<p><strong>Scientists at IOCB Prague are uncovering new details of gene transcription. They have identified a previously unknown molecular mechanism by which the transcription of genetic information from deoxyribonucleic acid (DNA) into ribonucleic acid (RNA) can be initiated. The researchers focused on a specific class of molecules known as alarmones, which are found in cells across a wide range of organisms and whose levels often increase under conditions of cellular stress. The results were published in the prestigious scientific journal </strong><i><strong>Nature Chemical Biology</strong></i><strong>.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_bc2bfffa6c.jpg" alt="IOCB Prague/Tomáš Belloň: Research from IOCB Prague reveals a previously unknown mechanism of genetic transcription: Dr. Hana Cahová, head of the Chemical Biology of Nucleic Acids research group." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_bc2bfffa6c.jpg 234w," sizes="100vw" width="2000" height="1335"></p><div><p>Molecules of ribonucleic acid can carry a variety of chemical modifications at one end, known as caps. In eukaryotic organisms, including human cells, the best-known cap plays an important role in RNA stability and in regulating its subsequent fate. In recent years, however, it has become clear that alternative, non-canonical RNA caps also exist, although their formation and the mechanisms by which they are attached to RNA remain only partially understood. These include alarmone caps, formed by dinucleoside polyphosphate molecules, that protect cellular RNA at moments when the cell is under threat.</p><p>In their study,<strong> </strong><a target="_blank" rel="noopener noreferrer" href="https://cahova.group.uochb.cz/en"><strong>Dr. Hana Cahová</strong></a> and colleagues examined bacterial RNA polymerase and investigated how this enzyme can initiate transcription using dinucleoside polyphosphates (NpNs) instead of the standard RNA building blocks. For the first time, the scientists described, at the atomic level, how RNA bearing an alarmone cap can be generated directly at the initiation of gene transcription. They also observed that NpNs bind through a different type of base pairing than is typically observed.</p><p>The work was significantly shaped by <strong>Valentina Serianni from Hana Cahová’s team</strong>, who demonstrated the ability of dinucleoside polyphosphates to initiate gene transcription, and by <strong>Jana Škerlová</strong>, who focused on the structural analysis of RNA polymerase. Using cryogenic electron microscopy data, she showed how dinucleoside polyphosphate molecules bind within the active site of RNA polymerase – that is, the core of the enzyme where genetic information is transcribed. Together, their findings help to elucidate the processes that accompany the transcription of genetic information from DNA to RNA.</p><p>Hana Cahová adds: <i>“We’re describing something that truly occurs in cells and that we’re now able to observe directly at the level of individual molecules. This allows us to answer fundamental questions about cellular processes, such as how cells adapt to stress. RNA plays a central role in this, as it carries the cascade of information underlying any cellular response – for example, to threatening conditions caused by nutrient deprivation or temperature shock.”</i></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_2_0fe88ff9e9.jpg" alt="IOCB Prague/Tomáš Belloň: Research from IOCB Prague reveals a previously unknown mechanism of genetic transcription: Dr. Tomáš Kouba, head of the Cryogenic Electron Microscopy core facility." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_2_0fe88ff9e9.jpg 234w," sizes="100vw" width="2000" height="1335"></p><div><p>Cryogenic electron microscopy (cryo-EM) was central to the project. This part of the work was led by<strong> Dr. Tomáš Kouba</strong>, one of the authors of the study. <i>“Cryogenic electron microscopy allows us to freeze biological molecules in a state very close to their natural form and then determine their three-dimensional structure. This makes it possible to look directly into the active centers of enzymes and observe their function down to the atomic level,”</i> he explains.</p><p>Last year, IOCB Prague opened a new <a target="_blank" rel="noopener noreferrer" href="https://cryoem.group.uochb.cz/en"><strong>cryo-EM center</strong></a><strong> </strong>that is unique within the Czech research environment. State-of-the-art cryogenic electron microscopes are housed in a specially designed building and enable the study of biological processes with exceptional structural precision.</p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_3_4d881e7a58.jpg" alt="IOCB Prague: Research from IOCB Prague reveals a previously unknown mechanism of genetic transcription: This work shows how NpnNs initiate bacterial transcription. Cryo-EM reveals how nucleobases pair with the template: one in canonical and one in reverse Watson–Crick mode, providing a structural basis for their function as non-canonical RNA caps." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_UOCHB_Vyzkum_z_UOCHB_odhaluje_dosud_nepoznany_zpusob_prepisu_geneticke_informace_3_4d881e7a58.jpg 245w," sizes="100vw" width="2000" height="1077"></p><blockquote><h4><strong>The Original article</strong></h4><h4><a target="_blank" rel="noopener noreferrer" href="https://www.nature.com/articles/s41589-025-02134-5"><strong>Molecular insight into 5′ RNA capping with Np<sub>n</sub>Ns by bacterial RNA polymerase</strong></a></h4><p>Valentina M. Serianni, Jana Škerlová, Anna Knopp Dubánková, Anton Škríba, Hana Šváchová, Tereza Vučková, Anatolij Filimoněnko, Milan Fábry, Pavlína Řezáčová, Tomáš Kouba &amp; Hana Cahova&nbsp;</p><p>Nat. Chem. Biol. 21 (2026)</p><p><a target="_blank" rel="noopener noreferrer" href="https://doi.org/10.1038/s41589-025-02134-5">https://doi.org/10.1038/s41589-025-02134-5</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>RNA capped with dinucleoside polyphosphates has been discovered in bacteria and eukaryotes only recently. The likely mechanism of this specific capping involves direct incorporation of dinucleoside polyphosphates by RNA polymerase as noncanonical initiating nucleotides. However, how these compounds bind into the active site of RNA polymerase during transcription initiation is unknown. Here, we explored transcription initiation in vitro, using a series of DNA templates in combination with dinucleoside polyphosphates and model RNA polymerase from <i>Thermus</i> <i>thermophilus</i>. We observed that the transcription start site can vary on the basis of the compatibility of the specific template and dinucleoside polyphosphate. Cryo-electron microscopy structures of transcription initiation complexes with dinucleoside polyphosphates revealed that both nucleobase moieties can pair with the DNA template. The first encoded nucleotide pairs in a canonical Watson–Crick manner, whereas the second nucleobase pairs noncanonically in a reverse Watson–Crick manner. Our work provides a structural explanation of how dinucleoside polyphosphates initiate RNA transcription.</p></div></div>

