Pharmaceutical Microbiology Quality Control
In a pharma QC microbiology laboratory, the primary objective is to identify viable microorganisms at every stage of the production process: in the air, raw materials, water and finished products. This enables lot release with the assurance of safety, efficacy and purity. To achieve this, regulatory compliant equipment and consumables are required and help to promote safe handling while minimizing the risk of false positives and false negatives.
Sartorius supports your contamination control strategy (CCS) with consumables and hardware for:
- Microbial enumeration - USP <61>, <62> and <1231>, Ph.Eur. 2.6.12, JP 4.05
- Growth-based sterility testing - USP <71>, EP 2.6.1, JP 4.06
- Rapid PCR sterility testing - EP 5.1.6, EP 2.6.27
- Microbial air monitoring – EN ISO 17141, EU GMP Annex 1
- Rapid PCR mycoplasma detection - EP 2.6.7
Microbial Enumeration | Bioburden Testing | Growth-based method
To ensure the safety of pharmaceutical products, it is crucial to monitor for and eliminate microbiological contaminants from laboratory and manufacturing environments. While membrane filtration is the preferred method for microbial enumeration, transferring the filter to the media can lead to contamination and false positives.
Microsart® filters provide a solution by combining a sterile filter unit with pre-filled agar media plate, enabling touch-free membrane transfer, and reducing the risk of secondary contamination. This innovative approach enhances the accuracy and reliability of microbial enumeration/bioburden testing, meeting regulatory requirements such as USP <61>, <62> and <1231>, Ph.Eur. 2.6.12, JP 4.05. Don't compromise when it comes to patient safety - choose Microsart® filters for reliable and accurate microbial enumeration.
- Touch-free membrane transfer
- Easy colony enumeration and picking
- Filter base and gridded membrane comes combined with a sterile 100 mL or 250 mL funnel
- Media comes pre-filled, sterile packaged, and ready-to-use
Sterility Testing | Compendial Growth-Based Method
Testing for sterility is required for sterile pharmaceutical, medical, and cosmetics products such as aqueous solutions, soluble solids, ointments, creams, parenteral preparations, and ophthalmics. According to USP <71>, EP 2.6.1, or JP 4.06, membrane filtration can be used for filterable aqueous preparations, alcoholic or oily preparations, and preparations that are miscible with or soluble in aqueous or oily solvents. To ensure accurate results, the test environment must be kept aseptic.
The Sterisart® family provides a range of closed, compatible, and cost-effective systems for sterility testing. These systems protect samples from secondary contamination and lab personnel from hazards associated with handling spikes and needles. The Sterisart® solution is more than just a product; it is a comprehensive customer solution:
- 20 different types of filtration units, canisters Sterisart® NF with specific adapters to best fit your specific product
- One system for the entire testing procedure, including Sterisart® Universal peristaltic pump
- Designed for use in laminar flow and isolators
- Backed by a global network of highly qualified service engineers and accredited local service centers
Alternative Rapid Sterility Testing | Real-Time PCR method
Compendial microbiological methods may be considered as slow, making proactive corrective action difficult at times. Alternative sterility testing methods offer near real-time results and an opportunity for earlier corrective action, improving testing quality for small volumes or non-filterable new cell-based therapeutics like ATMPs or CAR-T cells. These methods can be used for in-process samples or final product release. Even if methods described in the Pharmacopoeia are still reference methods, validation of new, rapid methods is described in EP 5.1.6, USP <1223> or USP <1071>, as they increase patient safety and reduce the need for prophylactic medication with antibiotics.
Microsart® ATMP Sterile Release Kit, a combination of Microsart® ATMP Bacteria and of Microsart® ATMP Fungi, detects bacterial and fungal contamination within three hours, using the proven real-time PCR method.
- Developed in close cooperation with authorities, and validated according to EP 5.1.6 and USP <1223> for sensitivity, specificity, and robustness
- TaqMan® probes ensure the highest level of qPCR specificity
- Lyophilized, inactivated or non-viable validation standards for required bacterial and fungal species
Microbial Air Monitoring
Monitoring air ambient in cleanrooms, isolators, or filling lines (i.e. RABS or BFS) for viable microorganisms is a routine task in the pharmaceutical, medical and cosmetics industries. Continuous monitoring of airborne viruses, bacteria, yeasts, and fungi permit trends to be recognized early. This helps in determining the root cause when contamination results are Out Of Tolerances (OOT), as a key requirement of standards and guidelines, such as the EN 17141 and the revision of the EU GMP Annex 1.
With MD8 Airscan®, you can continuously monitor the air environment for a minimum of 8 hours using just one gelatin membrane filter. However, gelatin filters have the retentive capacity of a depth filter (e.g. HEPA H14 filter) and facilitate near complete retention of microbes and viruses, due to the sieving and diffusion effect, with no loss of recovery.
