepare one airtight container containing water for injection. In the gelation method, the amount of water for injection stored in the airtight container is appropriately adjusted according to the display sensitivity λ and the dilution factor.

When an airtight container is filled with water for injection and an airtight container containing water for injection is manufactured, a sterilization environment is used. Specifically, the sterilization standard is D6 (the viable count is less than 10 ⁻⁶ cfu / mL). Is preferred. Moreover, it is preferable to manufacture with the aseptic filling system in which sterilization validation was implemented.

  The number of airtight containers is 3 or more, which is the same as the number of dilutions of the endotoxin standard stock solution. For example, when the endotoxin standard stock solution is diluted seven times stepwise as shown in FIGS. 1 (a) and 1 (b), the number of airtight containers is seven.

2. Dilution step In the dilution step of the present invention, the endotoxin standard stock solution is injected into one of the above airtight containers to dilute, the diluted endotoxin standard stock solution is injected into the other above airtight containers, and further diluted. This is a step of repeatedly diluting the endotoxin standard stock solution by the number.

The endotoxin standard stock solution is an endotoxin standard stock solution defined by the Japanese Pharmacopoeia, and is prepared by dissolving an endotoxin standard product in water for injection.
Endotoxin standards are endotoxins with quantitative guarantees established by the Japanese Pharmacopoeia for confirming the sensitivity of lysate reagents. For example, the Pharmaceuticals and Medical Devices Regulatory Science Foundation, the endotoxin detection reagent manufacturers Seikagaku Corporation, and Wako Jun. It can be obtained from drugs, Lonza, etc.

  After opening the airtight container and injecting the endotoxin standard stock solution or diluted endotoxin standard stock solution, it is preferable to quickly close the opening of the airtight container in order to prevent endotoxin contamination in the external environment. At this time, when the hermetic container has a lid, the hermetic container can be closed by the lid. Further, the opening of the airtight container may be closed with an aluminum cap or aluminum foil sterilized by dry heat.

Further, when diluting the endotoxin standard stock solution, the endotoxin standard stock solution or the diluted endotoxin standard stock solution is injected into an airtight container, and then stirred using a stirring device such as a test tube mixer or a vortex mixer.
The stirring is preferably performed with a 2 to 10 mL wide-mouth plastic ampoule. This is because the standard solution in the ampoule can be mixed uniformly and the desired endotoxin standard solution can be accurately prepared without overflowing the ampoule.

  The number of dilutions of the endotoxin standard stock solution is appropriately selected according to the endotoxin test method and the type of lysate reagent. For example, a kinetic colorimetric method (microplate method) using Pyrochrome manufactured by Associates of Cape Cod as a lysate reagent, and a kinetic colorimetric method (micromicrometer method) using Endospecy ES-50M manufactured by Seikagaku Corporation as a lysate reagent. Plate method), kinetic colorimetric method (single vial method) using Endspecy ES-24S manufactured by Seikagaku Corporation as the lysate reagent, and kinetic using Pyrotel-T manufactured by Associates of Cape Cod as the lysate reagent In the case of the turbidimetric method (microplate method), endotoxin standard stock solution is diluted seven times stepwise. In the gelation method, the number of dilutions is appropriately adjusted according to the display sensitivity λ.

3. Endotoxin standard solution In the present invention, three or more endotoxin standard solutions having different endotoxin concentrations are prepared.
The number of types of endotoxin standard solutions used for preparing a calibration curve is appropriately selected according to the endotoxin test method and the type of lysate reagent. For example, a kinetic colorimetric method (microplate method) using Endspecy ES-50M manufactured by Seikagaku Corporation as the lysate reagent, and a kinetic colorimetric method using Endspecy ES-24S manufactured by Seikagaku Corporation as the lysate reagent. (Single vial method), and kinetic turbidimetric method (microplate method) using Pyrotel-T manufactured by Associates of Cape Cod as a lysate reagent, at least three kinds of endotoxin standard solutions (0 0.1, 0.025, 0.00625 EU / mL). For example, in the case of a kinetic colorimetric method (microplate method) using Pyrochrome manufactured by Associates of Cape Cod as a lysate reagent, five kinds of endotoxin standard solutions (50, 5, 0.5, 0.05, 0.005 EU / mL). In the gelation method, four endotoxin standard solutions are prepared for preliminary tests.