Research from IOCB Prague reveals a previously unknown mechanism of genetic transcription

<p><strong>Food adulteration is a phenomenon with a long history that has gained increasing importance in recent years due to the globalization of food supply chains and strong economic pressures. Whether it involves diluting olive oil with cheaper oils, adding sugar to honey, or the hidden use of substitutes in protein supplements, the result is a loss of consumer trust and, in some cases, a potential risk to health. Analytical chemistry plays a key role in this area by providing methods and technologies capable of identifying and quantifying target compounds as well as unwanted impurities in a wide range of food matrices.</strong></p><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Praha_Falsovani_potravin_a_jak_ho_odhali_analyticka_chemie_6912db5492.jpg" alt="Watrex: Food Adulteration and How It Is Revealed by Analytical Chemistry" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Praha_Falsovani_potravin_a_jak_ho_odhali_analyticka_chemie_6912db5492.jpg 245w," sizes="100vw" width="1600" height="900"></p><h4><strong>The Role of Chromatography in Forensic Food Analysis</strong></h4><p>Chromatography, particularly liquid chromatography (HPLC), is one of the most effective analytical tools for separating complex mixtures of molecules. It enables:</p><ul><li>qualitative identification of individual components in a sample,</li><li>quantitative determination of their concentrations,</li><li>detection of impurities and unexpected components that may indicate<br>adulteration.</li></ul><p>In practice, HPLC is often combined with various types of detectors (UV/VIS, DAD, refractive index detectors) or with other techniques (e.g., mass spectrometry) to increase selectivity and sensitivity.</p><h4><strong>Isocratic vs. Gradient Elution</strong></h4><p>In the analysis of food samples, chromatographic methods are selected according to the analytical goal:</p><ul><li><strong>Isocratic HPLC </strong>uses a constant mobile phase composition throughout the analysis. It is simple, robust, and ideal for routine determinations of well-defined target analytes (e.g., sugars or vitamins).</li><li><strong>Gradient HPLC</strong> changes the mobile phase composition during separation, enabling better resolution of complex mixtures with a wide range of polarities (e.g., peptides or phenolic compounds).</li></ul><p>Gradient methods are particularly useful when analytes have very different properties and high separation efficiency is required.</p><h4><strong>Watrex Praha and the Contribution of Modern Systems to Food Analysis</strong></h4><p><a target="_blank" rel="noopener noreferrer" href="https://watrex.com/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_clanky_HPLCsystemy"><strong>Watrex Praha, s.r.o.</strong></a> is one of the leading Czech manufacturers and distributors of liquid chromatography systems and instruments, with a long tradition dating back to 1990. The company has successfully transformed from a distributor into a manufacturer of its own HPLC and related technologies, focusing on modular, robust systems suitable even for demanding analytical tasks.</p><p>Their <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products?manufacturerItems=93&amp;search=streamline"><strong>Streamline™ product line</strong></a><strong> </strong>includes several HPLC system concepts suitable for food analysis:</p><ul><li><strong>Isocratic HPLC system</strong> – a simple yet precise configuration with a single pump, column thermostat, and UV/VIS detector; suitable for routine determination of known analytes (e.g., basic carbohydrates, food additives).</li><li><strong>Binary gradient HPLC system</strong> – equipped with two pumps enabling mobile phase gradient formation, which enhances separation performance for complex mixtures<br>(e.g., phenolic profiles of wine, components of aromatic extracts).</li><li><strong>GPC/SEC system (Gel Permeation / Size Exclusion Chromatography)</strong> – a specialized solution for the determination of macromolecules and polymers, also relevant for the analysis of high-molecular-weight components in foods (e.g., polysaccharides).</li></ul><blockquote><p class="cke-user_style-important"><strong>👉</strong><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/Watrex_catalog_2024_ba97395f92.pdf"><strong>Product catalogue HERE</strong></a></p></blockquote><p>Thanks to the modular nature of these systems, analysts can easily adapt the configuration to a specific analytical problem, from simple routine analyses to complex characterization of food samples.</p><h4><strong>Practical Examples of Adulteration Revealed by HPLC</strong></h4><ul><li><strong>Detection of olive oil dilution: </strong>HPLC can separate and quantify characteristic triacylglycerols and fatty acids; unusual patterns indicate the presence of cheaper oils.</li><li><strong>Estimation of sugar addition in honey:</strong> Isocratic HPLC with a refractive index detector can determine sugar profiles (glucose, fructose, sucrose), revealing dilution or the addition of sugar syrups.</li><li><strong>Identification of aromatic adulterants in wine:</strong> Gradient HPLC with a PDA detector can reveal phenolic and aromatic profiles that are altered by the addition of synthetic compounds.</li></ul><h4><strong>Conclusion</strong></h4><p>Food adulteration remains a current issue that requires reliable and sensitive analytical methods. Liquid chromatography, and in particular modern HPLC systems, represent one of the most powerful tools for detecting unwanted additives, impurities, and unexpected components in complex food samples. Manufacturers such as <a target="_blank" rel="noopener noreferrer" href="https://watrex.com/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_clanky_HPLCsystemy"><strong>Watrex Praha</strong></a> provide flexible chromatographic platforms covering a wide range of analytical tasks from simple isocratic to advanced gradient and GPC/SEC applications thereby supporting analysts in ensuring food quality and consumer protection.</p><blockquote><p>Did you find this article interesting? Follow <a target="_blank" rel="noopener noreferrer" href="https://watrex.com/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_clanky_HPLCsystemy"><strong>Watrex Praha</strong></a> on their social media (<a target="_blank" rel="noopener noreferrer" href="https://www.linkedin.com/company/watrex/?viewAsMember=true"><strong>LinkedIn</strong></a> and <a target="_blank" rel="noopener noreferrer" href="https://www.facebook.com/WatrexPrague"><strong>Facebook</strong></a>) for more engaging topics.</p></blockquote>

