Sterility testing is a critical step in pharmaceutical manufacturing. It ensures that products meet the highest quality standards before reaching patients. As regulatory authorities encourage the adoption of Rapid Microbial Methods (RMMs), updates to USP <73> and <1071> are shaping the future of microbial detection. In this Q&A, we speak with Miriam Guest, Senior Principal Scientific Advisor at Charles River, about these changes and what they mean for sterility testing. Prior to joining Charles River, Miriam Guest spent 20+ years implementing global technologies and initiatives at AstraZeneca.

What are USP <73> and <1071>, and why are they important?

Miriam: USP <73> and <1071> are key chapters in the United States Pharmacopeia (USP) related to sterility testing. USP <73> (ATP Bioluminescence-Based Microbiological Methods for the Detection of Contamination in Short-Life Products) provides guidelines for alternative sterility methods that offer faster and reliable results compared to traditional growth-based methods. USP <1071> (Rapid Microbial Tests for Release of Sterile Short-Life Products: A Risk-based Approach) outlines the principles of risk assessment, sampling plans, and validation strategies for adopting RMMs across pharmaceutical applications.

These updates are important because they signal a growing regulatory acceptance of modern technologies, enabling manufacturers to transition from 14-day sterility testing to faster, more efficient methods for short-life products.

Regulatory Changes: USP 73 & 1071 Updates at a Glance
The latest chapters of USP <73> and <1071> introduce key updates impacting sterility testing and rapid microbiological methods (RMMs). Understanding these changes is crucial for compliance and optimizing your microbial testing strategy. Our expert guide breaks down what’s new and how Celsis^® can help you stay compliant and efficient.
Explore the Guide

Why is traditional sterility testing a challenge for pharmaceutical manufacturers?

Miriam: Advances in pharmaceutical development has seen many new modalities in recent years.  As our products evolve, our sterility assurance strategy must also evolve.  The USP <71> sterility test requires a 14-day incubation period; this is beyond the shelf-life of many products.  To that end, the adoption of robust rapid methods is essential.

How do the updates to USP <73> support the adoption of rapid sterility testing?

Miriam: USP <73> provides a framework for demonstrating method suitability of ATP-bioluminescence, ensuring they meet the same stringent reliability and accuracy standards as traditional tests. With technologies like ATP-bioluminescence, microbial contamination can be detected in just 4-6 days, significantly reducing the time required for sterility testing.

Regulatory bodies, including the FDA and EMA, are encouraging pharmaceutical companies to modernize their microbiological testing strategies, aligning with initiatives like Pharmaceutical Quality Systems (ICH Q10) and continuous process improvements. Ensuring a holistic contamination control strategy is key to sterility assurance, and rapid microbiological test methods are a key component of the contamination control strategy for products with short shelf-life.

Webinar: Accelerate Sterility Testing with USP <73>
In this webinar, Labor LS and Charles River industry experts discuss how ATP-bioluminescence is shaping the future of short-life products. Explore real-world insights and validation strategies.
Watch the Replay

What is the role of USP <1071> in implementing Rapid Microbial Methods?

Miriam: USP <1071> serves as a guidance document for pharmaceutical manufacturers looking to implement Rapid Microbial Methods, or RMT as referred to in the chapter title. It provides direction on key aspects such as sample size considerations, technology selection-including ATP-bioluminescence, flow cytometry, and PCR-validation requirements like specificity, detection limits, and robustness, as well as regulatory submission strategies to gain approval for alternative methods. This chapter helps companies navigate the method validation and regulatory approval process, addressing concerns about compliance with existing sterility testing standards.

How does ATP-bioluminescence technology align with USP <73> and <1071>?

Miriam: ATP-bioluminescence is a well-established growth-based detection method that can identify viable microorganisms within a few days, rather than waiting for visible microbial growth. ATP-bioluminescence can detect the presence of viable contamination at lower levels than visual inspection, this results in a shorter time to detect. This aligns with USP <73>’s push for sterility tests that offer faster and equally reliable results for short shelf life products.

When following USP <73>, ATP-bioluminescence provides a robust, compliant alternative to membrane filtration and direct inoculation, making it ideal for sterility testing in injectable drugs, biologics, and CGT products.

What are the key benefits of switching to a rapid sterility testing method like Celsis^® ATP-bioluminescence?

Miriam: When it comes to transitioning to ATP bioluminescence-based methods like Celsis^® for rapid sterility testing, there are several advantages. One of the most significant is the reduction in sterility testing time from 14 days to just 4-6 days, which accelerates product availability. It also enhances data integrity by generating automated digital reports that minimize human error and comply with 21 CFR Part 11. From a regulatory perspective, it aligns with USP <73> and <1071>, supporting a science-driven approach to RMM validation. Additionally, earlier microbial detection strengthens contamination control by enabling faster investigations and corrective actions, ultimately enhancing patient safety.

What steps should pharmaceutical companies take to implement rapid sterility testing?

Miriam: Adopting a rapid method requires regulatory strategy, validation planning, and cross-functional alignment. Companies should engage with regulators early to discuss validation plans, follow USP <73> and <1071> guidance to ensure method robustness, and digitally integrate RMM results into quality systems for streamlined data tracking.

Many manufacturers partner with regulatory experts and technology providers like Celsis^® to facilitate a smooth transition to RMM while ensuring compliance.

What’s next for sterility testing and RMM adoption?

Miriam: With the increasing regulatory push for modernized microbiology testing, we expect greater global acceptance of RMM by pharmacopoeias beyond USP, such as Ph. Eur. and JP, expanded use of ATP-bioluminescence in sterility and microbial limit testing, and more flexible regulatory pathways for validating and submitting RMM-based sterility methods.

