Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 15th July 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!
<blockquote>
<h4>👉 <a href="https://lcms.labrulez.com/library">SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</a></h4>
</blockquote>
<blockquote>
👉 Need info about different analytical techniques? Peek into <a href="https://gcms.labrulez.com/library">LabRulezGCMS</a> or <a href="https://icpms.labrulez.com/library">LabRulezICPMS</a> libraries.
</blockquote>
This week we bring to you poster presentations from <a href="https://lcms.labrulez.com/events/568">ASMS 2024</a> by Agilent Technologies, Shimadzu, Thermo Fisher Scientific, and Waters Corporation!
<h4>1. Agilent Technologies: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_fluoromatch_asms_2024_tp113_en_agilent_93ee360315.pdf">Leveraging the MS1 Dimension and Formula Prediction in Non-Targeted Analysis of PFAS using New FluoroMatch Algorithms: Assessing Confidence and Coverage</a></h4>
<ul>
<li>poster - ASMS 2024</li>
</ul>
FluoroMatch Covers the Entire Data-Processing Workflow, and Supports Various Acquisition Modes
<ul>
<li>Data was acquired on an Agilent 6546 LC/Q-TOF for this study to assess PFAS in dried blood spot cards (paper) and blood on dried blood spot cards</li>
</ul>
Conclusions:
<ul>
<li>FluoroMatch can be used to rapidly annotate PFAS in an automatic fashion, determine unknowns and expand annotation using an interactive visualizer. Formula prediction provides a significant benefit by providing evidence for annotation for molecules which are considered tentative using other lines of evidence, or for which the signal is too low to get quality MS/MS.</li>
<li>The FluoroMatch formula prediction algorithm is highly sophisticated and has several new or optimized approaches for formula prediction of fluorine containing compounds. With the additional use of homologous series assigning the predicted formula with the most common motifs for all members of a series, the resulting annotations are highly confident (0% false positives in this case for the four series with confident assignments).</li>
</ul>
<h4>2. Agilent Technologies: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_exd_6545xt_mabs_glycopeptide_asms_2024_mp665_en_agilent_01a5c15cdd.pdf">Peptide Mapping of Tryptic Digests for mAbs using a novel ECD cell on the 6545XT AdvanceBio LC/Q-TOF Mass Spectrometer</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
Introduction:
Post-translation modifications (PTMs) on monoclonal antibodies (mAb) play an important role in the safety, efficacy, and binding of a therapeutic to its target [1]. Common examples of PTMs that seek to be identified include glycosylation and phosphorylation. One challenge in the identification of these PTMs is that dissociation techniques such as collision-induced dissociation (CID) can break apart these fragile modifications. Here, we describe the use of an electron-based dissociation technique (ExD) that supplies low-energy electrons for the fragmentation of tryptic digests that contain glycosylation at asparagine consensus sites (NXS/T). The fragmentation of trastuzumab tryptic digest on a 6545XT AdvanceBio LC/Q-TOF using CID is compared to illustrate that CID alone will fragment these labile PTMs.
Conclusions: The Agilent 6545XT AdvanceBio LC/Q-TOF with ExD cell enables glycopeptide analysis with excellent sequence coverage assessed using MassHunter BioConfirm 12.1
<ul>
<li>LC/MS conditions were optimized to detect low-abundant glycopeptide species from NIST and trastuzumab</li>
<li>MS/MS spectra for glycopeptides using ExD show peptide fragments with intact glycans and excellent sequence coverage along the peptide backbone</li>
<li>For CID mode, no intact glycan fragments are observed. Instead, HexNAc glycan fragment at m/z 204.086 is observed</li>
</ul>
<h4>3. Shimadzu: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ASMS_2024_Th_P_178_e34df60086.pdf">Quantitation of free amino acids in human plasma by Single Quadrupole LC-MS</a></h4>
<ul>
<li>poster - ASMS 2024</li>
</ul>
Introduction: Amino acids are organic compounds that combine to form proteins. They are typically known as the building blocks of protein. Quantitative analysis of amino acids from biological fluids is of great importance in the clinical diagnosis of inborn errors of amino acid metabolism. The assessment of amino acids in body fluids is needed for the diagnosis and treatment of metabolic diseases. We have here performed a high-throughput method for analysis of amino acids in plasma samples. The method combines classical protein hydrolysis and derivatization with fast separation by UHPLC and detection by a single quadrupole mass spectrometer
Conclusion:
<ul>
<li>Liquid chromatography triple quadrupole mass spectrometry (LCMS/MS) analysis of amino acids is becoming the method of choice by more and more laboratories because of its speed, sensitivity, and increased specificity. Although LC-MS/MS instruments are more selective and sensitive than LC-MS single quadrupole instruments, LC-MS can also be used in the quantitative analysis of amino acids when the necessary conditions are provided.</li>
<li>The required sensitivity and chromatographic separation were achieved with the single quadrupole LC-MS.</li>
<li>The method can quantitatively analyze 37 Amino acids in biological fluids such as plasma in less than 20 minutes.</li>
</ul>
<h4>4. Shimadzu: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ASMS_2024_TP_198_Shimadzu_MALDI_IHC_Benchtop_bad9f3fdc3.pdf">Benchtop Matrix Assisted Laser Desorption Ionization Mass Spectrometry Imaging of Human Tonsil Proteins using MALDI HiPLEX-IHC Probes</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
Introduction:
<ul>
<li>MALDI mass spectrometry imaging (MSI) is a versatile technique capable of characterising lipid, polysaccharide, metabolite, pharmaceutical, protein and peptide distributions within a wide variety of samples. Recently, MSI has been combined with commercially available antibody probes containing photocleavable mass-tags (PC-MTs) which target specific proteins in what has been termed MALDIimmunohistochemistry (MALDI-IHC) MSI.</li>
<li>In this study, we show imaging of FFPE human tonsil sections on an entry level benchtop MALDI-8020 using both a multiplex of 6 antibodies and a cancer marker screening panel consisting of a multiplex of 20 antibodies at a stage step size of 30μm, demonstrating a reliable, lowcost approach with good quality results.</li>
<li>Analysis of a follicular hyperplasia tissue sample showed visible differences from normal human tonsil tissue relating to structural features and cell distributions (Fig. 1)</li>
