Jean-Luc Veuthey

University of Geneva | UNIGE

In the past few years, there has been significant interest within the forensic community regarding the deployment of portable solutions that provide real‐time results. This article introduces an innovative technology or technology architecture that enables the integration of a handheld device, specifically, Viavi MicroNIR , with a cloud‐based syste...

The purpose of this chapter is to provide a brief overview of the past, present, and future applications of supercritical fluid chromatography (SFC) in pharmaceutical drug discovery and development. In the first part of this chapter, various examples are provided, showing how SFC has been used in the past. Originally employed for the analysis of ap...

Recently, decellularized plant biomaterials have been explored for their use as tissue engineered substitutes. Herein, we expanded upon the investigation of the mechanical properties of these materials to explore their elasticity as many anatomical areas of the body require biomechanical dynamism. We first constructed a device to secure the scaffol...

Design of Experiments (DoE) is a well-established tool used for analytical methods robustness studies, because of its ability to assess the effect of a great number of factors in a minimal number of experiments. However, when assessing the robustness of an analytical method the analysis of the individual effect of each factor is not sufficient on i...

The aim of this study was to evaluate the potential of ultra-high performance supercritical fluid chromatography (UHPSFC) for peptide analysis by comparing its analytical performance to several chromatographic approaches based on reversed-phase liquid chromatography (RPLC), hydrophilic interaction liquid chromatography (HILIC) and mixed-mode liquid...

Modern supercritical fluid chromatography (SFC) is now a well-established technique, especially in the field of pharmaceutical analysis. We recently demonstrated the transferability and the reproducibility of a SFC-UV method for pharmaceutical impurities by means of an inter-laboratory study. However, as this study involved only one brand of SFC in...

The aim of the present study was to explore the feasibility of applying near-infrared (NIR) spectroscopy for the quantitative analysis of Δ⁹-tetrahydrocannabinol (THC) in cannabis products using handheld devices. A preliminary study was conducted on different physical forms (entire, ground and sieved) of cannabis inflorescences in order to evaluate...

Recent developments in supercritical fluid chromatography (SFC) have highlighted the applicability of this technique for the analysis of highly polar compounds. The combination of polar stationary phases and CO2-based mobile phases with an increasing presence of liquid co-solvent (up to 100%) has enabled to further expand the application field of S...

The aim of this work was to expand the applicability range of UHPSFC to series of synthetic and commercialized peptides. Initially, a screening of different column chemistries available for UHPSFC analysis was performed, in combination with additives of either basic or acidic nature. The combination of an acidic additive (13 mM TFA) with a basic st...

Due to the particular elution mechanism observed with large solutes (e.g., proteins) in liquid chromatography, column length has less impact in controlling their retention compared to small solutes. Moreover, long columns-in theory-just broaden the peaks of large solutes since a great part of the column only acts as void (extra) volume. Such a theo...

The aim of this study was to assess the interlaboratory reproducibility of ultra‐high performance supercritical fluid chromatography coupled with tandem mass spectrometry method for routine antidoping analyses. To do so, a set of 21 doping agents, spiked in urine and analyzed after dilute and shoot treatment, was used to assess the variability of t...

In the first part of the series, it was demonstrated that very fast (<30 s) separations of therapeutic protein species are feasible using ultra-short (5 × 2.1 mm) columns. In the second part, our purpose was to find the appropriate column length; therefore, a systematic study was performed using various custom-made prototype reversed-phase liquid c...

The full text of the paper entitled: The analysis of cannabinoids in cannabis samples by supercritical fluid chromatography and ultra-high-performance liquid chromatography: A comparison study is available at the following website permalink: http://hdl.handle.net/2268/253874 After accepting the ORBI license and if your request is accepted you will...

The use of unorthodox temperatures, ranging from -5°C up to 80°C, have been thoroughly investigated in supercritical fluid chromatography. To this purpose, an initial evaluation of the kinetic and thermodynamic performance has been made with a set of 4 analytes eluting at different percentages of organic co-solvent in the mobile phase (3% - 10% - 4...