The pharmaceutical industry is moving toward faster, more automated, and highly reliable microbiological quality control and rapid sterility testing is a key part of that transformation.

Final Thoughts?

Miriam: The updates to USP <73> and <1071> reflect a shift towards modern, science-driven sterility testing. By adopting rapid microbial detection technologies like ATP-bioluminescence, pharmaceutical manufacturers can achieve faster release times, better contamination control, and stronger regulatory compliance.

The post The Future of Sterility Testing for Short Shelf Life Products appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

Is a highly skilled, dedicated analyst needed to conduct a routine bacterial endotoxins test (BET)? By the year 2000, major parenteral drug firms were conducting the BET—which is designed to detect all environmental Gram-negative bacterial endotoxins—with the aid of an incubating microplate reader and endotoxin-specific software. The advent of the chromogenic substrate in 1982 produced a more robust color signal than did turbidity in kinetic LAL methods.

Despite these advances, many QC supervisors were frustrated with the high percentage of invalid BET results. Moreover, some companies operated with limited microbiological staff, forcing them to send their BET samples to contract labs to reduce invalidity.

So, Charles River Laboratories Microbial Solutions, a leader in LAL testing, began exploring more modern options to the early gel-clot method that, while widely accepted, posed more of a risk for human error and compromised data integrity.

The Age of LAL Cartridges Begins

Under the tutelage of Foster Jordan, who joined Charles River in 1994 following the acquisition of Endosafe^®, Charles River launched a revolutionary hand-held, rapid endotoxin detection test. Jordan sought to eliminate the technical flaws and weaknesses of early photometric BET methods by creating a new automated microfluidic test platform that utilized a single-use cartridge with dried, embedded reagents. But creating a completely automated and more robust LAL test would require a total redesign of existing LAL reagents and photometric readers. 

Recognizing there was simpler, faster, more automated solution, Foster began collaboration with a company that specialized in microfluidic test systems. Charles River acquired this technology in 2001 and thus began bringing forth the industry-changing LAL cartridge technology platform. Among the initial features like dried, pre-calibrated and pre-loaded stabilized reagents, Foster created an archived standard curve, established from known reference standard endotoxin values, eliminating the lengthy time and experience required by the technician of preparing a daily standard curve. Patents were filed and granted. Extensive conversations and negotiations with FDA began in 2003, primarily concerning the, at the time, novel features like the archived standard curve, its linearity, system suitability, negative and positive controls, manufacturing requirements, and software design.

The archived standard curve, a key part of the invention, represented a completely new approach for the field of microbial identification and more specifically endotoxin testing, and required multiple data sets to validate its accuracy to the regulators. Charles River needed to re-educate regulators on why this was important in reducing the variability of the control standards.

Why was the LAL Cartridge Such a Major Advance?

The uniqueness of the LAL cartridge could not be overstated. Among the advantages:

• Reduces the amount of LAL reagent up to 95% compared to the gel-clot assay
• A quantitative kinetic chromogenic assay accepted by FDA and complies with harmonized BET chapters (USP/Ph. Eur.)
• Real-time results in approximately 15 minutes
• Simple to use, cutting down on the time it took laboratories to train and qualify new analysts
• Eliminates complexity and subjectivity with the built-in archived standard curve, the main source of error in the gel-clot test. 
• Reduces manual pipetting, diluting, reconstituting steps, and consumables which requires less glassware, plastic accessories, and eliminates reagent waste.

A cartridge and associated Endosafe instrumentation that uniquely allow for unprecedented collection and electronic documentation of results; the cartridge and automated / robotic endotoxin technologies allowed laboratories to better utilize their highly trained, highly-educated scientists and analysts. In short, the lower variability achieved with the LAL cartridge meant that laboratory retest rates dropped dramatically along with the time needed to retest murky results. And, since the cartridge technology did not require water for BET to prepare the liquid reagents, the principal reason for the negative controls was eliminated.

Recognizing that the new approach to BET was a significant advance toward production of safer parenteral products, the FDA approved the cartridge in 2006 (see image below). The new system was named PTS, for Portable Test System, to emphasize the suitability of its ability to perform testing at the point of use or point of sampling, freeing it from strict performance in a firm’s QC lab.

Among the early users of the Endosafe^®-PTS™ product was in fact, NASA. They used the cartridge technology to monitor the environment for microbial contamination during the construction of the Mars exploration rovers, “Spirit” and “Opportunity” and deployed the Endosafe^®-PTS™ aboard the International Space Station (see image below) to assist in studies for an extended human presence in space. 

The Ever-Evolving Cartridge

Arguably, Charles River’s invention of the PTS was the most important innovation in BET testing since the introduction of the chromogenic substrate. But the innovation didn’t end there. Over the years, Charles River introduced new variations on the LAL cartridge including:  

• Endosafe^® nexgen-PTS™, which included an embedded operating system for true portability. It was designed for wireless capability for remote system access, and data export and printing. 
• Endosafe^® nexgen-MCS™, which provides a stackable, multi-cartridge system that allow you to test up to five samples simultaneously in approximately 15 minutes. The samples are run independently, allowing for random access, and eliminating the need for batch sampling.
• Endosafe^® Nexus 200™, a fully automated robotic system designed specifically for testing in the central quality control lab. It ties together LAL cartridge technology, full automation of liquid handling, and simple data management.

Recombinant Cascade Reagent (rCR) Cartridges

With last year’s launch of an animal-free, recombinant cascade reagent, Endosafe^® Trillium™, Charles River has introduced the next iterations of the flagship cartridges in 2024. The rapid and robust Trillium cartridges fully align with the existing suite of scalable, automated, Endosafe cartridge technology platforms, utilizing the same kinetic chromogenic testing method and sample preparation. Labs are still able to receive quantitative results in approximately 15 minutes, expediating their endotoxin testing program to keep pace with their manufacturing and production timelines. The beauty of this flexible technology allows for customers to transition from LAL to rCR cartridges seamlessly, as the LAL cartridges that users have trusted for decades will continue to be available.