Food Adulteration and How It Is Revealed by Analytical Chemistry

<p><strong>Join us for a fascinating conversation with Dr. Subhoja Chakraborty from the University of Central Florida, whose cutting-edge research targets the global challenge of malaria drug resistance. Dr. Chakraborty shares her journey from structure-based drug design during her PhD to her recent collaborative work repurposing human kinase inhibitors as rapid, selective antimalarials against drug-resistant Plasmodium falciparum strains.</strong></p><h4><strong>In this episode, you'll learn:</strong></h4><ul><li>How structure-guided approaches reveal potential drug targets, including cysteine proteases like falcipain-2, and kinases implicated in malaria's complex life cycle.</li><li>The power of repurposing kinase inhibitors originally developed for cancer and other diseases, focusing on compound 12, which blocks hemozoin formation and protein kinase 6—key steps in parasite survival.</li><li>Lab techniques such as affinity, gel filtration, and size exclusion chromatography for recombinant protein purification—and how troubleshooting is crucial for high-yield, stable complexes.</li><li>Advances in mass spectrometry for characterizing protein interactions, validating drug targets, and mapping molecular pathways pivotal to malaria therapeutics.</li><li>Tips and lessons for new researchers facing the challenges of sample preparation, complex protocols, and working as part of a collaborative team.</li></ul><p>Dr. Chakraborty’s insights bridge the gap between basic science and translational drug discovery, offering hope for more effective, targeted malaria treatments in a world where resistance to frontline therapies threatens millions.</p><h4><strong>Referenced Papers:</strong></h4><ul><li>"Plasmodium falciparum protein kinase 6 and hemozoin formation are inhibited by a type II human kinase inhibitor exhibiting antimalarial activity" (Cell Chemical Biology, 2025)</li><li>"New insights of falcipain 2 structure from Plasmodium falciparum 3D7 strain" (BBRC, 2022)</li><li>"Structure-Based Optimization of Protease−Inhibitor Interactions to Enhance Specificity of Human Stefin‑A against Falcipain‑2 from the Plasmodium falciparum 3D7 Strain" (Biochemistry, 2023)</li></ul><p>If you’re a student, scientist, or simply passionate about advances in infectious disease research, this episode offers actionable insights, inspiring advice, and a front-row seat to innovation in combating malaria.</p><h3><strong>Video Transcription</strong></h3><h4><strong>From Structural Biology to Drug Discovery: Chromatography and Mass Spectrometry in Malaria Research</strong></h4><h5><strong>Introduction</strong></h5><p>In this discussion, Subhoja shares insights into her research journey, focusing on malaria drug discovery and the role of modern analytical techniques in biological and biomedical research. The conversation is aimed primarily at undergraduate students and early-career researchers interested in chemistry, biotechnology, and life sciences.</p><h5><strong>Research Focus: Malaria Drug Discovery and Kinase Targets</strong></h5><p>Subhoja is currently involved in malaria drug discovery research at the University of Central Florida, where she has been working since beginning her postdoctoral position two years ago. Her earlier PhD research focused on structure-based drug design in malaria, using a combination of in silico approaches, recombinant protein expression, and X-ray crystallography to determine precise three-dimensional structures of target proteins and their inhibitors.</p><p>This structural perspective provided a foundation for her current work, which centers on drug discovery through compound screening and repurposing. A recent publication from her laboratory explores the use of repurposed mammalian CDK-like kinase inhibitors as potential antimalarial agents. While such kinase inhibitors are well known in cancer and cardiovascular research, their application to malaria targets represents a novel and promising approach.</p><p>Kinases are attractive drug targets because of their central role in cell signaling and growth pathways. By targeting specific kinases involved in different stages of the malaria parasite life cycle—whether asexual growth, gametocytogenesis, or differentiation—it may be possible to significantly inhibit parasite growth and progression.</p><h5><strong>Chromatography in Protein Expression and Purification Workflows</strong></h5><p>Chromatography plays a critical role throughout Subhoja’s research. Her experience spans from basic chromatography techniques during her early training to advanced protein purification workflows using affinity chromatography, gel filtration (size exclusion chromatography), and ion exchange chromatography.</p><p>Recombinant proteins are routinely purified using affinity resins such as nickel or cobalt, followed by further purification to remove contaminants. Fast Protein Liquid Chromatography (FPLC) enables compact, stepwise purification workflows suitable for producing proteins for in vitro assays.</p><p>A recurring challenge in this process is the expression of high-molecular-weight proteins, which often accumulate in insoluble fractions or inclusion bodies. Careful selection of expression vectors, host systems, and induction conditions is therefore essential to maximize yield and solubility.</p><h5><strong>Integrating Size Exclusion Chromatography into the Workflow</strong></h5><p>Size exclusion chromatography (SEC) is typically used after affinity purification to further improve protein purity. Following elution with imidazole or glutathione, desalting is required to prevent buffer components from interfering with downstream assays. This is often achieved using desalting columns such as G-25 or PD-10.</p><p>To remove lower-molecular-weight contaminants, finer SEC media—such as Superdex or Sephacryl columns—are employed. Concentration steps using molecular-weight cut-off filters are then applied to obtain the desired protein concentration for functional studies.</p><p>These iterative purification and troubleshooting steps are a routine part of protein biochemistry and reflect the importance of methodical optimization and careful validation.