Method for detecting sealing performance of medicine packaging system by using microbiological invasion method

Abstract

The invention provides a method for checking the sealing performance of a medicine packaging system by using a microbiological invasion method, which comprises the following steps of: filling a certain amount of sterile tryptone soybean broth culture medium into an injection package; the method comprises the following steps: placing a to-be-detected sample in a sterile culture medium, then inverting a packaging system filled with the sterile culture medium, immersing the packaging system in serratia marcescens suspension with the bacterial liquid concentration higher than 108cfu / ml, maintaining the to-be-detected sample in a vacuum environment of-67 kpa + / -5kpa for 30-35 minutes in the immersing process, then releasing the vacuum degree, maintaining the to-be-detected sample in a normal-pressure environment for 30-35 minutes, then taking out the to-be-detected sample, inversely culturing the to-be-detected sample at 30-35 DEG C for 7 days, and observing the to-be-detected sample, if microorganisms grow in the packaging system, leakage points exist. The method for detecting the sealing performance of the injection packaging system is high in sensitivity and beneficial for checking the sealing performance defect of injection packaging, and the product quality is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of pharmaceutical packaging materials, and in particular relates to a method for detecting the sealing performance of pharmaceutical packaging using a microbial invasion method. Background Art

[0002] With the rapid development of biotechnology in the medical field and the in-depth understanding of the pathogenesis at the cellular and molecular levels, tumor biotherapy has entered a new era. Targeted therapy refers to the means of blocking tumor growth and spread by interfering with specific molecules involved in tumor growth and development. Related drugs are called targeted drugs, which can be divided into two categories: monoclonal antibodies (abbreviated as mAbs) and small molecule compounds. Monoclonal antibody molecular targeted drugs act outside the cell membrane, compete with growth factors for binding to receptors, and block signal transduction. Molecular targeted therapy is a major progress in tumor biotherapy.

[0003] Most biological targeted drugs are sterile injections. After production and inspection, drugs must be properly and completely packaged during storage, transportation, distribution and use. Compared with ordinary product packaging, the packaging system standards for injections are more stringent. The packaging system for injections should be able to keep the contents of the product intact and prevent microbial invasion. The sealing of the packaging system refers to the ability of the packaging system to prevent content loss, microbial invasion, and gas (oxygen, air, water vapor, etc.) or other substances from entering, ensuring that the drug continues to meet safety and quality requirements. The sealing of the injection packaging system meets the requirements, which usually means that the packaging system has passed or can pass the microbial challenge test. In a broad sense, it means that there is no leakage that affects the quality of the drug. Based on scientific research and risk assessment, the maximum allowable leakage limit should be determined by considering the packaging composition and assembly, product contents, and the environment to which the product may be exposed during its life cycle. If the leakage of a packaging system does not exceed its maximum allowable leakage limit (MALL), the packaging system is considered to have good sealing.

[0004] The sealing inspection of injection packaging should take into account the type of packaging, the expected control requirements, and select appropriate sealing inspection methods based on risk assessment, according to the characteristics of the drug itself, the production process and the different stages of the drug life cycle, combined with the sensitivity and applicability of the inspection method. At present, the sealing inspection methods for injection packaging materials are divided into vacuum decay, high-voltage discharge method, laser headspace analysis method, color water method and microbial invasion method. Among the above methods, except for the microbial invasion method, all other methods are physical and chemical methods. Compared with the physical and chemical methods, the microbial invasion method is more appropriate in the following situations: 1) There is no appropriate physical and chemical leakage test method; 2) The product packaging proves to be incompatible with the known physical and chemical leakage test method; 3) The result of the required test method depends on the challenge of the microorganism. For example: a closed system for multi-dose, non-preserved products may include sterilizing filters or other mechanisms to ensure the delivery of sterile products. The validation of unpreserved multi-dose containers must ensure that the packaging can safely transport the product without being invaded by microorganisms.

[0005] The microbial invasion method for testing the sealing of injection packaging systems has been widely used in the pharmaceutical industry for decades because it uses live microorganisms to simulate the process of contamination of sterile preparations. However, the leak detection sensitivity limit of the microbial challenge method disclosed in the prior art is a leak hole ≥10μm, and a 5μm leak hole can only be detected probabilistically, and the detection rate data of a 3μm leak hole has not yet been disclosed. Therefore, it is of great significance to provide a microbial invasion method with high sensitivity and high detection rate for injection packaging systems with low leakage apertures to ensure the product quality of biological sterile injections. Summary of the invention

[0006] In view of the shortcomings of the prior art and to solve the problem of limited detection sensitivity of the existing microbial invasion method, the present invention aims to provide a microbial invasion method with higher sensitivity and higher detection rate for injection packaging systems with low leakage apertures.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides a method for detecting the sealing performance of a pharmaceutical packaging system using a microbial invasion method, the method comprising the following steps:

