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International Journal of Molecular... Jul 2023Although LC-MS with atmospheric pressure ionization (API) sources is the primary technique used in modern bioanalytical studies, electron ionization mass spectrometry... (Review)
Review
Although LC-MS with atmospheric pressure ionization (API) sources is the primary technique used in modern bioanalytical studies, electron ionization mass spectrometry (EI-MS) can provide some substantial advantages over it. EI-MS is a matrix effect-free technique that provides reproducible and comparable mass spectra, serving as a compound fingerprint for easy identification through automated comparison with spectral libraries. Leveraging EI-MS in biochemical studies can yield critical analytical benefits for targeted and untargeted analyses. However, to fully utilize EI-MS for heavy and non-volatile molecules, a new technology that enables the coupling of liquid chromatography with EI-MS is needed. Recent advancements in nanoLC have addressed the compatibility issues between LC and EI-MS, and innovative interfacing strategies such as Direct-EI, liquid electron ionization (LEI), and Cold-EI have extended the application of EI-MS beyond the determination of volatile organic molecules. This review provides an overview of the latest developments in nanoLC-EI-MS interfacing technologies, discussing their scope and limitations. Additionally, selected examples of nanoLC-EI-MS applications in the field of biochemical analysis are presented, highlighting the potential prospects and benefits that the establishment of this technique can bring to this field.
Topics: Spectrometry, Mass, Electrospray Ionization; Electrons; Chromatography, Liquid; Atmospheric Pressure; Technology
PubMed: 37511506
DOI: 10.3390/ijms241411746 -
Journal of Clinical Laboratory Analysis Jul 2023Vitamin D toxicity is rare in pediatric population. Falsely elevated levels of 25-hydroxyvitamin D have been reported as a major challenge with immunoassay methods for...
BACKGROUND
Vitamin D toxicity is rare in pediatric population. Falsely elevated levels of 25-hydroxyvitamin D have been reported as a major challenge with immunoassay methods for quantifying vitamin D metabolites.
CASE PRESENTATION AND METHOD
Here, we present two pediatric cases of falsely elevated 25-hydroxyvitamin D that resulted in unnecessary further testing. We also report significant same-day variation in the measurement of 25-hydroxyvitamin D using the Abbott i2000SR immunoassay. Samples were spun twice and their values were confirmed with the gold standard liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for confirmation.
CONCLUSION
The addition of a centrifugation step prior to sample testing resolved the variation observed in the measurement of 25-hydroxyvitamin D levels. The patient samples were confirmed with instruments from a different vendor and LC-MS/MS. Re-centrifugation of samples resolved the variation in the 25-hydroxyvitamin D values.
Topics: Humans; Child; Chromatography, Liquid; Tandem Mass Spectrometry; Vitamin D; Calcifediol; Vitamins; Immunoassay; 25-Hydroxyvitamin D 2
PubMed: 37526221
DOI: 10.1002/jcla.24950 -
Journal of Pharmaceutical and... Aug 2023Amatoxins are toxic bicyclic octapeptides found in certain wild mushroom species, particularly Amanita phalloides. These mushrooms contain predominantly α- and... (Review)
Review
Amatoxins are toxic bicyclic octapeptides found in certain wild mushroom species, particularly Amanita phalloides. These mushrooms contain predominantly α- and β-amanitin, which can lead to severe health risks for humans and animals if ingested. Rapid and accurate identification of these toxins in mushroom and biological samples is crucial for diagnosing and treating mushroom poisoning. Analytical methods for the determination of amatoxins are critical to ensure food safety and prompt medical treatment. This review provides a comprehensive overview of the research literature on the determination of amatoxins in clinical specimens, biological and mushroom samples. We discuss the physicochemical properties of toxins, highlighting their influence on the choice of the analytical method and the importance of sample preparation, particularly solid-phase extraction with cartridges. Chromatographic methods are emphasised with a focus on liquid chromatography coupled to mass spectrometry as one of the most relevant analytical method for the determination of amatoxins in complex matrices. Furthermore, current trends and future perspectives in amatoxin detection are also suggested.