</ul>
Conclusion:
<ul>
<li>We have successfully mapped up to 20 biomarkers with a stage step size of 30µm using a low cost, linear benchtop MALDI-TOF mass spectrometer in conjunction with the Miralys probe kit.</li>
<li>The spectra were well-resolved allowing for clear and specific imaging in both normal and clinically significant samples.</li>
<li>The Shimadzu MALDI-8020 and MALDI-8030 MALDI-TOF mass spectrometers are robust systems with class leading sensitivity and resolution capable of a wide variety of applications including, as demonstrated here, MALDI imaging. These systems can provide an invaluable contribution to any research laboratory.</li>
</ul>
<h4>5. Thermo Fisher Scientific: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_144_transformation_protac_metabolites_multi_stage_lc_hram_ms_po144_asms2024_en_4db8598a6b.pdf">Confident transformation site localization of PROTAC drug metabolites facilitated by multi-stage fragmentation LC-HRAM-MS</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
<h5>Abstract:</h5>
<ul>
<li>Purpose: Confident characterization of PROTAC drug metabolites for transformation site localization and soft-spot analysis.</li>
<li>Methods: Drug metabolites were analyzed using MSn fragmentation data acquisition on a Thermo Scientific Orbitrap Ascend Biopharma Tribrid mass spectrometer and data interpretation in Thermo Scientific Compound Discoverer 3.3 SP3 software.</li>
<li>Results: PROTAC drug metabolism was found to be extensive for the investigated compounds. Among the detected metabolites, several isomeric compounds were found where MSn fragmentation data could be used to reduce ambiguity in the transformation site localization, especially for metabolites with higher molecular weights.</li>
</ul>
Conclusions:
<ul>
<li>The utility of flexible MS data acquisition methods for metabolite identification could be demonstrated with two representative PROTAC compounds, MZ1 and dBET1, which exhibit extensive metabolism in vitro.</li>
<li>MSn fragmentation data helped to enhance the precision of transformation site localization, particularly for isomeric metabolites and those with high molecular weights.</li>
</ul>
<h4>6. Thermo Fisher Scientific: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_145_unleashing_power_ht_dia_orbitrap_exploris_240_ms_accurate_quantitation_po145_asms2024_en_2a2f215fdf.pdf">Unleashing the power of HT-DIA acquisition on Orbitrap Exploris 240 MS – Precise and accurate quantitation at 260 SPD</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
<h5>Abstract:</h5>
Assessing and demonstrating the qualitative and quantitative performance of fast data-independent acquisition (DIA) gradients (&lt;270SPD) on a Thermo Scientific Orbitrap Exploris 240 mass spectrometer (MS), Thermo Scientific Vanquish Neo UHPLC system and micropillar array-based 5.5 cm Thermo Scientific µPAC Neo high-throughput HPLC column.
Conclusions: The performance of a novel high throughput (HT) DIA workflow for label-free quantitation was demonstrated by combining an Orbitrap Exploris 240 mass spectrometer with fast capillary flow chromatography on a high throughput µPAC Neo HPLC column and Vanquish Neo UHPLC system. Key performance criteria of the setup/workflow include:
<ul>
<li>Excellent quantitation accuracy and precision for small amounts of bacterial and fungal proteomes from mammalian background proteomes</li>
<li>Increased sample throughput with minimal impact on the quality of the obtained data and proteome coverage </li>
<li>Outstanding instrument productivity at sample turnover rates up to 250 samples per day </li>
<li>Interlaboratory reproducibility, long-term stability, and robustness essential for large cohort and multi center studies PO002624</li>
</ul>
<h4>7. Waters Corporation: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/2024_asms_hatch_mushroom_coffee_9c919673f3.pdf">NON-TARGETED ANALYSIS OF MUSHROOM-CONTAINING COFFEE PRODUCTS USING ION MOBILITY-HIGH RESOLUTION MASS SPECTROMETRY</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
INTRODUCTION:
<ul>
<li>In recent years, the health benefits of mushrooms—including anti- inflammatory effects1 and improved cognitive function2—have led to huge growth in the number of food and supplement products containing so- called &quot;functional&quot; mushrooms.</li>
<li>There is a wide variety of coffee products currently on the market that are blended with extracts from an assortment of different mushroom species.</li>
<li>In this study, we characterized mushroom coffee products containing up to three mushroom species: chaga, cordyceps, and lion&#39;s mane. The chemical fingerprints were compared with that of traditional coffee.</li>
</ul>
Conclusion:
<ul>
<li>A series of oligosaccharides were observed in mushroom coffees and cordyceps capsules, but not chaga or lion’s mane capsules, nor in traditional coffee. These oligosaccharides may contribute to enhanced health benefits of mushroom coffee.</li>
<li>Trehalose disaccharide displayed variability among the mushroom coffees, but was relatively consistent across the cordyceps, chaga, and lion’s mane capsules.</li>
<li>Taken together, these observations suggest that the extraction process (and/or the parts of the mushrooms extracted) governs the oligosaccharide content more than the specific mushroom species included in a product, though further testing is needed.</li>
</ul>
<h4>8. Waters Corporation: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/2024_asms_sarah_characellides_828ec17fb4.pdf">Ion Mobility Separation Of Marine Natural Products: Characellides from the Deep-sea Sponge Characella pachastrelloides</a></h4>
<ul>
<li>Poster - ASMS 2024</li>
</ul>
Introduction: Characella pachastrelloides was first described in 1876 as part of a collection of deep- sea sponges.1 Deep-sea sponges are a source of unique natural products partly due to the extreme environment they inhabit. This study looks at a group of glycolipopeptides known as characellides that were isolated from C. pachastrelloides and found to be bioactive metabolites.2,3 While HRMS can be used for some structural characterization, full elucidation of natural product structures remains challenging. The ‘gold-standard’ method for structural characterization of natural product compounds is nuclear magnetic resonance (NMR). However, certain compounds can be difficult to isolate or at concentrations too low for quality NMR analysis. This study investigates if ion mobility (IMS) can be another tool for structural assignments of these compounds.
