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Food Chemistry Nov 2022Fish is an important nutrition source because its lipids, which are rich in ω-3 fatty acids, are beneficial for human health. However, studies focusing on their...
Fish is an important nutrition source because its lipids, which are rich in ω-3 fatty acids, are beneficial for human health. However, studies focusing on their detection, composition, and nutritional value are limited. In this study, we applied a non-targeted lipidomic approach based on ultra-high performance liquid chromatography coupled with linear-ion trap-Orbitrap mass spectrometry (UHPLC/LTQ-Orbitrap-MS) to comprehensively profile, compare, and detect unknown lipids in eleven types of dietary fish. A total of 287 molecular species from five major lipid classes were characterized by MS/MS analysis. Multivariate principal component analysis revealed the distinct lipid composition in shishamo smelt and Japanese sardine compared to other fish types. The assessment of nutritional indices based on the levels of free fatty acid suggested that among the eleven fish types, shishamo smelt is highly beneficial for health. Further, lipids such as N-acyl lysophosphatidylethanolamine were detected and characterized for the first time in fish fillets. Hierarchical cluster correlations indicated the predominance of glycerophospholipids (GPs) and sphingolipids in sardine, whereas fatty acyls and triacylglycerols (TAGs) were predominant in shishamo smelt. The high levels of polyunsaturated fatty acid-enriched GPs and TAGs in dietary fish endow it with great potential as a health-promoting food for human consumption. This study offers a comprehensive analysis of lipids and their compositions in fish fillets, demonstrating their potential use in the nutritional assessment of functional foods.
Topics: Animals; Chromatography, High Pressure Liquid; Chromatography, Liquid; Glycerophospholipids; Lipidomics; Lipids; Tandem Mass Spectrometry; Triglycerides
PubMed: 35751211
DOI: 10.1016/j.foodchem.2022.133402 -
Journal of Chromatography. A Sep 2023Today, reverse phase liquid chromatography (RPLC) analysis of proteins is almost exclusively performed on conventional columns (100-150 mm) in gradient elution mode.... (Review)
Review
Today, reverse phase liquid chromatography (RPLC) analysis of proteins is almost exclusively performed on conventional columns (100-150 mm) in gradient elution mode. However, it was shown many years ago that large molecules present an on/off retention mechanism, and that only a very short inlet segment of the chromatographic column retains effectively the large molecules. Much shorter columns - like only a few centimetres or even a few millimetres - can therefore be used to efficiently analyse such macromolecules. The aim of this review is to summarise the historical and more recent works related to the use of very short columns for the analysis of model and therapeutic proteins. To this end, we have outlined the theoretical concepts behind the use of short columns, as well as the instrumental limitations and potential applications. Finally, we have shown that these very short columns were also possibly interesting for other chromatographic modes, such as ion exchange chromatography (IEX), hydrophilic interaction chromatography (HILIC) or hydrophobic interaction chromatography (HIC), as analyses in these chromatographic modes are performed in gradient elution mode.
Topics: Chromatography, Reverse-Phase; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Proteins; Hydrophobic and Hydrophilic Interactions
PubMed: 37562104
DOI: 10.1016/j.chroma.2023.464285 -
Journal of Chromatography. A Apr 2024Two-dimensional liquid chromatography (2D-LC), and in particular comprehensive two-dimensional liquid chromatography (LC×LC), offers increased peak capacity, resolution... (Review)
Review
Two-dimensional liquid chromatography (2D-LC), and in particular comprehensive two-dimensional liquid chromatography (LC×LC), offers increased peak capacity, resolution and selectivity compared to one-dimensional liquid chromatography. It is commonly accepted that the technique produces the best results when the separation mechanisms in the two dimensions are completely orthogonal; however, the use of similar separation mechanisms in both dimensions has been gaining popularity as it helps avoid difficulties related to mobile phase incompatibility and poor column efficiency. The remarkable advantages of using reversed phase in both dimensions (RPLC×RPLC) over other separation mechanisms made it a promising technique in the separation of complex samples. This review discusses some physical and practical considerations in method development for 2D-LC involving the use of RP in both dimensions. In addition, an extensive overview is presented of different applications that relied on RPLC×RPLC and 2D-LC with reversed phase column combinations to separate components of complex samples in different fields including food analysis, natural product analysis, environmental analysis, proteomics, lipidomics and metabolomics.
