“Within our comprehensive end-to-end CDMO services, our clients rely on us for insights into impurities, including nitrosamines. As we cater to a global clientele subject to diverse regulatory frameworks, we must meticulously navigate the nuances of each. Notably, the U.S. FDA and the EMA employ distinct approaches. While we endeavor to establish a universal methodology encompassing all requirements, occasionally achieving this harmonization proves challenging.”
Mr Mariusz Kurowski – Analytical Expert at Polpharma API Business Unit
Focused on method development and testing, Polpharma API continuously adapts to evolving regulatory guidelines, notably addressing the detection of nitrosamines in pharmaceutical products. The analytical team, part of the R&D Department at Polpharma API, collaborates within the ‘control strategy’ group and interfaces with other sections and departments. Leveraging advanced analytical instrumentation, particularly liquid chromatography coupled with mass spectrometry (LC-MS), Polpharma API ensures compliance and quality in its pharmaceutical processes.
Backed by a long history of support from Waters, Polpharma API has continued to upgrade its Waters analytical instruments to achieve the sensitivity and robustness necessary for impurity analysis.
Meeting global nitrosamine requirements
Polpharma API supplies products to highly regulated global markets, such as the US, Europe, Korea and Japan. Its product range encompasses baclofen, aripiprazole, tadalafil, vardenafil, alendronate, risedronate, sildenafil, and hydrochlorothiazide. Additionally, the company vertically integrates 19 substances to manufacture ready‑made pharmaceutical forms, including sildenafil citrate, piracetam, pentoxifylline, metronidazole, and hydrochlorothiazide. Managing this diverse portfolio necessitates advanced analytical instrumentation to support Polpharma API’s R&D, quality control, and method development efforts.
Like pharmaceutical companies globally, Polpharma API faced the consequences of regulatory guidance following the 2018 discovery of nitrosamines in pharmaceutical products, posing potential health risks. Nitrosamines, which are carcinogenic mutagenic impurities, can be found in various sources, including groundwater, treated water, foods, beverages, and consumer products. Detection of nitrosamine impurities in specific drug products led to recalls, prompting regulatory agencies to issue guidance on preventative measures for API and drug product manufacturers.
The potential presence of nitrosamine impurities is linked to specific processes and materials that can generate these impurities during API synthesis. Factors such as chemistry selection, contaminated solvents, and water contribute to nitrosamine formation. Besides APIs, nitrosamines may also be present in the final product due to degradation during formulation processing or storage, involving contaminated excipients and printing inks.
Reflecting on five years of quality control for nitrosamine impurities.
Regulatory agencies have imposed strict limits on nitrosamine presence in medications due to growing concerns. However, Polpharma API’s global experience as a CDMO allows it to address regulatory challenges effectively, considering variations in the regulatory landscape across different countries. This experience positions Polpharma API to navigate and comply with diverse regulatory requirements worldwide.
Polpharma API follows a reactive approach based on risk assessment for nitrosamines control, as per regulatory guidance. When an external authority has questions or sends a letter, the team must react appropriately. These interactions with regulatory agencies are common in the API development and testing field, and the CDMO’s extensive experience serves as an advantage when they need to respond quickly.
Mr Kurowski explains: “Using risk analysis, we know the general possibility of forming an impurity. Our first workflow focuses on testing and proving the absence of contamination, which is relatively easy. However, if a regulatory agency has a question, our second workflow requires developing a method and analysing batches of API samples to answer those questions, usually in a relatively short timeframe of 50–90 days. That’s where our experience really helps us support our customers.”
LC-MS/MS for identification and quantification of nitrosamines
LC-MS/MS stands out as a preferred analytical technique for quantifying nitrosamines due to its ability to separate complex impurity mixtures from the API, excipients, and other matrix components in drug formulations. This technique enables sensitive detection and robust quantification of trace impurities beyond regulatory requirements.
“Adapting to evolving regulatory requirements is an intrinsic part of our journey. With a rich history of engagement in nearly every corner of the globe, we have amassed invaluable expertise in API development and testing, successfully navigating shifting regulatory landscapes. A decade ago, our focus was on genotoxic impurity testing, whereas today, we must diligently monitor a spectrum of contaminants, including nitrosamines. Change is the constant in our domain, and we remain steadfast in our commitment to addressing emerging challenges as they arise.”
Dr Paweł Olszowy – Head of Analytical Development Section at Polpharma API
To address the challenge of controlling nitrosamine impurities in both API and final drug products, regulatory bodies establish Acceptable Intake (AI) limits for N-nitrosamine impurities in the final product. These limits often demand a level of sensitivity achievable only with a triple quadrupole instrument. While acquiring such advanced instrumentation and expertise can be challenging for many companies, Polpharma API prioritised investing in more sensitive and robust equipment to meet these limits and support their supply chain effortlessly.