Conclusions:
<ul>
<li>The structurally similar characellide isomers A and B were able to be separated in ion mobility space by utilizing multiple passes in the Cyclic IMS device</li>
<li>The added separation power of IMS led to the discovery of two new compounds with the same molecular formula as Characellides C and D.</li>
<li>Additional related compounds were identified with molecular networking and their CCS values were calculated to give insights into the structural diversity of this unique set of compounds.</li>
</ul>

News from LabRulezLCMS Library Week 29, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 19th August 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!
<blockquote>
<h4>👉 <a href="https://lcms.labrulez.com/library">SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</a></h4>
</blockquote>
<blockquote>
👉 Need info about different analytical techniques? Peek into <a href="https://gcms.labrulez.com/library">LabRulezGCMS</a> or <a href="https://icpms.labrulez.com/library">LabRulezICPMS</a> libraries.
</blockquote>
This week we bring to you posters from <a href="https://gcms.labrulez.com/events/568">ASMS 2024</a> by Agilent Technologies, Shimadzu, Thermo Fisher Scientific, and Waters Corporation!
<h4>1. Agilent Technologies: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_6475_spe_pfas_semiconductor_asms_2024_wp496_en_agilent_cf6e3f6528.pdf">Quantitative Analysis of Legacy &amp; Emerging PFAS in Semiconductor Lubricant using Triple Quadrupole LC/MS</a></h4>
<ul>
<li>poster / <a href="https://gcms.labrulez.com/companies/296">ASMS 2024</a></li>
</ul>
Introduction: Per- and polyfluoroalkyl substances (PFAS) are known to play an important role in the semiconductor industry. For example, PFAS-containing lubricants used in manufacturing processes mainly include perfluoropolyether (PFPE), polytetrafluoroethylene (PTFE) and others that are believed to be of very low toxicity.¹ With environmental regulations &amp; manufacturing restrictions continuing to expand in their scope, the semiconductor industry is facing the challenge of sustainability.² Reliable workflows are needed to ensure effective monitoring and control of PFAS in upstream processes. This study established a workflow based on solid phase extraction (SPE) and LC/MS/MS for analyzing over 100 PFAS from semiconductor lubricant matrix. 
Conclusions:
<ul>
<li>A reliable workflow for quantitation of trace levels of PFAS in semiconductor lubricant, based on SPE cleanup and LC/MS/MS was developed.</li>
<li>The Agilent Bond Elut PFAS WAX cartridge offered selective and effective extraction for matrix cleanup and preconcentration of PFAS analytes in lubricant.</li>
<li>The workflow performance confirmed the applicability of the Agilent PFAS MRM Database and eMethod, for quantitation of PFAS using the Agilent 6475 LC/TQ system.</li>
<li>This workflow enables a ready-to-use protocol for lubricant suppliers and semiconductor manufactures to monitor trace levels of critical PFAS in lubricants.</li>
</ul>
<h4>2. Shimadzu: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/MP_220_Identification_of_Double_Bond_Position_in_Cyclic_Ester_using_OAD_TOF_system_5918fdc1f4.pdf">Identification of Double Bond Position in Cyclic Ester using OAD TOF system</a></h4>
<ul>
<li>poster / <a href="https://gcms.labrulez.com/companies/296">ASMS 2024</a></li>
</ul>
<h5>Overview</h5>
Oxygen Attachment Dissociation (OAD) (x1 000 000) is a novel method for fragmentation providing additional fragmentation to CID on the LCMS 9050 To enhance identification of cyclic ester(lactone) which is one of the aromatic ingredients OAD-MS/MS has been applied to provide double bond specific dissociation enabling C=C position assignments.
Introduction: Lactones Lactones, which are aromatic aromatic ingredients find wide applications in perfumes detergents, and foods The aroma is derived from a combination of various compounds rather than single compound The presence of different types and amounts of impurities in these compounds whether synthesized or purified from natural products can result in distinct aromas To accurately evaluate the aroma it is crucial to identify the precise structural information of the compound including impurities However when the compound has a stable cyclic structure such as lactone direct access to the double bond position within the ring by the neutral radical generated through OAD becomes challenging. In this study, various lactones were hydrolyzed to convert the cyclic ester into a hydroxycarboxylic acid, enabling the identification of the double bond position.
<h5>Conclusions:</h5>
<ul>
<li>Hydrolyzation combined with OAD MS/MS was worked to identify C=C bond positions of cyclic ester.</li>
<li>It was suggested that there are several isomers of Lactone B other than the position of the C=C bond.</li>
<li>OAD-MS/MS can distinguish isomers that cannot be identified by CID.</li>
</ul>
<h4>3. Waters Corporation: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/2024_asms_stevens_pahs_micro_plastic_bf33e7cfcd.pdf">ANALYSIS OF HIGH MASS POLYCYCLIC AROMATIC HYDROCARBONS (PAHS) EXTRACTED FROM MICROPLASTICS SPILLED IN THE MARINE ENVIRONMENT</a></h4>
<ul>
<li>poster / <a href="https://gcms.labrulez.com/companies/296">ASMS 2024</a></li>
</ul>
INTRODUCTION
In 2021 the container ship M/V X-Press Pearl caught fire and later sank off the coast of Sri Lanka. Over 1600 tons of polyethylene pellets, aka nurdles, were aboard the ship along with 25 tons of nitric acid which began to leak and is suspected to be the cause of the fire that lead to the sinking. To add to the witches brew of ingredients in this fire were fertilizers and explosives being transported and flame retardants deployed to douse the fire. When combined with abundant, naturally-occurring compounds found in the marine environment, the need arises to characterize the composition of intact, burned, and broken pieces of the spilled plastic later collected from kilometers of Sri Lankan beaches¹.
Determining the distribution and abundance of PAHs in the samples is important for characterizing the source of the microplastics (MPs) and in selecting appropriate handling, disposal and/or recycling methods. Both traditional and advanced GC/MS methods such as 2D GC are suitable for the determination of PAHs up to 302 Da which is useful for such purposes as the differentiation of pyrogenic versus petrogenic inputs2. However, the toxicity characteristics of samples has demonstrated poor correlation with low mass (&lt;302 Da) PAHs in past studies3. For that reason this work was aimed at investigating the routine analysis of higher mass PAHs in the range of 314 to 424 Da to provide access to additional analytes that may better correlate with toxicity as well as to provide the possibility of additional source markers.