[0009] (1) Pour a certain amount of sterile tryptone soy broth (TSB) into the injection packaging system to be tested and seal it for later use;

[0010] (2) inverting the packaging system filled with sterile culture medium in step (1) and immersing it in a high-concentration bacterial suspension, wherein the bacterial suspension is prepared from bacteria with strong motility and small size;

[0011] (3) During the immersion process, the sample to be tested is maintained in a vacuum environment for a period of time, and then the vacuum is released and maintained in a normal pressure environment for a period of time;

[0012] (4) After taking out, place it upside down and culture it at 30-35°C for 7 days and then observe it. If microorganisms grow in the packaging system, it indicates that there is a leak.

[0013] In some embodiments of the present invention, the injection packaging system to be tested in step (1) is a vial, preferably with a packaging size of 3-20 ml, more preferably 3 ml, 10 ml, 20 ml, and more preferably 20 ml.

[0014] In some embodiments of the present invention, the injection packaging system to be tested in step (1) is a vial, and the thickness of the vial wall is preferably 0.96 mm-1.04 mm, 1.15 mm-1.25 mm, and more preferably 1.15 mm-1.25 mm.

[0015] In some embodiments of the present invention, the amount of sterile tryptone soy broth medium injected in step (1) is one-half to one-third of the volume of the sample to be tested.

[0016] In some embodiments of the present invention, the bacterial species of the bacterial suspension is selected from one or more of Serratia marcescens, Clostridium spore-forming bacteria, Pseudomonas aeruginosa, Staphylococcus epidermidis and Brevundimonas diminuta; preferably, the bacterial species of the bacterial suspension is Serratia marcescens or Brevundimonas diminuta; more preferably, the bacterial species of the bacterial suspension is Serratia marcescens.

[0017] In some embodiments of the present invention, the concentration of the bacterial suspension in step (2) is higher than 10 8 cfu / ml; preferably, the concentration of the bacterial suspension is 1×10 8 cfu / ml to 5×10 9 cfu / ml; more preferably, the concentration of the bacterial suspension is 4.625×10 8 cfu / ml, 1.43×10 9 cfu / ml and 1.43×10 8 cfu / ml.

[0018] In some embodiments of the present invention, in step (3), the sample to be tested is maintained in a vacuum environment of -67 kPa±5 kPa for 30-35 min, preferably 30-31 min, and more preferably 30 min.

[0019] In some embodiments of the present invention, the step (3) is maintained at normal pressure for another 30-35 min, preferably 30-31 min, more preferably 30 min.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] For common vial and rubber stopper injection packaging systems, the packaging system sealing detection method of the present invention can increase the detection sensitivity to 3-5 μm. The present invention provides a simple and reliable injection packaging system sealing inspection method with high sensitivity and high detection rate for packaging systems with low leakage apertures. DETAILED DESCRIPTION

[0022] In order to make the purpose, technical scheme and advantages of the embodiments of the present invention clearer, the technical scheme in the embodiments of the present invention will be described clearly and completely below. If the specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0023] The Serratia marcescens used in the embodiments of the present invention is Serratia marcescens CMCC (B) 41002, which was purchased from the China Food and Drug Inspection Institute and originated from the China Medical Bacteria Culture Collection Administration Center.

[0024] Example 1

[0025] Take a vial with a specification of 3ml and a thickness of 0.96mm-1.04mm, and use laser drilling to punch holes in the upper half of the vial. Prepare 11 samples of three standard leak holes of φ3μm, φ5μm, and φ10μm. The standard apertures of the prepared samples should all be tested to confirm that the apertures meet expectations. Add TSB at half to one-third of the vial capacity to the prepared sample vial in a Class A clean environment, and seal it with a stopper to prepare it into a positive sample. At the same time, take three non-punched sample bottles and prepare them in the same way as the positive samples as negative controls. All punched bottles should be carefully avoided from contacting the punched parts during transfer and operation.

[0026] Take 11 positive samples of the three standard leak holes of φ3μm, φ5μm and φ10μm prepared above and 3 negative control samples without holes, invert them and immerse them in the bacterial solution with a concentration of 4.625×10 8 cfu / ml of Serratia marcescens suspension. During the immersion process, the positive and negative samples were maintained in a vacuum environment of -67kPa for 30 minutes, and the vacuum was released and maintained for another 30 minutes in a normal pressure environment. After being taken out, they were inverted and cultured at 30-35℃ for 7 days and observed. If the negative control had no sterile growth, the experiment was valid. If microorganisms grew in the packaging system of the positive sample, it indicated that there was a leak.

