bims-metlip 2024-06-16 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2024‒06‒16
thirty papers selected by
Sofia Costa, Matterworks

Gel-assisted mass spectrometry imaging enables sub-micrometer spatial lipidomics.
High-throughput, low-cost quantification of 11 therapeutic antibodies using caprylic acid precipitation and LC-MS/MS.
A new chemical derivatization reagent sulfonyl piperazinyl for the quantification of fatty acids using LC-MS/MS.
Quantifying acyl-chain diversity in isobaric compound lipids containing monomethyl branched-chain fatty acids.
Evaluation of a Reference-Free Collision Cross Section Calibration Strategy for Proteomics Using SLIM-Based High-Resolution Ion Mobility Spectrometry-Mass Spectrometry.
Dilute-and-shoot approach for the high-throughput LC-MS/MS determination of illicit drugs in the field of wastewater-based epidemiology.
Overcoming the challenge of potent endogenous interferences in limaprost quantification: An innovative methodology combining differential mobility spectrometry with LC-MS/MS for ultra-high sensitivity, selectivity and significantly enhanced throughput.
Quantification of multiple steroid hormones in serum and human breast cancer tissue by liquid chromatography-tandem mass spectrometry analysis.
Detection of 13 emerging soil pollutant compounds using a dual extraction method (QuEChERS and solid phase extraction) and a liquid chromatography/mass spectrometry LC-MS/MS method.
SPE-UHPLC-ESI-MS/MS Analysis of Cocaine and Its Metabolites in Conventional and Alternative Biological Specimens: Application to Real Samples.
Advances in Chromatographic and Mass Spectrometric Techniques for Analyzing Reducing Monosaccharides and Their Phosphates in Biological Samples.
Simultaneous measurement of COVID-19 treatment drugs (nirmatrelvir and ritonavir) in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study.
Pillar Array Mixer for Postcolumn Derivatization Integrated into Liquid Chromatography-Based Microfluidic Device.
A sensitive bioanalytical ultra-high-performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantitation of lactone and carboxylate forms of topotecan in plasma and vitreous.
Advanced Method for the Detection of Saturated Monoglycerides in Biodiesel Using GC-EI-MS/MS.
Combining Gas Exchange and Rapid Quenching of Leaf Tissue for Mass Spectrometry and NMR Analysis Using an External Chamber.
Solving the retention time repeatability problem of hydrophilic interaction liquid chromatography.
Metabolite profiling of liquiritin in acute myocardial infarction model rat after intragastric administration using an information-dependent acquisition-mediated ultra-high-performance liquid chromatography-tandem mass spectrometry method.
Ultrafast, Selective, and Highly Sensitive Nonchromatographic Analysis of Fourteen Cannabinoids in Cannabis Extracts, Δ8-Tetrahydrocannabinol Synthetic Mixtures, and Edibles by Cyclic Ion Mobility Spectrometry-Mass Spectrometry.
Untargeted metabolomics of blood plasma samples of patients with hepatocellular carcinoma.
Impact of the delay in cryopreservation timing during biobanking procedures on human liver tissue metabolomics.
Deep profiling of plasmalogens by coupling the Paternò-Büchi derivatization with tandem mass spectrometry.
Quantification of Efavirenz Hydroxymetabolites in Human Plasma Using LC-HRMS/MS.
Biomonitoring urinary organophosphorus flame retardant metabolites by liquid-liquid extraction and ultra-high performance liquid chromatography-tandem mass spectrometry and their association with oxidative stress.
Wideband PRM: Highly Accurate and Sensitive Method for High-Throughput Targeted Proteomics.
Chain electrospray ionization mass spectrometry for ultra-low volume sample analysis.
Development and validation of a method for analysis of 25 cannabinoids in oral fluid and exhaled breath condensate.
Development and validation of LC-MS/MS method for trace analysis of acrylamide, acrylic acid and N, N-methylene bis acrylamide in sandy loam soil.
Low-Cost Alternative for Online Analysis of Volatile Organic Compounds.
The Pre- and Post-column Derivatization on Monosaccharide Composition Analysis, a Review.