CONCLUSIONS
<ul>
<li>Analysis of an expanded range of high molecular weight PAHs in unfractionated extracts of MPs was achieved using a standard GC-APCI QqQ configuration combined with asimple, generic METHODS microextraction</li>
<li>Charge transfer, atmospheric pressure chemical ionization facilitated the implementation of a class-specific MRM acquisition scheme</li>
<li>Only minor modification to the GC method used for lower mass PAHs is required</li>
<li>Nitrogen carrier gas achieved elution of HWM PAHs with symmetric, narrow peaks through the use of flow programming</li>
<li>Further investigation of HMW PAHs for potential source markers and toxic compounds is enabled with this approach</li>
</ul>
<h4>4. Thermo Fisher Scientific: <a href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_carbon_nitrogen_isotopomer_orbitrap_stellar_ms_hybrid_po_en_0e1e253794.pdf">Carbon and Nitrogen Positional Isotopomer Determination in Metabolites using Orbitrap IQ-X and Stellar MS - a novel hybrid nominal mass instrument</a></h4>
<ul>
<li>poster / <a href="https://gcms.labrulez.com/companies/296">ASMS 2024</a></li>
</ul>
<h5>Abstract</h5>
Purpose: Development of a targeted MS²/MS³ approach for the determination of positional isotopomers.
Methods: Initial data acquired on Thermo Scientific Orbitrap IQ-X Tribrid Mass Spectrometer was used for the structural annotation. A targeted MSn based assay utilizing the above data to get the structural information of the metabolites was developed on a novel hybrid nominal mass instrument.
Results: A targeted MS²/MS³ method was established for the determination of positional isotopomers in Adenosine Monophosphate (AMP). The method was used for evaluation of the effect of hypoxia on cellular metabolism in an in-vitro model.
<h5>Conclusions</h5>
<ul>
<li>A workflow for targeted stable isotope analysis of isotopomer is described.</li>
<li>The use of product scans with MSn and alternate fragmentation can give positional information of label using nominal mass resolution instrument.</li>
</ul>

News from LabRulezLCMS Library - Week 34, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 26th August 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!<blockquote><h4>👉 <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library">SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</a></h4></blockquote><blockquote>👉 Need info about different analytical techniques? Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library">LabRulezGCMS</a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library">LabRulezICPMS</a> libraries.</blockquote>This week we bring to you posters from <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/events/568">ASMS 2024</a> and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/events/592">52nd HPLC</a> by Agilent Technologies, Shimadzu, Thermo Fisher Scientific, and Waters Corporation!<h4>1. Agilent Technologies: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/enhancing_high_throughput_preparative_hplc_poster_06b9abadd0.pdf">Enhancing High Throughput Preparative HPLC: Offline Regeneration of Identical Superficially Porous Particle Preparative Columns Utilizing The 1290 Infinity II Preparative LC System and Agilent InfinityLab Poroshell 120 Preparative LC Columns to extend Column Lifetime, Minimize Sample Carryover and Increase Throughput</a></h4><ul><li>poster / <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/303">HPLC</a></li></ul>Abstact: High throughput sample purification and analysis are important tasks in many industries. With the increasing number of samples synthesized– HPLC purification should not be the bottleneck in discovery and development. The throughput on the Prep system can be increased by optimizing the chromatographic method and by optimizing the instrument hardware. In this poster we show how the throughput can be enhanced even further by using two identical Poroshell Preparative HPLC columns in the system and operating them alternately optimizing the 1290 Infinity II Preparative LC System for Off-Line Column Regeneration. The greatest benefit of the offline column cleanup and equilibration is the increase in sample throughput, but added benefits include extended column lifetime and a reduction in sample carryover.Conclusions: Faster run times and higher throughput is desirable when doing preparative HPLC. Using higher flow rates, shorter and lower pressure columns as well as optimized equipment can help to achieve these goals. Higher backpressure limits of the Agilent Inf II Prep pumps along with high pressure valves further reduce cycle times by allowing configuration of automated offline column regeneration. This means two columns are alternatively used via a 2-position/14-port valve. Cleaning and equilibration time can be excluded from the cycle time and 50% more runs were performed.<h4>2. Waters Corporation: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/2024_hplc_maziarz_hydrophilic_hilic_f04a258724.pdf">Scaling Reverse-Phase and Hydrophilic Interaction Liquid Chromatography (HILIC) Methods to HPLC</a></h4><ul><li>poster / <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/303">HPLC</a></li></ul>INTRODUCTION:Achieving identical chromatographic separation and equivalent results are critical when scaling and migrating methods between different liquid chromatography (LC) instruments.In this work, two different methods developed on 1.7 μm particle size columns using a low dispersion system were scaled to HPLC. In first example, a reverse-phase method for naphazoline hydrochloride, pheniramine maleate and associated related substances was scaled from 1.7 μm to a 2.5 μm particle column. A second method for the analysis of ribavirin and related compounds, operated under HILIC conditions, was scaled from 1.7 μm to 2.5 and 3.5 μm particle columns and run on different HPLC platforms. The scaled methods generated equivalent chromatographic performance and assay results as the original method.CONCLUSION:<ul><li>Reverse-phase and HILIC methods were successfully scaled from 1.7 µm particle size columns to HPLC.</li><li>The target LC systems including ACQUITY Arc, Arc HPLC and Alliance iS HPLC Systems generated equivalent performance as the original method run on an ACQUITY UPLC H-Class Plus System.</li><li>Dwell and extra-column volumes were accounted for to ensure quality of the chromatographic data for method scaling and migration across different LC platforms.</li></ul><h4>3. Thermo Fisher Scientific: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/po_003231_hplc_peptide_level_analysis_monoclonal_antibodies_evaluation_po003231_na_en_31f6f13cf1.pdf">Performance evaluation of a new reversed-phase column for peptide mapping and monitoring of monoclonal antibodies</a></h4><ul><li>poster / <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/303">HPLC</a></li></ul><h5>Abstract</h5>Purpose: Demonstrating the benefits of the newly introduced Thermo Scientific Hypersil GOLD Peptide C18 column with 1.9 µm particle and 175 Å average pore size, in the dimension of 150 x 2.1 mm (L x ID) for the analysis of tryptic mAb digest samples.Methods: UHPLC-MS/MS peptide mapping and UHPLC-UV-MS peptide monitoring.Results: The Hypersil GOLD Peptide column enables to uniquely meet the demands of peptide level analysis where consistent lot-to-lot and column-to-column performance is required.