Validation of bioanalytical chromatographic methods

A strategy is discussed for the validation of chromatographic methods that are developed to quantify drugs in biological matrices. Both the validation terminology and the hypothesis testing are briefly reviewed. The emphasis is on the design of the experiments required to allow a reliable conclusion about acceptance or rejection of the bioanalytical method. In particular, it is explained how to evaluate the calibration line, devise experiments to estimate precision and bias and how to determine the stability of the analyte between the time of the sample collection and the analysis of the processed sample.     

Capsule review on bioanalytical method transfer: opportunities and challenges for chromatographic methods

With the globalization of drug development activities, transferring a validated bioanalytical procedure to a different site within a pharmaceutical company, or to one or multiple contract research organizations has been dramatically increased in recent years. Undeniably, bioanalytical method transfer is the needed step prior to routine sample analysis at the receiving laboratory. It is clearly stated in the 2001 US FDA Guidance on Bioanalytical Method Validation that a partial validation is needed for method transfer between laboratories. In the current EMA draft guidelines on method validation, the necessity of a method transfer is also emphasized. However, the above guidelines do not give many details on how and when a method transfer validation should be conducted. There is a need for a step-by-step deliberation on the overall strategies, procedures and even technical details for a successful bioanalytical method transfer. In this article, we review the contemporary information available in the scientific literature on method transfer and illustrate various bioanalytical method transfer scenarios using case studies. A 'flexible and fit-for-purpose' bioanalytical method transfer strategy is proposed.     

EBF recommendation on the validation of bioanalytical methods for dried blood spots

Over the last few years bioanalysts, pharmacokineticists and clinical investigators have rediscovered the technique of dried blood spots. The revival has provided pharmaceutical R&D a wealth of opportunities to optimize the drug-discovery and development process with respect to animal and patient ethics, new scientific insights and costs savings. On the bioanalytical front, multiple experiments have been performed and a lot of experience has been gained. Nevertheless, the technique still has a number of bioanalytical challenges. The European Bioanalysis Forum discussed the advantages and hurdles of the technique and summarized their current thinking in a recommendation on the validation of bioanalytical methods for dried blood spots, which can be used as a cornerstone for further discussions and experiments.     

Advances in validation, risk and uncertainty assessment of bioanalytical methods

Bioanalytical method validation is a mandatory step to evaluate the ability of developed methods to provide accurate results for their routine application in order to trust the critical decisions that will be made with them. Even if several guidelines exist to help perform bioanalytical method validations, there is still the need to clarify the meaning and interpretation of bioanalytical method validation criteria and methodology. Yet, different interpretations can be made of the validation guidelines as well as for the definitions of the validation criteria. This will lead to diverse experimental designs implemented to try fulfilling these criteria. Finally, different decision methodologies can also be interpreted from these guidelines. Therefore, the risk that a validated bioanalytical method may be unfit for its future purpose will depend on analysts personal interpretation of these guidelines. The objective of this review is thus to discuss and highlight several essential aspects of methods validation, not only restricted to chromatographic ones but also to ligand binding assays owing to their increasing role in biopharmaceutical industries. The points that will be reviewed are the common validation criteria, which are selectivity, standard curve, trueness, precision, accuracy, limits of quantification and range, dilutional integrity and analyte stability. Definitions, methodology, experimental design and decision criteria are reviewed. Two other points closely connected to method validation are also examined: incurred sample reproducibility testing and measurement uncertainty as they are highly linked to bioanalytical results reliability. Their additional implementation is foreseen to strongly reduce the risk of having validated a bioanalytical method unfit for its purpose.     

Development and validation of a high-performance liquid chromatographic method for bioanalytical application with rimonabant

A simple and feasible high-performance liquid chromatographic method with UV detection was developed and validated for the quantification of rimonabant in human plasma. The chromatographic separation was carried out in a Hypersil BDS, C₁₈ column (250 mm × 4.6 mm; 5 μm). The mobile phase was a mixture of 10 mM phosphate buffer and acetonitrile (30:70, v/v) at a flow rate of 1.0 ml/min. The UV detection was set at 220 nm. The extraction recovery of rimonabant in plasma at three quality control (QC) samples was ranged from 84.17% to 92.64%. The calibration curve was linear for the concentration range of 20–400 ng/ml with the correlation coefficient (r²) above 0.9921. The method was specific and sensitive with the limit of quantification of 20 ng/ml. The accuracy and precision values obtained from six different sets of QC samples analyzed in separate occasions ranged from 88.13% to 106.48% and 0.13% to 3.61%, respectively. In stability tests, rimonabant in human plasma was stable during storage and assay procedure. The method is very simple, sensitive and economical and the assay was applied to human plasma samples in a clinical (pharmacokinetic) study of rimonabant.     