Nat Commun. 2024 Jun 12. 15(1): 5036

Gel-assisted mass spectrometry imaging enables sub-micrometer spatial lipidomics.

Yat Ho Chan, Koralege C Pathmasiri, Dominick Pierre-Jacques, Maddison C Hibbard, Nannan Tao, Joshua L Fischer, Ethan Yang, Stephanie M Cologna, Ruixuan Gao.

A technique capable of label-free detection, mass spectrometry imaging (MSI) is a powerful tool for spatial investigation of native biomolecules in intact specimens. However, MSI has often been precluded from single-cell applications due to the spatial resolution limit set forth by the physical and instrumental constraints of the method. By taking advantage of the reversible interaction between the analytes and a superabsorbent hydrogel, we have developed a sample preparation and imaging workflow named Gel-Assisted Mass Spectrometry Imaging (GAMSI) to overcome the spatial resolution limits of modern mass spectrometers. With GAMSI, we show that the spatial resolution of MALDI-MSI can be enhanced ~3-6-fold to the sub-micrometer level without changing the existing mass spectrometry hardware or analysis pipeline. This approach will vastly enhance the accessibility of MSI-based spatial analysis at the cellular scale.

DOI: https://doi.org/10.1038/s41467-024-49384-w
Anal Chim Acta. 2024 Jul 18. pii: S0003-2670(24)00590-7. [Epub ahead of print]1313 342789

High-throughput, low-cost quantification of 11 therapeutic antibodies using caprylic acid precipitation and LC-MS/MS.

Erik I Hallin, Trond Trætteberg Serkland, Tormod K Bjånes, Silje Skrede.

  BACKGROUND: Therapeutic drug monitoring of treatment with therapeutic antibodies is hampered by the application of a wide range of different methods in the quantification of serum levels. LC-MS based methods could significantly improve comparability of results from different laboratories, but such methods are often considered complicated and costly. We developed a method for LC-MS/MS based quantification of 11 therapeutic antibodies concomitantly measured in a single run, with emphasis on simplicity in sample preparation and low cost.RESULTS: After a single-step sample purification using caprylic acid precipitation to remove interfering proteins, the sample underwent proteolysis followed by LC-MS/MS analysis. Infliximab is used as internal standard for sample preparation while isotope-labeled signature peptides identified for each analyte are internal standards for the LC-MS/MS normalization. The method was validated according to recognized guidelines, and pipetting steps can be performed by automated liquid handling systems. The total precision of the method ranged between 2.7 and 7.3 % (5.1 % average) across the quantification range of 4-256 μg/ml for all 11 drugs, with an average accuracy of 96.3 %. Matrix effects were xamined in 55 individual patient samples instead of the recommended 6, and 147 individual patient samples were screened for interfering compounds.
SIGNIFICANCE AND NOVELTY: Our method for simultaneous quantification of 11 t-mAb in human serum allows an unprecedented integration of robustness, speed and reduced complexity, which could pave the way for uniform use in research projects and clinical settings alike. In addition, the first LC-MS protocol for signature peptide-based quantification of durvalumab is described. This high throughput method can be readily adapted to a drug panel of choice.

Keywords:  Liquid chromatography-mass spectrometry; Multiplex; Protein precipitation; Quantification; Signature peptide; Therapeutic antibody

DOI:  https://doi.org/10.1016/j.aca.2024.342789
Talanta. 2024 Jun 06. pii: S0039-9140(24)00757-4. [Epub ahead of print]277 126378

A new chemical derivatization reagent sulfonyl piperazinyl for the quantification of fatty acids using LC-MS/MS.