Reversed-phase liquid chromatography (RPLC) is one of the most widely used chromatographic approaches for the characterization of therapeutic proteins because it is able to answer several analytical queries and allow the fine characterization of the structural heterogeneity of these drugs. However, some issues may possibly occur when analysing ther...

In this work, the impact of biological matrices, such as plasma and urine, was evaluated under SFC-HRMS in the field of metabolomics. For this purpose, a representative set of 49 metabolites were selected. The assessment of the matrix effects (ME), the impact of biological fluids on the quality of MS/MS spectra and the robustness of the SFC-HRMS me...

Background: Cetuximab (CTX) is a glycosylated anti-EGFR monoclonal antibody of great interest in the treatment of non-melanoma skin cancers. Its intravenous administration is associated with severe side effects. This is the first report on the non-invasive iontophoretic targeted topical delivery of CTX to skin. Methods: Iontophoretic transport of C...

A novel column-coupling approach is suggested to improve both the selectivity and efficiency of protein separations in liquid chromatography. Protein separations often suffer from limited selectivity or not appropriate resolving power. For a new biopharmaceutical product, the identification of the main and minor variant species is required. For tha...

Along with surgery and chemotherapy, radiation therapy (RT) is an important modality in cancer treatment, and the development of radiosensitizers is a current key challenge in radiobiology to maximize RT efficiency. In this study, the radiosensitizing effect of a natural compound from the withanolide family, withanolide D (WD), was assessed. Clonog...

Reversed phase liquid chromatography (RPLC) is a widely used technique for the analytical characterization of proteins biopharmaceuticals, due to its inherent compatibility with mass spectrometry (MS). However, this chromatographic mode suffers from a limited selectivity when analyzing large molecules. Due to the on/off mechanism observed with larg...

Supercritical fluid chromatography (SFC) has recently experienced renovated impulse from research groups. Its hyphenation to mass spectrometers (MS) proved to be of significant importance in catalysing interest from researchers. In contrast to liquid chromatography (LC), the coupling of SFC-MS requires the use of an interface in order to deal effic...

Along with surgery and chemotherapy, radiation therapy (RT) is an important modality in cancer treatment, with about half of cancer patients receiving RT during their treatment. However, there are still a lot of challenges to improve its efficiency by minimizing normal tissue toxicity while maximizing tumor control. In this perspective, radiosensit...

In this paper, the benefits of using columns packed with particles of decreasing size (particle size gradient) in liquid chromatography was investigated from a theoretical point of view. It is indeed well known that such columns may be useful in gradient elution, since the decrease of particle size along the chromatographic column can provide extra...

There is an extensive body of literature linking ADHD to overweight and obesity. Research indicates that impulsivity features of ADHD account for a degree of this overlap. In comparison, the neural and polygenic correlates of this association are less examined. In 874 participants of IMAGEN study, we examined whether phenotypic and genetic liabilit...

This chapter describes the application of modern supercritical fluid chromatography (SFC), coupled with mass spectrometry (MS), for the analysis of various classes of doping agents. Even if liquid chromatography (LC) and gas chromatography (GC) remain the reference techniques for doping control analysis, SFC appears as a promising strategy for the...

The pharmaceutical industry is one of the most regulated industries worldwide, since the drug products have to be safe and effective. The levels of impurities and degradation products in active pharmaceutical ingredients (APIs) have to be strictly controlled and meet the specifications required by international authorities. Chromatography has been...

This review paper discusses the success of columns packed with superficially porous particles (SPP) in liquid chromatography for the analysis of peptides and proteins. First, it summarizes the history of SPP, including the development of different SPP generations from particles of 50 μm to sub-2 μm. It also critically discusses the improved kinetic...

Modern size exclusion chromatography (SEC) can be defined by the use of relatively small columns (e.g., 150 × 4.6 mm) packed with sub-3 μm particles, allowing a 3- to 5-fold increase in method throughput compared to that of conventional SEC. The quick success of the first sub-2 μm SEC column introduced in 2010 led to the development of numerous ult...