Topics: Chromatography, Liquid; Proteomics; Chromatography, Reverse-Phase
PubMed: 38522405
DOI: 10.1016/j.chroma.2024.464824 -
Journal of Veterinary Diagnostic... Jul 2023There is growing interest in using hemp materials as animal feed ingredients, which may raise safety concerns because of the potential transfer of active cannabinoids to...
There is growing interest in using hemp materials as animal feed ingredients, which may raise safety concerns because of the potential transfer of active cannabinoids to the resultant products of animal origin. Hence, the detection and identification of cannabinoids in feeds would be useful. We developed a simple, fast, and sensitive method for simultaneous quantification of 4 major cannabinoids in animal feeds by liquid chromatography-tandem mass spectrometry (LC-MS/MS). We used a simple solvent extraction and dilution approach to extract cannabinoids from the feed matrix. We validated the method in 2 types of cattle feeds with acceptable intra-day and inter-day accuracy (87.5-116%) and precision (< 15%). The limit of detection was 0.05 µg/g, and the limit of quantification was 0.1 µg/g. Furthermore, the method was able to identify and quantify cannabinoids in cattle feeds mixed with hempseed cake as well as in several different hempseed materials, demonstrating its potential in veterinary laboratory applications.
Topics: Animals; Cattle; Cannabinoids; Chromatography, Liquid; Tandem Mass Spectrometry; Cannabis; Animal Feed
PubMed: 37067036
DOI: 10.1177/10406387231169290 -
Journal of Chromatography. B,... Dec 2022For large-scale and long-term metabolomics studies that involve a large batch or multiple batches of analyses, batch effects cause nonbiological systematic biases that...
For large-scale and long-term metabolomics studies that involve a large batch or multiple batches of analyses, batch effects cause nonbiological systematic biases that may lead to false positive or false negative findings. Quantitative monitoring and correction of batch effects is critical to the development of reproducible and robust metabolomics platforms either for untargeted or targeted analyses. To achieve sufficient retention and separation of a broad range of metabolites with diverse chemical structures and physicochemical properties, LC-MS/MS based targeted metabolomics often involves 3 complemented chromatographic separation methods, including reversed-phase liquid chromatography (RP-LC), hydrophilic interaction liquid chromatography (HILIC), and ion-pair liquid chromatography (IP-LC). The purpose of this study is to quantitatively evaluate intra-batch variations or injection order effects of the RP-LC, HILIC, and IP-LC methods for targeted metabolomics analyses, and develop strategies to minimize intra-batch variations and correct injection order effects for problematic metabolites. Both RP-LC and HILIC methods exhibit robust intra-batch reproducibility in 0.2 µM standard mix QC, with ∼96 % of the measured metabolites showing acceptable intra-batch variations (<20 %); whereas, the intra-batch reproducibility for some metabolites in cell matrix QC may be compromised due to stability issue, suboptimal chromatographic retention, and/or matrix effects causing ionization suppression and/or retention instability. The IP-LC method exhibits significant injection order effects, which could be effectively corrected by the developed exponential models of signal drift trends as a function of injection order for individual targeted metabolites.
Topics: Chromatography, Liquid; Reproducibility of Results; Metabolome; Tandem Mass Spectrometry; Metabolomics; Hydrophobic and Hydrophilic Interactions
PubMed: 36283260
DOI: 10.1016/j.jchromb.2022.123513 -
Journal of Chromatography. A Aug 2014The challenges in HPLC are fast and efficient separation for a wide range of samples. Fast separation often results in very high operating pressure, which places a huge... (Review)
Review
The challenges in HPLC are fast and efficient separation for a wide range of samples. Fast separation often results in very high operating pressure, which places a huge burden on HPLC instrumentation. In recent years, core-shell silica microspheres (with a solid core and a porous shell, also known as fused-core or superficially porous microspheres) have been widely investigated and used for highly efficient and fast separation with reasonably low pressure for separation of small molecules, large molecules and complex samples. In this review, we firstly show the types of core-shell particles and how they are generally prepared, focusing on the methods used to produce core-shell silica particles for chromatographic applications. The fundamentals are discussed on why core-shell particles can perform better with low back pressure, in terms of van Deemter equation and kinetic plots. The core-shell particles are compared with totally porous silica particles and also monolithic columns. The use of columns packed with core-shell particles in different types of liquid chromatography is then discussed, followed by illustrating example applications of such columns for separation of various types of samples. The review is completed with conclusion and a brief perspective on future development of core-shell particles in chromatography.