Siqi Li, Qinwen Xiao, Jiarui Sun, Zhaoqian Li, Mengting Zhang, Yuan Tian, Zunjian Zhang, Haijuan Dong, Yu Jiao, Fengguo Xu, Pei Zhang.

  In our previous study, a chemical derivatization reagent named 5-(dimethylamino) naphthalene-1-sulfonyl piperazine (Dns-PP) was developed to enhance the chromatographic retention and the mass spectrometric response of free fatty acids (FFAs) in reversed-phase liquid chromatography coupled with electrospray ionization-mass spectrometry (RPLC-ESI-MS). However, Dns-PP exhibited strong preferences for long-chain FFAs, with limited improvement for short- or medium-chain FFAs. In this study, a new series of labeling reagents targeting FFAs were designed, synthesized, and evaluated. Among these reagents, Tmt-PP (N2, N2, N4, N4-tetramethyl-6-(4-(piperazin-1-ylsulfonyl) phenyl)-1,3,5-triazine-2,4-diamine) exhibited the best MS response and was selected for further evaluations. We compared Tmt-PP with Dns-PP and four commonly used carboxyl labeling reagents from existing studies, demonstrating the advantages of Tmt-PP. Further comparisons between Tmt-PP and Dns-PP in measuring FFAs from biological samples revealed that Tmt-PP labeling enhanced the MS response for about 80 % (30/38) of the measured FFAs, particularly for short- and medium-chain FFAs. Moreover, Tmt-PP labeling significantly improved the chromatographic retention of short-chain FFAs. To ensure accurate quantification, we developed a stable isotope-labeled Tmt-PP (i.e., d12-Tmt-PP) to react with chemical standards and serve as one-to-one internal standards (IS). The method was validated for accuracy, precision, sensitivity, linearity, stability, extraction efficiency, as well as matrix effect. Overall, this study introduced a new chemical derivatization reagent Tmt-PP (d12-Tmt-PP), providing a sensitive and accurate option for quantifying FFAs in biological samples.

Keywords:  Chemical derivatization; Free fatty acids; LC−MS; Submetabolome; Targeted metabolomics

DOI:  https://doi.org/10.1016/j.talanta.2024.126378
bioRxiv. 2024 May 28. pii: 2024.05.28.596332. [Epub ahead of print]

Quantifying acyl-chain diversity in isobaric compound lipids containing monomethyl branched-chain fatty acids.

C R Green, M J Kolar, G H McGregor, A T Nelson, M Wallace, C M Metallo.

Compound lipids comprise a diverse group of metabolites present in living systems, and metabolic- and environmentally-driven structural distinctions across this family is increasingly linked to biological function. However, methods for deconvoluting these often isobaric lipid species are lacking or require specialized instrumentation. Notably, acyl-chain diversity within cells may be influenced by nutritional states, metabolic dysregulation, or genetic alterations. Therefore, a reliable, validated method of quantifying structurally similar even-, odd-, and branched-chain acyl groups within intact compound lipids will be invaluable for gaining molecular insights into their biological functions. Here we demonstrate the chromatographic resolution of isobaric lipids containing distinct combinations of straight-chain and branched-chain acyl groups via ultra-high-pressure liquid chromatography (UHPLC)-mass spectrometry (MS) using a C30 liquid chromatography column. Using metabolically-engineered adipocytes lacking branched-keto acid dehydrogenase A (Bckdha), we validate this approach through a combination of fatty acid supplementation and metabolic tracing using monomethyl branched-chain fatty acids and valine. We observe resolution of numerous isobaric triacylglycerols and other compound lipids, demonstrating the resolving utility of this method. This approach strengthens our ability to quantify and characterize the inherent diversity of acyl chains across the lipidome.

DOI: https://doi.org/10.1101/2024.05.28.596332
J Am Soc Mass Spectrom. 2024 Jun 12.

Evaluation of a Reference-Free Collision Cross Section Calibration Strategy for Proteomics Using SLIM-Based High-Resolution Ion Mobility Spectrometry-Mass Spectrometry.

