Small Steps to Giant Leaps: Advancing Data Integrity in Sterility Testing

Webinar

With Sterility Testing serving as the final critical data point for product release to the public, the Pharmaceutical Industry is continually looking to methods which will increase efficiencies and reduce errors.

What if there was a software that could work with your existing Steritest^® Symbio pump and also allow you to digitally capture every step of the Sterility test? 

You may have already about the M-Trace^® 21 CFR Part 11 compliant solution, giving step-by-step handling instructions, while collecting all relevant data in realtime. The system has launched its second version, which is now Microsoft Windows compatible, providing easier integration in your company IT infrastructure.

Agenda:

• M-Trace^® solution existing and new features that enable seamless integration with your Steritest^® Symbio pump and with your current IT environment
• Importance of capturing every action during Sterility Testing to improve operational efficiency and reduce errors
• Practical examples of how digitalization can transform your facility's testing procedures and support compliance with Standard Operating Procedures (SOPs)
• Live Q&A

Speakers:

• Michel van Musschenbroek: Regional Application & Commercial Tactics Manager, North America, Merck KGaA
• Nicolas Lelièvre: Regional Application & Commercial Tactics Manager, Western Europe, Merck KGaA

Language:

• English, but live translation in Japanese and Spanish (subtitles)

Time and date

May 13 at 10:00am CEST (Paris) / 4:00pm SGT (Singapore) / 5:00pm JST (Tokyo)     

Link to register: https://event.on24.com/wcc/r/4903610/5BDE8D3C78E8CDB11007DF9A05049616?partnerref=PMRWeb

 

Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (http://www.pharmamicroresources.com/)

Improving the sterility assurance of the sterility test: How the Steritest® System leads to enhanced controls

Image: Sterility test incubation (by Tim Sandle)

For conducting the pharmacopeia test for sterility the membrane filtration is the method of choice because all of the contents of a small volume product are filtered or at least half the contents of a large volume product are passed through a membrane filter. Therefore, a much larger sample size is evaluated than is so for the direct inoculation method (where the amount of product can vary from 1- 2 ml to half the container contents). 

By Tim Sandle

Furthermore the method is more adept at overcoming interfering factors; for any microorganisms present are far more likely to be separated from potentially inhibitory substances in the product through the act of filtration or, should they remain, they can be eliminated by rinsing the filter.



READ MORE: Raising the bar with sterility test compliance: Merck’s M-Trace® System


However, not all approaches to sterility testing are the same and there are differences in relation to the test method, test environment, test controls and so on. These variances, if not controlled, can adversely impact upon the sterility assurance of the test1. By considering these variances, as discussed in this article, the sterility assurance of the test can be strengthened.

Membrane filtration method

Membrane filtration is the appropriate method for all aqueous, alcoholic, oily and solvent products that can pass through a sterile filter with a porosity of 0.45mm. The standard filter is manufactured from cellulose esters or other similar plastics. The filter acts to separate the product from any microorganisms so that the product passes through the filter and any microorganisms present in the product are trapped within the filter matrix. A rinse solution (such as phosphate buffered saline, saline or Ringer’s solution) is used to remove any product residues (due to the risk of antimicrobial activity posed from the residues)2. This washing process is normally performed three times. The maximum number of rinses permitted in the pharmacopeia is 5 x 100 mL.


To support the membrane filtration method, demonstrably effective culture media must be used. This requires the use of soya-bean casein digest (SCD) and fluid thioglycolate media (FTM) that can support the growth of a range of microorganisms, and which can be introduced into the test environment safely, to avoid cross-contamination and with the containers being sufficiently integral to avoid the seepage of any decontamination agents3.

Achieving improved sterility assurance with the Steritest® System

To avoid cross-contamination from the operator and test environment, the membrane filtration systems needs to be enclosed, such as with the Merck Steritest® System (first introduced in 1974). This system minimises risk of contamination by reducing transfer steps and by minimising the possibility of contamination deposition (Van Doorne et al, 1998). Furthermore, there are no open containers or membrane manipulations, which could increase the risk of contamination, supported by an effective ergonomic design .


