[Skip to Content](https://www.sigmaaldrich.com#main-content) [](https://www.sigmaaldrich.com/HK/en) Products Cart0 HKEN Products ProductsApplicationsServicesResourcesSupport [Login / Register](https://www.sigmaaldrich.com/oidc-sign-in) [Home](https://www.sigmaaldrich.com/HK/en)[Applications](https://www.sigmaaldrich.com/HK/en/applications)[Microbiological Testing](https://www.sigmaaldrich.com/HK/en/applications/microbiological-testing)Sterility Testing # Sterility Testing Sterility testing is a critical GMP requirement to confirm that pharmaceutical products, medical devices, biologics, and cell & gene therapies are free from viable microorganisms prior to patient administration. A failed sterility test — whether a true positive or a false result — carries significant consequences: product recalls, patient safety risks, and costly regulatory investigations. As pharmacopoeia standards evolve and regulatory scrutiny intensifies, modern sterility testing programs must address three interconnected challenges: __methodological accuracy__, __data integrity__, and __speed-to-release__. Understanding the available sterility testing methods, their regulatory frameworks, and the latest advances in digital workflows and rapid microbiological methods is essential for any quality control laboratory operating under GMP. From selecting the right testing method for your product and packaging format, to meeting evolving regulatory expectations around data integrity and speed-to-release, this guide walks you through __every key aspect of sterility testing.__  - [Pharmaceutical Sterility Testing Methods](https://www.sigmaaldrich.com#pharma-sterility) - [Membrane Filtration Sterility Testing](https://www.sigmaaldrich.com#membrane) - [Direct Inoculation Sterility Testing](https://www.sigmaaldrich.com#direct-inoculation) - [Rapid Microbiological Methods (RMM) for sterility testing](https://www.sigmaaldrich.com#rmms) - [Data Integrity and Governance in Sterility Testing](https://www.sigmaaldrich.com#data-integrity) - [Sterility Testing Applications: From Pharmaceuticals to Cell & Gene Therapies](https://www.sigmaaldrich.com#sterility-testing-applications) - [Sterility Testing Workflows](https://www.sigmaaldrich.com#workflows) [Speak to a specialist](https://www.sigmaaldrich.com/HK/en/campaigns/sterility-testing-consumables) * * * ## Related Products Slide 1 of 16 1 of 4 [ \ Millipore \ TZHALA210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhala210) Quick View [ \ Millipore \ STBMTSB12 \ Tryptic Soy Broth Ready-to-use Bottles](https://www.sigmaaldrich.com/HK/en/product/mm/stbmtsb12) Quick View [ \ Millipore \ SYMBLFH01 \ Steritest® Symbio LFH Pump Kit](https://www.sigmaaldrich.com/HK/en/product/mm/symblfh01) Quick View [ \ Millipore \ SYMBABR01 \ Steritest® Glass Ampoule Breaker](https://www.sigmaaldrich.com/HK/en/product/mm/symbabr01) Quick View [ \ Millipore \ STBMFTM12 \ Fluid Thioglycollate Medium](https://www.sigmaaldrich.com/HK/en/product/mm/stbmftm12) Quick View [ \ Millipore \ STBMRFK34 \ Fluid K - Ready-to-use Rinse Fluid](https://www.sigmaaldrich.com/HK/en/product/mm/stbmrfk34) Quick View [ \ Millipore \ TZHALV205 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhalv205) Quick View [ \ Millipore \ TZHASY210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhasy210) Quick View [ \ Millipore \ TZHASV210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhasv210) Quick View [ \ Millipore \ TZHAPC210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhapc210) Quick View [ \ TZHACA210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhaca210) Quick View [ \ Millipore \ TZHVAB210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhvab210) Quick View [ \ Millipore \ TZHVMD210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhvmd210) Quick View [ \ Millipore \ TZHVDV210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhvdv210) Quick View [ \ Millipore \ TZHVSL210 \ Steritest® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzhvsl210) Quick View [ \ Millipore \ TZVC00010 \ Steridilutor® NEO Device](https://www.sigmaaldrich.com/HK/en/product/mm/tzvc00010) Quick View * * * ## Featured categories [](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/sterility-testing-consumables-media-and-filtration) [Sterility Testing Consumables, Media & Instruments](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/sterility-testing-consumables-media-and-filtration) Laboratory instruments and consumables for sterility testing in microbiological quality control: Sterility test media, pumps, hardware and accessories. [Shop Products](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/sterility-testing-consumables-media-and-filtration) [](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/pyrogen-testing) [Pyrogen Testing](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/pyrogen-testing) Ensure safety: Pyrogen testing is vital for pharmaceuticals, devices. Monocyte Activation Test (MAT) detects endotoxin, non-endotoxin pyrogens. [Shop