Dylan H Ross, Jung Yun Lee, Yuqian Gao, Adam L Hollerbach, Aivett Bilbao, Tujin Shi, Yehia M Ibrahim, Richard D Smith, Xueyun Zheng.

Ion mobility spectrometry (IMS) is a gas-phase analytical technique that separates ions with different sizes and shapes and is compatible with mass spectrometry (MS) to provide an additional separation dimension. The rapid nature of the IMS separation combined with the high sensitivity of MS-based detection and the ability to derive structural information on analytes in the form of the property collision cross section (CCS) makes IMS particularly well-suited for characterizing complex samples in -omics applications. In such applications, the quality of CCS from IMS measurements is critical to confident annotation of the detected components in the complex -omics samples. However, most IMS instrumentation in mainstream use requires calibration to calculate CCS from measured arrival times, with the most notable exception being drift tube IMS measurements using multifield methods. The strategy for calibrating CCS values, particularly selection of appropriate calibrants, has important implications for CCS accuracy, reproducibility, and transferability between laboratories. The conventional approach to CCS calibration involves explicitly defining calibrants ahead of data acquisition and crucially relies upon availability of reference CCS values. In this work, we present a novel reference-free approach to CCS calibration which leverages trends among putatively identified features and computational CCS prediction to conduct calibrations post-data acquisition and without relying on explicitly defined calibrants. We demonstrated the utility of this reference-free CCS calibration strategy for proteomics application using high-resolution structures for lossless ion manipulations (SLIM)-based IMS-MS. We first validated the accuracy of CCS values using a set of synthetic peptides and then demonstrated using a complex peptide sample from cell lysate.

DOI: https://doi.org/10.1021/jasms.4c00141
Water Res. 2024 Jun 04. pii: S0043-1354(24)00765-6. [Epub ahead of print]259 121864

Dilute-and-shoot approach for the high-throughput LC-MS/MS determination of illicit drugs in the field of wastewater-based epidemiology.

Elisa Gracia-Marín, Félix Hernández, María Ibáñez, Lubertus Bijlsma.

  The determination of illicit drugs in urban influent wastewater (IWW) enables the monitoring of spatial and temporal drug usage trends and assessment of community lifestyle habits. The increasing number of wastewater surveillance studies has emphasized the necessity for the development of rapid, high-throughput methods that maintain high quality data. This work evaluates the use of a dilute-and-shoot methodology, based on direct injection (DI) of centrifuged samples, as an alternative approach to the widely applied sample pre-treatment based on solid-phase extraction, for the liquid chromatography-tandem mass spectrometry determination of seven widely consumed illicit drugs and their metabolites in IWW (amphetamine; cocaine metabolite, benzoylecgonine; ketamine; 3,4-methylenedioxymethamphetamine (MDMA); methamphetamine; cannabis metabolite, 11-nor-9-carboxy-delta-9-tetrahydrocannabinol (THCCOOH); heroin metabolite, 6-acetylmorphine (6-MAM)). Comparison of both approaches in terms of matrix effects, sensitivity and accuracy, demonstrates the DI method suitability to correctly quantify these analytes in IWW, with a limit of quantification lower than 30 ng L-1 for most compounds. After validation of the method and participation in an interlaboratory exercise, the DI method was applied to the analysis of 54 IWW samples collected from different Spanish wastewater treatment plants. Additionally, quality controls were incorporated in each analysis batch to support the DI method applicability and robustness. The use of a 10 μL-DI reduces time-consuming sample preparation, analysis time and measurement uncertainty. Moreover, it supports green chemistry by reducing the consumption of organic solvents and it facilitates logistics by collecting, transporting, and storing less sample volume. The methodology is therefore especially appropriate for monitoring illicit drugs in large wastewater-based epidemiology sampling campaigns or when fast near real-time results are needed.