</p><h5><strong>The Role of Mass Spectrometry in Biomolecular Characterization</strong></h5><p>Mass spectrometry is a key analytical tool in Subhoja’s research pipeline, particularly for target identification and validation. In both malaria and cancer research, MS is used alongside chromatography, immunoprecipitation, and pull-down assays to study protein interactions and molecular recognition events.</p><p>Affinity-based mass spectrometry enables the identification of binding partners and interactors, whether studying proteins, long non-coding RNAs, or protein complexes. Techniques such as native MS, tandem affinity purification, and proximity labeling (e.g., TurboID-based workflows) provide deeper insights into molecular networks and signaling pathways.</p><p>These approaches are essential for understanding how candidate drug targets function within complex biological systems.</p><h5><strong>Sample Preparation Challenges in Biological Research</strong></h5><p>Sample preparation remains one of the most challenging aspects of biological experimentation. Whole-cell lysates contain proteases, nucleases, and other components that can rapidly degrade target molecules. Preventing degradation requires minimizing preparation time, maintaining low temperatures, and using appropriate inhibitor cocktails.</p><p>Reproducibility is also critical: biological experiments require multiple replicates, and variability is often unavoidable. Careful handling, protocol optimization, and continuous troubleshooting are essential to ensure reliable results, whether preparing samples for chromatography, mass spectrometry, sequencing, or immunoprecipitation.</p><h5><strong>Advances in Chromatography and MS for Malaria Research</strong></h5><p>Modern advances in chromatography and mass spectrometry significantly enhance malaria research by enabling precise target identification, quantification, and functional characterization. Drug discovery workflows begin with construct design and recombinant expression, followed by purification and interaction studies.</p><p>In malaria research, stage-specific parasite biology requires careful extraction and analysis of proteins from different life cycle stages. Tandem affinity MS, immunoprecipitation, and in vivo validation studies help confirm target relevance and therapeutic potential.</p><p>Recent work on repurposed kinase inhibitors has shown promising results, including inhibition of hemozoin formation—a key detoxification pathway for the malaria parasite—highlighting the potential of such compounds for further development.</p><h5><strong>Malaria, Drug Resistance, and Global Impact</strong></h5><p>Malaria remains a major global health burden, with over 600,000 deaths annually and widespread drug resistance complicating treatment. Artemisinin resistance, particularly associated with mutations in the K13 propeller domain, has driven the search for new targets and therapies.</p><p>During her PhD, Subhoja focused on falcipain-2, a cysteine protease involved in hemoglobin degradation within the parasite. This pathway is critical for parasite survival and has emerged as a promising drug target. Her work revealed that falcipain-2 operates within a larger protein complex responsible for hemoglobin processing and amino acid release.</p><p>Understanding these molecular mechanisms is essential for developing robust, stage-specific antimalarial strategies.</p><h5><strong>Advice for Early-Career Researchers</strong></h5><p>For researchers just starting out, Subhoja emphasizes the importance of hands-on experience. Working directly with cells, lysates, and analytical tools is essential for truly understanding experimental systems. Failure is an inherent part of research, and perseverance is key.</p><p>Motivation, teamwork, careful sample handling, and thorough preparation all contribute to long-term success. Reading extensively, critically evaluating one’s own work, and appreciating incremental progress are vital aspects of a sustainable research career.</p><p>Every successful experiment—no matter how small—represents meaningful progress and contributes to the broader goals of science and medicine.</p><h5><strong>Closing Remarks</strong></h5><p>The discussion highlights how interdisciplinary approaches combining chromatography, mass spectrometry, structural biology, and biochemistry are driving advances in malaria drug discovery. Through careful experimental design, analytical rigor, and collaborative teamwork, researchers can address complex biological challenges and contribute to global health solutions.</p><blockquote><p><strong>This text has been automatically transcribed from a video presentation using AI technology. It may contain inaccuracies and is not guaranteed to be 100% correct.</strong></p></blockquote><blockquote><h4><strong>Concentrating on Chromatography Podcast</strong></h4><p>Dive into the frontiers of chromatography, mass spectrometry, and sample preparation with host David Oliva. Each episode features candid conversations with leading researchers, industry innovators, and passionate scientists who are shaping the future of analytical chemistry. From decoding PFAS detection challenges to exploring the latest in AI-assisted liquid chromatography, this show uncovers practical workflows, sustainability breakthroughs, and the real-world impact of separation science. Whether you’re a chromatographer, lab professional, or researcher you'll discover inspiring content!</p><p>You can find <strong>Concentrating on Chromatography Podcast </strong>in podcast apps:</p><ul><li><a target="_blank" rel="noopener noreferrer" href="https://open.spotify.com/show/1wZg67rCjoGju4yhqxI3tL"><strong>Spotify</strong></a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podcasts.apple.com/tz/podcast/concentrating-on-chromatography/id1788429185"><strong>Apple podcast</strong></a></li><li><a target="_blank" rel="noopener noreferrer" href="https://podscan.fm/podcasts/concentrating-on-chromatography"><strong>podscan</strong></a></li></ul><p>and on <a target="_blank" rel="noopener noreferrer" href="https://www.youtube.com/@ChromatographyTalk/videos"><strong>YouTube channel</strong></a></p></blockquote>