Topics: Humans; Animals; Chromatography, High Pressure Liquid; Mushroom Poisoning; Chromatography, Liquid; Mass Spectrometry; Toxins, Biological
PubMed: 37146495
DOI: 10.1016/j.jpba.2023.115421 -
Biosensors Jun 2023Rapid and efficient detection of mycotoxins is of great significance in the field of food safety. In this review, several traditional and commercial detection methods... (Review)
Review
Rapid and efficient detection of mycotoxins is of great significance in the field of food safety. In this review, several traditional and commercial detection methods are introduced, such as high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS), enzyme-linked immunosorbent assay (ELISA), test strips, etc. Electrochemiluminescence (ECL) biosensors have the advantages of high sensitivity and specificity. The use of ECL biosensors for mycotoxins detection has attracted great attention. According to the recognition mechanisms, ECL biosensors are mainly divided into antibody-based, aptamer-based, and molecular imprinting techniques. In this review, we focus on the recent effects towards the designation of diverse ECL biosensors in mycotoxins assay, mainly including their amplification strategies and working mechanism.
Topics: Mycotoxins; Chromatography, Liquid; Biosensing Techniques; Chromatography, High Pressure Liquid; Enzyme-Linked Immunosorbent Assay; Luminescent Measurements
PubMed: 37367018
DOI: 10.3390/bios13060653 -
Toxins Sep 2023In this study, aflatoxins (AFs) and ochratoxin A (OTA) were analyzed in grains, specifically wheat and corn, from Albania. To summarize, 71 wheat and 45 corn samples...
In this study, aflatoxins (AFs) and ochratoxin A (OTA) were analyzed in grains, specifically wheat and corn, from Albania. To summarize, 71 wheat and 45 corn samples from different growing areas were collected. The multi-toxin analytical procedure involved sample extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS). The incidence of AF was 18% in the analyzed wheat and 71% in the corn samples. The concentration of AFs was much higher in the corn samples than in the wheat samples. The maximum permitted levels for aflatoxin B1 (AFB1) and total AFs were not exceeded in the wheat samples, while they were exceeded in 36% of the corn samples. In the wheat samples, the AFB1 concentration varied between 0.2 and 0.4 µg kg. However, the highest concentrations in the corn samples were 2057, 2944, and 3550 µg kg. OTA was present in only three corn samples and one wheat sample. However, all contaminated samples exceeded the maximum permitted levels. This report reveals the presence of AFs and OTA in grain commodities, specifically wheat and corn, grown in Albania.
Topics: Aflatoxins; Zea mays; Triticum; Chromatography, Liquid; Albania; Incidence; Food Contamination; Tandem Mass Spectrometry; Ochratoxins; Aflatoxin B1; Chromatography, High Pressure Liquid
PubMed: 37755993
DOI: 10.3390/toxins15090567 -
Environmental Science & Technology Sep 2023Non-targeted analysis (NTA) has emerged as a valuable approach for the comprehensive monitoring of chemicals of emerging concern (CECs) in the exposome. The NTA approach... (Review)
Review
Non-targeted analysis (NTA) has emerged as a valuable approach for the comprehensive monitoring of chemicals of emerging concern (CECs) in the exposome. The NTA approach can theoretically identify compounds with diverse physicochemical properties and sources. Even though they are generic and have a wide scope, non-targeted analysis methods have been shown to have limitations in terms of their coverage of the chemical space, as the number of identified chemicals in each sample is very low (e.g., ≤5%). Investigating the chemical space that is covered by each NTA assay is crucial for understanding the limitations and challenges associated with the workflow, from the experimental methods to the data acquisition and data processing techniques. In this review, we examined recent NTA studies published between 2017 and 2023 that employed liquid chromatography-high-resolution mass spectrometry. The parameters used in each study were documented, and the reported chemicals at confidence levels 1 and 2 were retrieved. The chosen experimental setups and the quality of the reporting were critically evaluated and discussed. Our findings reveal that only around 2% of the estimated chemical space was covered by the NTA studies investigated for this review. Little to no trend was found between the experimental setup and the observed coverage due to the generic and wide scope of the NTA studies. The limited coverage of the chemical space by the reviewed NTA studies highlights the necessity for a more comprehensive approach in the experimental and data processing setups in order to enable the exploration of a broader range of chemical space, with the ultimate goal of protecting human and environmental health. Recommendations for further exploring a wider range of the chemical space are given.