Keywords:  Direct injection; Drugs of abuse; Green chemistry; Liquid chromatography-tandem mass spectrometry; Solid phase extraction; Wastewater surveillance

DOI:  https://doi.org/10.1016/j.watres.2024.121864
Talanta. 2024 Jun 10. pii: S0039-9140(24)00790-2. [Epub ahead of print]277 126411

Overcoming the challenge of potent endogenous interferences in limaprost quantification: An innovative methodology combining differential mobility spectrometry with LC-MS/MS for ultra-high sensitivity, selectivity and significantly enhanced throughput.

Xiangjun Meng, Chong Su, Zhi Zhang, Yaoshuang Li, Yuyao Zhang, Jingtao Li, Leling Gan, Jingkai Gu.

  Limaprost, an orally administered analogue of prostaglandin E1, possesses potent vasodilatory, antiplatelet, and cytoprotective properties. Due to its extremely low therapeutic doses and exceedingly low plasma concentrations, the pharmacokinetic and bioequivalence studies of limaprost necessitate a highly sensitive quantitative method with a sub-pg/mL level of lower limit of quantification. Moreover, the intensity of endogenous interferences can even exceed the maximum concentration level of limaprost in human plasma, presenting further challenge to the quantification of limaprost. As a result, existing methods have not yet met the necessary level of sensitivity, selectivity, and throughput needed for the quantitative analysis of limaprost in pharmacokinetic and bioequivalence investigations. This study presents a new methodology that combines differential mobility spectrometry (DMS) with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and utilizes a distinctive strategy to achieve more accurate DMS conditions. This integration yields a method that is currently the most sensitive and features the shortest analytical time, making it the sole technique capable of meeting the requirements for limaprost pharmacokinetic and bioequivalence investigations. This method demonstrates robustness and is successfully employed in a pharmacokinetic investigation of limaprost in human subjects, underscoring that the combination of DMS with LC-MS/MS serves as an efficacious strategy for overcoming the challenges inherent in analyzing biological samples afflicted by multiple interferences.

Keywords:  Differential mobility spectrometry; Enhanced selectivity; LC-MS/MS; Limaprost; Ultra-high sensitivity

DOI:  https://doi.org/10.1016/j.talanta.2024.126411
Front Oncol. 2024 ;14 1383104

Quantification of multiple steroid hormones in serum and human breast cancer tissue by liquid chromatography-tandem mass spectrometry analysis.

Feng Wang, Eline Eikeland, Randi J Reidunsdatter, Lars Hagen, Monica J Engstrøm, Jürgen Geisler, Mikko Haanpää, Esa Hämäläinen, Guro F Giskeødegård, Tone F Bathen.

  Introduction: Systemic and local steroid hormone levels may function as novel prognostic and predictive biomarkers in breast cancer patients. We aimed at developing a novel liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous measurement of multiple, biologically pivotal steroid hormones in human serum and breast cancer tissue.Methods: The quantitative method consisted of liquid-liquid extraction, Sephadex LH-20 chromatography for tissue extracts, and analysis of steroid hormones by liquid-chromatography-tandem mass spectrometry. We analyzed serum and tissue steroid hormone levels in 16 and 40 breast cancer patients, respectively, and assessed their correlations with clinical parameters.
Results: The method included quantification of nine steroid hormones in serum [including cortisol, cortisone, corticosterone, estrone (E1), 17β-estradiol (E2), 17α-hydroxyprogesterone, androstenedione (A4), testosterone and progesterone) and six (including cortisone, corticosterone, E1, E2, A4, and testosterone) in cancer tissue. The lower limits of quantification were between 0.003-10 ng/ml for serum (250 µl) and 0.038-125 pg/mg for tissue (20 mg), respectively. Accuracy was between 98%-126%, intra-assay coefficient of variations (CV) was below 15%, and inter-assay CV were below 11%. The analytical recoveries for tissue were between 76%-110%. Tissue levels of E1 were positively correlated with tissue E2 levels (p<0.001), and with serum levels of E1, E2 and A4 (p<0.01). Tissue E2 levels were positively associated with serum E1 levels (p=0.02), but not with serum E2 levels (p=0.12). The levels of tissue E2 and ratios of E1 to A4 levels (an index for aromatase activity) were significantly higher in patients with larger tumors (p=0.03 and p=0.02, respectively).
Conclusions: The method was convenient and suitable for a specific and accurate profiling of clinically important steroid hormones in serum. However, the sensitivity of the profile method in steroid analysis in tissue samples is limited, but it can be used for the analysis of steroids in breast cancer tissues if the size of the sample or its steroid content is sufficient.