[0027] Example 2

[0028] Take a 10ml vial with a thickness of 0.96mm-1.04mm, prepare 11 positive samples and 3 negative control samples with three standard leak holes of φ3μm, φ5μm, and φ10μm according to the method of Example 1, then invert and immerse it in a bacterial solution with a concentration of 1.43×10 9 cfu / ml of Serratia marcescens suspension. During the immersion process, the positive and negative samples were maintained in a vacuum environment of -67kPa for 30 minutes, and the vacuum was released and maintained for another 30 minutes in a normal pressure environment. After being taken out, they were inverted and cultured at 30-35℃ for 7 days and observed. If the negative control had no sterile growth, the experiment was valid. If microorganisms grew in the packaging system of the positive sample, it indicated that there was a leak.

[0029] Example 3

[0030] Take a 20ml vial with a thickness of 1.15mm-1.25mm, prepare 11 positive samples and 3 negative control samples with three standard leak holes of φ3μm, φ5μm, and φ10μm according to the method of Example 1, then invert and immerse them in a bacterial solution with a concentration of 1.43×10 8 cfu / ml of Serratia marcescens suspension. During the immersion process, the positive and negative samples were maintained in a vacuum environment of -67kPa for 30 minutes, and the vacuum was released and maintained for another 30 minutes in a normal pressure environment. After being taken out, they were inverted and cultured at 30-35℃ for 7 days and observed. If the negative control had no sterile growth, the experiment was valid. If microorganisms grew in the packaging system of the positive sample, it indicated that there was a leak.

[0031] The results of missed detection rates of vials with leak holes of different sizes in Examples 1-3 are shown in Table 1 below.

[0032] Table 1 Results of sealing test of vials with different sizes of leak holes

[0033]

[0034]

[0035] It can be seen from the above table that when the Serratia marcescens of the present invention is selected, the φ5μm and φ10μm leakage apertures of commonly used specifications of syringe bottles can all be detected, and the detection rate of the φ3μm leakage aperture is high, which can reach more than 70%, especially for the 20ml specification and the thickness of the syringe bottle of 1.15mm-1.25mm, the detection rate of the φ3μm leakage aperture is the highest, which can reach 90%.

[0036] The embodiments described above are part of the embodiments of the present invention, rather than all of the embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

Claims

1. A method for detecting the sealing of a pharmaceutical packaging system using a microbial invasion method, characterized in that: The following steps are involved: (1) Pour a certain amount of sterile tryptone soy broth (TSB) into the injection packaging system to be tested and seal it for later use; (2) inverting the packaging system filled with sterile culture medium in step (1) and immersing it in a high-concentration bacterial suspension, wherein the bacterial suspension is prepared from bacteria with strong motility and small size; (3) During the immersion process, the sample to be tested is maintained in a vacuum environment for a period of time, and then the vacuum is released and maintained in a normal pressure environment for a period of time; (4) After taking out, place it upside down and culture it at 30-35°C for 7 days and then observe it. If microorganisms grow in the packaging system, it indicates that there is a leak.

2. The method according to claim 1, characterized in that The injection packaging system to be tested in step (1) is a vial, and the preferred packaging specification is 3-20 ml, and the more preferred packaging specification is 20 ml.

3. The method according to claim 1, characterized in that The injection packaging system to be tested in step (1) is a vial, and the wall thickness of the vial is preferably 0.96 mm-1.04 mm, 1.15 mm-1.25 mm, and more preferably 1.15 mm-1.25 mm.

4. The method according to any one of claims 1 to 3, characterized in that The amount of sterile tryptone soy broth medium injected in step (1) is one-half to one-third of the volume of the sample to be tested.

5. The method according to any one of claims 1 to 4, characterized in that The bacterial species of the bacterial suspension in step (2) is selected from one or more of Serratia marcescens, Clostridium spore-forming bacteria, Pseudomonas aeruginosa, Staphylococcus epidermidis and Brevundimonas diminuta; preferably, the bacterial species of the bacterial suspension is Serratia marcescens or Brevundimonas diminuta; more preferably, the bacterial species of the bacterial suspension is Serratia marcescens.

6. The method according to any one of claims 1 to 5, characterized in that The concentration of the bacterial suspension in step (2) is higher than 10 8 cfu / ml; preferably, the concentration of the bacterial suspension is 1×10 8 cfu / ml to 1×10 9 cfu / ml.

7. The method according to any one of claims 1 to 6, characterized in that In the step (3), the sample to be tested is maintained in a vacuum environment of -67 kPa±5 kPa for 30-35 min, preferably 30-31 min.

8. The method according to any one of claims 1 to 7, characterized in that In the step (3), the mixture is kept at normal pressure for another 30-35 min, preferably 30-31 min.

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