Affinity, Gel Filtration, Size Exclusion Chromatography: Repurposing Kinase Inhibitors for Malaria

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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

WATREX Prague is the distributor and producer of HPLC techniques in the Czech Republic and Slovakia. Main Chromatografic and Related Techniques Supported: LC, GPC, IC, IEC, Automated SPE, LC Columns and special column packings, LC method development, Software development, Laboratory Robots.

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Bioanalytic Development of LC-MS/MS method

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Bioanalytic Individual LC-MS/MS training

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<h4><strong>BASIC INFORMATION</strong></h4><ul><li>TITLE: <strong>Customized LC-MS/MS Training</strong></li><li>CATEGORY: <strong>LC-MS/MS training</strong></li><li>LOCATION: <strong>Lodz, PL</strong> or other</li><li>TRAINING TIME: <strong>1-5 days</strong> or more</li><li>NUMBER OF PARTICIPANTS: <strong>4-8 people</strong></li><li>EQUIPMENT:<strong> SCIEX QTRAP/QTOF LC-MS/MS</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Individual_LC_MS_MS_training_8a73408c56.jpg" alt="Bioanalytic: Bioanalytic Individual LC-MS-MS training." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Individual_LC_MS_MS_training_8a73408c56.jpg 245w," sizes="100vw" width="1536" height="930"></p><h4><strong>TRAINING DESCRIPTION</strong></h4><p>Customized LC-MS/MS training we are ready to prepare for the needs of participants, discussing:</p><ul><li>theoretical basics of chromatography, mass spectrometry,</li><li>practical operation of the LC-MS/MS system,</li><li>qualitative and quantitative analysis,</li><li>DDA &amp; DIA screening methods,</li><li>“omics” analyses,</li><li>validation of analytical methods,</li><li>solving basic methodological and technical problems,</li><li>software,</li><li>calibration,</li><li>“tips&amp;tricks”,</li><li>etc...</li></ul><p>We offer training for both novices starting the fascinating adventure of LC-MS/MS analytics, as well as for those who are LC-MS/MS users and are interested in expanding and consolidating their knowledge.</p><h4><strong>Framework training program</strong></h4><ul><li>individual LC-MS/MS training is developed according to the customer's demand</li><li>conducting theoretical and/or practical training at the customer's workplace</li><li>carrying out theoretical and/or practical training in a laboratory in Lodz</li></ul>

Bioanalytic Basic LC-MS/MS training

Basic LC-MS/MS training covers the theoretical basics of mass spectrometry and familiarization with the basic operation of the instrument.

