Briana Nunez, Hayden Skalski, and Kaitlyn Vap Veolia-Water Technologies & Solutions, Sievers Instruments
For years pharmaceutical waters have been released at risk due to waiting for QC results. This is because the testing of water for pharmaceutical production can be labor and time intensive, requiring analysts to isolate samples from a water loop for evaluation in the lab, with test results such as bioburden not being known for days. Even compendial tests that don’t take days – such as endotoxin, total organic carbon (TOC), and conductivity testing – still leave a lot to be desired in terms of efficiency and reducing opportunities for human error. Waiting on test results can force the choice of releasing at risk or delaying production, both of which can be costly. Pharmaceutical manufacturers need simpler and more efficient analytical testing solutions to lean out water testing and drive process efficiencies. With process analytical technology (PAT) initiatives and the introduction of innovative instrumentation and software, a Lean Lab is now within reach.
Compendial water testing and PAT
Compendial pharmaceutical water testing requires measuring four parameters: conductivity, TOC, endotoxin, and bioburden. Controlling these four parameters ensures the purity of water used in all areas of pharmaceutical production. Recently, technologies have been developed to better support and streamline the release of water for production and improve the adoption of PAT for greater efficiencies. For example, real-time release testing (RTRT) for TOC and conductivity, microfluidic technology for bacterial endotoxins testing (BET), and rapid microbiological methods (RMMs) for bioburden are all ways in which QC laboratories can lean out processes and reduce human interference associated with water quality testing. Through the adoption of lean laboratory practices/PAT, pharmaceutical manufacturers benefit from improved process efficiencies, faster product release to market, reduced workload for analysts, and maximized sustainability – all while maintaining data integrity and compliance.
Leaning out TOC, conductivity, endotoxin, and bioburden testing
Lab, at-line, and online testing
If you are seeking practical and achievable steps to lean out your water testing processes, you’ll want to consider various aspects of testing such as sample handling, instrumentation capabilities, data review, process, and sustainability. Lean Lab can come in the form of laboratory testing, at-line testing, or online testing depending on the available technology, with each of these monitoring approaches offering advantages and disadvantages.
The downside of lab-based sampling is the potential to introduce contaminants and delaying the release of water into production or introducing risk by conditionally releasing water. Alternatives to lab-based monitoring include at-line and online testing. If validated appropriately, online monitoring is used for real-time release testing (RTRT), the process of releasing water into production in real time with a validated online instrument of record. RTRT maintains a closed loop system, ensuring process and sample integrity by eliminating human factors. As you can imagine, transitioning from lab, to at-line, to online is leaning out the labor and consumables required for water testing, in the long run saving time and money on fewer resources and materials and optimizing efficiencies.
TOC & Conductivity
It is most common for TOC and conductivity measurements to be taken from the lab with a TOC analyzer and a conductivity probe. This requires isolating samples from various points-of-use to be analyzed in the lab. Not only is the process of isolating samples, transferring them to the lab, and analyzing labor intensive and costly, the process can also introduce contaminants that can cause over-reporting or an out-of-specification (OOS) result. To eliminate the need to routinely sample for conductivity and TOC, many end-users are transitioning to RTRT.
For TOC and conductivity analysis, there are three ways in which online instruments can be used: (1) for process/compendial monitoring; (2) for process control and understanding; (3) for compendial monitoring, release, and process control and understanding. RTRT involves using online instruments for all three and allows for the monitoring and release of pharmaceutical grade water into production in real time. This requires additional validation, inherently improving confidence in all three ways online instrumentation can be used.
Endotoxin
How can you lean out the lab with endotoxin testing? Until recently, there has been little innovation to Limulus amebocyte lysate (LAL) testing in the last 40 years, and most testing today still consists of time consuming, traditional methods. But now, there’s a better way. Automation using a centripetal microfluidic platform offers the simplest form of BET automation available, providing significant time savings and reducing opportunities for error. With the introduction of this technology in the Sievers Eclipse, BET assays are automated while remaining fully compliant with the compendia.
