Review_Recent advances in chromatography for pharmaceutical analysis

Recent Advances in Chromatography for Pharmaceutical Analysis

Authors: Valentina D’Atri, Szabolcs Fekete, Adrian Clarke, Jean-Luc Veuthey, Davy Guillarme
Affiliations: University of Geneva and University of Lausanne, Novartis Pharma AG

The pharmaceutical industry is highly regulated, mandating strict control over impurities and degradation products in active pharmaceutical ingredients (APIs).
Chromatography remains the gold standard for assessing chemical purity in drug substances and products, widely used from R&D to QC laboratories.
Reversed-phase liquid chromatography (RPLC) holds a pivotal position, particularly for substances with specific log P values identified by Lipinski’s rule.
Advances in methodologies aim to enhance throughput and resolution in response to the complexities of drug development.

Ultrahigh Performance Liquid Chromatography (UHPLC)
- Developed in 2004, allows for faster analysis (1-4 minutes) using sub-2 μm particles and high-pressure instruments (up to 1500 bar).
- Benefits include increased throughput and reduced costs, critical during drug discovery and development.
- Techniques such as superficially porous particles (SPP) technology enhance efficiency with lower back pressure.

2D-LC enhances peak capacity to resolve complex mixtures, utilizing two different separation mechanisms.
Key considerations include compatibility of mobile phase solvents and the speed of the second dimension separation (less than 2 minutes).
Applications include simultaneous achiral-chiral analysis and pharmacokinetic studies.

SFC combines liquid-like solvating power with gas-like diffusivity, enhancing kinetic performance while reducing solvent usage.
Recent advancements in mobile phases and stationary phases expand SFC’s applicability for both polar and non-polar drugs.
Key applications include chiral separations and drug formulations analysis.

Hydrophilic Interaction Liquid Chromatography (HILIC): Effective for analyzing polar and ionizable substances that are poorly retained in RPLC.
The method utilizes a hydrophilic stationary phase with a polar aprotic solvent, commonly acetonitrile.
Notable advantages include better retention of polar substances and the ability to interface easily with MS.

The growth of biopharmaceuticals (monoclonal antibodies, ADCs, etc.) has created unique challenges in terms of analytical methodologies.
Innovations in chromatographic technology target the specific needs of protein analysis, including size exclusion and ion-exchange techniques.
Key qualities addressed include ensuring sample integrity and minimizing interactions with analytical surfaces.

Mass spectrometry is critical in pharmaceutical analysis despite high costs and operational complexity.
Recent advances in compact, affordable MS devices enable practical applications within traditional laboratories.
These devices allow direct and efficient integration with chromatographic systems for routine monitoring and quality assurance.

PAT is a strategy for monitoring pharmaceutical manufacturing processes through the measurement of critical quality attributes.
Techniques such as online HPLC and advanced spectroscopic tools enhance real-time process monitoring, aiming for improved product quality and reduced costs.

Software-assisted tools streamline method development, enhancing efficiency and supporting QbD principles.
Automated screening and optimization contribute to faster innovation cycles and improved analytical protocol robustness.

Increased focus on chiral drug development drives innovations in chiral separations in both LC and SFC modes.
New columns and methods facilitate the development of efficient and robust manufacturing processes.

Identification and control of genotoxic impurities in pharmaceuticals are critical to ensure patient safety.
Advanced chromatographic techniques coupled with sophisticated MS instruments provide necessary sensitivity and specificity for trace analysis.

Establishing validated cleaning procedures is essential in preventing cross-contamination in drug manufacturing.
Analytical challenges involve developing sensitive methods for detecting residual APIs and cleaning agents at trace levels.

Efforts to minimize the environmental impact of chromatographic methods are gaining momentum, focusing on reducing solvent usage and waste.
Strategies include the adoption of greener solvents, shorter columns, and innovative separation techniques.

GC continues to play a vital role in pharmaceutical analysis, particularly in identifying volatile compounds and impurities.
Technological advancements in fast and ultrafast GC improve efficiency and performance within the pharmaceutical sector.

Chromatography will remain a cornerstone of pharmaceutical analysis, evolving through advancements in technology and methodologies.
The future of LC/MS integration, online monitoring, and further innovations in analytical techniques like miniaturization highlight exciting prospects for the industry in the face of increasing complexity in drug analysis.