With 20 years of cartridge technology behind us, what will the next two decades bring? Stay tuned!

The post 20 Years of Trailblazing Science with the LAL Cartridge appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

As industrialization continues to advance, an increase in pollution and waste threatens our water, air, and soil. We have a corporate responsibility as well as a personal responsibility to reduce our impact on the environment and strive for more sustainability. In 2019, Charles River announced goals to reduce Scope 1 and 2 greenhouse gas emissions by 50%, Scope 3 gas emissions by 15%, and achieve 100% renewable electricity globally by 2030. In the past three years, we’ve reduced gas emissions by 25% and achieved 100% renewable energy in the US.

But the progress doesn’t end there!

To further meet this initiative, our laboratories have invested in a robust recycling program, reduction of hazardous chemicals and waste, and automated systems that require fewer resources.

For instance, in 2022 Charles River’s Accugenix labs identified over 600,000 customer samples. These samples are shipped to our laboratories following national and international shipping regulations, which have strict standards for packaging. All cardboard shipping material is recycled unless it is obviously contaminated with biological matter. We also recycle our pipette tip boxes, consumable packaging, and office papers.

Fewer plastics, less travel

The COVID-19 pandemic highlighted potential weaknesses in supply chains for just about every company. Accugenix made considerable efforts to move away from single use plastics and find reusable alternatives where appropriate. We are also part of the Pharmaceutical Supply Chain Initiative. Charles River is committed to conducting business ethically, responsibly, and with integrity, and we expect the same from our suppliers. Each are required to comply with applicable laws and regulations, as well as the standards set forth in our Supplier Code of Conduct, which mirrors the Pharmaceutical Supply Chain Initiative’s (PSCI) Principles for Responsible Supply Chain Management.

Communication technology is also helping us to reduce our carbon footprint. In this global operation, there is an increased need to communicate in real time. Thanks to the pandemic, most people are comfortable with modern communication technology, and we can teleconference anywhere. This reduces the need to travel and the associated environmental impact.

Our microbial identification assays are also designed to minimize the use of hazardous chemicals. We’ve optimized our proprietary DNA extraction and purification techniques to use materials that are safer than what is commonly used in commercial DNA sequencing kits. We’ve also been able to reduce reaction sizes, which results in less chemicals being used overall. Because chemical waste can be dangerous, Accugenix has taken these reduction steps to keep both our environment and technicians safe.

To further increase our sustainability efforts, we have implemented several automated systems. Our laboratory processes are controlled through a custom laboratory information management system (LIMS), which allows us to document sample processing electronically. The LIMS not only prevents errors from occurring and bolsters our data integrity, but also prevents the need for hundreds of thousands of pages of batch records per year! This reduces our paper consumption and the carbon footprint of off-site transportation and archival. We also have liquid handling machines and automated visual aids that ensure efficient processing the first time, preventing the need for retests and additional use of chemicals and consumables.

Charles River’s microbial identification laboratories strive to make responsible and environmentally conscious decisions. We prioritize sustainability and conservation. These changes go a long way toward making a positive impact on the environment and preserving our planet.

The post Sustainability and Conservation in Microbial ID Labs appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

Dangerous microorganisms are everywhere. Humans do their best to get rid of them, but this labor is, unfortunately, not always successful. Even though we do understand the importance of microorganisms in some contexts, we are extremely aware of the consequences their presence has in a manufacturing environment. Many regulations, including the latest publication of the EU GMP Annex 1, mention the importance of an effective Contamination Control Strategy (CCS) to control ingress, transport, and proliferation of these bugs.

Despite all the efforts, studies have demonstrated that there are some microorganisms that stubbornly resist radiation and disinfectants. To learn more about useful strategies to combat these bugs, we turned to one of Charles River’s experts, Sujan Timilsina, PhD. Sujan is a Bioinformatics Scientist II with the Microbial Solutions business and has over 10 years of experience in microbial detection and identification, and high throughput sequencing. Sujan is also knowledgeable about the development and application of bioinformatic pipelines that can be used to phylogenetically analyze microbial evolution and movement across a wide range of microorganisms.

What do you think are the best technologies or platforms to use to identify these resistant microorganisms? 

Resistance in microorganism is an associated phenotype and usually strain specific. Some are resistant to antibiotics, UV and high temperatures, though not all species strains carry that trait. The distribution of these organisms varies as each resistance bug may have different niches where they flourish and where selection pressure against other microorganisms is high. Nonetheless, for identification purposes, we use different phenotypic, proteotypic, and genotypic methods that help identify the organism at various levels. Identifying specific resistance will need in-depth analysis, but for general taxonomic identification purposes, we can use proteotypic and genotypic methods. The microbial identification methods need a robust database that strains can be compared against. For example, we can use sequencing platform to sequence genes used for identification, and if that organism of interest is not described in the database we are comparing against, we may not be getting the correct result. Among the platforms we use, these resistant organisms can be identified by comparing against a well curated microbial database, such as the one maintained by Accugenix^®.

What would be the right path to identify them? Are there any best practices you would like to share? 

