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

A method for detecting the sealing performance of pharmaceutical packaging systems using microbial invasion method Technical Field

[0001] The invention belongs to the technical field of pharmaceutical packaging materials, and particularly 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 pathogenesis at the cellular and molecular levels, tumor biotherapy has entered a new era. Targeted therapy refers to the method of blocking tumor growth and spread by interfering with specific molecules involved in tumor growth and development. Related drugs are called targeted drugs and can be divided into two major categories: monoclonal antibodies (abbreviated as mAbs) and small molecule compounds. Monoclonal antibody molecular targeted drugs act on the outside of the cell membrane, competing with growth factors for binding to receptors and blocking signal transduction. Molecular targeted therapy is a major advancement in tumor biotherapy.

[0003] Most targeted biological drugs are sterile injectables. After production and testing, drugs must be properly and completely packaged during storage, transportation, distribution, and use. Compared to packaging for general products, the standards for injectable packaging systems are more stringent. Injectable packaging systems must maintain the integrity of the product contents while preventing microbial intrusion. The sealing performance of a packaging system refers to its ability to prevent content loss, microbial intrusion, and the ingress of gases (oxygen, air, water vapor, etc.) or other substances, ensuring that the drug continues to meet safety and quality requirements. A packaging system that meets the sealing requirements for an injectable packaging system generally means that the packaging system has passed or is capable of passing microbial challenge testing. Broadly speaking, it means that there are no leaks that could affect drug quality. Based on scientific research and risk assessment, the maximum allowable leakage limit (MALL) should be determined, taking into account the packaging composition and assembly, the product contents, and the environment to which the product may be exposed during its lifecycle. A packaging system is considered to have good sealing performance if its leakage does not exceed its maximum allowable leakage limit (MALL).

[0004] The seal integrity inspection of injectable packaging should consider the packaging type and expected control requirements. Based on a risk assessment, an appropriate seal integrity inspection method should be selected, taking into account the characteristics of the drug, the manufacturing process, and the different stages of the drug lifecycle, in combination with the sensitivity and applicability of the inspection method. Currently, the seal integrity inspection methods for injectable packaging materials are categorized into vacuum decay, high-voltage discharge, laser headspace analysis, water colorimetry, and microbial intrusion. Of these methods, all but the microbial intrusion method are physicochemical. Microbial intrusion methods are more appropriate than physicochemical methods in the following situations: 1) no appropriate physicochemical leak testing method exists; 2) the product packaging demonstrates incompatibility with known physicochemical leak testing methods; or 3) the results of the required test method depend on a microbial challenge. For example, closed systems for multi-dose, non-preservative products may include sterilizing filters or other mechanisms to ensure sterile product delivery. Validation of unpreserved multi-dose containers must ensure that the packaging can safely transport the product without microbial intrusion.

[0005] Microbial challenge testing for leak tightness in injectable packaging systems has been widely used in the pharmaceutical industry for decades because it uses live microorganisms to simulate the contamination process of sterile preparations. However, the leak detection sensitivity limit of currently available microbial challenge methods is limited to leaks ≥10μm. Leaks of 5μm can only be detected probabilistically, and data on the detection rate of leaks of 3μm is not yet available. Therefore, developing a microbial challenge method with high sensitivity and a high detection rate for injectable packaging systems with small leak diameters is of great significance for ensuring the product quality of biologically sterile injectables.

[0006] Summary of the Invention

[0007] In order to address the shortcomings of the existing technology and solve the problem of limited detection sensitivity of existing microbial invasion methods, 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.

[0008] To achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] 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:

[0010] (1) Pour a certain amount of sterile tryptone soy broth (TSB) into the packaging system of the injection to be tested and seal it for later use;

[0011] (2) inverting the packaging system filled with sterile culture medium in step (1) and immersing it in a high-concentration bacterial suspension prepared from a bacterial species with strong motility and small size;

[0012] (3) During the immersion process, the sample to be tested is kept in a vacuum environment for a period of time, and then the vacuum is released and kept in a normal pressure environment for a period of time;

[0013] (4) After taking out, place it upside down and culture at 30-35℃ for 7 days and observe. If microorganisms grow in the packaging system, it indicates that there is a leak.

[0014] 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.

[0015] 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.

[0016] 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.

[0017] 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.

[0018] 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.

[0019] 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.

[0020] In some embodiments of the present invention, the step (3) is maintained at normal pressure for another 30-35 minutes, preferably for 30-31 minutes, and more preferably for 30 minutes.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] For common vial and stopper injection packaging systems, the present invention's packaging system sealing detection method can increase detection sensitivity to 3-5 μm. This invention provides a simple and reliable method for testing the sealing of injection packaging systems, with high sensitivity, a high detection rate for packaging systems with low leakage apertures, and a high degree of accuracy. DETAILED DESCRIPTION

[0023] To make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are described clearly and completely below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be purchased commercially.

[0024] The Serratia marcescens used in the examples of the present invention is Serratia marcescens CMCC(B)41002, which was purchased from the China Food and Drug Administration and the source of which is the China Center for the Collection of Medical Bacteria.

[0025] Example 1

[0026] Take a 3ml vial with 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: φ3μm, φ5μm, and φ10μm. The standard apertures of the prepared samples should all be tested to confirm that the apertures meet expectations. After preparation, add TSB at a volume of one-half to one-third of the vial's capacity in a Class A clean environment, and seal with a stopper to prepare a positive sample. At the same time, take three unpunched sample bottles and prepare them in the same way as the positive samples as negative controls. All punched bottles should be carefully avoided from contact with the punched area during transfer and operation.

[0027] 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 During the immersion process, the positive and negative samples were kept in a vacuum environment of -67kPa for 30 minutes, and then the vacuum was released and kept in a normal pressure environment for another 30 minutes. After being taken out, they were placed in an inverted culture at 30-35℃ and observed for 7 days. If the negative control showed 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.

[0028] Example 2

[0029] Take a 10ml vial with a thickness of 0.96mm-1.04mm, prepare 11 positive samples and 3 negative control samples of 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 9 During the immersion process, the positive and negative samples were kept in a vacuum environment of -67kPa for 30 minutes, and then the vacuum was released and kept in a normal pressure environment for another 30 minutes. After being taken out, they were placed in an inverted culture at 30-35℃ and observed for 7 days. If the negative control showed 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.

[0030] Example 3

[0031] Take a 20ml vial with a thickness of 1.15mm-1.25mm, and prepare 11 positive samples and 3 negative control samples of 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 During the immersion process, the positive and negative samples were kept in a vacuum environment of -67kPa for 30 minutes, and then the vacuum was released and kept in a normal pressure environment for another 30 minutes. After being taken out, they were placed in an inverted culture at 30-35℃ and observed for 7 days. If the negative control showed 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.

[0032] The results of the missed detection rates of the vials with leak holes of different sizes in Examples 1-3 are shown in Table 1 below.

[0033] Table 1. Test results of sealing performance of vials with different sizes of leak holes

[0034] As can be seen from the above table, when the Serratia marcescens of the present invention is selected, all leak pore sizes of φ5μm and φ10μm for commonly used syringes can be detected. The detection rate for leak pores of φ3μm is high, reaching over 70%. In particular, the detection rate for leak pores of φ3μm for syringes with a specification of 20ml and a thickness of 1.15mm-1.25mm is the highest, reaching 90%.

[0035] The embodiments described above are some, but not all, of the embodiments of the present invention. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention as claimed, but rather merely represents selected embodiments of the present invention. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are also 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.