B. Endotoxin Standard Solution Preparation Kit The endotoxin standard solution preparation kit of the present invention is characterized by containing a predetermined amount of water for injection and having three or more airtight containers that can be closed again.

The endotoxin standard solution preparation kit of the present invention will be described with reference to the drawings.
FIG. 2 is a schematic diagram showing an example of a kit for preparing an endotoxin standard solution of the present invention. As illustrated in FIG. 2, the endotoxin standard solution preparation kit 10 includes five airtight containers 1 containing 0.9 mL of water for injection 11 and airtight containers 1 containing 0.6 mL of water for injection 11. I have two. This hermetic container 1 is a plastic ampule, and has a main body 2, a head 3, a main body 2 and a head 3, and a cutting part 4 that can be cut. The head 3 is integrally formed.

  FIGS. 3A to 3B are schematic views showing an example of a method for using an airtight container in the kit for preparing an endotoxin standard solution of the present invention. As illustrated in FIG. 3A, the airtight container 1 can be separated by a cutting portion 4 by twisting, pulling, or pressing the head portion 3 toward the main body portion 2, and also in FIG. 3B. As illustrated, the head 3 is a lid and can be closed again by the head 3. First, as shown in FIG. 3 (a), the head 3 is twisted to open the airtight container 1, and the endotoxin standard stock solution 12 or the diluted endotoxin standard stock solution is injected into the airtight container 1 containing the water 11 for injection. To dilute. In this case, after the endotoxin standard stock solution 12 or the diluted endotoxin standard stock solution 12 is injected into the airtight container 1 in order to prevent endotoxin contamination from the external environment, as shown in FIG. 3 closes the airtight container 1.

  FIGS. 1A to 1B are process diagrams showing an example of a method for using the kit for preparing an endotoxin standard solution of the present invention, and an example using the kit for preparing an endotoxin standard solution shown in FIG. 1 (a) and 1 (b) have been described in detail in the above "A. Preparation method for endotoxin standard solution", description thereof is omitted here.

  In the present invention, a predetermined amount of water for injection is previously stored in an airtight container, and the endotoxin standard stock solution can be diluted stepwise by injecting the endotoxin standard stock solution or the diluted endotoxin standard stock solution into the airtight container. . Therefore, by using the endotoxin standard solution preparation kit of the present invention, an endotoxin standard solution can be prepared by a simple process, and the preliminary test process in the endotoxin test method can be simplified.

  Moreover, in this invention, it is possible to apply to the preliminary test of various endotoxin test methods by adjusting the quantity of the water for injection accommodated in an airtight container suitably.

  In the present invention, since the airtight container is reclosable, when the endotoxin standard solution is prepared using the endotoxin standard solution preparation kit of the present invention, the endotoxin in the external environment is mixed into the endotoxin standard solution. Can be suppressed.

  Hereinafter, each structure in the endotoxin standard solution preparation kit of this invention is demonstrated.

1. Airtight container The airtight container used in the present invention contains a predetermined amount of water for injection and is reclosable.

  The hermetic container is described in “A. Preparation method of endotoxin standard solution” above and may be reclosable, but it is preferable to have a lid. The lid allows the airtight container to be easily reclosed, and when the endotoxin standard solution is prepared using the endotoxin standard solution preparation kit of the present invention, the endotoxin standard solution is prevented from contaminating the endotoxin in the external environment. Because it can be done.

  As an airtight container having a lid, for example, ampoules and vials, screw caps and pill fur proof caps, flip tops, containers with lids having a disk top, containers having an inner lid having an open valve, containers having a seal part, Examples include a container having an opening seal tab.

Specifically, examples of the airtight container include a main body part, a head part, a main body part and a cutting part that connects the head part and can be cut, and the head part is a lid part.
2 and 3 (a) and 3 (b) are schematic views showing an example of an airtight container in the present invention. Since FIG. 2 and FIGS. 3A and 3B have been described above, description thereof is omitted here. Such an airtight container can be opened by twisting, pulling, or pressing the head portion toward the main body portion and separating it by the cutting portion, and the head portion in which a sterilized state is ensured can be used as the lid portion.
Fig.4 (a) is a schematic diagram which shows the other example of the airtight container in this invention. As illustrated in FIG. 4A, the airtight container 1 includes a main body portion 2, a head portion 3, a main body portion 2 and a head portion 3, and a cutting portion 4 that can be cut. 2, the cutting part 4 and the head part 3 are integrally formed. The airtight container 1 can be opened by twisting the head 3 or separating it with a pull cutting portion 4 as illustrated in FIG. 4B, and as illustrated in FIG. 4C. Functions as a plug and can be reclosed by the head 3.