<h5>Conclusions</h5>The two studies of peptide mapping and peptide monitoring demonstrate the benefits of the Hypersil GOLD Peptide column for peptide level biopharma analysis based on a comprehensive dataset obtained from rituximab and adalimumab digest samples.<ul><li>SET and SST criteria of retention time and peak area precision are easily met when using the Hypersil GOLD Peptide column.</li><li>The Hypersil GOLD Peptide column provides<ul><li>peaks with good asymmetry values, strong retention time and peak area reproducibility.</li><li>retains and separates hydrophilic peptides and has very good chromatographic resolution values for deamidated species in the PENNYK region.</li><li>excellent column lot-to-lot reproducibility with % RSD RT ≤ 0.6, and % RSD peak area ≤ 6.2 for peptides across the entire elution window.</li><li>consistent column-to-column results for the quantification of glycopeptides with % RSD of relative abundance ≤ 3.0.</li></ul></li></ul><h4>4. Shimadzu: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/ASMS_2024_LEIPP_WP_119_45a784b1d5.pdf">Development of a blood-based multiplex quantitative LC-MS method for neurodegenerative diseases patient stratification : towards an innovative diagnostic tool</a></h4><ul><li>poster / <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/companies/296">ASMS 2024</a></li></ul>Introduction and objectives: Current neurodegenerative biomarkers measurements for diagnosis purposes are performed using CSF samples which requires invasive procedures for the patient. Besides, AD and other associated dementias are very complex pathologies that can have intricate symptoms and pathological deleterious effects. Therefore, the developed quantitative multiplex method includes 22 blood neuroinflammatory, brain function-linked, genetic and general state biomarkers for disease state evaluation and monitoring, patient stratification and differential diagnosis purposes.Conclusions and Perspectives: Further analytical validation would be performed to fully assess method performances (stabilities, intra-laboratory reproducibility). Then, large patient cohorts would be analyzed to evaluate and clinically validate this innovative tool and its capacity to differentiate diseases, severity and co-pathologies. Such tool would bring dementia diagnosis within the landscape of personalized medicine fostering better patient care.

News from LabRulezLCMS Library - Week 35, 2024

Our Library never stops expanding. What are the most recent contributions to LabRulezLCMS Library in the week of 16th September 2024? Check out new documents from the field of liquid phase, especially HPLC and LC/MS techniques!<blockquote>👉 <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/library">SEARCH THE LARGEST REPOSITORY OF DOCUMENTS ABOUT LCMS AND RELATED TECHNIQUES</a></blockquote><blockquote>👉 Need info about different analytical techniques? Peek into <a target="_blank" rel="noopener noreferrer" href="https://gcms.labrulez.com/library">LabRulezGCMS</a> or <a target="_blank" rel="noopener noreferrer" href="https://icpms.labrulez.com/library">LabRulezICPMS</a> libraries.</blockquote>This week we bring to you applications, technical notes, posters, and other documents by Agilent Technologies, Shimadzu, Thermo Fisher Scientific, Waters Corporation, Metrohm, and <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/313">Galochrom</a>!<h4>1. Galochrom: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/Chiral_ION_QN_and_Chiral_ION_QD_Brochure_10c44e1448.pdf">Galochrom Chiral ION-QN &amp; Chiral ION-QD</a></h4><ul><li>Brochure</li></ul><h5>Weak Anion-Exchanger (WAX) stationary phases for reliable chiral separation of acidic compounds</h5>Transform Your Chiral AnalysisWeak Anion-Exchanger (WAX) stationary phases for reliable chiral separation of acidic compounds One of the main challenges of today’s liquid chromatography is chiral separation of ionised and ionisable analytes, e.g., organic acids, amino acids, N-protected amino acids, amino acid mimics, etc. The most efficient approach to purify and resolve such troublesome racemates utilises ion exchange-type stationary phases that facilitate strong interactions between the chiral selector and the analyte.Galochrom introduces its own generic series of weak anion-exchangers, based on tert-butyl-carbamoylated Cinchona alkaloids quinine (QN) and quinidine (QD). These chiral phases have demonstrated excellent selectivity and improved peak resolution, resulting in highly accurate and replicable analyses. Our innovative manufacturing and strict quality control ensure that these generic columns not only outperform commercial alternatives, but also meet your technical requirements.Chiral <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1241">ION-QN</a> &amp; Chiral <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1240">ION-QD</a> represent the most efficient approach to purify and resolve even the most challenging mixtures.<h5>Applications</h5>Chiral ION-QN &amp; Chiral ION-QD are ideal for enantioseparations of:<ul><li>N-protected amino acids</li><li>Aminophosphonic &amp; Aminosulfonic acids</li><li>Lactic &amp; Thiolactic acids</li><li>Clenbuterol &amp; Thyroxine</li></ul><h4>2. Metrohm: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_eis_004_download_4e13c9c1a9.pdf">Electrochemical Impedance Spectroscopy - Equivalent circuit models</a></h4><ul><li>Technical note</li></ul><h5>Electrochemical Impedance Spectroscopy</h5>Part 4 – Equivalent circuit modelsThe circuit elements which are described in Application Note AN-EIS-003 can be combined in series and in parallel to build equivalent circuit models which can then be used to model the various phenomena occurring at the electrochemical interface.This seven-part series introduces EIS and covers basic theory, experimental setups, common equivalent circuits used for fitting data, and tips for improving the quality of the measured data and fitting. This Application Note (part 4) provides example equivalent circuit models produced from the elements discussed in part 3. These models are among the most commonly encountered within electrochemistry research. The associated Nyquist plot is also shown for each model discussed.CONCLUSIONSThis Application Note shows how electrical elements can be arranged in order to build both simple and more complex equivalent circuits that can be used for fitting EIS data. The resulting Nyquist plots for all the equivalent circuits are shown as well.