In scenarios where we are interested in identifying the organisms and determining its source, a multi-prong approach will be valuable. As a first step, it is critical to identify the organism. Using 16S for prokaryotic bacteria and ITS gene sequences for eukaryotic fungal organisms and comparing them against a larger database, we can get a good idea about the organism of interest. As the resistance among organisms could be strain specific, it will be ideal to use strain typing methods that will help to piece together the puzzle on where the organism could have been potentially introduced. Initial organism identification is critical, as the subsequent process of the strain typing could be implemented accordingly by selectively using 4-10 housekeeping genes (genes associated with microbial metabolism) to determine the strain diversity. With the application of whole genome sequencing and utilizing the larger genomic region for comparison, we can further advance our capacity to effectively trace and track resistant microorganisms.

What makes some microorganisms so hard to identify and, ultimately, kill? 

Tricky question. There are a multitude of reasons why these organisms are hard to identify. Sometimes we don’t have any existing knowledge of the organism. Some organisms do not grow in a growth media, which limits our understanding of these organisms. We have greatly improved our ability to learn about these microbes after being able to sequence such organisms without the need of growing them in a growth media. New organisms are described very frequently, and this frequency has increased significantly, for prokaryotes since 2012 due to whole genome sequencing tools. 

For the ones, that we recognize and have studied the challenges are different. Some fungi may have unique reproductive and dormant structures that limit rapid growth or extraction of genetic material to help in species identification. For example, if we consider an Aspergillus that produces ascus spores, it can remain dormant for a considerable length of time, making the identification process challenging. 

Correct identification is important to evaluate methods for eliminating the organisms from their unwanted areas. Microorganisms are quick to adapt, and the population could be selected in a different direction depending on their exposure and selection pressure. Continuous exposure to a certain antibiotic or fumigation could lead to selection of strains that are relatively resistant to the methods we use to eradicate them. So, it is important to understand what we are dealing with in addition to applying general control procedures. We may have to implement organism or niche specific approaches to control them.

Based on your experience, are there specific regions where we “encounter” these organisms the most? 

Initial assumptions are that resistant organisms appear where there is high selection. Nonetheless, with more study and identification of organisms, we have found significant diversity in microbe populations and their strategies to adapt to unfavorable environment even when they are not exposed to selective conditions. Some organisms are ubiquitous in distribution and additional identifications are being made with the availability of central databases to compare and describe them phylogenetically and taxonomically. Deinococcus is one example. Some of the species of this genus are resistant to high radiation and have unique DNA repair mechanisms that help them to thrive in these conditions. It is likely that certain population within the species group might be favored in such harsh conditions. 

However, the species can be encountered frequently in areas outside the radiation zone. Same goes for multiple antibiotic resistant and heat tolerant organisms. Several of these species have been identified during the microbial identification process through Accugenix^®. This shows that these organisms are out there; the strains may not be resistant to these control mechanisms, but the species could be present in our surroundings.

Are there any other recommendations you would like to share?

In cases of microbial contamination preventative measures are always better. However, if contamination did occur, initial identification is the key to successful mitigation of microbial issues. Identifying the microorganisms and tracking the strains in production facilities becomes critical. My recommendation would be to use a validated library for identification and keep track of your organisms. Sudden changes in microbial populations could indicate contamination issues. Accugenix^® provides an in-depth evaluation of organisms reported from the facility through Tracking and Trending features via AccuPedia^®, which  could be a good starting resource for understanding these microbes.

For more details and interesting facts on these radio- and disinfectant- resistant microorganisms, watch our exclusive on-demand video series (only available for a limited time) presented by Ziva Abraham, President and Founder of Microrite, Inc. and Duncan Barlow, Technology and Market Development Manager for Charles River Laboratories’ Microbial Solutions division.

Source: www.criver.com

The post Hard to Kill Bugs appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

The home and personal care market is a fast-growing segment. Manufacturers face many challenges to release their products to the market safely while keeping up with increased consumer demands. To provide some insights and topics in this exciting industry, we are chatting with Anne Wagner, PhD, Technology & Market Development Manager for home and personal care for Charles River. She has 18 years of diverse scientific research experience with eight years specifically focused on home and personal care product formulation. Anne has a passion for science advocacy and communicating scientific concepts to a variety of audiences along with teaching and mentoring. She values research that impacts people’s daily lives and is committed to making high quality and ethical scientific contributions.

Chiara: From your perspective, are there any opportunities where companies can improve their lab efficiencies that will help streamline their operations process?

Anne: Great question! In my opinion, we should consider their context and if they are using traditional microbial testing, then they are missing out on what is probably the single greatest opportunity to take days out of their manufacturing process. Using the traditional plating methods, it takes days, even up to 7 days, to get microbial results while inventory is on “QA hold.” A rapid microbial method or RMM for short gives them the same results in a fraction of the time. Moving to a rapid method can have significant impact in both their lab and operations. RMMs give them automated results, increased data integrity, and in case of a contamination event, faster corrective action. When manufacturers release products faster, held inventory and working capital requirements are reduced – saving them time and money…which are really appreciated by stakeholders and financial teams. This allows for improvement and optimization of warehouses. Why waste time and money when there is a better way to get your products to market faster? The key is to think Smarter. Faster. Better.

Chiara: What does the word “Rapid” mean for Home & Personal Care Manufacturers?

Anne: The “rapid” in rapid microbial method refers to faster time to result. For manufacturers it means faster and higher product throughput. Some rapid methods can process a large number of samples with minimal effort, reducing labor, physical lab space and other resources needed for testing. This allows manufacturers’ labs to run at maximum efficiency. Faster results enable companies to detect contamination earlier by testing upstream in their process, like raw materials and in-process samples. However, not all rapid methods are created equal. Some may require extra preparation steps, have system complexity, or need multiple systems to get the results they need. Understanding the real value of a rapid method starts with knowing your options, and the different types of technologies out there to find the best fit for your company.

Chiara: One of the most common trends lately is transparency. Would you mind telling us a bit more about this?