Application issues in bioanalytical method validation,sample analysis and data reporting

Although some degree of consensus has been reached concerning the requirements for acceptable method validation, the procedures used to establish them vary significantly between laboratories. Also, issues arising from application of these requirements during validation and subsequent sample analysis need to be addressed. The purpose of this paper is to discuss application issues concerning prerequisites to method validation, and all validation criteria for evaluation of method reliability and overall performance. Other poorly addressed issues such as re-validation, cross-validation, partial sample volume, multicomponent analysis and reporting will also be discussed. Although many issues discussed are of a general nature, the scope of this presentation is primarily to address issues arising from the validation and routine application of chromatographic methods.     

单克隆抗体药物药代动力学特征、分析方法以及体内分析方法学研究进展

近年来,单克隆抗体药物发展迅速,在肿瘤、免疫、血液等系统疾病领域应用日益广泛。截至2019年其全球处方药物市场占比已达15.3%(1400亿美元)。单克隆抗体药物作为一种大分子蛋白,因其特殊的结构和生理性质,在体内的吸收、分布、代谢及排泄均与小分子药物存在较大差异,具有靶点介导的药物处置、非线性药动学代谢、时间依赖性、较长的半衰期等独特的药代动力学特征,充分了解这些特征有益于该类药物分析方法的开发。单克隆抗体药物在生物体内的处置具有特殊性和复杂性,极大地增加了生物检测的难度,因此必须建立专属、灵敏、准确、可重复的测定分析方法。本文旨在论述单克隆抗体药物的药代动力学特征、常用分析方法及其优缺点、体内分析方法学验证要求等,并逐点与小分子药物进行对比讨论,以期为单克隆抗体药物的分析技术开发提供部分参考。     

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--part II

As reported in a previous paper [1], the main objective of the new commission of the Société Française des Sciences et Techniques Pharmaceutiques (SFSTP) was the harmonisation of approaches for the validation of quantitative analytical procedures. In a series of meetings, members of this Commission have first tried to review the objectives of analytical methods and the objectives of validation methods and to recommend the use of two-sided β-expectation tolerance intervals for total error of validation samples (accuracy profile) in the acceptance/rejection of analytical method in validation phase.     

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--Part I

This paper is the first part of a summary report of a new commission of the Société Fran?aise des Sciences et Techniques Pharmaceutiques (SFSTP). The main objective of this commission was the harmonization of approaches for the validation of quantitative analytical procedures. Indeed, the principle of the validation of theses procedures is today widely spread in all the domains of activities where measurements are made. Nevertheless, this simple question of acceptability or not of an analytical procedure for a given application, remains incompletely determined in several cases despite the various regulations relating to the good practices (GLP, GMP, ...) and other documents of normative character (ISO, ICH, FDA, ...). There are many official documents describing the criteria of validation to be tested, but they do not propose any experimental protocol and limit themselves most often to the general concepts. For those reasons, two previous SFSTP commissions elaborated validation guides to concretely help the industrial scientists in charge of drug development to apply those regulatory recommendations. If these two first guides widely contributed to the use and progress of analytical validations, they present, nevertheless, weaknesses regarding the conclusions of the performed statistical tests and the decisions to be made with respect to the acceptance limits defined by the use of an analytical procedure. The present paper proposes to review even the bases of the analytical validation for developing harmonized approach, by distinguishing notably the diagnosis rules and the decision rules. This latter rule is based on the use of the accuracy profile, uses the notion of total error and allows to simplify the approach of the validation of an analytical procedure while checking the associated risk to its usage. Thanks to this novel validation approach, it is possible to unambiguously demonstrate the fitness for purpose of a new method as stated in all regulatory documents.     

Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--part III

In the first two documents [Ph. Hubert, J.J. Nguyen-Huu, B. Boulanger, E. Chapuzet, P. Chiap, N. Cohen, P.A. Compagnon, W. Dewé, M. Feinberg, M. Lallier, M. Laurentie, N. Mercier, G. Muzard, C. Nivet, L. Valat, J. Pharm. Biomed. Anal. 36 (2004) 579-586; Ph. Hubert, J.J. Nguyen-Huu, B. Boulanger, E. Chapuzet, P. Chiap, N. Cohen, P.A. Compagnon, W. Dewé, M. Feinberg, M. Lallier, M. Laurentie, N. Mercier, G. Muzard, C. Nivet, L. Valat, E. Rozet, J. Pharm. Biomed. Anal., in press], a recent SFSTP Commission on the validation of analytical procedure has introduced a harmonized approach for the validation of analytical procedures. In order to complete this guide, the statistical methodology allowing to correctly conclude about the validity of a procedure is proposed in this third part of the guide. Indeed all the steps to obtain the decision tool namely the accuracy profile are described and illustrated step by step by a numerical example. This tool, based on the concept of total error (bias+standard deviation) build with a beta-expectation tolerance interval, allows to easily take the right decision and simultaneously minimizing the risk of the future use of the analytical procedure.     

Calibration and validation of linearity in chromatographic biopharmaceutical analysis.

Calibration in chromatographic biopharmaceutical analysis is a major determinate of method performance and many methods have been proposed to evaluate an appropriate calibration model, to determine the linear range and to evaluate the goodness of fit. Ten chromatographic bioanalytical methods have been evaluated in this work by observation of concentration-response curves, linearity plots, calculation of concentration residuals, correlation coefficients and lack of fit analysis. These methods were applied to univariant linear regression, weighted regression, polynomial regression and power fit models in order to determine the most appropriate way to establish and evaluate calibration functions. It was found that weighted linear regression provided the most appropriate calibration function for eight of the 10 methods studied, whereas unweighted regression and the power fit model proved appropriate for one each of the other two methods. The choice of calibration function was best accomplished through observation of calculated concentration residuals. Linearity and sensitivity plots were of little value for assessment of linearity through the selected calibration range if conventional (+/- 5%) tolerance limits are employed. Validation of the calibration model can be accomplished by demonstrating the concentration residuals and the slope of the log concentration-log response plots are within reasonable tolerance limits or by lack of fit analysis. Correlation coefficients were demonstrated to be of little value for this purpose and the quadratic approach to linearity validation was in disagreement with other methods in four of the 10 methods evaluated.     

Matrix metalloproteinase inhibitors: a review on bioanalytical methods, pharmacokinetics and metabolism

Enzymes are major drug targets in drug discovery and development processes in the pharmaceutical and biotechnology industry. A recent survey found that nearly half of all the marketed small-molecule drugs are inhibitors of enzymes. Matrix metalloproteinases (MMPs) are a family of 28 enzymes capable of degrading the constituents of the extracellular matrix (ECM) and the basement-membrane. MMPs play an essential role in several normal physiological processes including growth, wound healing and tissue repair. Over-expression and activation of MMPs has been linked to a range of diseases which include osteoarthritis, tumor metastasis, angiogenesis and cardiovascular diseases. The development of MMP inhibitors as therapeutic agents has kept an important place in drug discovery. Therefore, there is also an increasing need for robust analytical methods for evaluation of inhibitory potency and for the analysis of MMP inhibitors and their metabolites which can even play a more significant role than the parent drug. Modern analytical techniques and hyphenated instrumentations such as liquid chromatography-mass spectrometry with a function of structure elucidation can provide a profound insight into the research of MMP inhibitors and also serve as a complementary method to zymographic techniques for the analysis of biological samples. This review mainly summarizes bioanalytical methods, pharmacokinetics and related metabolites of MMP inhibitors over the last 12 years.     

色谱法在麻黄生物碱手性分析中的研究进展

生物碱为麻黄中的主要活性兼毒性成分,以3对互为光学非对映异构体的苯异丙胺类生物碱形式存在,这些生物碱不仅对与对之间的药理作用存在显著差异,而且互为非对映异构体的一对生物碱之间作用也不同.此外,通过合成途径获得的几种麻黄生物碱,总是掺杂少量的相应对映体,这些对映体由于失去活性而以杂质的形式存在.因此,为有效合理地控制麻黄及麻黄制剂的质量,应对所存在的异构体分别进行测定.近几年,麻黄生物碱的手性分析成为研究的热点.将近年来麻黄生物碱手性分析的色谱方法进行综述,主要包括高效液相色谱法、气相色谱法、毛细管电泳法,并对各方法的优劣进行讨论.