Topics: Humans; Biological Assay; Chromatography, Liquid; Environmental Health; Mass Spectrometry
PubMed: 37704971
DOI: 10.1021/acs.est.3c03606 -
Journal of Separation Science Sep 2023Ultralow flow LC employs ultra-narrow bore columns and mid-range pL/min to low nL/min flow rates (i.e., ≤20 nL/min). The separation columns that are used under these... (Review)
Review
Ultralow flow LC employs ultra-narrow bore columns and mid-range pL/min to low nL/min flow rates (i.e., ≤20 nL/min). The separation columns that are used under these conditions are typically 2-30 μm in inner diameter. Ultralow flow LC systems allow for exceptionally high sensitivity and frequently high resolution. There has been an increasing interest in the analysis of scarce biological samples, for example, circulating tumor cells, extracellular vesicles, organelles, and single cells, and ultralow flow LC was efficiently applied to such samples. Hence, advances towards dedicated ultralow flow LC instrumentation, technical approaches, and higher throughput (e.g., tens-to-hundreds of single cells analyzed per day) were recently made. Here, we review the types of ultralow flow LC technology, followed by a discussion of selected representative ultralow flow LC applications, focusing on the progress made in bioanalysis of amount-limited samples during the last 10 years. We also discuss several recently reported high-sensitivity applications utilizing flow rates up to 100 nL/min, which are below commonly used nanoLC flow rates. Finally, we discuss the path forward for future developments of ultralow flow LC.
Topics: Chromatography, Liquid
PubMed: 37528733
DOI: 10.1002/jssc.202300440 -
Molecules (Basel, Switzerland) Mar 2024The analysis of enantiomers in food has significant implications for food safety and human health. Conventional analytical methods employed for enantiomer analysis, such... (Review)
Review
The analysis of enantiomers in food has significant implications for food safety and human health. Conventional analytical methods employed for enantiomer analysis, such as gas chromatography and high-performance liquid chromatography, are characterized by their labor-intensive nature and lengthy analysis times. This review focuses on the development of rapid and reliable biosensors for the analysis of enantiomers in food. Electrochemical and optical biosensors are highlighted, along with their fabrication methods and materials. The determination of enantiomers in food can authenticate products and ensure their safety. Amino acids and chiral pesticides are specifically discussed as important chiral substances found in food. The use of sensors replaces expensive reagents, offers real-time analysis capabilities, and provides a low-cost screening method for enantiomers. This review contributes to the advancement of sensor-based methods in the field of food analysis and promotes food authenticity and safety.
Topics: Humans; Chromatography, High Pressure Liquid; Food; Pesticides; Chromatography, Gas; Biosensing Techniques; Stereoisomerism
PubMed: 38474618
DOI: 10.3390/molecules29051106 -
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 -
Nature Communications Jul 2023Peptide identification in liquid chromatography-tandem mass spectrometry (LC-MS/MS) experiments relies on computational algorithms for matching acquired MS/MS spectra...
Peptide identification in liquid chromatography-tandem mass spectrometry (LC-MS/MS) experiments relies on computational algorithms for matching acquired MS/MS spectra against sequences of candidate peptides using database search tools, such as MSFragger. Here, we present a new tool, MSBooster, for rescoring peptide-to-spectrum matches using additional features incorporating deep learning-based predictions of peptide properties, such as LC retention time, ion mobility, and MS/MS spectra. We demonstrate the utility of MSBooster, in tandem with MSFragger and Percolator, in several different workflows, including nonspecific searches (immunopeptidomics), direct identification of peptides from data independent acquisition data, single-cell proteomics, and data generated on an ion mobility separation-enabled timsTOF MS platform. MSBooster is fast, robust, and fully integrated into the widely used FragPipe computational platform.
Topics: Chromatography, Liquid; Tandem Mass Spectrometry; Deep Learning; Peptides; Algorithms; Databases, Protein
PubMed: 37500632
DOI: 10.1038/s41467-023-40129-9