Anne: Consumers are demanding that future beauty products have a clear stance on environmental protections and sustainability. Consumers want transparency on how products are manufactured, purchased, and tested. Already, there are products with traceable ID numbers that can be entered on websites to see manufacture date, full ingredient details, a supplier list, and test results for their purity, potency, and quality. It will be interesting to see if the ingredient transparency trend will spread globally across all consumer products. Upcoming regulation in the US — from the passing of the California Cruelty-Free Cosmetics Act to more recent proposals for sunscreen reform — suggests that greater ingredient transparency is on the horizon, so manufacturers should take this into account and be ready for this upcoming change.

Chiara: Are there other regulations that manufacturers should be aware of?

Anne: The quality of your product is essential to the reputation of your brand, and therefore, the trust that consumers have in your company. Manufacturers are held accountable for these standards of quality. Some personal care products are classified as over-the-counter drugs (called OTCs), such as sunscreen and fluoride toothpastes. Additional regulatory oversight for OTC drugs requires companies to adopt Good Manufacturing Practices (known as GMP) that enforce quality control standards similar to the pharma industry. GMP provides guidelines for quality manufacturing of products through standardized procedures and regular oversight of facilities, records, staff, and overall operations. Though GMP is not currently required by law for cosmetics, that is changing, too. The Modernization of Cosmetics Regulations Act of 2022 will require cosmetics manufacturers that sell products in the US to follow GMP. The FDA is planning to publish new rulemaking for cosmetic GMP by 2025 with implementation by 2026. To learn more about GMP standards in the US go to FDA.gov.

Chiara: When we talk about good practices, we hear a lot about Environmental Monitoring. What does that mean?

Anne: Environmental monitoring programs or EM is essential to the manufacturing quality control process. The true value of EM comes down to understanding what microorganisms are in your environment in the first place. If you don’t have a clear picture of your environment, you don’t know your risks. EM is a lot like the impressionism technique pointillism, a process using lots of little dots to create a picture. When you get close to the picture all you see is dots, but as you step away and see the picture as a whole, you can truly appreciate all that is there. You can see the lady in the sun hat by the river. When you are too close you can’t see everything. This is a great analogy for EM, if you are only testing your final product, you are only getting a snapshot, only see the dots, but when you monitor your air system, water system, and equipment; you can see what is really going on…the full picture. EM programs are early warning systems that will save you time and money with faster and more effective microbial investigations, changing your manufacturing processes from reactive to proactive.

Chiara: Another topic to explore with you is preservatives. Why do cosmetics need preservatives?

Anne: Preservatives play a crucial role in keeping our cosmetics safe. Many cosmetics are water-based; however, water is also a good medium for microbial growth, some of which can be harmful. The most common strategy to keep your products free from harmful microorganisms is using preservatives and anti-microbial agents that can be synthetic, natural or even multi-functional ingredients. Preservative regulation can vary by geographical region. The FDA doesn’t have specific rules only applying to cosmetic preservatives, but the EU’s EC regulation 1223/2009 Annex 5 sets limits on preservatives allowed in formulations. It’s one of the most comprehensive regulatory cosmetic guidelines, setting the standard for many regulatory agencies around the world.

Chiara: You mention that water is a good medium for microbial growth, would you mind telling us more?

Anne: Sure! With water being an essential ingredient in most home and personal care products, it can have an impact on the product quality. If manufacturers are not testing their water systems frequently, they could miss harmful organisms like Burkholderia cepacia. Since Burkholderia cepacia is a waterborne microorganism, this contaminant can be a reoccurring problem in a facility. Monitoring the environment allows quality control professionals to detect and analyze any microbial deviations. Although routine water system monitoring is not required in cosmetics, water remains one of the top contamination sources found in manufacturing. It’s important to choose a robust system of identification to stay informed of the microbial presence in your environment.

For more information on Home & Personal Care Trends :  www.criver.com/cosmetics and download our brochure.

The post What’s Hot in 2023: Home & Personal Care Manufacturing appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

The product insert, which accompanies the LAL product, is regulated under the FDA BLA and provides details on instructions for handling and usage, storage conditions, safety precautions, limitations, warnings, and other relevant information for use. Off-label usage, including utilizing any other volume, would be outside of current FDA licensing. It’s critical that users thoroughly read and understand the intended use and instructions to remain compliant.

The harmonized pharmacopoeia for the Bacterial Endotoxin Test including the United States Pharmacopoeia (USP), the European Pharmacopoeia (EP), and the Japanese Pharmacopoeia (JP) do not specify LAL reconstitution volumes or ratios for testing, but each state that the LAL manufacturers product insert must be followed for intended use as follows:

USP <85>1 Photometric Quantitative Techniques:

• Listed under assurance of criteria for the standard curve – ‘according to the manufacturer’s instructions for the lysate (volume ratios, incubation time, temperature, pH, etc.)’.
•  Listed under test for interfering factors – ‘according to the instructions for the lysate employed, for example, concerning volume of sample solution and lysate TS, volume ratio of sample solution to lysate TS, incubation time, etc’.

EP 2.6.142 Photometric Quantitative Techniques (Methods C, D, E, and F):

• Listed under assurance of criteria for the standard curve – ‘as recommended by the lysate manufacturer (volume ratios, incubation time, temperature, pH, etc.)’
• Listed under test for interfering factors – ‘as recommended by the lysate manufacturer (volume of test solution and lysate solution, volume ratio of test solution to lysate solution, incubation time, etc.)’. 

The end user must follow the manufacturers product insert to ensure compliance with both the FDA and the Harmonized Pharmacopoeia. For Charles River, our LAL product inserts state to use the lysate at 100 µl lysate to 100 µl sample but other LAL manufacturers may vary. If a user wishes to deviate from what is listed in the product insert, they must validate the deviation fully by following an alternate method validation on the lysate to ensure comparability and submitting all relevant data to the competent regulatory authority before using the altered method in routine use.