Products](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/pyrogen-testing) [](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/microbial-filtration) [Microbial Filtration](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/microbial-filtration) Speed up your time-to-result with rapid microbial testing using our complete membrane filtration systems; used in bioburden testing, food and beverage analysis, and cannabis microbiology testing. [Shop Products](https://www.sigmaaldrich.com/HK/en/products/industrial-microbiology/microbial-filtration) * * * Overview Related articles and protocols Support ## [](https://www.sigmaaldrich.com)Pharmaceutical sterility testing methods Compendial sterility testing for pharmaceutical products is governed by __USP <71>__, __Ph. Eur. 2.6.1__, and __JP <4.06>__, which require samples to be cultured in two separate media over a __14-day incubation period__. Two culture media are used to ensure comprehensive microorganism coverage: [Fluid Thioglycolate Medium (FTM)](https://www.sigmaaldrich.com/HK/en/search/ftm-steritest?focus=products&page=1&perpage=30&sort=relevance&term=ftm%20steritest&type=product) for anaerobic and aerobic bacteria (incubated at 32.5°C), and __Soybean Casein Digest Medium (SCDM)__ also called [Trypticase Soy Broth (TSB)](https://www.sigmaaldrich.com/HK/en/search/tsb-steritest?focus=products&page=1&perpage=30&sort=relevance&term=tsb%20steritest&type=product) for aerobic bacteria and fungi (incubated at 22.5°C). Any turbidity observed after incubation indicates potential microbial contamination and must be investigated. Two compendial methods are recognized: [membrane filtration](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/steritest-neo-sterility-test) and [direct inoculation](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-direct-inoculation). The choice of method depends on the nature of the product, its packaging format, and its physicochemical properties. Where the nature of the product permits, __membrane filtration is the method of choice__ as stated by USP <71>, Ph. Eur. 2.6.1, and JP <4.06>, owing to its superior sensitivity and ability to remove inhibitory substances. Direct inoculation remains applicable where filtration is not feasible. While the 14-day incubation period remains the pharmacopoeial standard, [Rapid Microbiological Methods (RMMs)](https://www.sigmaaldrich.com#rmms) based on ATP bioluminescence technology are increasingly accepted by regulators for short shelf-life products, including cell and gene therapies. The new __USP <73>__ chapter — "ATP Bioluminescence-Based Microbiological Methods for the Detection of Contamination in Short-Life Products" — provides a clear regulatory framework for RMM adoption and for the validation and implementation of this technology, effective __August 2025__. Regardless of the method selected, [data integrity](https://www.sigmaaldrich.com#data-integrity) requirements apply equally to all sterility testing workflows: all test data must be fully traceable, audit-ready, and compliant with __ALCOA++__, __21 CFR Part 11__, and __EU Annex 11__ — from sample receipt to final result recording. ## [](https://www.sigmaaldrich.com)Membrane filtration sterility testing [](https://www.sigmaaldrich.com/documents-search) [Looking for more specific information?](https://www.sigmaaldrich.com/documents-search) Visit our document search for data sheets, certificates and technical documentation. [Find documents](https://www.sigmaaldrich.com/documents-search)  With 50+ years of experience as the industry pioneer, the [Steritest® solution](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/steritest-neo-sterility-test) delivers robust and safe sterility testing for trusted results. Membrane filtration is the __regulatory method of choice__ for filterable pharmaceutical products under __USP <71>__, __Ph. Eur. 2.6.1__, and __JP <4.06>__. The sample is passed through a __0.45 µm membrane filter__, where microorganisms are retained and subsequently cultured in appropriate media. This method offers key advantages over direct inoculation: - __Higher sensitivity__ — the entire sample, or a composite, is filtered through a single membrane - __Inhibitor removal__ — antimicrobial compounds such as antibiotics or preservatives can be rinsed away before incubation, preventing false negative results - __Closed-system protection__ — modern closed-system filtration devices ensure the product is never exposed to the environment during testing, significantly reducing the risk of extraneous contamination and invalid test results The use of __closed-system, single-use membrane filtration devices__ is considered best practice in GMP sterility testing environments, both in laminar flow hoods and isolators. ## [](https://www.sigmaaldrich.com)Direct inoculation sterility testing In [direct inoculation sterility testing](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-direct-inoculation), a small volume of sample is aseptically transferred into a suitable growth medium and incubated. While