EU Regulation 396/2005: Pesticide Residue Testing and the Critical Role of QuEChERS Evaporation in Food Safety

<div><div><p><strong>Researchers led by </strong><a target="_blank" rel="noopener noreferrer" href="https://kudova.group.uochb.cz/en"><strong>Eva Kudová</strong></a><strong> from IOCB Prague and Jitka Viktorová from UCT Prague have identified new androstane-based steroid derivatives that can significantly improve the effectiveness of existing antibiotics against multidrug-resistant </strong><i><strong>Staphylococcus aureus</strong></i><strong>.</strong></p></div></div><div><p>The team screened dozens of natural steroidal hormones and synthetic derivatives to see whether they could block bacterial efflux pumps – mechanisms that bacteria use to eject antibiotics and survive treatment. Two compounds emerged as particularly promising: they inhibited efflux pumps and lowered the effective doses of antibiotics such as ciprofloxacin and erythromycin.</p><p>Further analysis showed that one of the compounds also suppressed genes linked to bacterial virulence. Another key finding is that the most active molecules are non-toxic to human immune cells and showed no detectable endocrine activity, suggesting a favorable safety profile.</p><p>These findings support the idea that rationally designed steroid-based molecules could act as antibiotic adjuvants, helping existing drugs overcome bacterial resistance.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/IOCB_Prague_Steroidal_hormones_and_neurosteroids_novel_therapeutic_strategies_in_bacterial_infections_8e5d2a7b48.jpg" alt="IOCB Prague: Steroidal hormones and neurosteroids – novel therapeutic strategies in bacterial infections" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_IOCB_Prague_Steroidal_hormones_and_neurosteroids_novel_therapeutic_strategies_in_bacterial_infections_8e5d2a7b48.jpg 222w," sizes="100vw" width="1258" height="886"></p><blockquote><h4><strong>Original Article</strong></h4><h4><a target="_blank" rel="noopener noreferrer" href="https://www.sciencedirect.com/science/article/pii/S0223523426001613?via%3Dihub"><strong>Steroidal hormones and neurosteroids - novel therapeutic strategies in bacterial infections: Design, synthesis, and biological evaluation</strong></a></h4><p>Daniela Brdová, Bára Křížkovská, Jan Špaček, Zdeněk Míchal, Eva Jablonská, Ondřej Strnad, Hana Chodounská, Eszter Szánti-Pintér, Marina Morozovová, Václav Hanžl, Jan Tkadlec, Martijn Riool, Jan Lipov, Jitka Viktorová, Eva Kudová&nbsp;</p><p><i><strong>Eur. J. Med. Chem., 2026, 308, 118716</strong></i></p><p><a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1016/j.ejmech.2026.118716">https://doi.org/10.1016/j.ejmech.2026.118716</a></p><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/"><strong>CC-BY 4.0</strong></a></h6></blockquote><h4><strong>Abstract</strong></h4><p>The global rise of antibiotic resistance necessitates novel therapeutic strategies for infectious diseases. Inhibition of bacterial efflux pumps, which contribute to multidrug resistance, represents a promising approach to restore or even increase the efficacy of existing antibiotics. Using fluorescence-based ethidium bromide accumulation, broth microdilution, and checkerboard assays, we evaluated 26 endogenous steroidal hormones and neurosteroids, along with 30 synthetic derivatives, for their ability to enhance antibiotic susceptibility in multidrug-resistant <i>Staphylococcus aureus</i>. Structure-activity relationship analysis identified compounds <strong>13</strong> and <strong>16</strong> as lead candidates, exhibiting strong efflux pump inhibition and marked reductions in the minimum inhibitory concentrations of ciprofloxacin and erythromycin. Both compounds showed additive effects in checkerboard assays. Modifications at C-3 (polar substitution) and C-17 (3α,5β-stereochemistry and nonpolar substitution) were essential for potent efflux inhibition and sensitization, although these modifications were not additive when combined. Transcriptome analysis further revealed that compound <strong>13</strong> significantly downregulated <i>S. aureus</i> virulence-associated genes when administered alone or in combination with antibiotics. Cytotoxicity assessment in human peripheral blood mononuclear cells and receptor transactivation assays for estrogen, androgen, and progesterone receptors indicated that the most active derivatives were non-toxic and lacked detectable endocrine activity, suggesting a favorable safety profile. Overall, these findings support the concept that rationally designed andostane-based steroidal scaffolds can function as competitive bacterial efflux pump inhibitors and serve as potential antibiotic adjuvants to mitigate efflux-mediated resistance.</p><h4><strong>Instrumentation</strong></h4><h5><strong>Structural characterization</strong></h5><ul><li><strong>AVANCE III 400 MHz NMR spectrometer</strong> (Bruker)</li><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?mainauthorItems=3&amp;search=LTQ+Orbitrap+XL"><strong>LTQ Orbitrap XL HRMS</strong></a> (Thermo Fisher Scientific, ESI)</li></ul></div>