<h4>3. Thermo Fisher Scientific: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/wp_002641_ic_dionex_inuvion_ionic_small_polar_compounds_wp002641_na_en_9c0213a96b.pdf">Developing an ion chromatography system</a></h4><ul><li>Technical note</li></ul>IntroductionIon chromatography (IC) has become essential for analyzing ionic and small polar compounds. It has been applied in many areas, ranging from the analysis of drinking water, food and beverages, and cooling water to lithium-ion battery research, process quality assurance, and the presence of corrosives in liquified petroleum gas.IC can trace its origins to work done at what would become Dionex Corporation in the mid-1970s with the introduction of the first ion chromatograph in 1975, the Dionex™ Model 10 IC system. This system had the essential elements of an IC system: a pump, separating column, eluent suppressor, conductivity detector, and data recorder (Figure 1).This basic system provided adequate data, but several factors restricted sample throughput and required a considerable investment in labor and significant expertise to maintain. Column capacity and packed bed suppressor capacity were limited, and the instrument configuration was inflexible, constraining the type of samples that could be analyzed. Offline suppressor column regeneration was needed, the pressure tolerance was low, and the background conductivity was comparatively high. Furthermore, manual peak integration was required, eluent usage was significant, and there was substantial run-to-run variability. Since the Dionex Model 10 IC system, many innovations have been developed and incorporated into new systems to address these deficiencies. Additional advancements have enhanced robustness, reliability, user-friendliness, functional flexibility, and integrated data analysis. Ideally, an ion chromatograph will meet current customer demands while being adaptable to address potential future analytical needs.ConclusionThe progression from the Dionex Model 10 IC system to the sophisticated Dionex Inuvion IC system highlights the significant advancements that have been made in the field of IC. These advancements have not only addressed previous limitations but also introduced new features and capabilities that significantly enhance the performance, reliability, and user-friendliness of IC systems. As a result, the Dionex Inuvion IC system represents the current pinnacle in the ongoing development of IC technology, setting a new standard for what can be achieved in the analysis of ionic and small polar compounds.The Dionex Inuvion IC system is the result of state-of-the-art techniques and processes used to manufacture, design, and optimize essential hardware components such as pumps, instrument electronics, range-free detectors, and conductivity cells. This integration of advanced technology has resulted in a system that is not only reliable and robust but also flexible and user-friendly.As we look to the future, the field of ion chromatography will continue to evolve with new challenges and opportunities on the horizon. The Dionex Inuvion IC system is well-positioned to meet these challenges, providing laboratories with a reliable and efficient solution for their analytical needs. In conclusion, the Dionex Inuvion IC system represents a significant achievement in ion chromatography, solidifying Thermo Fisher Scientific's position as a leader in the industry.<h4>4. Waters Corporation: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/2024_asms_alelyunas_cellculturemediabioprocessing_27380a2953.pdf">A High Throughput LC-MS Method for Cell Culture Media Nutrient and Metabolite Analysis Supporting Upstream Bioprocessing</a></h4><ul><li>poster / <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/296">ASMS</a></li></ul>INTRODUCTIONA 9-minute rapid and direct liquid chromatography and mass spectrometry (LC-MS) analysis method for cell culture media (CCM) nutrients and metabolites covering 220+ compounds is described. The method is based on reversed phase chromatography where amino acids are directly detected without derivatization. This rapid method reduces turn-around time for monitoring and decision making using previously established user-friendly data review workflows and access to multivariate data analytics (MVDA). The method has been applied for the qualitative and quantitative determination of cell culture media nutrient and metabolite analysis in commercial cell culture media and spent media from process optimization in cNISTmAb production using NISTCHO cells. Monitoring of glucose directly from media using this method was also investigated and described.CONCLUSION<ul><li>A rapid throughput method based on 9 min data acquisition is described for cell culture media nutrient and metabolite monitoring.</li><li>The method uses less than 10 μL of spent media sample and has coverage of 220 compounds.</li><li>Comparison with the previous 20 min method suggests the same high-quality data and method robustness, even when the analysis time is shortened by 50%.</li><li>Combination of the rapid throughput method with Waters bioprocess walk-up solution2 can enable analytical scientist and bioprocess engineers to obtain high quality data easily and routinely for upstream bioprocess optimization.</li></ul><h4>5. Shimadzu: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_01_00767_en_135939ebe6.pdf">Automatic Optimization of Gradient Conditions by AI Algorithm for Impurity Analysis</a></h4><ul><li>Application</li></ul>User Benefits<ul><li>The AI algorithm of LabSolutions MD can automatically optimize gradient conditions to greatly reduce labor of LC method development.</li><li>Anyone can optimize gradient conditions, regardless of their experience in chromatography.</li><li>Gradient conditions that meet the resolution criteria for specified peaks are automatically searched (e.g., principal component and its related impurities).</li></ul>IntroductionIn the typical LC method development, the process begins with “preparation” which includes mobile phase preparation, column installation, and creation of analysis schedules, then the analysis is started. After that, the acquired data is analyzed and “preparation” for the subsequent analysis is carried out, followed by starting the next analysis again. The method development progresses by repeating these processes, but in addition to the significant time required to repeatedly create analysis schedules, expertise in chromatography is necessary to explore optimal conditions based on data analysis. In other words, typical method development requires “human intervention”. Therefore, eliminating human involvement and automating such method development processes would be desirable to improve labor efficiency. This article introduces an example of automatic optimization of gradient conditions to separate montelukast (a small molecule drug) and its related impurities using LabSolutions MD (<a target="_blank" rel="noopener noreferrer" href="https://www.shimadzu.com/an/sites/shimadzu.com.an/files/pim/pim_document_file/technical/technical_reports/22105/c190-e309.pdf">Technical Report C190-E309</a>), a dedicated software for supporting method development.ConclusionAutomatic optimization of gradient conditions using AI algorithm of LabSolutions MD was applied to montelukast and its related impurity. As a result, gradient conditions that met the criteria (“resolution for montelukast and related impurity &gt; 3.0” and “elution time of the last peak &lt; 15 minutes”) were successfully explored. This result indicates that significant labor saving in method development can be expected by LabSolutions MD. This article introduces an automatic optimization of gradient conditions in method development while LabSolutions MD also supports a series of workflow of method development, including the screening phase and robustness evaluation phase. For details, please refer to the Technical Report “<a target="_blank" rel="noopener noreferrer" href="https://www.shimadzu.com/an/sites/shimadzu.com.an/files/pim/pim_document_file/technical/technical_reports/14629/c190-e284.pdf">Efficient Method Development Based on Analytical Quality by Design with LabSolutions MD Software (C190-E284)</a>”.