The determination of mAb critical quality attributes (CQA) is crucial for their successful application in health diseases. A generic CZE method was developed for the high‐resolution separation of various mAb charge variants, which are often recognized as important CQA. A dynamic coating of the capillary was obtained with polyethylene oxide (PEO), w...

The aim of this study was to evaluate the suitability of SFC-MS for the analysis of a wide range of compounds including lipophilic and highly hydrophilic substances (log P values comprised between -6 and 11), for its potential application toward human metabolomics. For this purpose, a generic unified chromatography gradient from 2 to 100% organic m...

The MS hyphenation performance of UniSpray®, a new design of atmospheric pressure ionization source was evaluated in both SFC and RPLC modes. Sensitivity, stability, versatility and matrix effects offered by the UniSpray were assessed in positive and negative ionization modes and systematically compared to an electrospray source (ESI) using 120 nat...

Mass spectrometry (MS) is considered today as one of the most popular detection methods, due to its high selectivity and sensitivity. In particular, this detector has become the gold standard for the analysis of complex mixtures such as biological samples. The first successful SFC-MS hyphenation was reported in the 80's, and since then, several ion...

A generic liquid chromatographic method development workflow was developed and successfully applied to the analysis of phytocannabinoids and Cannabis sativa extracts. Our method development procedure consists in four steps: i) The screening of primary parameters (i.e. stationary phase nature, organic modifier nature and approximate mobile phase pH)...

Size exclusion chromatography (SEC) is a historical technique, routinely applied for the separation of species possessing different molecular masses (sizes). It is considered as a reference method for the qualitative and quantitative analysis of protein aggregates. In the last few years, several improvements were brought to high performance SEC (HP...

Matrix effects (ME) is acknowledged as being one of the major drawbacks of quantitative bioanalytical methods, involving the use of liquid chromatography coupled to mass spectrometry (LC-MS). In the present study, the incidence of ME in SFC-MS/MS and LC-MS/MS in the positive mode electrospray ionization (ESI+) was systematically compared for the an...

Due to its high sensitivity and selectivity, mass spectrometry (MS) is considered today as one of the most versatile and powerful detection methods. This detector is considered as the gold standard for analytical separations dealing with complex mixtures and biological matrices. Since the first report of supercritical fluid chromatography-mass spec...

A liquid chromatography with tandem mass spectrometry method was developed for the determination of 27 glutamyl di- and tripeptides in food ingredients. Such compounds are of importance for the food industry, as they can modulate the perception of basic tastes (sweet, salty, and umami). Due to their high polarity, the hydrophilic interaction chroma...

Ion mobility spectrometry is an analytical technique known for more than 100 years, which entails separating ions in the gas phase based on their size, shape and charge. While ion mobility spectrometry alone can be useful for some applications (mostly security analysis for detecting certain classes of narcotics and explosives), it becomes even more...

Natural products are an important reservoir for the discovery of pharmacologically active drugs. Concomitantly, interest for natural products has gained importance in the food industry thanks to the better understanding of both absorption and metabolism of phytochemicals in humans. Current efforts are invested to develop functional foods and nutrac...

This paper focuses on the possibility to inject large volumes (up to 10 μL) in ultra-high performance supercritical fluid chromatography (UHPSFC) under generic gradient conditions. Several injection and method parameters have been individually evaluated (i.e. analyte concentration, injection volume, initial percentage of co-solvent in the gradient,...

Antibody Drug Conjugates (ADCs) are innovative biopharmaceuticals gaining increasing attention over the last two decades. The concept of ADCs lead to new therapy approaches in numerous oncological indications as well in infectious diseases. Currently, around 60 CECs are in clinical trials indicating the expanding importance of this class of protein...

Plant secondary metabolites are an almost unlimited reservoir of potential bioactive compounds. In view of the wide chemical space covered by natural compounds, their comprehensive analysis requires multiple and complementary approaches. In this study, numerous chromatographic conditions were tested for the analysis of a set of 120 representative n...

The separation performance of a chromatographic system is often described in terms of column efficiency and peak capacity. Thanks to the new developments in column technology over the past few years, the achievable peak capacity drastically improved and the analysis time can be significantly shortened. Indeed, highly efficient wide-pore reversed-ph...