Benefits of microfluidic testing:
- 5–10 minute setup time
- 89% fewer pipetting steps than 96-well plate (reduced from 242 to less than 30), improving employee sustainability
- Significantly less training than traditional methods
- 90% reduction in LAL reagent
- Automated creation and loading of standard curve
- Automated creation and loading of positive product controls (PPCs)
A microfluidic system enables you to carry out the same biochemistry that is performed in traditional 96-well plate assays but with minimal manual eff ort, greater consistency, and reduced reagent consumption. Preloaded standards and PPCs are used to automate standard curves and PPC spikes, saving you significant amounts of time and reducing pipetting steps and opportunities for error.
By introducing microfluidic technology, you can also reduce cold room storage and improve the footprint in your lab. Eclipse microplates can be stored at room temperature, therefore reducing the need to take up extra space in a typical 2-8 °C cooler. Eclipse analyzers are smaller and less bulky than typical 96-well plate readers or robotic systems, so more benchtop space will be available. The Eclipse BET testing software also has the functionality of including a client server installation so endotoxin data can be reviewed and signed off remotely, minimizing the need to physically be in the lab.
Bioburden
Growing and enumerating microbes has remained largely unchanged since the late 19th century when agar began to be used as a medium for growth. Bioburden testing has historically relied on agar plates to quantify the microorganisms within pharmaceutical grade water because of its reliability and accuracy. Although compendial plate counts to determine viable microorganisms is dependable, it is also time consuming and labor intensive, often requiring at least two analysts. Bioburden testing for ultrapure pharmaceutical water requires multiple days in incubation before an agar plate can be read. Results are typically manually recorded, leaving opportunity for data integrity gaps. Due to the time duration it takes for accurate plate counts, most pharmaceutical water is released at risk before bioburden results come back.
To mitigate risk and reduce the time component of bioburden testing, the bioburden industry is seeing a rise in Rapid Microbiological Methods, or RMMs. RMMs return a microbiology result faster than compendial plate counts. Some RMMs can return a result in less than an hour.
By introducing a RMM in a laboratory, you can lean out your process by:
- Reducing time to a result
- Reducing the risk of a contamination event
- Monitoring your process through every stage
- Releasing water with higher confidence
Conclusion
Pharmaceutical water testing doesn’t have to be so time consuming and difficult. As organizations implement PAT and drive toward leaner processes, compendial tests can be optimized and simplified without compromising on regulatory requirements. Important shifts in the transition to a Lean Lab include adopting PAT technologies, reducing human factors and opportunities for errors, and initiating more efficient ways of working – within the lab, at-line, or online. With the right tools and support, leaning out the lab and moving toward real-time release testing are readily achievable, saving you significant time and resources.
Author Biographies
Briana Nuñez is the Lead Researcher within Sievers Instruments’ Microbiology Center of Excellence in Tucson, Arizona. She is responsible for supporting research and development of Rapid Microbiological Methods for bioburden analysis. Briana also plans and conducts laboratory experiments, provides training, delivers presentations and product demonstrations at customer sites, and assists in customer support. Briana holds a B.S. from State University of New York at Purchase College in Biology.
Hayden Skalski is the Life Sciences Product Application Specialist for the Sievers Instruments product line, specializing in bacterial endotoxins testing (BET). Hayden has over 8 years of experience in the pharmaceutical industry and Quality Control Microbiology and has presented on numerous topics surrounding endotoxin testing. Previously, Hayden held roles at Charles River Laboratories, Regeneron and Novartis, validating and executing method development protocols for endotoxin testing, providing customer support, troubleshooting and supporting high-volume product testing. Hayden has a B.S. from the University at Albany (SUNY) in Biology.
Kaitlyn Vap is the Life Sciences Lead Product Application Specialist responsible for supporting Sievers total organic carbon (TOC) customers. She works with companies in the pharmaceutical industry to implement Sievers analytical instruments for the validation of laboratory water and drug products in accordance with global pharmacopeia regulatory standards. As an application specialist, she also explores new and/or developing applications to streamline the process of water and drug product testing. Kaitlyn holds a B.S. in Chemical Engineering from the University of Wyoming.
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