<h4><strong>BASIC INFORMATION</strong></h4><ul><li>TITLE: <strong>Basic LC-MS/MS training</strong></li><li>CATEGORY: <strong>LC-MS/MS training</strong></li><li>LOCATION: <strong>Lodz, PL</strong></li><li>TRAINING TIME: <strong>3 days</strong></li><li>NUMBER OF PARTICIPANTS: <strong>up to 5 people</strong></li><li>EQUIPMENT:<strong> SCIEX QTRAP </strong><span class="ts-pf-details-content"><strong>&amp; </strong></span><strong>QTOF LC-MS/MS</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Basic_LC_MS_MS_training_fb1382a55b.jpg" alt="Bioanalytic: Bioanalytic Basic LC-MS-MS training." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Basic_LC_MS_MS_training_fb1382a55b.jpg 205w," sizes="100vw" width="1293" height="987"></p><h4><strong>TRAINING DESCRIPTION</strong></h4><p>Basic LC-MS/MS training covers the theoretical basics of mass spectrometry and familiarization with the basic operation of the instrument. Basic LC-MS/MS training is primarily aimed at those who are new to LC-MS/MS equipment or who wish to become familiar with this analytical technique. During the training, the following will be presented:</p><ul><li>principles of developing and optimizing quantitative and qualitative methods on low (triple quadrupole, QTRAP) and high resolution (QTOF) instruments,</li><li>basic functionality of the software,</li><li>procedures and objectives of instrument calibration,</li><li>principles of maintaining the cleanliness of mass spectrometer operation.</li></ul><h5><strong>Basic LC-MS/MS training - the program</strong></h5><p><span style="font-size:20px;"><strong>Day I</strong></span></p><p><strong>Theoretical classes:</strong></p><ul><li>Introduction to LC</li><li>Introduction to mass spectrometry (basics)</li><li>Quantitative analysis.</li></ul><p><strong>Practical part:</strong></p><p>QTRAP:</p><ul><li>QTRAP scanning modes on the instrument - Q1, , MS2, MRM, demonstration and discussion on the example of a selected pesticide of the fragmentation spectrum and its fragmentation pathway.</li><li>MRM pair formation (manual, semi-automatic, automatic).</li><li>optimization of ion source parameters -T-conector</li><li>explanation of the main parameters related to LC (column selection, gradient vs isocrat, autosampler)</li><li>LC-MS/MS method setup and optimization</li></ul><p>QTOF:</p><ul><li>manual and automatic optimization of MRM<sup>HR</sup></li><li>ion source and LC parameters (transfer from method developed above)</li></ul><p>Sequence of quantitative analysis on both instruments.</p><p><span style="font-size:20px;"><strong>Day II</strong></span></p><p><strong>Quantitative analysis:</strong></p><ul><li>data processing and analysis,</li><li>reporting - Sciex OS (QTRAP, QTOF).</li><li>Discussion of types of integration algorithms (MQ4, autopeak) and integration parameters.</li><li>independent selection of integration parameters and plotting of master curves.</li><li>independent data analysis and generation of analysis report.</li></ul><p><strong>Qualitative analysis:</strong></p><ul><li>Theoretical introduction - introduction to qualitative analysis:</li><li>qwadrupole modes (Prec/NL),</li><li>trap modes (EMS, EPI, MS3),</li><li>QTOF modes (TOF MS, TOF MS/MS).</li><li>mixed methods,</li><li>targeted vs. untargeted analysis,</li><li>DDA (IDA) - discussion of method development,</li></ul><p><strong>Practical part:</strong></p><ul><li>creation of qualitative methods on QTRAP (Prec-IDA, MRM-IDA), other parameters (LC, ion source) as the previous day,</li><li>LightSight - discussion of operation, creation of MRM-IDA multimethod for selected pesticide and analysis of samples (night).</li><li>QTOF - creation of IDA method, other parameters (LC, ion source) as the previous day.</li></ul><p>Sequence of qualitative analysis overnight.</p><p><span style="font-size:20px;"><strong>Day III</strong></span></p><p><strong>Qualitative data analysis:</strong></p><p>QTRAP:</p><ul><li>Discussion of Prec-IDA, MRM-IDA results - what we see in different types of MS and MS/MS scans. Manual review of data and via IDA Explorer (SciexOS).</li><li>Analysis of results in LightSight, comparison with Prec-IDA, for example - advantages/disadvantages</li></ul><p>QTOF:</p><ul><li>search for potentially present compounds in the sample,</li><li>manual comparison of blank vs. test sample,</li><li>manual by XIC extraction (by sum formula),</li><li>Formula finder - derivation of sum formula on TOF MS data and verification of fragmentation on TOF MS/MS data</li><li>search in IDA Explorer mode - application of filters: similarity, isotopic distribution, mass defect,</li><li>database searching,</li><li>Molecule Profiler - demonstration of software capabilities on selected pesticide example,</li></ul><p><strong>Instrument calibration:</strong></p><ul><li>resolution, mass-to-charge ratio, intensity,</li><li>Q1 and Q3, LIT (MS and MS/MS), TOF MS and TOF-MS/MS</li><li>Automation vs. manual calibration,</li></ul><p><strong>LC-MS/MS application examples</strong></p><p><strong>Summary</strong></p><p><strong>Discussion</strong></p>

Bioanalytic Quantitative LC-MS/MS analysis training

The goal of the training is to introduce users to the procedures and capabilities of the LC-MS/MS system and software to achieve sensitivity, selectivity, precision and reproducibility in quantitative analysis.