straightforward to perform, this method presents several limitations that laboratories should consider: - __Reduced sensitivity__ — only small sample volumes can be inoculated, increasing the risk of missing low-level contamination - __Turbidity interference__ — inherently turbid or colored products can mask microbial growth during visual examination - __Antimicrobial inhibition__ — products with antimicrobial properties must be neutralized prior to inoculation to avoid false negative results  ## [](https://www.sigmaaldrich.com)Rapid Microbiological Methods (RMM) for sterility testing As regulatory frameworks evolve to accommodate innovation, __Rapid Microbiological Methods (RMMs)__ are gaining acceptance as scientifically validated alternatives to traditional compendial sterility testing. RMMs are particularly relevant for __short shelf-life products__ — such as cell and gene therapies, radiopharmaceuticals, and short-life biologics — where a 14-day incubation period is incompatible with product release timelines or patient treatment schedules. Among the most established RMM technologies, __ATP bioluminescence__ detects light emitted by viable microorganisms following reaction with a bioluminescence reagent, enabling rapid, sensitive, and quantitative contamination detection. Unlike traditional incubation-based methods, ATP bioluminescence replaces subjective visual turbidity assessment with a __quantitative, instrument-based measurement__, enabling earlier and more reproducible result interpretation. The introduction of __USP <73>__ — "ATP Bioluminescence-Based Microbiological Methods for the Detection of Contamination in Short-Life Products", effective __August 2025__ — marks a significant regulatory milestone, providing laboratories with a clear pharmacopoeial framework to implement and validate RMM-based sterility testing. This is complemented by __USP <1071>__, which provides broader guidance on the implementation and validation of rapid microbial methods. RMMs are suitable not only for __end-product sterility testing__ but also for __in-process bioburden monitoring__ throughout the manufacturing workflow, offering laboratories a versatile and compliant solution across multiple quality control applications. The [Milliflex® Rapid System 2.0](https://www.sigmaaldrich.com/HK/en/campaigns/rapid-microbial-testing-with-milliflex-rapid-2) is fully aligned with __USP <73>__ requirements and enables your laboratory to implement a validated, compliant RMM-based sterility testing workflow — delivering results in __5 days or less__.  ## [](https://www.sigmaaldrich.com)Data integrity and governance in sterility testing A sterility test result is only as reliable as the data behind it — and regulatory agencies worldwide are placing data integrity at the heart of GMP microbiology compliance. Beyond methodology, equal importance is placed on how sterility test data is generated, recorded, and maintained throughout the entire test lifecycle. Regulatory bodies including the FDA, EMA, and PMDA require that all laboratory data meets ALCOA++ principles: data must be Attributable, Legible, Contemporaneous, Original, and Accurate, with additional expectations around completeness, consistency, and availability. In the US, 21 CFR Part 11 defines the requirements for electronic records and electronic signatures, making digital documentation legally equivalent to paper records. In Europe, EU Annex 11 establishes equivalent expectations for computerized systems in GMP environments. Together, these regulations set a clear framework for Data Integrity for QC lab data, including Sterility Testing: real-time data capture, automatic audit trail generation, and complete end-to-end digital traceability — from operator identification and instrument data, to sample information and final results — are increasingly expected by regulators during GMP inspections and considered industry best practice.  The [M-Trace® software & Mobile App](https://www.sigmaaldrich.com/HK/en/services/software-and-digital-platforms/m-trace-electronic-test-record-software) helps your laboratory meet these regulatory expectations.  [Speak to a specialist](https://www.sigmaaldrich.com/HK/en/campaigns/sterility-testing-consumables) ## [](https://www.sigmaaldrich.com)Sterility Testing Applications: From Pharmaceuticals to Cell & Gene Therapies Sterility testing is a routine quality control requirement across a broad range of GMP-regulated industries. While the fundamental principles remain consistent, the choice of method, regulatory framework, and speed-to-result requirements vary significantly by segment: - Pharmaceutical products — including injectables, ophthalmic preparations, and infusion fluids — represent the largest volume of sterility testing, governed by USP <71>, Ph. Eur. 2.6.1, and JP <4.06> - Biologics, biosimilars & vaccines — sterile parenteral products requiring rigorous batch release testing under GMP - Medical devices — sterility testing adapted to device format, using direct transfer or product flush methods - Cell & Gene Therapies (CGT) and ATMPs — short or ultra-short shelf-life products where rapid microbiological methods are often the only viable option for same-day or next-day release decisions - Radiopharmaceuticals — products with half-lives measured in hours, for which traditional 14-day incubation is not feasible and RMMs are increasingly mandated - Tissue & regenerative medicine — sterility confirmation prior to transplantation or administration" ## [](https://www.sigmaaldrich.com)Sterility testing workflows ### __Traditional compendial workflow — membrane filtration__ The following step-by-step workflow describes the membrane filtration sterility test as performed under __USP <71>__, __Ph. Eur. 2.6.1__, and __JP <4.06>.