  In the airtight container having the main body, the cutting portion, and the head, the lid configured by the head may be any one that can close the opening of the main body, for example, FIG. 2 and FIG. ) And (b) may be merely placed on the opening of the main body, or may function as an inner plug as shown in FIG. It may be one that covers. In the case where the lid portion constituted by the head functions as an inner plug, the lid portion is difficult to be removed, and thus the airtight container after reclosing can be easily handled. In addition, even when the lid portion constituted by the head covers the opening of the main body portion and can be locked to the main body portion, the lid portion is difficult to be removed. Easy handling of the container.

When an airtight container has a main-body part, a cutting part, and a head, it is preferable that the head is connected with the main-body part via the hinge. This is because it is possible to prevent the head, which is a lid, from falling during work.
Fig.5 (a) is a schematic diagram which shows the other example of the airtight container in this invention. As illustrated in FIG. 5A, the airtight container 1 includes a main body 2, a head 3, the main body 2 and the head 3, and a cutting part 4 that can be cut, and the main body 2 and the head. The main body 2, the cutting part 4, the hinge 5, and the head 3 are integrally formed with the hinge 3 having a thickness larger than that of the cutting part 4. As illustrated in FIG. 5B, the airtight container 1 can be opened by pulling the head 3 or pressing the head 3 toward the main body 2 and separating it by the cutting portion 4, as illustrated in FIG. 5C. The head 3 can be closed again. When opening and closing, the head 3 can be prevented from dropping by the hinge 5.
As the hinge, any body may be used as long as it connects the main body and the head and is not cut when cut by the cutting part. For example, the hinge may be a thick part thicker than the cutting part. There may be. When the hinge is a thick part, the head 3 is not completely separated as illustrated in FIG. 5B, but when the hinge is a strap-like hinge, the head is completely separated although not shown. It may be.

  As an airtight container which has a main-body part, a cutting part, and a head, it can select from a well-known plastic ampule, for example, and can apply.

The airtight container includes a main body, an inner lid that is disposed in the opening of the main body and has an open valve, a knob, and a cutting part that connects the inner lid and the knob and can be cut. The inner lid is a lid portion. Such an airtight container can be opened and closed by an opening valve.
Fig.6 (a) is a schematic diagram which shows the other example of the airtight container in this invention. As illustrated in FIG. 6A, the airtight container 1 includes a main body 2, an inner lid 7 that is disposed in the opening of the main body 2, and has an opening valve, a knob 8, an inner lid 7, and a knob. The main body 2, the inner lid 7, the cutting part 4, and the knob part 8 are integrally formed. As illustrated in FIG. 6B, the airtight container 1 is twisted, pulled, or pressed toward the main body 2 by the knob 8, and separated by the cutting unit 4, and as illustrated in FIG. 6C. The opening valve 6 of the inner lid 7 can be opened by pushing the side surface of the main body 2. Moreover, the opening valve 6 can be closed by returning the side surface of the main body 2 to the original position as illustrated in FIG.
In this case, by making the pressure in the airtight container higher than the atmospheric pressure, it is possible to effectively prevent endotoxin in the external environment from being mixed when the opening valve is opened.

In a container having a seal portion, the seal portion becomes a lid portion. In this case, the hermetic container can be opened by peeling off a part of the seal part, and the hermetic container can be closed again by returning the peeled part of the seal part.
As a seal part, it can select from a general thing suitably and can be used, for example, an aluminum seal is mentioned.

  In a container with a lid, for example, a screw cap, a pilfer proof cap, or the like serves as a lid. Moreover, the container with a lid may have a seal part as an inner lid. In this case, when the airtight container is opened, all the seal portions are peeled off.