<h4><strong>BASIC INFORMATION</strong></h4><ul><li>TITLE: <strong>Quantitative LC-MS/MS analysis training</strong></li><li>CATEGORY: <strong>LC-MS/MS training</strong></li><li>LOCATION: <strong>Lodz, PL</strong></li><li>TRAINING TIME: <strong>3 days</strong></li><li>NUMBER OF PARTICIPANTS: <strong>up to 5 people</strong></li><li>EQUIPMENT:<strong> SCIEX QTRAP LC-MS/MS</strong></li></ul><figure class="image"><img style="aspect-ratio:1200/675;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_Quantitative_LC_MS_MS_analysis_training_d13b5d8127.jpg" alt="Bioanalytic: Bioanalytic Quantitative LC-MS-MS analysis training." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_Quantitative_LC_MS_MS_analysis_training_d13b5d8127.jpg 245w," sizes="100vw" width="1200" height="675"></figure><h4><strong>TRAINING DESCRIPTION</strong></h4><p>Quantitative LC-MS/MS analysis - the training is designed for laboratory personnel using LC-MS/MS systems for quantitative analysis. The following topics are covered during the training:</p><ul><li>Triple quadrupole spectrometer calibration</li><li>MRM pair formation and optimization</li><li>Optimization of ion source parameters</li><li>Optimization of chromatographic parameters</li><li>Sample preparation using the example of blood drawn on <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1283"><strong>Mitra®</strong></a> samplers (VAMS)</li><li>Matrix effect, internal standards (ISTD), matrix curves</li><li>Validation of the quantitative method</li><li>Reporting of results</li></ul><p>The goal of the training is to introduce users to the procedures and capabilities of the LC-MS/MS system and software to achieve sensitivity, selectivity, precision and reproducibility in quantitative analysis while covering the widest possible linear range of concentrations.</p><h3><strong>Training program</strong></h3><h4><span style="font-size:20px;"><strong>Day I</strong></span></h4><h5><strong>Theoretical part:</strong></h5><p>Triple quadrupole structure, ionization, MRM and sMRM scanning, chromatography, multimethods</p><h5><strong>Practical part:</strong></h5><ul><li>Q1/Q3 calibration (10 Da/s), MS/MS - resolution, m/z, intensity.</li><li>MRM pair formation and optimization (manual, semi-automatic, automatic).</li><li>Ion source optimization - T-concector and FIA</li><li>Gradient vs isocrat, dosing, column selection, temperature selection, other important parameters</li><li>Establishment and optimization of LC-MS/MS method.</li><li>How to take care of MS cleanliness - ionization time programming, multi-way valve, cleaning of ion source and instrument front end</li></ul><p>Sample preparation in pure solvent (standard curve, QC, solvent, solvent+ISTD, test samples), overnight analysis sequence.</p><h4><span style="font-size:20px;"><strong>Day II</strong></span></h4><h5><strong>Practical part - preparation of whole blood samples (Mitra®):</strong></h5><ul><li>Preparation of matrix standard curves</li><li>Preparation of quality control (QC) samples in three concentrations</li><li>Preparation of blank samples (without ISTD and with ISTD).</li><li>Preparation of samples for recovery determination (“pre-spiked” and “post-spiked”)</li><li>Preparation of test samples</li></ul><h5><strong>Theoretical part:</strong></h5><ul><li>Samples in the matrix - discussion of the matrix effect and ways to level it.</li><li>Optimization of sample preparation method and LC-MS/MS method</li></ul><h5><strong>Practical part:</strong></h5><ul><li>Sample extraction from Mitra® samplers for LC-MS/MS analysis.</li><li>Sequence of analysis of prepared samples overnight</li></ul><h4><span style="font-size:20px;"><strong>Day III</strong></span></h4><h5><strong>Theoretical part:</strong></h5><ul><li>Validation of the quantitative LC-MS/MS method</li></ul><h5><strong>Practical part:</strong></h5><ul><li>Software - data integration algorithms, signal smoothing, weighing.</li><li>Matrix curves vs curves in solvent, influence of ISTD (Sciex OS).</li><li>Method validation - validation criteria, determination of LOD, LLOQ, ULOQ, linearity, working range of the method, repeatability, recovery.</li><li>Preparation of summary validation reports</li><li>Reporting of results from test samples - report vs export of data to Excel.</li><li>Summary and discussion.</li></ul>

Bioanalytic QTRAP vs QTOF Quality analysis training

The training includes a presentation of the basic differences in application and a comparison of the capabilities of QTRAP and QTOF instruments in qualitative analysis.

<h4><strong>BASIC INFORMATION</strong></h4><ul><li>TITLE: <strong>QTRAP vs QTOF Quality analysis training</strong></li><li>CATEGORY: <strong>LC-MS/MS training</strong></li><li>LOCATION: <strong>Lodz, PL</strong></li><li>TRAINING TIME: <strong>3 days</strong></li><li>NUMBER OF PARTICIPANTS: <strong>up to 5 people</strong></li><li>EQUIPMENT:<strong> SCIEX QTRAP </strong><span class="ts-pf-details-content"><strong>&amp; </strong></span><strong>QTOF LC-MS/MS</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_QTRAP_vs_QTOF_Quality_analysis_training_18ca66163f.jpg" alt="Bioanalytic: Bioanalytic QTRAP vs QTOF Quality analysis training." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_QTRAP_vs_QTOF_Quality_analysis_training_18ca66163f.jpg 221w," sizes="100vw" width="1000" height="707"></p><h4><strong>TRAINING COURSE DESCRIPTION</strong></h4><p><strong>QTRAP vs QTOF qualitative analysis</strong> - the training is aimed at people who perform analyses related to substance identification or metabolite search in their research work. The training includes a presentation of the basic differences in application and a comparison of the capabilities of QTRAP and QTOF instruments in qualitative analysis. In addition to the theoretical session, participants will undergo a course in the development and optimization of <strong>DDA (IDA)</strong> (QTRAP &amp; QTOF) and <strong>DIA (SWATH) </strong>(QTOF) methods. Analysis and comparison of results will be conducted using <a target="_blank" rel="noopener noreferrer" href="https://sciex.com/products/software/sciex-os-software">SciexOS</a>, <a target="_blank" rel="noopener noreferrer" href="https://sciex.com/products/software/markerview-software">MarkerView</a>, <a target="_blank" rel="noopener noreferrer" href="https://sciex.com/products/software/lightsight-software">LightSight</a> and <a target="_blank" rel="noopener noreferrer" href="https://sciex.com/products/software/molecule-profiler-software">MoleculeProfiler</a> software.</p><h5><strong>Training program</strong></h5><h4><span style="font-size:20px;"><strong>Day I</strong></span></h4><h5><strong>Theoretical part</strong></h5><ul><li>Principle of operation of MS/MS systems</li><li>Differences and capabilities of QTRAP and QTOF</li><li>Comparison of typical analytical procedures and their application</li><li>DDA (IDA) vs DIA (SWATH) methodology.</li><li>Supporting software - LightSight, Molecule Profiler, ACD Labs, MarkerView</li><li>General principles of compound fragmentation</li></ul><h4><span style="font-size:20px;"><strong>Day II</strong></span></h4><p><strong>Objective - to develop methods to search for metabolites of a known substance.</strong></p><h5><strong>QTRAP</strong></h5><ul><li>Development of IDA screening methods (pMRM, Prec/NL, EMS)</li><li>Optimization of LC-MS/MS IDA method</li><li>Data acquisition</li></ul><h5><strong>QTOF</strong></h5><ul><li>Development and optimization of LC-MS/MS IDA and SWATH methods</li><li>Data acquisition</li></ul><h4><span style="font-size:20px;"><strong>Day III</strong></span></h4><p><strong>Objective - identification and profiling of metabolites of a known substance</strong></p><h5><strong>QTRAP data analysis</strong></h5><ul><li>Manual search and identification</li><li>LightSight, MarkerView</li></ul><h5><strong>QTOF data analysis</strong></h5><ul><li>Search and data analysis methods</li><li>Manual search and identification</li><li>IDA vs SWATH</li><li>Automated search and identification - SciexOS, databases</li><li>Molecule Profiler, MarkerView</li></ul><p>Summary and discussion<br>&nbsp;</p>