__  __Test preparation__ — Assemble the [membrane filtration device](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/steritest-neo-sterility-test) and connect tubing to the [sterility testing pump](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-hardware-accessories) inside the controlled environment (laminar flow hood or isolator)  1. __Rinsing of the membrane/filter prewetting__ — Prewet the membrane to optimize filtration efficiency and minimize product adsorption to the filter 2. __Sample filtration__ — Filter equal volumes of product into both canisters; microorganisms are retained on the 0.45 µm membrane 3. __Rinsing__ — Remove inhibiting compounds using a validated rinsing solution to prevent false negative results 4. __Culture media filling__ — Fill membrane filtration devices with [SCDM (TSB) and FTM sterile culture media](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-membrane-culture-media-rinsing-fluids) 5. __Incubation__ — 14 days at 32.5°C (FTM) and 22.5°C (SCDM). These temperatures represent the midpoints of the pharmacopoeial ranges of 30–35°C and 20–25°C respectively, as specified under USP <71>, Ph. Eur. 2.6.1, and JP <4.06> 6. __Result examination__ — Visual turbidity check; any turbidity must be investigated as potential microbial contamination 7. __Digital test record__ — Contemporaneous, automatic capture of all test steps, operator actions, and instrument data into a single, auditable electronic record supports compliance with [21 CFR Part 11, ALCOA++](https://www.sigmaaldrich.com/HK/en/services/software-and-digital-platforms/m-trace-electronic-test-record-software) data integrity principles, and regulatory inspection readiness Step descriptions are aligned with compendial requirements of USP <71>, Ph. Eur. 2.6.1, and JP <4.06>. Step 8 reflects current regulatory expectations for data integrity under 21 CFR Part 11 and EU Annex 11, beyond pharmacopoeial requirements. ### Rapid sterility testing workflow — ATP bioluminescence For __short shelf-life products__ — including __cell and gene therapies__, radiopharmaceuticals, and short-life biologics — the traditional 14-day incubation period is often incompatible with product release timelines. [Rapid Microbiological Methods (RMMs)](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/early-microbial-detection-enumeration-milliflex) offer a scientifically validated, regulatory-accepted alternative. __ATP bioluminescence__ is one of the most established RMM technologies for sterility testing. It detects light emitted by viable microorganisms following reaction with a bioluminescence reagent, enabling rapid, sensitive, and quantitative contamination detection. The new __USP <73>__ chapter—"ATP Bioluminescence-Based Microbiological Methods for the Detection of Contamination in Short-Life Products"—provides a clear regulatory framework for RMM adoption, effective __August 2025__. A typical ATP bioluminescence rapid sterility testing workflow proceeds as follows: 1. __Sample filtration__ — The sample is filtered through a dedicated membrane, retaining viable microorganisms 2. __ATP reagent application__ — Bioluminescence reagents are applied to the membrane, targeting viable cells 3. __Automated detection and Quantification__ — A highly sensitive detection camera captures and quantifies light signals from viable microorganisms 4. __Results in 5 days or less__ — Delivering results up to 75% faster than the traditional 14-day compendial method. Actual time-to-result depends on the validated method and product type. 5. __Electronic test record__ — All results are captured in a __21 CFR Part 11__-compliant, fully auditable digital record This workflow is based on ATP bioluminescence technology as described in USP <73>. Step 5 reflects current regulatory expectations for data integrity and electronic recordkeeping under 21 CFR Part 11 and EU Annex 11. RMMs using ATP bioluminescence are suitable for both __sterility testing__ and __in-process bioburden monitoring__, providing laboratories with a versatile, fast, and compliant solution across the manufacturing workflow. ## Related Articles - [Steritest® Symbio Sterility Testing Pump Guide](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-hardware-accessories) The user-friendly Steritest® Symbio