While there are different closed systems on the market, not all systems have proven container integrity. The Steritest® System has been developed with robust sealing technology and verified not to allow any ingress from the external environment. It is additionally important that a supplier can provide supporting information for the integrity of the membrane filter. When assessing the filter, the ability of the filter to cope with the expected pressure applied to the system is important (and once established, the selected system should have the ability to carefully control and to program the pump speed). The control of the pump speed not only allows proper product splitting between the 2 pathways, and hence the 2 media for incubation, but it also helps to guard against products from foaming and it reduces the physical stress applied to any microorganisms that might be present to aid their subsequent cultivation.

Strengthening sterility assurance: Test controls

During testing it is prudent to include a negative control. There are different ways to perform a negative control and in a sense there is no right or wrong approach. One common way, with the membrane filtration test, is to filter a bottle of the rinse medium through the same batch of the filtration kit used to conduct the test, and then to add culture media. This provides an indication of the rinse fluid, culture media, test kit and testing environment.


Negative controls are most useful when a sterility test failure occurs. The result of the control may indicate a false positive (in a circumstance where both the sterility test and the negative control fail). Controls should also be trended and out of trend situation should be reacted to even where the sterility test is satisfactory for negative control failures in themselves are indicators of a potential loss of environmental control or poor technique.

Strengthening sterility assurance: Environmental monitoring

During the sterility test, environmental monitoring should be conducted. This is in order to show that the test environment was satisfactory during the time of testing and, should a sterility test fail, to provide a means for assessing whether the contaminant arose from the test environment rather than from the manufacturing process (a complicated process, requiring genotypic microbial identification)4.



The types of environmental monitoring samples will be the same as per the monitoring of pharmaceutical facilities in relation to the viable counting methods:

• Settle plates
• Active (volumetric) air-samplers

Which are taken at some stage during the test session.

• Contact plates. Swabs
• Finger plates of the operator’s gloves (or of the gloveport gauntlet, should an isolator be used)

Which are taken at the end of the test session.

The limit, given the grade of the environment (ISO class 5 or the equivalent PIC/S GMP Grade A) should be ‘no growth’ (effectively 1 CFU - colony forming unit) per sample. The results should be trended. Repeated occurrences of microbial counts from environmental monitoring should trigger an investigation and such results may indicate a loss of environmental control.


Media should be assessed prior to release by conducting growth promotion testing. When testing media, the microbial challenge must be of a low level, at less than 100 colony forming units (CFU). The inoculation challenge must be verified at the time of testing via a plate count. At the same time as conducting the test the use of a previously released batch of the same media as a control provides a useful comparator should anomalous results be seen with the medium under test5.

Particle counting is not normally conducted during the test session, and it not recommended here due to the nature of the activities, which are likely to generate particle counts. It is prudent, however, to periodically ascertain the particle levels within the isolator or mobile unidirectional airflow cabinet within which the sterility test is conducted when such devices are not being used for testing.

Strengthening sterility assurance: Controlled environments

There are two types of environments within which the sterility test can be conducted. These are:


a) A unidirectional airflow (UDAF) cabinet contained within a classified cleanroom;

b) An isolator.


Of the two, the isolator affords better control by virtue of it being a barrier device the analyst is separated from the product under test. However, both types of environments, if they are not properly maintained, or if the methods for transferring materials in and out are not validated, can lead to cross contamination and hence lead to the risk of false positives occurring. Nonetheless, the use of isolators, when used correctly, has reduced incidents of false positive6. Isolators are most commonly sanitised using surface contact disinfectants: hydrogen peroxide vapour (an alternative is ionized hydrogen peroxide) or peracetic acid.

Membrane filtration systems can that be permanently situated inside of the isolator help to minimise the risks of transferring equipment into and out of the controlled environment.

The equipment entering the controlled environment ,including the membrane filtration apparatus and pump, needs to be of an appropriate design (such as the quality of the surface finish) an easily decontaminated using disinfectant agents.