Bioanalytic QTOF quantitative and qualitative analysis training

The purpose of the training is to introduce users to the capabilities of the LC-HR-MS/MS (QTOF) system in quantitative and qualitative analysis.

<h4><strong>BASIC INFORMATION</strong></h4><ul><li>TITLE:&nbsp;<strong> QTOF quantitative and qualitative analysis training</strong></li><li>CATEGORY: <strong>LC-MS/MS training</strong></li><li>LOCATION: <strong>Lodz, PL</strong></li><li>TRAINING TIME: <strong>3 days</strong></li><li>NUMBER OF PARTICIPANTS: <strong>up to 5 people</strong></li><li>EQUIPMENT:<strong> SCIEX</strong><span class="ts-pf-details-content"><strong> </strong></span><strong>QTOF LC-MS/MS</strong></li></ul><p><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_Bioanalytic_QTOF_quantitative_and_qualitative_analysis_training_af1620ae8c.jpg" alt="Bioanalytic: Bioanalytic QTOF quantitative and qualitative analysis training." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_Bioanalytic_QTOF_quantitative_and_qualitative_analysis_training_af1620ae8c.jpg 221w," sizes="100vw" width="1000" height="707"></p><h4><strong>TRAINING COURSE DESCRIPTION</strong></h4><p><strong>Qualitative and quantitative analysis of QTOF</strong> - the training is designed for laboratory personnel using <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library?instrumentationItems=30&amp;mainauthorItems=55"><strong>high-resolution (HR) LC-MS/MS</strong></a> systems for qualitative and quantitative analysis. The following topics are covered during the training:</p><ul><li><strong>QTOF spectrometer calibration</strong></li><li><strong>Targeted and untargeted methods</strong></li><li><strong>Quantitative analysis (TOF MS, MRMhr)</strong></li><li><strong>Qualitative and quantitative analysis - DDA (IDA) vs DIA (SWATH) methodology</strong></li><li><strong>Software and its use in high-resolution data analysis</strong></li><li><strong>Reporting of results</strong></li></ul><p>The purpose of the training is to introduce users to the capabilities of the LC-HR-MS/MS (QTOF) system in quantitative and qualitative analysis. To point out the advantages of high resolution in identifying or confirming the presence of substances in a sample and also the strengths of modern QTOF systems in parallel quantitative analysis.</p><h3><strong>Training program</strong></h3><h4><span style="font-size:20px;"><strong>Day I</strong></span></h4><h5><strong>Theoretical part</strong></h5><ul><li>The principle of operation and capabilities of the QTOF system</li><li>Comparison of typical analytical procedures and their application</li><li>Targeted and quantitative analysis (TOF MS vs MRMhr)</li><li>DDA (IDA) vs DIA (SWATH) methodology.</li><li>Supporting software - Molecule Profiler, ACD Labs, MarkerView</li><li>General principles of compound fragmentation</li></ul><h4><span style="font-size:20px;"><strong>Day II</strong></span></h4><p><strong>Objective</strong> - to develop methods for quantitative analysis, confirmation or search for metabolites of a known substance, general characterization of the sample.</p><h5><strong>Practical part:</strong></h5><ul><li>Creation and optimization of the MRMhr targeted method</li><li>Creation and optimization of a non-targeted method IDA</li><li>Creation and optimization of the non-targeted method SWATH</li><li>Data acquisition (overnight)</li></ul><h4><span style="font-size:20px;"><strong>Day III</strong></span></h4><p><strong>Objective</strong> - quantitative analysis, confirmation of the presence or identification of metabolites of a known substance, general characterization of the sample (identification of compounds present in the sample), profiling.</p><h5><strong>Practical part:</strong></h5><ul><li>Quantitative analysis - TOF MS vs MRMhr</li><li>Methods for searching and analyzing data</li><li>IDA vs SWATH data</li><li>Manual search and identification</li><li>Automated search and identification - SciexOS, databases</li><li>Molecule Profiler, MarkerView</li><li>Summary and discussion</li></ul>

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