pumps and accessories are designed for easy and reliable sterility testing. The system improves your workflow and maximizes safety, confidence, and convenience. - [Steritest® NEO Membrane Filtration Sterility Test](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/steritest-neo-sterility-test) Sterility testing is one of the most crucial steps in pharmaceutical product release. Regulatory-compliant membrane filtration sterility testing devices ensure the safety of pharmaceutical products. - [Sterility Testing Culture Media & Rinsing Fluids](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-membrane-culture-media-rinsing-fluids) Our sterility testing media and fluids are formulated and tested to meet EU, US, and JP pharmacopeia regulations, and are supplied with full certification containing pH, sterility, and growth promotion information. - [Sterility Testing by Direct Inoculation](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-direct-inoculation) Direct inoculation of culture media is the method of choice for sterility testing of non-filterable samples. Select from our range of sterility testing media including soybean casein digest medium (TSB), and Fluid Thioglycollate medium (FTM standard and clear). - [Sterility Testing of Products in Isopropyl Myristate](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-increased-chemical-compatibility) Irradiated Isopropyl Myristate (IPM) enhances filterability for sterility testing of viscous products using the Steritest® NEO device. - [See All (8)](https://www.sigmaaldrich.com/HK/en/search/facet-search?focus=sitecontent&term=facet-search) ## Related Protocols - [Testing Cells for Bacterial and Fungal Contamination](https://www.sigmaaldrich.com/HK/en/technical-documents/protocol/cell-culture-and-cell-culture-analysis/mammalian-cell-culture/testing-for-bacteria) Cell culture protocol for testing cell lines for bacterial and fungal contamination. ### Find More Articles and Protocols Enter Keywords Search ## How Can We Help In case of any questions, please submit a [customer support request](https://www.sigmaaldrich.com/HK/en/support/customer-support) or talk to our customer service team: Email [custserv@sial.com](mailto:custserv@sial.com) or call +1 (800) 244-1173 ## Additional Support - [Chromatogram Search](https://www.sigmaaldrich.com/chromatogram-search) Use the Chromatogram Search to identify unknown compounds in your sample. - [Calculators & Apps](https://www.sigmaaldrich.com/HK/en/support/calculators-and-apps) Web Toolbox - science research tools and resources for analytical chemistry, life science, chemical synthesis and materials science. - [Customer Support Request](https://www.sigmaaldrich.com/HK/en/support/customer-support) Customer support including help with orders, products, accounts, and website technical issues. - [FAQ](https://maestro.my.site.com/knowledgeportal/s/) Explore our Frequently Asked Questions for answers to commonly asked questions about our products and services. * * * [ \ No missing data. No scissors. No paper. No waiting 14 days. \ Our innovations help you improve data traceability and safety, accelerate results, and digitize workflows. Discuss with our expert to explore our comprehensive portfolio designed to boost your lab's efficiency and compliance. \ I want to upgrade my sterility test](https://www.sigmaaldrich.com/GB/en/campaigns/sterility-testing-consumables) * * * __Featured Articles__ [Steritest® Symbio Sterility Testing Pump Guide](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-hardware-accessories) The user-friendly Steritest® Symbio pumps and accessories are designed for easy and reliable sterility testing. The system improves your workflow and maximizes safety, confidence, and convenience. [Steritest® NEO Membrane Filtration Sterility Test](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/steritest-neo-sterility-test) Sterility testing is one of the most crucial steps in pharmaceutical product release. Regulatory-compliant membrane filtration sterility testing devices ensure the safety of pharmaceutical products. [Sterility Testing Culture Media & Rinsing Fluids](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-membrane-culture-media-rinsing-fluids) Our sterility testing media and fluids are formulated and tested to meet EU, US, and JP pharmacopeia regulations, and are supplied with full certification containing pH, sterility, and growth promotion information. [Sterility Testing by Direct Inoculation](https://www.sigmaaldrich.com/HK/en/technical-documents/technical-article/microbiological-testing/sterility-testing/sterility-testing-direct-inoculation) Direct inoculation of culture media is the method of choice for sterility testing of non-filterable samples. Select from our range of sterility testing media including soybean casein digest medium (TSB), and Fluid Thioglycollate medium (FTM standard and clear). Top __Sign In To Continue__ To continue reading please sign in or create an account. Sign In__Don't Have An Account?__Register