Reversed phase liquid chromatography (RPLC) is the gold standard analytical strategy in the pharmaceutical industry. In this contribution, we will describe two alternative chromatographic approaches, namely hydrophilic interaction chromatography (HILIC) and supercritical fluid chromatography (SFC), which are more and more commonly used in pharmaceu...

The free spreadsheet-based program HPLC Teaching Assistant was developed for effective and innovative learning and teaching of liquid chromatography. This software allows teachers to illustrate the basic principles of high performance liquid chromatography (HPLC) using virtual chromatograms (simulated chromatograms) obtained under various analytica...

To separate proteins solely based on their difference in hydrodynamic volume in size exclusion chromatography (SEC), the ionic strength of the mobile phase has to be increased in order to avoid secondary ionic interactions between proteins and the stationary phase. However, adding salts to the mobile phase can have a serious effect on protein aggre...

The goal of this study was to better understand the chromatographic conditions in which monoclonal antibodies (mAbs) of broad hydrophobicity scale and a cysteine conjugated antibody-drug conjugate (ADCs), namely brentuximab-vedotin, could denaturate. For this purpose, some experiments were carried out in HIC conditions using various organic modifie...

This review summarizes the use of computer assisted liquid chromatographic method development for the analytical characterization of protein biopharmaceuticals. Several modes of chromatography including reversed-phase liquid chromatography (RPLC), ion exchange chromatography (IEX), hydrophobic interaction chromatography (HIC) and some perspectives...

This work describes the development of two methods involving supported liquid extraction (SLE) sample treatment followed by ultra-high performance liquid chromatography or ultra-high performance supercritical fluid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS and UHPSFC-MS/MS) for the screening of 43 anabolic agents in human urin...

Hydrophobic interaction chromatography (HIC) is a historical strategy used for the analytical purification and characterization of proteins. Similarly to what can be done in reversed-phase liquid chromatography (RPLC), HIC is able to separate protein species based on their hydrophobicity, but using different conditions. Compared to RPLC, the main b...

A new stationary phase based on wide-pore hybrid silica bonded with amide ligand has been used to explore the utility of HILIC for the analytical characterization of protein biopharmaceuticals. Various, highly-relevant samples were tested, including different insulins, interferon α-2b and trastuzumab. This work shows that HILIC can be successfully...

Secondary metabolites are an almost unlimited reservoir of potential bioactive compounds. In view of the wide chemical space covered by natural compounds, their comprehensive analysis requires multiple cutting-edge approaches. This study evaluates the applicability of ultra-high performance supercritical fluid chromatography coupled to quadrupole-t...

Similar to reversed phase liquid chromatography, basic compounds can be highly challenging to analyze by supercritical fluid chromatography (SFC), as they tend to exhibit poor peak shape, especially those with high pKa values. In this study, three new stationary phase ligand chemistries available in sub –2 μm particle sizes, namely 2-picolylamine (...

This study shows the possibility offered by modern ultra-high performance supercritical fluid chromatography combined with tandem mass spectrometry in doping control analysis. A high throughput screening method was developed for 100 substances belonging to the challenging classes of anabolic agents, hormones and metabolic modulators, synthetic cann...

Three novel hydrophilic interaction chromatography columns packed with bare silica 2.6 μm superficially porous particles were evaluated. These stationary phases undergo a different pretreatment temperature (400, 525 and 900°C) that might influence their kinetic performance and thermodynamic properties. In the first instance, we demonstrated that th...

The free spreadsheet-based program HPLC Teaching Assistant was developed for effective and innovative learning and teaching of liquid chromatography. This software allows teachers to illustrate the basic principles of high performance liquid chromatography (HPLC) using virtual chromatograms (simulated chromatograms) obtained under various analytica...

Part II of this series describes additional features of the HPLC Teaching Assistant software, including the capability to simulate the impact of the mobile-phase temperature on high performance liquid chromatography (HPLC) separations, understand the chromatographic behavior of a mixture of diverse compounds in both isocratic and gradient elution m...