There has been much discussion about using LAL at different lysate/volume ratios. Charles River LAL is only validated under our BLA at the 100 µl lysate:100 µl of sample ratio and, as discussed above, it is not advisable to use any other ratio without performing alternate method validation. If this extra validation is not performed, the user would be using the LAL off label from the BLA, and this could lead to considerable ramifications from a regulatory perspective. 

Charles River Endosafe® LAL Cartridges utilize a substantially smaller volume of LAL to sample and have been validated for use as per the expectations from the FDA and submitted all relevant data under our BLA. The cartridges are accepted by the FDA for routine use in bacterial endotoxin testing. Charles River cartridges offer rapid, easy to use testing, in addition to utilizing 95% less LAL than a standard microplate or gel-clot method, providing users a sustainable LAL test method. 

It’s imperative that the end-users of an FDA licensed product understand and comply with the manufacturer’s product insert and recommendations. If you have any questions, you can reach out to your local LAL experts to discuss further.

References:

1.  United States Pharmacopoeia 44 2021, chapter <85> Bacterial Endotoxin Test

2.  European Pharmacopoeia (Ph. Eur.) 11th Edition, chapter 2.6.14 Bacterial Endotoxins

Author: Nicola Swift Reid, Associate Director Product Management, Endotoxin Products at Charles River Laboratories

The post The Risk of Non-compliance with Off-label LAL Usage appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

Lessons from three pharma quality investigations. Let’s call them the tub, the tubes, and the sledgehammer

Ceilings are often ignored in cleanrooms. Best case, they are cleaned once a quarter. In fact, if it wasn’t for the location of critical HEPA filters, ceilings would garner little attention at all.

When walking through your cleanroom, try to look at the stars knowing the ceiling is in the way and that’s what you need to inspect.

Here are three quick stories to give you another reason to thoroughly walk through your cleanrooms.

The Tub . . .

In 2013, my employer, a large contract manufacturer of sterile parenterals, announced they were closing. Almost all manufacturing stopped immediately. We needed to make dozens of batches of one last critical product on the FDA drug shortage list to fulfill a customer contract.

Due to all the site issues that led to this decision, we implemented extra controls to ensure product was safe. One of those controls included testing rinse water from every Clean-Out-of-Place (COP) equipment cleaning cycle. The COP washer was responsible for cleaning equipment with direct product contact. Soon after this testing started, we recovered an increasing variety of gram-negative and gram-positive Rods.

We swabbed all over the COP washer clean room to find the contamination source. We sampled the usual dirty suspects—corners, crevices, underneath the washer, etc. The swabs were surprisingly clean.

However, we found the root cause thanks to the keen eye of my supervisor. She looked up during her walk-through. She noticed a slightly discolored ceiling tile above the COP washer. Upon further inspection, we found leaking exhaust piping in the mechanical space. The leaks dripped through the ceiling directly into the COP washer tub while the cycle was running!

Through all the corrective action fall-out from this event, I was responsible for removing the ceiling tile from the cleanroom. I did it too quickly. I wanted to avoid the “extra work” of swabbing and matching identifications to what we recovered from the rinse sample. It was obvious this was the root cause. Were ID’s necessary?

Absolutely.

For a site with a history of mold recovery, ID’s would have been critical to tracking and trending impact to the rest of our facility’s EM results.

. . . The Tubes . . .

One event that contributed to this site closing happened a few years earlier. An operator was waiting for parts to arrive so they could assemble the filling line. During the wait, the operator casually looked around the room. Through the haze of their frequently irradiated clean-room goggles, the operator noticed some spots on the HEPA filter grates. What were those spots? Cleaning agent residue? Mold? They discussed these observations with their supervisor.

In these rooms, product transferred from formulation vessels to filling lines through pressurized, flexible, tubes. The tubes were disconnected after filling cycles. They were often still pressurized, causing residual product to spray into the room, unnoticed by the operators. The HEPA grate spots were the buildup of product residue from years of these disconnections.

The site made a variety of products in these rooms. Most were clear, but some were bright red, blue, yellow, or even cloudy white. The inspection team could only see the colored product residue. However, it’s inevitable clear products splashed like this too. The possibility of cross contamination between products necessitated a temporary shut-down to replace every HEPA filter in the entire building. Processes were also updated to prevent product splashing in the future*.

On behalf of patients receiving these drugs, thank you operator for looking up!

. . . And the Sledgehammer . . .

I previously wrote about a Pseudomonas Contamination Event . I inspected centrifuges that were used for lots contaminated with Pseudomonas. I didn’t mention the sledgehammer because it didn’t contribute to the root cause. However, it left recognizable marks on the cleanrooms.

Centrifuges are very active pieces of machinery for a manufacturing process. Constant vibration can loosen pieces leading to spills. Their lids need to be screwed on tight! A specialized tool to tighten and loosen industrial grade centrifuge lids didn’t exist. The site used a simple, cleanable sledgehammer. After screwing on the lid, the operators would bang a specific spot to fully tighten it.

My attention went to the sledgehammer when it was used to open the centrifuges. To the operator, it was the same process, but in reverse. However, the disassembly process was messy; the equipment was still covered in product residue. This residue was essentially a micro broth paradise. With every BANG and SLAM, residue splashed around the room. Operators and cleaning crew were great cleaning the floors and walls, but the ceiling looked like a monotone Jackson Pollock.

It was disgusting! Can you imagine what regulators would think if they walked in and looked up? Who wants micro broth hanging over their manufacturing equipment? We immediately upped our cleaning frequency. Unfortunately, we didn’t have an affordable work-around for the sledgehammers.