Topics: Chromatography, High Pressure Liquid; Chromatography, Liquid; Microspheres; Particle Size; Porosity; Silicon Dioxide
PubMed: 24856904
DOI: 10.1016/j.chroma.2014.05.010 -
Journal of Chromatography. A Aug 2022Nucleotide sugars and more specifically UDP-sugars, represent a major source of energy, key components of extracellular matrix, glycosylation and glucuronidation...
Nucleotide sugars and more specifically UDP-sugars, represent a major source of energy, key components of extracellular matrix, glycosylation and glucuronidation reactions, and emerge as important signaling molecules through P2Y14 purinergic receptor. Despite their pivotal role in a variety of physiological and pathological processes and their potential as biomarkers, UDP-sugar composition of biological fluids remains poorly studied. We developed a liquid chromatography electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode for the simultaneous quantification of UDP-glucose, UDP-galactose, UDP-glucuronic acid, UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine in human blood and urine. Relative to existing methods, UDP-sugar recovery was enhanced with perchloric acid and ammonium formate during sample preparation that also significantly improved chromatographic stability. Performance of the assay was validated and allowed the absolute quantification of UDP-sugars with a wide dynamic range (0.1 to 200 ng/mL) using stable deuterated isotopes as internal standards. We report a fast (13 min run) and sensitive method (limit of detection: 10-30 pg/mL; lower limit of quantification ≤ 0.2 ng/ml) to simultaneously quantify five UDP-sugars in a low volume (100 µL) of plasma and urine. Findings identified sex-specific profiles in both plasma and urine of healthy subjects. Applicability was also successfully demonstrated in specimens collected from individuals displaying a variety of medical conditions. This validated method was optimized for a high-throughput assessment of UDP-sugars in specimens of clinical importance and enabled an accurate and reliable absolute quantification of important UDP-sugars in diverse clinical contexts.
Topics: Chromatography, High Pressure Liquid; Chromatography, Liquid; Female; Humans; Male; Nucleotides; Sugars; Tandem Mass Spectrometry; Uridine Diphosphate Glucose
PubMed: 35820232
DOI: 10.1016/j.chroma.2022.463296 -
Analytical Chemistry Jun 2022Many chromatographers have observed that the operating pressure can dramatically change the chromatographic retention of solutes. Small molecules show observables...
Many chromatographers have observed that the operating pressure can dramatically change the chromatographic retention of solutes. Small molecules show observables changes, yet even more sizable effects are encountered with large biomolecules. With this work, we have explored the use of pressure as a method development parameter to alter the reversed-phase selectivity of peptide and protein separations. An apparatus for the facile manipulation of column pressure was assembled through a two-pump system and postcolumn flow restriction. The primary pump provided an eluent flow through the column, while the secondary pump provided a pressure-modulating flow at a tee junction after the column but ahead of a flow restrictor. Using this setup, we were able to quickly program various constant pressure changes and even pressure gradients. It was reconfirmed that pressure changes impact the retention of large molecules to a much greater degree than small molecules, making it especially interesting to consider the use of pressure to selectively separate solutes of different sizes. The addition of pressure to bring the column operating pressure beyond 500 bar was enough to change the elution order of insulin (a peptide hormone) and cytochrome C (a small serum protein). Moreover, with the proposed setup, it was possible to combine eluent and pressure gradients in the same analytical run. This advanced technique was applied to improve the separation of insulin from one of its forced degradation impurities. We have referred to this method as pressure-enhanced liquid chromatography and believe that it can offer unseen selectivity, starting with peptide and protein reversed-phase separations.