Mr Kurowski describes the impact: “As an API producer, we must always prove that our products are free of contaminants – not only nitrosamines, but all kinds of genotoxins, carcinogens, and other impurities. About 10 years ago, we were focused on testing for genotoxic impurities, which could be done on a single quadrupole mass spectrometer. Nitrosamines require more sensitivity, so when we brought our analysis in‑house, we moved to triple quads and MS/MS techniques. These MS systems have become more robust and more precise in terms of the sample integration, injection, and other factors, which offers the capabilities we need for these types of analyses.”
The shift from outsourced analysis to the creation of their in-house centre of excellence for impurity analysis brought positive results for the team at Polpharma API. In the past, the company outsourced part of genotoxic impurity testing as well as some method development but, as the nitrosamine situation developed, a strategic decision was made to bring all testing in house. The expertise that the Polpharma API team has built up is now relied on by customers in their supply chain. Mr Kurowski explains: “We simply cannot afford any delays. Not just in answering regulatory queries regarding nitrosamine or other contaminants, but also because even a short delay in getting a result can mean an API or a finished product not being able to be released for sale.”
Polpharma API expanded its analytical capabilities by investing in two Waters Xevo™ TQ-S micro Mass Spectrometers, enhancing consistent low-level quantitation across a broad dynamic range alongside existing Waters instrumentation. MassLynx™ Security software is employed for 21CFR11 compliance, ensuring data integrity and documenting audit trails throughout the data lifecycle.
The Xevo TQ-S micro MS consistently delivers high-quality analytical performance, even in complex sample matrices. Beyond these capabilities, Polpharma API’s positive experiences with Waters’ comprehensive instrumentation portfolio influenced their decision for in-house expansion. The satisfaction derived from using Waters ACQUITY™ UPLC™ I-Class PLUS Systems further solidified their choice to stick with the same vendor for achieving desired results.
“I think 50% of success in LC-MS is due to the chromatography. The quality of the peaks gives you an advantage right from the start. The compatibility of the Waters Xevo TQ-S micro with our Waters ACQUITY UPLC I-Class PLUS Systems was very appealing. Of course, the performance of the mass spectrometer also provides a lot of benefits in the heavily regulated environment of pharma testing, particularly compared to instruments from some other vendors.”
Mr Mariusz Kurowski – Analytical Expert at Polpharma API Business Unit
Waters Service and Support plays an important role in keeping things moving smoothly in both busy departments. Prompt and reliable service from Waters helps minimize downtime and ensures that the instruments are running at optimal performance. Dr Olszowy continues: “Service is vital. We only have two triple quads now, and we have many methods to be developed and validated. At the same time, our QC department is sometimes waiting to release the product, so with our regulatory time pressures we can’t afford for our instrumentation to not be working. We need Waters to help us get up and running as soon as possible.”
Next steps
In the dynamic field of API development and testing, the Polpharma API team remains proactive in adapting to evolving trends. Recent regulatory developments have adjusted initial AI limits for specific nitrosamines (NDSRIs), expanding the acceptable range from parts per billion (ppb) to parts per million (ppm) levels in certain cases. This shift is supported by versatile instrumentation accommodating detection sensitivity, including QDaTM Detector or other single quadrupole MS where higher detection limits are acceptable. Despite this, UPLCs with tandem mass spectrometry systems maintain a crucial role for Polpharma API in method development, impurity identification, and other trace‑level applications.
Looking ahead, Mr Kurowski anticipates a significant change in the CDMO’s future, emphasizing the rising focus on Analytical Quality by Design (AQbD). AQbD is a systematic approach endorsed by regulatory agencies, ensuring consistent and high-quality analytical results throughout the method’s life cycle. The Polpharma API team is proactively preparing for AQbD implementation, particularly in terms of software development. The objective is to automate the AQbD method development process as much as possible.
Of course, the CDMO’s recent investments in the Waters analytical instrumentation will also play a role. The capabilities of the Waters Xevo TQ-S micro mass spectrometers will likely help the Polpharma API team navigate the constant changes in API development and testing. Mr Kurowski explains:
“We plan on using the Waters Xevo TQ-S micro mass spectrometer in future projects. The capabilities of the triple quad will be needed as we solve new problems. It will be one of our main R&D resources. In addition, designing methods that run on the simpler QDa detector systems for routine QC will remain a focus for our work too.”
However, Polpharma API’s extensive experience, including navigating the evolving nitrosamines situation, makes them very confident that when new challenges emerge, the team will be prepared to meet them. Dr. Olszowy summarizes: “In our field, we know that something else is always around the corner. I remember 10 years ago the focus was on genotoxic impurities and since 2018 we have been dealing with the challenges around nitrosamines. I don’t have a crystal ball to see the future but, whatever emerges in the future we can be calm because we know we are ready.”
Amy Bartlett
Amy’s role in Pharmaceutical Market Development, within Waters’ Global Testing and Lab Services business, centres primarily on analytical solutions and workflows to support the Pharmaceutical Industry’s quantitative analysis needs. Specifically, Amy is focused on Waters’ Tandem Mass Spectrometry solutions that support the industry’s most pressing issues, including the challenging regulated analysis of genotoxic impurities.