Summary

There are a number of factors that can aid the sterility assurance of the sterility test. The article has looked at the controlled environment, environmental monitoring, negative controls, and the design of membrane filtration equipment and methods. These factors should be considered when building in quality by design into the sterility test process. To support this, there are considerable advantages to be drawn from technology like the Merck Steritest® System.

References

1. Sandle, T. (2013) Sterility Testing of Pharmaceutical Products, PDA / DHI, River Grove, IL, USA

2. Proud, D. W. and Sutton, S.V.W. (1992): Development of a Universal Diluting Fluid for Membrane Filtration Sterility Testing, Applied and Environmental Microbiology, March 1992, Vol. 58, No.3, p1035 – 1038.

3. Ohresser, S., Griveau, S. and Schann, C. (2004): Validation of Microbial Recovery from Hydrogen Peroxide-Sterilised Air, PDA J Pharm Sci Tech. 58 (2): 75-80.

4. Sandle, T. (2012). ‘Environmental Monitoring: a practical approach’ In Moldenhauer, J. Environmental Monitoring: a comprehensive handbook, Volume 6, PDA/DHI: River Grove, USA, pp29-54

5. Sandle, T. (2010): The Media Kitchen: Preparation and Testing of Microbiological Culture Media in Sutton, S. (ed.): Laboratory Design: Establishing the Facility and Management Structure, DHI / Parenteral Drug Association, Bethesda, MD, United States, ISBN 1-933722-46-0, pp269-293.

6. Ackers, J., Agalloco, J. and Kennedy, K. (1995): Experience in the Design and Use of Isolator Systems for Sterility Testing, PDA Journal of Pharmaceutical Science and Technology, 49(3): 140-144


Posted by Dr. Tim Sandle, Pharmaceutical Microbiology Resources (http://www.pharmamicroresources.com/)

Raising the bar with sterility test compliance: Merck’s M-Trace® System

Maintaining compliance for the sterility test, demonstrating the validated state, enabling testing consistency, and showing traceability of consumables – each an important component of laboratory data integrity – has been made easier through the introduction of Merck’s M-Trace^® system. This article looks at the importance of sterility test compliance and how M-Trace^® can deliver the solution.

By Tim Sandle

The test for sterility

 

The pharmacopeial test for sterility is one of the most important laboratory tests and a regulatory requirement for aseptically filled pharmaceutical products. A test recording a ‘pass’ result, together with evidence of sterility assurance during manufacturing, provides evidence of the quality of the manufactured product.

 

The most common form of the test is the membrane filtration method, where the presence or absence of microorganisms is assessed through an examination of turbidity, following incubation, of microbiological culture media.

 

Maintaining the validated state

 

The criticality of the test places it under regular audit scrutiny, from the validation of the test to its routine performance. Sterility test validation is performed to develop a test method through which it can be demonstrated that the product does not have a microstatic or microbicidal effect.

 

To show that the test has been conducted effectively and following the initial validation, a series of parameters need to be controlled (such as the use of the correct membrane filter; the speed at which the product is passed through the membrane filter; and the number of rinses required to remove any potentially antimicrobial product residues). These parameters often vary between products and include:

 

• The test kit to be used
• The rinse solution to be used
• If a reconstitution fluid is required
• The pump speed
• The number and volume of rinses required.

 

Many of these parameters need to be submitted as part of the regulatory filing for new products. It is also important that the validated conditions are accurately captured in the test procedure and replicated by each analyst who performs the test.

 

Traceability

 

It is also important to carefully record the culture media used (typically soya-bean casein digest and fluid thioglycolate media). These media need to undergo quality control testing and release, before being used in the sterility test. The release process includes growth promotion testing (using pharmacopeia strains and isolates from the facility’s own manufacturing environment) and a ‘sterility test’ (incubating some of the test bottles to show the absence of contamination). Use of the incorrect culture medium will lead to test invalidation.