In addition, examples of the container with a lid include a container in which a main body part and a lid part are connected via a thin-walled part and can be initially opened by twisting the lid part and can be resealed. Can do.
FIG. 7 is a schematic sectional view showing another example of an airtight container in the present invention. In FIG. 7, from the opening part of an airtight container main body to the shoulder part is shown. FIG. 7A is a schematic cross-sectional view showing an example before opening the airtight container, and FIG. 7B is a schematic cross-sectional view showing an example when the airtight container is resealed.
As illustrated in FIG. 7A, in the airtight container 1, the main body portion 2 and the lid portion 13 are connected via the thin wall portions 14 and 14 ′, and the thin wall portions 14 and 14 ′ are formed by twisting the lid portion 13. Is a container with a lid that can be initially opened, and has a check valve-like taper 15 at the top of the lid.
Moreover, as illustrated in FIG. 7B, the airtight container 1 is a container that can be resealed by pushing the lid 13 into the opening of the main body 2 after opening. At the time of resealing, the check valve-like taper 15 inside the lid part abuts on the inner peripheral surface of the opening part of the main body part 2, and the claw part 16 formed on the lower outer periphery of the lid part 13 and the main body part 2 By engaging the flange portion 17 formed on the neck portion, it becomes possible to maintain the airtightness of the resealed container.

  Moreover, it is preferable that a cover part is reclosable with one hand. It is because work efficiency can be improved and contamination of endotoxin from the external environment can be further suppressed. For example, when the airtight container has a main body part, a cutting part and a head part, the head part is connected to the main body part via a hinge so that the head part can be closed again with one hand. Can do. In addition, when the airtight container has a main body, an inner lid having an opening valve, a cutting portion and a knob, the opening valve can be opened and closed with one hand by pushing or returning the side of the main body. is there. Moreover, when an airtight container is a container with a lid | cover, it can be made recloseable with one hand by using the screw cap which can be closed by one push, for example.

The material of the airtight container is not particularly limited as long as it has sufficient strength to withstand a stirring operation so that sufficient mixing can be performed when adjusting the standard solution. Examples thereof include plastic, glass, and metal. it can. Of these, plastic is preferable. This is because a disposable plastic airtight container is convenient. Examples of the plastic include polyethylene, polypropylene, and polyethylene terephthalate.
As the plastic airtight container, for example, a plastic ampule, a plastic bottle, or the like can be used.

  The hermetic container may be transparent, translucent, or opaque, but is preferably transparent or translucent because water for injection and endotoxin standard stock solution are easily visible.

In the case of a plastic airtight container, it is preferable that a barrier layer is formed on at least one of the inside and the outside of the airtight container. This is because evaporation of moisture in the hermetic container can be prevented.
The material of the barrier layer is not particularly limited as long as it can be formed on the surface of a plastic hermetic container, and examples thereof include ethylene-vinyl alcohol copolymer, silicon oxide, and alumina.
Examples of the method for forming the barrier layer include a coating method and a vapor deposition method.

Moreover, the airtight container in which a predetermined amount of water for injection is accommodated may be hermetically packaged. This is because evaporation of moisture in the hermetic container can be suppressed.
As a package, what has an aluminum foil or an aluminum vapor deposition film, for example can be used.
In this case, a humidity adjusting agent capable of storing predetermined moisture and absorbing and discharging moisture, such as clean cotton containing moisture, may be disposed in the package. The humidity adjusting agent may be in the form of a sheet or a block, for example.

  The capacity of the airtight container is appropriately selected according to the endotoxin test method, but preferably has a height that does not overflow from the inlet and outlet during stirring. For example, it is preferably within the range of 1 mL to 20 mL, and particularly preferably within the range of 2 mL to 10 mL. Conventionally, when preparing an endotoxin standard solution, for example, water for injection is dispensed from a 20 mL to 1000 mL plastic bottle into a few mL test tube. However, in the present invention, such a step can be omitted.

  Three or more hermetic containers may be connected. When the airtight container is connected, when the endotoxin standard solution preparation kit of the present invention is used, it is usually separated into individual airtight containers.

  As a manufacturing method of the hermetic container, a general molding method can be applied.

  The number of airtight containers and the like are described in “A. Method for preparing endotoxin standard solution”, and thus the description thereof is omitted here.

2. Water for injection In the present invention, a predetermined amount of water for injection is stored in the airtight container.
The water for injection was described in “A. Method for preparing endotoxin standard solution” above, and thus the description thereof is omitted here.

3. Method of Use The endotoxin standard solution preparation kit of the present invention can be used in the above-mentioned endotoxin standard solution preparation method.