*Interesting side note here: Recently, I talked to one of the quality managers responsible for fleshing out the corrective actions for this event. She moved to a manufacturer of a popular OTC product. This product has a very distinctive color. Do you know what that site did to prevent this issue? They painted all the walls, floors, and ceilings the color of that product.

The post Look Up! appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

Or how to avoid getting nipped in the BUD

“They’re cutting our BUDs!” I heard this from an exasperated compounding pharmacist looking for any way to make the most out of his product’s shelf life. BUDs, or the Beyond-Use Dates assigned to compounded products, determine when a product must be destroyed if it has not yet been used.  Wasted products are not good for business or patients.  

The draft chapter for USP <797> “Pharmaceutical Compounding – Sterile Preparations” has been available for almost a year. The massive overhaul of the parameters used to assign Beyond-Use Dates should no longer be a surprise. In most cases, compounded products will have shorter shelf lives. This heavily impacts batch planning and availability to patients.  

There are some ways to extend the BUDs that are outlined in the new revision. Most notably, performing a sterility test on each lot can often double the BUD window. But waiting for a sterility test takes at least two weeks, which significantly cuts into that window.  

There’s another seemingly minor change that could help alleviate this issue. In the current version of USP <797>, there’s a single line buried in the sterility test section that briefly mentions alternative methods:

“A method not described in the USP may be used if verification results demonstrate that the alternative is at least as effective and reliable as the USP Membrane Filtration method or the USP Direct Inoculation of the Culture Medium method where the Membrane Filtration method is not feasible”

Unfortunately, the current reference offers no direction for demonstrating method effectiveness.  Without direction as to what will be accepted by relevant regulatory bodies, the reference is nearly useless.  It’s difficult to invest in a project without a clear road map for approval.

Rapid Acceptance of Alternative Methods

The simple change in the USP <797> revision is the addition of three cross-references to another USP chapter, <1223> “Validation of Alternative Microbiological Methods”. One reference is in the new introduction section, which signals alternative methods are front of mind for the standard setters. The other two references are in the Sterility test section, highlighting the test with such a pressing need for faster results. USP <1223> is the roadmap hopeful rapid method users were looking for. Using a rapid sterility test boosts the BUD timeline while minimizing the impacts of the traditional incubation period.  

I’ve written about <1223> on this Eureka blog before. I am proud of what my team at Charles River did, putting together a package that could be used by Celsis® rapid method users to validate their system and gain regulatory approval. Prominent service laboratories that work with compounding labs, including ARL Bio Pharma and Pharmetric Laboratory, have also completed their own independent Celsis validation per <1223>. There’s not much time left before <797> becomes official, but rapid methods in good regulatory standing are available to you today.

Personal Reflections on Shelf-life

It’s difficult to make this post without acknowledging the importance of the compounding industry for meeting unique patient needs. The shift in regulatory standards around BUDs undoubtedly makes it more difficult to meet those needs. I also understand how regulators, who are acting on behalf of patients, want these products to be safe.  

In 2009 I was working in a QC department for a large CMO. We received a warning letter. Included in it was a minor observation related to reagent expiration dates in the QC labs. Most of these test reagents were in multi-use containers. Analysts opened a container, weighed out the material they needed, then returned the container to storage. All material was removed from storage and disposed once it exceeded the manufacturer-assigned expiration date.

But the date assigned by the manufacturer is based on unopened material held in ideal storage conditions. Repeatedly opening the container may or may not impact the material’s effectiveness over time. How could we justify using the material for as long as we did?

That’s a similar question USP <797> writers asked themselves when materials with defined expiration dates are compounded into a new product. Unless proven otherwise, it’s safer to assume a mixture of these materials will lose effectiveness prior to any of the individual material expiration dates.

USP <797> writers took an approach like our observation response. We assigned shorter expiration dates to all open material, similar to how <797> set maximum BUDs based on product category. There’s room for pushback because the dates seem arbitrary, which is why they allow for extensions of BUDs with appropriate stability data. It’s a difficult position to be in when you must standardize a broad market, but I can appreciate the work that was done.

With that in mind, let’s remember this industry challenge with BUDs exists for a good reason – patient safety.  With rapid methods, we have a way to minimize the impact of this challenge.   

The post Compounding Pharmacies and the Need for Rapid Methods appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

Another reason why it is important to identify the BBC species is that one preventive measure is not necessarily sufficient to eradicate all member species, as they are unique and may respond differently to remediation attempts. Some species like Burkholderia multivorans can cause additional concern and require different action or a broadened scope of investigation. With that in mind, manufacturers have begun to revisit their microbiological testing protocols to detect BCC contamination. Not all members of the complex are designated as pathogens so it’s important to accurately identify to the species-level. However, this has proven problematic using traditional DNA sequencing methods and has required advances in microbial identification strategies.

Going beyond the species
There are times when you need to go beyond the species level. In an investigation, you may find multiple contamination events that identified the same species. Did they come from the same origin? Strain typing is a microbial characterization technique that can help distinguish members of the same species to the strain level using well-established, highly accurate DNA sequence analysis. For example, a company identifies a microorganism of the same species in two separate samples, one was from environmental monitoring of the water system and the other was recovered from routine quality testing of a finished product. If the microbial strains are identical, according to strain typing, that means that their DNA is the same and the microorganism contaminating the finished product is the same one you’re finding in your water. This is the reason why strain typing is critical to resolving and remediating contamination events. Strain typing has been traditionally done by analyzing multiple coding genes in independent steps using Multi-Locus Strain Typing (MLST). Now, there is a technology that is being utilized called Next Generation Sequencing (NGS) that can analyze the entire DNA genome in one test producing the same results in less steps.
Test early and regularly for microbial contamination
Catching microbes, like BCC, before they become a problem and working to implement a microbial contamination control strategy means monitoring your environment with accurate technology. Testing earlier and regularly in the manufacturing process and implementing good manufacturing practices can reduce contamination in your raw materials, water, and finished products. These practices ensure that products are consistently produced and controlled according to quality standards designed to aid in the quality control process.
Any time part of a facility is offline due to a microbial contamination event, it equates to lost profits. If the contamination originated from your water system, no amount of cleaning the manufacturing vessels will solve the problem because that is not the source. An accurate species level identification and, when necessary, strain typing can help put you on the right path to finding the microorganism source.