Topics: Chromatography, High Pressure Liquid; Chromatography, Liquid; Insulins; Peptides; Pressure; Proteins
PubMed: 35607711
DOI: 10.1021/acs.analchem.2c00464 -
Journal of Chromatography. A Oct 2021Pycnometric and homologous series retention methods are used to determine the volume and mean composition of the water-rich layers partially adsorbed on the surface of...
Volume and composition of semi-adsorbed stationary phases in hydrophilic interaction liquid chromatography. Comparison of water adsorption in common stationary phases and eluents.
Pycnometric and homologous series retention methods are used to determine the volume and mean composition of the water-rich layers partially adsorbed on the surface of several hydrophilic interaction liquid chromatography (HILIC) column fillings with acetonitrile-water and methanol-water as eluents. The findings obtained in this work confirm earlier studies using direct methods for measuring the stationary phase water content performed by Jandera's and Irgum's research groups. Water is preferentially adsorbed on the surface of the HILIC bonded phase in hydroorganic eluents containing more than 40% acetonitrile or 70% methanol, and a gradient of several water-rich transition layers between the polar bonded phase and the poorly polar bulk mobile phase is formed. These layers of reduced mobility act as HILIC stationary phases, retaining polar solutes. The volume of these layers and concentration of adsorbed water is much larger for acetonitrile-water than for methanol-water mobile phases. In hydroorganic eluents with less than 20-30% acetonitrile or 40% methanol the amount of preferentially adsorbed water is very small, and the observed retention behavior is close to the one in reversed-phase liquid chromatography (RPLC). In eluents with intermediate acetonitrile-water or methanol-water compositions a mixed HILIC-RPLC behavior is presented. Comparison of several HILIC columns shows that the highest water enrichment in the HILIC retention region for acetonitrile-water mobile phases is observed for zwitterionic and aminopropyl bonded phases, followed in minor grade for diol and polyvinyl alcohol functionalizations. Pentafluorophenyl bonded phase, usually considered a HILIC column, does not show significant water adsorption, nor HILIC retention.
Topics: Adsorption; Chromatography, Liquid; Chromatography, Reverse-Phase; Hydrophobic and Hydrophilic Interactions; Water
PubMed: 34571282
DOI: 10.1016/j.chroma.2021.462543 -
Therapeutic Drug Monitoring Aug 2021Therapeutic drug monitoring of hydroxychloroquine (HCQ) has been recommended to optimize the treatment of patients with COVID-19. The authors describe an...
Development and Validation of a Simple and Rapid Ultrahigh-Performance Liquid Chromatography Tandem Spectrometry Method for the Quantification of Hydroxychloroquine in Plasma and Blood Samples in the Emergency Context of SARS-CoV-2 Pandemic.
Therapeutic drug monitoring of hydroxychloroquine (HCQ) has been recommended to optimize the treatment of patients with COVID-19. The authors describe an ultrahigh-performance liquid chromatography tandem spectrometry method developed in a context of emergency, to analyze HCQ in both human plasma and blood samples. After adding the labeled internal standard and simple protein precipitation, plasma samples were analyzed using a C18 column. Blood samples required evaporation before analysis. The total chromatographic run time was 4 minutes (including 1.5 minutes of column equilibration). The assay was linear over the calibration range (r2 > 0.99) and up to 1.50 mcg/mL for the plasma samples (5.00 mcg/mL for the blood matrix). The limit of quantification was 0.0150 mcg/mL for plasma samples (0.05 mcg/mL blood matrix) with accuracy and precision ranging from 91.1% to 112% and from 0.750% to 11.1%, respectively. Intraday and interday precision and accuracy values were within 15.0%. No significant matrix effect was observed in the plasma or blood samples. This method was successfully applied to patients treated for COVID-19 infection. A simple and rapid ultrahigh-performance liquid chromatography tandem spectrometry method adapted to HCQ therapeutic drug monitoring in the context of SARS-CoV-2 infection was successfully developed and validated.
Topics: Antirheumatic Agents; COVID-19; Chromatography, High Pressure Liquid; Chromatography, Liquid; Drug Monitoring; Emergency Medical Services; Humans; Hydroxychloroquine; Pandemics; Tandem Mass Spectrometry; COVID-19 Drug Treatment
PubMed: 33165216
DOI: 10.1097/FTD.0000000000000836