The goal of this study was to evaluate the possibilities offered by a prototype HPLC column packed with ∼2.5μm narrow size distribution sphere-on-sphere (SOS) silica particles bonded with C4 alkyl chains, for the analytical characterization of large biomolecules. The kinetic performance of this material was evaluated in both isocratic and gradient...

The fight against doping in sports has been governed since 1999 by the World Anti-Doping Agency (WADA), an independent institution behind the implementation of the World Anti-Doping Code (Code). The intent of the Code is to protect clean athletes through the harmonization of anti-doping programs at the international level with special attention to...

The recent trends in column technology for reversed-phase liquid chromatography (LC), size-exclusion chromatography (SEC), ion-exchange chromatography (IEX), and hydrophobic interaction chromatography (HIC) for analysis of biopharmaceuticals at the protein level is critically discussed.

Reversed phase liquid chromatography (RPLC) coupled to mass spectrometry (MS) is the gold standard technique in bioanalysis. However, hydrophilic interaction chromatography (HILIC) could represent a viable alternative to RPLC for the analysis of polar and/or ionizable compounds, as it often provides higher MS sensitivity and alternative selectivity...

Latanoprost is a practically insoluble prostaglandin F2α analog considered a first-line agent for glaucoma treatment. From a pharmaceutical point of view, latanoprost is challenging to be formulated as an eye drop due to its poor water solubility and the presence of an ester bond which needs to be cleaved in vivo but maintained unchanged during sto...

Water solubility is a key parameter in drug formulation since it highly influences drug pharmacokinetics and pharmacodynamics. In the past decades, the challenge with poorly water soluble drugs has been growing continuously. As a matter of fact, poorly soluble compounds represent 40% of the top 200 oral drugs marketed in the US, 33% of drugs listed...

Ion-exchange chromatography (IEX) is a historical technique widely used for the detailed characterization of therapeutic proteins and can be considered as a reference and powerful technique for the qualitative and quantitative evaluation of charge heterogeneity. The goal of this review is to provide an overview of theoretical and practical aspects...

The development of greener analytical techniques is a topic of great interest and in this context, supercritical fluid chromatography (SFC) is making an interesting comeback. The present review focuses on the latest developments of this technique. We discuss improvements brought to the chromatographic systems and requirements to achieve full compat...

Since its commercial introduction in 2004, there has been a considerable interest in ultra-high-performance (pressure) liquid chromatography (UHPLC), which dramatically increases the throughput of regular HPLC methods. Although the ability to achieve fast separations and good resolution are the main drivers for the increasing use of UHPLC, we descr...

Antidoping procedures are expected to greatly benefit from untargeted metabolomic approaches through the discovery of new biomarkers of prohibited substances abuse. Endogenous steroid metabolites were monitored in urine samples from a controlled elimination study of testosterone undecanoate after ingestion. A platform coupling ultra-high pressure L...

A capillary isoelectric focusing (cIEF) method was developed for the determination of the ionization constants (pKa) of small molecules. Two approaches used to decrease the electroosmotic flow (EOF) were compared: (i) a hydroxypropylcellulose (HPC) coated capillary in aqueous medium and (ii) the addition of glycerol to act as a viscosifying agent....

Size exclusion chromatography (SEC) is a historical technique widely employed for the detailed characterization of therapeutic proteins and can be considered as a reference and powerful technique for the qualitative and quantitative evaluation of aggregates. The main advantage of this approach is the mild mobile phase conditions that permit the cha...

The recent market release of a new generation of supercritical fluid chromatography (SFC) instruments compatible with state-of-the-art columns packed with sub-2μm particles (UHPSFC) has contributed to the reemergence of interest in this technology at the analytical scale. However, to ensure performance competitiveness of this technique with modern...

Biomarkers in amniotic fluid (AF) include both non-modified and phosphorylated proteins and can be used in the diagnosis of pregnancy-associated pathologic conditions. In this work, an integrated LC-MS method for selective, sensitive and reproducible analysis of phosphorylation in proteins has been applied to AF. Online digestion of (phospho) prote...