<h4>6. Agilent Technologies: <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/labrulez-bucket-strapi-h3hsga3/an_polar_nitrosamines_1260_infinity_ii_multisampler_5994_7589en_agilent_22cb1563ee.pdf">Improved LC/MS Performance to Determine Polar Nitrosamines Using the Agilent 1260 Infinity II Hybrid Multisampler</a></h4><ul><li>Application</li></ul>AbstractThis application note demonstrates the use of the Agilent 1260 Infinity II Hybrid Multisampler for the analysis of polar nitrosamines injected from organic solvents of high elution strength. The 1260 Infinity II Hybrid Multisampler is used under optimized feed speed conditions to trap and enrich the polar nitrosamines on the column. This avoids breakthrough and provides better peak shapes with more reliable quantification and improved detection limits.IntroductionWith the chemical synthesis of active pharmaceutical ingredients (APIs), impurities occur during chemical reactions. Genotoxic impurities (GTIs) are especially important because of their carcinogenic potential. To control the amount of GTIs, the U.S. Food and Drug Administration and the European Medicine Agency released guidelines for the maximum intake of GTIs with the daily dose of a drug depending upon the duration of application. According to these guidelines, the exposure to an individual genotoxic impurity must be below 1.5 μg/day for an application time longer than ten years.1 In 2018, the drug Diovan was withdrawn from the market due to an increased amount of the nitrosamine impurity N-nitrosodimethylamine (NDMA) in the included API valsartan.2,3 This application note describes the results achieved by the Agilent 1260 Infinity II hybrid injector for the analysis of nitrosamines injected from high organic solvents. The peak performance, like area precision, linearity, and sensitivity, will be discussed.ConclusionThis application note describes the comparative measurement of nitrosamines using the Agilent 1260 Infinity II Hybrid Multisampler in feed injection mode and classical flow through injection mode. With the application of an optimized feed injection method, highly polar nitrosamines in a solvent of high eluting strength can be separated with good peak shapes. With classical flow through injection, massive peak broadening or even breakthrough were observed. Peak area RSD and LOQs are typically lower by a factor of 3 to 5 with feed injection.

News from LabRulezLCMS Library - Week 38, 2024

<blockquote>Maciej Stopa 1,2, Julia Mironenka1, Rafał Szewczyk1,2, Anna Lenartowicz1, Adrian Soboń1,2, Katarzyna Krupczyńska-Stopa1,2, Andrzej Kwaśnica31. LabExperts sp. z o. o., Gdańsk, Poland; 2. <a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/companies/315">Bioanalytic</a> sp. z o. o., Gdańsk, Poland; 3. Lab4Tox sp. z o. o., Wrocław, Poland</blockquote><h4>Introduction</h4>The precision, reproducibility and trueness of the result in forensic analysis is crucial. The accuracy of sample preparation during the psychoactive compounds quantitation is one of the key aspects. Automatic sample preparation minimize mistakes caused by poor performance of the analysis, discrepancies between analysts habits or different days of analysis. Accurate analytes detection relies on efficient extraction, low detection limits and maintaining the efficiency and cleanliness of LC-MS/MS system. Liquid-liquid extraction, by default, are less amenable for automation, and may be challenging because of large amount of sample preparation steps. This work shows the effective translation from manual method to semi-automated procedure of determination of 34 psychoactive compounds from human plasma using LC-MS/MS method.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_1_Semi_automated_sample_07f1c50562.jpg" alt="Bioanalytic: Figure 1 - Semi-automated sample preparation protocol using Hudson SOLO automatic pipette station with Hudson SOLO Soft and SoftLink V protocol Editor." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_1_Semi_automated_sample_07f1c50562.jpg 123w," sizes="100vw" width="805" height="1021"><h4>Methods</h4>Liquid-liquid extraction is sufficient method to determine the amount of psychoactive compounds in human plasma. &nbsp;Sample preparation procedure developed using Hudson SOLO automatic pipette station with Hudson SOLO Soft and SoftLink V Protocol Editor include deproteinization and extraction, dissolution and transfer to a plate compatible with an LC autosampler in 100 minutes. Chromatography run was performed using ExionAC equipped with Fortis H2O C18 column. The method was run in reverse phase mode, using as mobile phases acidified with formic acid both, water and acetonitrile. Mass spectrometry was performed using &nbsp;ZenoTOF 7600 (SCIEX), with following conditions: positive electrospray scheduled MRMhr mode acquiring full MS/MS spectra. The data analysis including compound identification and quantification was performed with SciexOS 3.3 software.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Table_1_Part_A_The_comparison_136f3d2667.jpg" alt="Bioanalytic: Table 1. Part A - The comparison of manual vs-semi automated sample preparation precision; Part B: The results of method of semi-automated sample preparation." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Table_1_Part_A_The_comparison_136f3d2667.jpg 129w," sizes="100vw" width="1008" height="1217"><h4>Preliminary data</h4>Blood is&nbsp; easy to obtain information-rich matrix. It have only one disadvantage – it cannot be easily analyzed with LC-MS/MS. Preparation of a multimethod for psychoactive compounds determination which &nbsp;characterized by a different chemical properties requires complex and strict sample preparation to achieve accurate detection and compound presence confirmation. Unfortunately, in complex extraction steps are difficult to postpone due to various activities that has to be undertaken including – mixing, centrifugation, sample aspirations and evaporation. In this work, we show the possibility to transfer steps of aspiration, dilution, heating and mixing during the sample preparation on an automatic high precision pipette station. Other steps including centrifugation and evaporation were done manually. Beside the shortening of sample preparation time, which allows to perform the analysis of 96 samples in less than 2 hours, the results are subjected to less preparation error. Such configurations allow a substantial increase of sample throughput and convenience when compared to standard protocols. As a result of this translation, better repeatability (%CV ≤ 15%), maintaining a wide range of linearity (LLOQ-50 ng/ml) which met the validation criteria (R ≥ 0.995) of the method was achieved. An additional advantages of using automation – the analyst’s exposure to blood obtained from contractors is reduced. Also, automated protocol minimize the risk of losing valuable samples avoiding cases of human error.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_2_Chromatographic_separation_1f0e513084.jpg" alt="Bioanalytic: Figure 2: Chromatographic separation of analytes included in multi-method of psychoactive compounds quantitative determinantion [c=25 ng/ml]." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_improve_the_precision_of_analysis_semi_automated_sample_preparation_for_psychoactive_compounds_quantification_in_human_plasma_Figure_2_Chromatographic_separation_1f0e513084.jpg 245w," sizes="100vw" width="2841" height="923"><h4>Novel aspect</h4>Automated method offers excellent repeatability and high accuracy of psychoactive compounds determination in human blood.