The relative cost of accurate microbial IDs is small especially compared to the loss of inventory or, worst case, the cost of a recall. Charles River Labs recognizes that accurate and reliable microbial identification data is critical not only for certain organisms such as the Burkholderia cepacia complex, but for all microbial isolates that are part of quality control practices. Our unique combination of Celsis® rapid microbial detection and Accugenix® microbial identification keeps your manufacturing operations running efficiently and smoothly.

For more information contact Sopex d.o.o. distributor.

The post Your water may not be as clean as you think…meet Burkholderia cepacia appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.

If you’re reading this, it’s likely you have a clear understanding of the benefits and need for rapid microbiological methods (RMMs). You may also find it easy to understand the technology, and how to use the instrument and its key features.

At the same time, you may be at a complete loss on how to actually implement the rapid method into your organization’s lab processes. Don’t worry, you’re not alone. Selecting and implementing an alternative or rapid method is often the first major project many QC microbiologists face that’s outside their comfort zone. Selection, evaluation, procurement, finances, and presenting a business case are much different than performing assays. If understanding the RMM adoption process sounds hard, keep reading. It’s about to get a lot easier. 

Dr. Paul J. Newby, with 30 years of experience in the pharma industry and former Microbiology Modernization Lead at GlaxoSmithKline, understands this challenge. His approach to developing an efficient implementation process is detailed in his eBook “Guidance on Implementation of Alternative and Rapid Methods,” where he dissects the process for implementing an alternative or rapid microbial method in a simplistic, easy-to manage framework. 

 The framework focuses on key stages in the implementation process, which is fundamentally a cycle. You can take years off of the process of implementing a rapid microbial method by simply having a plan prior to beginning the journey.  This same framework can be repeated for any new technology, whether it be a rapid detection, identification, or preparation system. The three main phases are identified as Seek, Investigate, and Industrialize, with sub-tasks within each one. Below is a brief outline of each phase, and key questions to ask:

Phase 1: Seek

• Clarify needs. In this step, Dr. Newby recommends articulating a clear understanding of your organization’s end goal. Is it to save warehousing costs, or maybe to address your data integrity concerns? 
• Review technologies. When it comes to rapid methods, the number of on-market technologies can seem daunting. You can reduce the need to understand every technology by focusing on commercial readiness.  
• Identify candidates. Pick out the top contenders that match your business needs. Dr. Newby recommends choosing a Commercial Off-the-Shelf System to avoid lengthy validation and evaluation efforts. Speaking of finding the right system, it’s also best to research whether the chosen vendor has validation support and materials offerings to aid in the process with regulatory authorities.  

Phase 2: Investigate

• Business & technical acceptance criteria. Do you know what your current cost per test is, and how that compares to the cost per test with a new system? Comparison of labor costs and savings, as well as current and potential bottlenecks, are great places to start in evaluating your operational and financial factors. Most importantly, how does the new method compare in terms of performance? Before implementing any system, you have to first prove its non-inferiority to the compendial method of testing in terms of accuracy, limit of detection, specificity, and other areas. 
•  Data integrity. Dr. Newby lays out a comprehensive set of  elements to ensure that data integrity and archiving needs are met. Labs need to understand the importance of accuracy, consistency, and availability of data. Recording results automatically with most rapid method technologies will ensure the integrity of your data over time when looking at these characteristics.
• Proof of Concept. This serves as an evaluation of the suitability of various test systems, and an investigation of claims made by suppliers. Dr. Newby recommends including a concise summary report of these findings to aid in the go/no-go decision-making process for subject matter experts. 

Phase 3: Industrialize

• Validation. PDA Technical Report No. 33, as well as USP <1223> and Ph. Eur. 5.1.6 provide invaluable information for this stage in the implementation process. While the end user is mainly responsible for carrying out the procedures of the validation process, the vendor of the new method may also provide support in terms of necessary documentation and testing services. 
• Registration. Contacting regulatory authorities early-on in the processes of selection, validation, and implementation of your RMM ensure your alignment with the guidance provided by these authorities. It’s important to remain in contact with subject matter experts during all stages of the process to allow for an easier validation undertaking. 
• Implementation. Once integration into laboratory operations is underway, advocacy of laboratory innovation across industries and applications through shared knowledge is important. Many organizations would benefit greatly from a cost perspective and the ability to incorporate the method into global systems of operation. This would also benefit the patients and consumers in need of therapies and products tested with RMMs. 

Download the eBook to receive more details on this guidance and insightful tips for getting started.

A similar approach backed by quality control microbiology experts from some of the largest names in pharma, was used by organizations such as Roche Diagnostics and Merck & Co. The case studies recorded exemplify the planning, strategizing and execution of an RMM, but they  also offer a comparison of technology types to aid you in your decision. 

For more resources in the implementation journey, check out our How to Select an RMM web page.

The post Incorporating Quality Control Technologies in the Lab appeared first on Sopex d.o.o. - specialized services in medicine and the pharmaceutical industry.