 

In addition, the consumables used in the sterility test also need to be captured, including the test kit and rinse fluids. Different test kits and rinse fluids may be required for different product families, and knowing which consumables were used is essential should a sterility test failure investigation be required.

 

Data integrity

 

The adherence to the validated state and traceability requirements must conform to data integrity expectations. This is especially critical for a final product release test like the sterility test. A useful acronym when considering data integrity, and one that serves as an appropriate summary point, is ALCOA. Here: data must be attributable, legible (permanent), contemporaneous, original and accurate. This means:

• Attributable: The identity of the person completing a record should be unambiguous.

• Legible (permanent): It should not be possible to modify or recreate data without an audit trail that preserves the original record. For paper records, what has been written needs to be understandable.
  

• Contemporaneous: System design impacts upon contemporaneous record keeping. For instance, the availability of records in the right place at the right time removes the need for staff to remember things to write down or entering into a computer later on.
  

• Original: Original records must preserve data accuracy, completeness, content and meaning.
  

• Accurate: Automated data capture, with the required controls, provides greater control over the accuracy of a record.

From the above, where a computerised system is used, the system must:

 

•  Permit access by authorised personnel only.
•  Be password controlled, with different access levels.
•  Allow for the creation of backup copies.
•  Operate an accurate and secure audit trail.

 

M-Trace^® Solution

 

To help address the above best practice requirements, Merck has developed a means of ensuring that sterility tests are conducted as designed and validated (through the use of electronic standard operating procedures) and that all consumables are captured and connected with the relevant sterility test. This is through M-Trace^®, a system that ensures the sterility test achieves an improved state of compliance.

 

The use of the electronic procedure also ensures that operational steps are not missed or performed out of order and that any ambiguity or different interpretation between analysts is minimised.

 

M-Trace^® Software

 

The first part of the system is the M-Trace^® Software. This enables semi-automated control of the Merck Steritest^® Symbio Pump (used for the sterility test using the membrane filtration method). The analyst can view the software and control the operational steps via a headset and voice commands or directly from the Symbio Pump display. This streamlines the workflow and, through the avoidance of errors or delays to verify the required steps, it can boost productivity.

 

Automating the workflow: M-Trace^® Touch Mobile Computer

 

The second part of the system is a mobile device, which resembles a smartphone, used to scan and digitally capture the data matrix codes of consumables. With this, Merck has developed a digital companion called the M-Trace^® Touch Mobile Computer.

 

Capturing test consumables accurately is not only important; this is an area that can be prone to error within a busy laboratory, especially within the sterility test suite here personnel will be wearing cleanroom-style gowns and gloves. The M-Trace^® Touch Mobile Computer enables consumable data to be captured accurately and the automation element ensures that no consumables are missed, and that traceability is achieved through automation. Where scanning is undertaken within the sterility test facility, the M-Trace^® Touch Mobile Computer is positioned outside of the Grade A zone (outside of an isolator or unidirectional airflow device), using the M-Trace^® Mobile Computer Mount, enabling the analyst to perform the scanning hands-free.

 

Laboratory compliance: M-Trace^® Electronic Test Record Software

 

When the user is within range of the M-Trace^® All-in-One PC (MTRACEPC1) the data is transmitted to the electronic test record. Hence, the M-Trace^® Mobile App connects to M-Trace^® Electronic Test Record Software. This enables the analyst performing the sterility test to digitally capture records of consumables used in the sterility test together with test results.

 

In addition, the digital information can be integrated into a Laboratory Information Management System (LIMS), here a laboratory supervisor or manager can review the test parameters and consumables prior to releasing the result.

 

Compliant solution

 

The M-Trace^® system aids the laboratory in improving compliance by reducing the potential for error (in terms of testing and data capture) and provides an electronic repository to enable the accurate release of test results and a digital archive should test problems or failures be encountered, as well as providing a means for an auditor to examine historical test results.

 

The life science business of Merck operates as MilliporeSigma in the U.S. and Canada.

 

Dr. Tim Sandle, Pharmaceutical Microbiology Resources (http://www.pharmamicroresources.com/)