Steroidal hormones and neurosteroids – novel therapeutic strategies in bacterial infections

<p><strong>Metformin is currently the first-line oral drug of choice for the treatment of type 2 diabetes. It reduces glucose production in the liver, improves tissue sensitivity to insulin, and does not cause weight gain or hypoglycemia, which sets it apart from some other antidiabetic drugs. Moreover, its long-term beneficial effects on the risk of cardiovascular complications and mortality in people with type 2 diabetes have been well established. Researchers from the Institute of Physiology CAS (IPHYS) have identified a new group of blood metabolites that could help physicians objectively monitor whether patients with type 2 diabetes are taking their metformin as prescribed.</strong></p><h4><strong>New biomarkers for metformin use</strong></h4><p>A team led by Tomáš Čajka from the Laboratory of Translational Metabolism at IPHYS analyzed plasma samples from <strong>637 patients with type 2 diabetes treated with metformin</strong> and <strong>143 non-diabetic controls</strong> using advanced metabolomics and lipidomics. Among 614 detected metabolites, they identified five from the group of <strong>N-lactoyl-amino acids </strong>(e.g. N‑lactoyl-phenylalanine), which had previously been seen as unknown signals due to missing reference spectra in databases. The levels of these metabolites in the blood closely reflected the plasma concentrations of metformin. In patients with the highest metformin exposure, the concentrations of these metabolites were up to 7.2 times higher than in controls.</p><h4><strong>A sensitive “fingerprint” of the drug’s effect</strong></h4><p>N-Lactoyl-amino acids are formed in cells by combining lactate and amino acids, thus reflecting the metformin-induced changes in cellular energy metabolism. “<i>Importantly, even though they occur in plasma only at nanomolar concentrations, they reliably distinguish patients who take metformin regularly from those who take it irregularly or not at all</i>,” emphasizes T. Čajka.</p><h4><strong>Metabolomics in practice and international collaboration</strong></h4><p>The study also provides a detailed description of broader metabolic changes in type 2 diabetes, including increases in lactate, organic acids, and branched-chain amino acids, as well as extensive remodelling of the lipid profile. In addition to T. Čajka’s team, collaborators from the Institute for <strong>Clinical and Experimental Medicine in Prague</strong> and the <strong>West Coast Metabolomics Center</strong> (University of California, Davis) also participated in the study.</p><p>The results demonstrate that advanced metabolomics approaches developed at IPHYS within the <strong>CarDia project</strong> can find highly practical applications both in clinical studies and in personalized monitoring of treatment in patients with diabetes.</p><blockquote><h4><strong>The original article</strong></h4><h4><a target="_blank" rel="noopener noreferrer" href="https://ascpt.onlinelibrary.wiley.com/doi/10.1002/cpt.70205"><strong>Untargeted Metabolomics Identifies N-Lactoyl-Amino Acids as Dose-Responsive Plasma Biomarkers of Metformin Adherence in Type 2 Diabetes</strong></a></h4><p>Tomas Cajka, Jiri Hricko, Lucie Rudl Kulhava, Michaela Paucova, Michaela Novakova, Veronika Hola, Stanislava Rakusanova, Oliver Fiehn, Vojtech Skop, Ivana Lankova, Iva Miskova, Terezie Pelikanova, Martin Haluzik</p><p>Clin Pharmacol Ther, 119: 1057-1069</p><p><a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1002/cpt.70205">https://doi.org/10.1002/cpt.70205</a></p><h6>licensed under <a target="_blank" rel="noopener noreferrer" href="https://creativecommons.org/licenses/by/4.0/"><strong>CC-BY 4.0</strong></a></h6></blockquote><h4><strong>Abstract</strong></h4><p>Metformin is the most widely prescribed antidiabetic drug, yet adherence remains difficult to objectively assess. Using untargeted metabolomics and lipidomics, we analyzed plasma from 637 patients with type 2 diabetes (T2D) with confirmed metformin use and 143 nondiabetic controls, annotating 614 metabolites. Patients were stratified by plasma metformin into sub-therapeutic, therapeutic, and supra-therapeutic groups, and associations were evaluated by multiple linear regression and composite metabolite ranking. Five previously unannotated features were structurally identified as N-lactoyl-amino acids, whose levels correlated strongly with plasma metformin (ρ = 0.42–0.55, P &lt; 0.0001) and increased up to 7.2-fold in the supra-therapeutic group (&gt; 2000 ng/mL). While N-lactoyl-amino acids were consistently detected in the nanomolar range, they still displayed robust and dose-dependent associations with metformin. Broader metabolic changes in T2D included elevated lactate, organic acids, and branched-chain amino acids, together with reduced urea cycle metabolites. Lipidomics showed increases in saturated triacylglycerols and diacylglycerols and decreases in cholesteryl esters, sphingomyelins, and phospholipids. These findings establish N-lactoyl-amino acids as robust, dose-responsive plasma biomarkers of metformin exposure. Despite being up to four orders of magnitude less abundant than their amino acid precursors, they sensitively reflect mitochondrial lactate overflow and pharmacodynamic adaptation, offering objective assessment of adherence.</p>