Keywords:  LC-MS/MS; breast cancer; breast cancer tissue; quantification; steroid hormone

DOI:  https://doi.org/10.3389/fonc.2024.1383104
MethodsX. 2024 Jun;12 102771

Detection of 13 emerging soil pollutant compounds using a dual extraction method (QuEChERS and solid phase extraction) and a liquid chromatography/mass spectrometry LC-MS/MS method.

Lara Maria Wakim, Amandine Descat, Florent Occelli, Annabelle Deram, Jean-François Goossens.

  Emerging pollutants derived from human and animal sources, are present in soils and pose significant environmental and health impacts, even at low concentrations. Their detection in soil is analytically complex due to soil interference and the rapid degradation of compounds in the matrix. In this study, a protocol was optimized for quantifying hormonal steroids (n = 7), human drugs (n = 3), and antibiotics (n = 3) by a dual-phase extraction using QuEChERS and Solid Phase Extraction (SPE), followed by analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The double extraction phase allows an accurate and effective purification of the target compounds while eliminating the interference in the soil matrix. The method is optimized to detect environmental concentrations of these pollutants, to suit large-scale sampling campaigns and to maintain the efficiency of extraction while reducing analysis time. The limits of detection (LODs) of these compounds ranged between 0.0043 and 0.13 ng/g and recovery rates between 75.9 % and 105.39 %.•Enhanced Analyte Purification: Implements QuEChERS and SPE for robust removal of matrix interferences, optimizing target compound isolation.•Precision at Trace Levels: Secures LODs as minimal as 0.0043 ng/g, enabling accurate detection of low-concentration contaminants.•Adapted for Broad-scale sampling: Modifies extraction and analysis durations to accommodate large-scale environmental assessments.

Keywords:  Antibiotics; Bisphenol A; Detection of emerging pollutants in soil using a dual extraction of QuEChERS and SPE followed by analysis by LC-MS/MS; Emerging pollutants; Hormones; Human drugs; Soil contamination

DOI:  https://doi.org/10.1016/j.mex.2024.102771
ACS Omega. 2024 Jun 04. 9(22): 23355-23363

SPE-UHPLC-ESI-MS/MS Analysis of Cocaine and Its Metabolites in Conventional and Alternative Biological Specimens: Application to Real Samples.

Humera Shafi Makhdoom, Saira Afzal, Kishwar Sultana, Syed Nisar Hussain Shah, Majida Mujahid, Zulfiqar Ul Hassan, Farida Munir, Faryal Jahan, Zeerak Abbas, Ali Imran Abid, Najm Ul Hassan Khan.