How to improve the precision of analysis – semi-automated sample preparation for psychoactive compounds quantification in human plasma

<blockquote>Julia Mironenka1, Rafał Szewczyk1,2, Katarzyna Krupczyńska-Stopa1,2, Maciej Stopa 1,2&nbsp;LabExperts sp. z o. o., Gdańsk, Poland; 2. Bioanalytic sp. z o. o., Gdańsk, Poland;</blockquote><h4>Introduction</h4>Antibiotics are commonly used in therapy of infectious diseases. Their overuse to increase animal productivity, including honey production, has become a serious problem in food safety regulations. High doses of antibiotics used by farms, find their way into the end products that enter human consumption. Honey is a valuable product, often used in medicine and as a dietary ingredient. All the sweetness of honey disappears when it reaches the laboratory as a matrix for LC-MS/MS analysis, due to the great diversity of components content between each, or even the same kind of honey. In this work we have focused on the development of highly reproducible and precise method of 57 antibiotics determination by LC-MS/MS in honey, regardless of its origin.<h4>Methods</h4>Sample preparation is based on modified QuEChERS method. The honey samples after enrichment with known concentrations of analytes were hydrated using acidified water, heated and sonicated until sugar dissolved. Next, after mixing with acidified acetonitrile solution, samples were desalted using ready to use QuEChERS sachets and filtered with PVDF syringe filter. &nbsp;Before the LC-MS/MS analysis samples were diluted with water. Data were acquired on QTRAP 5500+ (SCIEX) mass spectrometer coupled with ExionAC LC (SCIEX) and processed with SciexOS 2.2 software. The LC-MS/MS analysis was performed in positive ionization sMRM mode during reversed phase 12-min separation on Fortis H2O C18 chromatographic column. Mobile phases consisted of water, acetonitrile and formic acid additive.<figure class="image image_resized" style="width:50%;"><img style="aspect-ratio:362/1175;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Fig_1_Sample_preparation_procedure_efcd25086b.jpg" alt="Bioanalytic: Fig 1. Sample preparation procedure." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Fig_1_Sample_preparation_procedure_efcd25086b.jpg 48w," sizes="100vw" width="362" height="1175"></figure><h4>Preliminary data</h4>Honey is a complex matrix that consists of over 180 components. Depending on the origin, type and bees, the ingredients are present in different amounts, which undoubtedly affects its properties. Developing a repeatable extraction method for one compound becomes a challenge. In the present work a multimethod for 57 analytes from 10 different classes of antibiotics was developed. The optimization of the sample preparation method consisted of: selection of solvents and pH of the solutions, concentration of additives, time of sonication and extraction, dissolution ratio and method of cleaning the sample before LC-MS/MS analysis. The intensity and shape of signals of samples subjected to evaporation under nitrogen concentration and dissolved in the initial phase of the chromatographic method were compared to samples analyzed immediately after extraction with different water dilutions. The present method allows sample preparation without time-consuming stage of evaporation followed by residues dissolving.The efforts devoted to method development resulted in obtaining averaged recovery of tetracyclkines, cefalosporins, sulfonamides and fluorochinones was found to be 106.4%. The linearity (R&lt;0.995) covered wide concentration range 0.5-500 µg/kg. LLOQ for most macrolides (tylosin, spiramycin, tilmicosin) and tetracyclines (doxycycline, chlorotetracycline, 4-epi-chlorotetracycline) was 0.5 µg/kg, for some of beta-lactams (nafcillin, cloxacillin, oxacyllin) was 0.1 µg/kg and for some sulfonamides (sulfadimethoxine, sulfachinoxaline, sulfchloropyrazine) reached 0.05 µg/kg. Thanks to a laborious optimization process excellent repeatability for 10 different honeys obtained from local beekeepers was achieved. The accuracy and precision for the whole set of honeys was lower than 20% in the working range of the assay.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Table_1_The_results_of_method_validation_6a3be83107.jpg" alt="Bioanalytic: Table 1. The results of method validation." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Table_1_The_results_of_method_validation_6a3be83107.jpg 245w," sizes="100vw" width="2126" height="1166"><figure class="image"><img style="aspect-ratio:897/575;" src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Table_2_Table_3_Table_4_58dd61a3ef.jpg" alt="Bioanalytic: Table 2. Ion source parameters; Table 3. sMRM scan parameters; Table 4. General LC-MS parameters." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Bioanalytic_How_to_deal_with_sweet_matrix_a_method_for_LC_MS_MS_analysis_of_antibiotics_in_honey_regardless_of_its_kind_Table_2_Table_3_Table_4_58dd61a3ef.jpg 244w," sizes="100vw" width="897" height="575"></figure><h4>Novel aspect</h4>The effective analytical procedure for the determination of 57 antibiotics residues from different honey samples in one LC-MS/MS run.

How to deal with sweet matrix? – a method for LC-MS/MS analysis of antibiotics in honey, regardless of its kind

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