Advancements in lc-ms/ms bioanalytical method validation

Lucia Romano

doi:10.35841/aacbc-9.3.225

Rapid Communication - Journal of Clinical and Bioanalytical Chemistry (2025) Volume 9, Issue 3

Advancements in lc-ms/ms bioanalytical method validation

Lucia Romano^*

Department of Pharmaceutical Analysis, University of Milan, Milan, Italy

*Corresponding Author:: Lucia Romano
Department of Pharmaceutical Analysis
University of Milan, Milan, Italy.
E-mail: lucia.romano@unimi.it

Received : 03-Sep-2025, Manuscript No. aacbc-225; Editor assigned : 05-Sep-2025, PreQC No. aacbc-225(PQ); Reviewed : 25-Sep-2025, QC No aacbc-225; Revised : 06-Oct-2025, Manuscript No. aacbc-225(R); Published : 15-Oct-2025 , DOI : 10.35841/aacbc-9.3.225

Citation: Romano L. Advancements in lc-ms/ms bioanalytical method validation. aacbc. 2025;09(03):225.

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Introduction

Bioanalytical method development and validation are critical steps in drug discovery and clinical pharmacology, ensuring reliable quantification of drugs, their metabolites, and biomarkers in biological matrices. Modern approaches frequently employ Liquid Chromatography–Mass Spectrometry (LC-MS/MS) due to its high sensitivity, selectivity, and throughput capabilities. Recent efforts focus on establishing robust and validated LC-MS/MS methods for a diverse range of compounds. For example, a method was developed for the simultaneous quantification of bosutinib and its N-desmethyl metabolite in human plasma. This method used acetonitrile protein precipitation for sample preparation and showed excellent linearity, precision, accuracy, and stability, making it ideal for pharmacokinetic studies in clinical settings [1].

Similarly, a robust LC-MS/MS method was presented for quantifying lenalidomide in human plasma. This method employed solid-phase extraction for efficient sample clean-up and underwent thorough validation according to regulatory guidelines, demonstrating excellent selectivity, sensitivity, and reproducibility for clinical pharmacokinetic evaluations [2].

The broader landscape of bioanalytical method development and validation for small molecule drugs in various biological matrices has also been reviewed. This review covered crucial aspects such as sample preparation techniques, chromatographic separation, mass spectrometric detection, and regulatory compliance, offering vital insights into best practices [3].

Further specialized work has focused on the unique challenges of quantifying biomarkers. One paper explored best practices and future directions for bioanalytical methods aimed at quantifying biomarkers, highlighting issues like low concentrations and matrix effects while outlining strategies for method reliability and regulatory acceptance [4].

In the realm of quantifying multiple analytes, a robust LC-MS/MS method was developed and validated for the simultaneous quantification of five non-benzodiazepine hypnotics in human plasma. This involved protein precipitation, optimized chromatographic conditions, and rigorous validation to ensure accuracy, precision, and applicability in clinical pharmacology studies [5].

Beyond human studies, preclinical pharmacokinetic assessments also rely on highly sensitive methods. A validated LC-MS/MS method for the simultaneous quantification of three active metabolites of a novel antidepressant in rat plasma demonstrated the importance of meticulous sample preparation and chromatographic separation to achieve high sensitivity and selectivity [6].

As technology evolves, so do the techniques for sample handling. The specific challenges and opportunities in bioanalytical method validation when using microsampling techniques have been discussed. This work emphasized the need for adapted validation parameters to ensure data quality and regulatory compliance for small sample volumes, which is crucial for preclinical and pediatric studies [7].

The regulatory environment is constantly evolving, shaping the future of bioanalytical method validation in drug development. An overview of the evolving regulatory landscape and future outlook covered guidelines from major regulatory bodies and discussed harmonizing global requirements, stressing the importance of robust validation strategies for clinical trial success [8].

Efficiency in drug discovery also drives innovation, leading to the development and validation of high-throughput bioanalytical methods for plasma samples. These methods often utilize automated sample preparation coupled with LC-MS/MS, significantly improving efficiency, reducing turnaround times, and maintaining data quality in large-scale bioanalysis [9].

Ultimately, the foundational work often involves the development and thorough validation of an LC-MS/MS method for quantifying a single small molecule drug in human plasma. Such methods detail chromatographic separation, mass spectrometric parameters, and validation outcomes concerning linearity, precision, accuracy, and stability, affirming their suitability for clinical studies [10].

Conclusion

The presented data highlights diverse advancements and applications in bioanalytical method development and validation, primarily using Liquid Chromatography–Mass Spectrometry (LC-MS/MS). The collection spans specific drug quantifications, such as bosutinib and its N-desmethyl metabolite in human plasma using protein precipitation [1], and lenalidomide in human plasma via solid-phase extraction [2]. It also includes methods for multiple compounds, like five non-benzodiazepine hypnotics in human plasma [5], and active antidepressant metabolites in rat plasma [6]. The articles collectively underscore the critical importance of rigorous validation, ensuring methods meet stringent criteria for linearity, precision, accuracy, stability, selectivity, and sensitivity to support pharmacokinetic and clinical pharmacology studies. Beyond specific drug assays, the data covers broader methodological aspects, including comprehensive reviews of best practices for small molecule drugs [3], strategies for quantifying biomarkers [4], and adapting validation for microsampling techniques [7]. Innovations in high-throughput analysis through automated sample preparation are also discussed [9]. A significant theme is the evolving regulatory landscape and future perspectives in bioanalytical method validation, emphasizing harmonized global requirements and the necessity of robust validation strategies for successful drug development [8]. These studies showcase the continuous effort to enhance the reliability, efficiency, and regulatory compliance of bioanalytical techniques.