New blood biomarkers reveal how faithfully diabetes patients take metformin

Article

Product

Environment

Food

Health

Scientific article

Science and research

Laboratories

The Czech Academy of Sciences (CAS) was established by Act No. 283/1992 Coll. as the Czech successor of the former Czechoslovak Academy of Sciences. It is set up as a complex of 54 public research institutions.

Czech Academy of Sciences

The Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences (IOCB Prague) is a leading scientific institution in the Czech Republic, recognized internationally. Its primary mission is basic research in the fields of chemical biology and medicinal chemistry, organic and material oriented chemistry, chemistry of natural compounds, biochemistry and molecular biology, physical chemistry, theoretical chemistry, and analytical chemistry. 

Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences

Organomation, founded in 1959, designs and manufactures high-quality nitrogen evaporators and extraction systems. Known for innovation and durability, their lab instruments are used globally for efficient sample preparation with strong customer support.

Organomation

Modern HPLC separations in theory and practice

This book is a unique combination of theory and practice, but also overviews of stationary phases, sample prep materials, and vendor’s brief presentations.

The Fitness for Purpose of Analytical Methods

The third edition of Eurachem’s flagship guide provides updated, practical guidance on method validation, verification, sampling, and fitness for purpose, aligned with ISO/IEC 17025:2017.