- Agar-free gelatin membrane filters do not dry out during long-term sampling nor influence microbial growth
- Adaptable sampling head based on your requirements: installation in your production lines and in lab isolators or stand-alone benchtop unit
- Optional aseptic transfer of gelatin membrane filters in isolators or RABS with single-use Biosafe® Bags
Rapid Mycoplasma Detection | Real-Time PCR method
Mycoplasmas are small bacteria that can reproduce independently and cause problems in cell cultures. Routine QC and in-process testing are required for master cell bank (MCB), working cell bank (WCB), as well as for cell substrates that are used in the manufacture of biotechnological/biological products. Nucleic acid amplification techniques (NAT) alternative methods can provide faster results than traditional methods and are well described in USP, JP and EP. They must detect 10 CFU/mL of Mycoplasma, when viable but non-culturable (VBNC) Mycoplasma may go undetected even after 28 days of culture with the traditional method of mycoplasma, risking false-negative results.
Microsart® Mycoplasma qPCR detection kits offer a method for early detection of Mycoplasma contamination. The easy-to-follow protocol is aided by color-coded reagents that are both clear and concise. Additionally, the lyophilized reagents guarantee consistent quality without requiring freezer storage.
- Validated according to EP 2.6.7and USP 63 for sensitivity, specificity, and robustness
- TaqMan® probes ensure the highest level of qPCR specificity
- Lyophilized, non-infectious Mycoplasma validation standards, each containing 10 CFU of the chosen Mycoplasma species
- Applicable on any qPCR cycler suitable to detect FAM™ and ROX™ dyes
Sartorius Pharmaceutical Features
Broad range of solutions to support your Contamination Control Strategy and batch release decisions, with compendial growth-based and rapid NAT methods.
Validated solutions and services to comply with the main international regulations and standards.
German-engineered manufacturing of microbiology membranes, consumables, and hardware for over 50 years and continuous improvement to serve our customer needs in highly regulated environments.
Cell-based Therapies - Be on the Safe Side
Watch how Microsart® ATMP Sterile Release and Mycoplasma Kits detect bacteria, fungi and mycoplasma in your cell therapy products in only 3 hours.
Frequently Asked Questions
The sterility test is performed on the product in the final container. This happens in microbiological laboratories operated by the Quality Control (QC) department, before the batch of the given pharmaceutical product can be released into the market. When following USP <71>, EP 2.6.1, or JP 4.05, after a period of 14 days plus the day on which the test is being performed, a final visual inspection concludes if the product batch can be released. If turbidity and/or colonies are observed during the 14-day examination, the result does not comply with the pharmacopeia. Investigations are conducted to identify the source of biocontamination before the batch is rejected.
Why are rapid microbiological methods even more relevant for ATMP and Cell and Gene Therapies (CGT)?
Traditional microbiological methods often require several days to obtain results, which can be detrimental for time sensitive ATMP and CGT products with limited stability. Rapid microbiological methods (RMMs), like PCR-based methods, offer quicker results, increased safety, cost-effectiveness, and compliance with regulations. RMMs enable faster release of the products and reducing the risk of contamination during the manufacturing process.
It is important to monitor the bioburden because it refers to the number of microorganisms present in a given sample or environment. High bioburden can indicate contamination, which can lead to product spoilage, reduced efficacy, or even harm to patients or consumers. By monitoring the bioburden, it is possible to detect and address contamination early, ensuring the safety and quality of products. Additionally, monitoring the bioburden can help identify trends and patterns, allowing for the implementation of preventative measures to reduce the risk of contamination in the future.
Instrument qualifications (IQ|OQ|PQ) are extremely important in the pharma industry. They ensure that the instruments used are functioning properly and producing accurate and reliable results. This is critical in ensuring the safety and efficacy of pharmaceutical and medical products. Instrument qualifications also help to comply with regulatory requirements and guidelines, such as those set by the FDA and other governmental bodies. Without proper instrument qualifications, there is a risk of invalid results or fail to meet regulatory standards.
According to EU GMP Annex 1, a contamination control strategy is a comprehensive plan that outlines the measures and procedures to be taken to prevent, control, and monitor contamination in pharmaceutical manufacturing facilities. The strategy should include a risk assessment of potential sources of contamination, such as personnel, equipment, and materials, and identify critical control points where contamination must be prevented or controlled. The strategy should also include procedures for cleaning and disinfection, environmental monitoring, and personnel training. The goal of a contamination control strategy is to ensure that pharmaceutical products are manufactured in a clean and controlled environment, and are free from contaminants that could compromise their safety, quality, or efficacy.
Viscous and oily products may be diluted with a sterile diluent like isopropyl myristate (IPM). Attached study compares the time-saving efficacy of elevating the sample temperature versus diluting with IPM.
Incubating Microsart® @filter membranes on Microsart® @media provides robust recoveries at both 20°C and 35°C over a seven-day incubation period. This complies with the minimum and maximum requirements for incubation time and temperature according to Ph. Eur. (0008/0169) and USP <1231>.