  The present invention is not limited to the above embodiment. The above-described embodiment is an exemplification, and the present invention has any configuration that has substantially the same configuration as the technical idea described in the claims of the present invention and that exhibits the same effects. Are included in the technical scope.

  Hereinafter, the present invention will be specifically described with reference to examples.

[Example 1]
(Preparation process)
Prepare 7 plastic ampules with 20 mL of water for injection manufactured by Otsuka Pharmaceutical Co., Ltd., and cut the ampoule head and main body with a dry heat sterilized cutter in the clean bench. Was discharged.
Place the plastic ampoule in a test tube inversion for 10 minutes, suck out the water for injection accumulated in the head using an endotoxin-free pipette tip made by Seikagaku Corporation, and invert it for 5 minutes. It was confirmed that there was no adhesion.
A plastic ampule with 20 mL of water for injection manufactured by Otsuka Pharmaceutical was newly prepared, and water for injection was dispensed into the above seven plastic ampules using an endotoxin-free pipette tip (0.6 mL: 2 tubes, 0.9 mL: 5).
(Dilution process)
In accordance with the standard operation method of kinetic colorimetric method using Endspecy ES-24S manufactured by Seikagaku Corporation, the Japanese Pharmacopoeia standard endotoxin standard product (10,000 EU / mL) in the dilution series shown in FIG. Were diluted to prepare three endotoxin standard solutions (0.1, 0.025, 0.00625 EU / mL).
During the above operation, in order to prevent endotoxin from entering from outside, the ampule prepared in the above process is pressed to open the notch, and a predetermined amount of endotoxin standard solution is dispensed into a test tube mixer. Mixed vigorously for 1 minute.
(Calibration curve reliability confirmation process)
The absorbance of the endotoxin standard solution prepared in the above step was measured at a wavelength of 545 nm in accordance with a kinetic colorimetric method (single vial method) using Endospecy ES-24S.

[Example 2]
An endotoxin standard solution was prepared and a calibration curve was prepared in the same procedure as in Example 1 using a 5 mL plastic ampoule manufactured by Niprofer Co., Ltd.

[Evaluation]
In Example 1 and Example 2, the capacity of the plastic ampule was changed, a calibration curve was prepared, and linear regression analysis was performed. As a result, the correlation coefficient r was a value within a range of 0.980 or more. A high calibration curve was obtained.
In addition, the 5 mL ampoule has better operability than the 20 mL ampoule, and the endotoxin standard solution can be suitably prepared without overflowing the solution outside the ampoule during stirring. did it.

DESCRIPTION OF SYMBOLS 1 ... Airtight container 2 ... Main-body part 3 ... Head 4 ... Cutting | disconnection part 5 ... Hinge 6 ... Opening valve 7 ... Inner lid 8 ... Knob part 10 ... Endotoxin standard solution preparation kit 11 ... Water for injection 12 ... Endotoxin standard undiluted solution 13 ... Lid part 14, 14 '... Thin part 15 ... Check valve-like taper 16 ... Claw part 17 ... Gutter part

Claims (7)

A preparation step of preparing three or more airtight containers containing a predetermined amount of water for injection;
Endotoxin standard stock solution is injected into one airtight container for dilution, diluted endotoxin standard stock solution is injected into another airtight container for further dilution, and the endotoxin standard stock solution is diluted by the number of the airtight containers. A method for preparing an endotoxin standard solution, comprising the step of repeatedly performing three or more types of endotoxin standard solutions having different endotoxin concentrations.   A kit for preparing an endotoxin standard solution, containing a predetermined amount of water for injection and having three or more airtight containers that can be closed again.   The kit for preparing an endotoxin standard solution according to claim 2, wherein the airtight container has a lid.   The airtight container has a main body part, a head part, a cutting part that connects the main body part and the head part and can be cut, and the head part is the lid part. 4. A kit for preparing an endotoxin standard solution according to 3.   The kit for preparing an endotoxin standard solution according to claim 4, wherein the head is connected to the main body via a hinge.   The airtight container has a main body, an inner lid having an opening valve, a knob, and a cutting part that connects the inner lid and the knob and can be cut. The kit for preparing an endotoxin standard solution according to claim 3, wherein the inner lid is the lid portion.   The kit for preparing an endotoxin standard solution according to any one of claims 2 to 6, wherein the airtight container is made of plastic.

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