<h3><strong>The Fitness for Purpose of Analytical Methods: A Laboratory Guide to Method Validation and Related Topics: Third edition (2025)</strong></h3><p><strong>Author: </strong>H. Cantwell</p><p>This guide was first issued in 1998 and the second edition published in 2014. Over the years it has been one of the most popular of the Eurachem guides. Since the release of the first edition, however, there have been many changes in terminology, working practices, reference documents and requirements. This third edition, produced by the Eurachem Method Validation Working Group, forms a thorough revision of the 2014 edition.</p><p>This third edition includes a section on sampling and sample handling as <strong>ISO/IEC 17025:2017</strong> introduced a focus on sampling associated with the testing or calibration being carried out by the laboratory and further guidance on assessing the calibration function has been included. The Guide is also now supported by a number of supplementary guidance documents – please see the section on supplementary guidance below.</p><h4><strong>Contents</strong></h4><p>This third edition includes guidance on:</p><ul><li>The importance of fitness for purpose;</li><li>The concept of method validation and verification;</li><li>How a method validation or verification study should be performed and how much should be done;</li><li>Sampling and sample handling in relation to method validation;</li><li>A thorough explanation of the various validation parameters (performance characteristics) and related topics;</li><li>Follow-up on the validation study (reporting, use of performance data in Internal Quality Control);</li><li>Documentation of analytical methods.</li></ul><p>Annexes also describe the statistical basis of detection limits and analysis of variance for precision studies, and add a check list to aid users in selecting and determining the validity of a test kit.</p><p>In preparing this guidance, the Working Group has aimed at the right balance between giving a solid theoretical background for method validation studies and providing practical guidelines on how to plan, perform and evaluate such studies in the laboratories.</p><p>The approach is generic in the sense that the Guide does not focus on particular fields of application (food, environment, pharmaceuticals etc.). However, the guide includes references to a number of specific guides in fields where different practices have been developed and become common.</p><p>The guide also includes a full bibliography. Further literature references on method validation can be found in the section "Validation of analytical methods" in the Reading list on this website.</p>

Terminology in Analytical Measurement: Introduction to VIM 3

This Eurachem guide introduces VIM 3 terminology relevant to analytical laboratories, explaining key metrological concepts for chemical, biological, and clinical measurements with practical examples.

<h3><strong>Terminology in Analytical Measurement: Introduction to VIM 3 (Second Edition 2023)</strong></h3><p><strong>Author: </strong>V. J. Barwick (Ed.)</p><h4><strong>Contents</strong></h4><p>This guide has been produced by members of the Eurachem Education and Training Working Group and others co-opted to the Project group for this task.</p><p>In the world of metrology – the science of measurement and its application – there is a language which has to be learned. The International Vocabulary of Metrology (VIM) was produced to provide a common language, primarily for physical measurements. The VIM 3 is a consistent set of concepts each described by a unique term, the ‘label’ of the concept. VIM 3 is applicable across all scientific disciplines thus making it relevant to those involved in performing measurements in chemistry and biology. Consistent definitions of concepts with their associated terms and symbols are essential if analysts and customers across the globe are to understand each other.</p><p>This Eurachem Guide discusses a selection of the concepts in VIM 3, focusing on those most likely to be encountered in analytical laboratories. It aims to cover chemical, biological and clinical measurements and gives examples over and above those in the notes accompanying the definitions in VIM 3. The guide is intended for laboratory staff, accreditation bodies, for those commissioning measurements and for those using measurement results. Lecturers and trainers may also find this guide useful when teaching aspects of metrology.</p><p>In this second edition the scope and structure, and the terms and concepts discussed, remain unchanged from the first edition. However, all sections have been reviewed and, where necessary, the text has been revised to improve clarity and ensure consistency with current international guidance at the time of publication.</p>

Products from category Educational literature - page 1

Related content

New blood biomarkers reveal how faithfully diabetes patients take metformin

Steroidal hormones and neurosteroids – novel therapeutic strategies in bacterial infections

EU Regulation 396/2005: Pesticide Residue Testing and the Critical Role of QuEChERS Evaporation in Food Safety