An increase in cocaine abuse has been observed globally since the past decade. Cocaine is among the commonly abused stimulants used for recreational purposes. In this study, the SPE-UHPLC-MS/MS method was developed and validated to be applied on real specimens of 20 chronic cocaine abusers to quantify cocaine/metabolites in conventional as well as alternative biological matrices. Cocaine was extracted from biological specimens using solid-phase extraction followed by liquid chromatography tandem mass spectrometry analysis. Chromatographic separation was achieved on a Poroshell120EC-18 column (2.1 mm × 50 mm, 2.7 μm particle size) using water-acetonitrile in 0.1% formic acid as a mobile phase in gradient elution mode. The flow rate of the mobile phase was 0.5 mL/min with a gradient varying the percentage of acetonitrile linearity ranging 15-95% in 6.0 min acquisition time, and the injection volume was set at 5 μL. Positive electrospray ionization with multireaction ion monitoring mode using two ion transitions for cocaine/metabolites and one for cocaine-d3 was employed. The quantification method demonstrated good linear ranges of 0.025-250 ng/mL in blood, urine, and oral fluid (ng/mg for hair and nail) with a ≥0.991% determination coefficient. The detection limit and lower quantification limit were 0.005 and 0.025 ng/mL in all matrices, respectively. The mean extraction recovery and ionization suppression ranged from 89.3 to 99.8% and -4.6 to -14.4% in the studied matrices. Within-run and between-days precisions were 1.8-7.2% and 1.9-6.1%, respectively. This study will not only help in quantifying cocaine/metabolites in alternative specimens (hair, nail, and oral fluid) but also guide clinical and forensic toxicologists in interpretation of exhumation cases. Furthermore, multiple specimens' analyses can be of significance in estimating the time/manner of drug exposure, in confirming the results of laboratories in cases of doubtful clinical histories, or in aiding medico-legal investigations.

DOI: https://doi.org/10.1021/acsomega.3c09669
Crit Rev Anal Chem. 2024 Jun 10. 1-23

Advances in Chromatographic and Mass Spectrometric Techniques for Analyzing Reducing Monosaccharides and Their Phosphates in Biological Samples.

Jiaqi Chen, Yifeng Lou, Yuwei Liu, Bowen Deng, Zheng Zhu, Sen Yang, Di Chen.

  Reducing monosaccharides and their phosphates are critical metabolites in the central carbon metabolism pathway of living organisms. Variations in their content can indicate abnormalities in metabolic pathways and the onset of certain diseases, necessitating their analysis and detection. Reducing monosaccharides and their phosphates exhibit significant variations in content within biological samples and are present in many isomers, which makes the accurate quantification of reducing monosaccharides and their phosphates in biological samples a challenging task. Various analytical methods such as spectroscopy, fluorescence detection, colorimetry, nuclear magnetic resonance spectroscopy, sensor-based techniques, chromatography, and mass spectrometry are employed to detect monosaccharides and phosphates. In comparison, chromatography and mass spectrometry are highly favored for their ability to simultaneously analyze multiple components and their high sensitivity and selectivity. This review thoroughly evaluates the current chromatographic and mass spectrometric methods used for detecting reducing monosaccharides and their phosphates from 2013 to 2023, highlighting their efficacy and the advancements in these analytical technologies.

Keywords:  Reducing monosaccharides; biological samples; chromatography; chromatography-mass spectrometry; detection techniques; mass spectrometry; sugar phosphates

DOI:  https://doi.org/10.1080/10408347.2024.2364232
Heliyon. 2024 Jun 15. 10(11): e32187

Simultaneous measurement of COVID-19 treatment drugs (nirmatrelvir and ritonavir) in rat plasma by UPLC-MS/MS and its application to a pharmacokinetic study.

Chen-Jian Zhou, Ya-Nan Liu, Anzhou Wang, Hualu Wu, Ren-Ai Xu, Qiang Zhang.

  PAXLOVID™ (Co-packaging of Nirmatrelvir with Ritonavir) has been approved for the treatment of Coronavirus Disease 2019 (COVID-19). The goal of the experiment was to create an accurate and straightforward analytical method using ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to simultaneously quantify nirmatrelvir and ritonavir in rat plasma, and to investigate the pharmacokinetic profiles of these drugs in rats. After protein precipitation using acetonitrile, nirmatrelvir, ritonavir, and the internal standard (IS) lopinavir were separated using ultra performance liquid chromatography (UPLC). This separation was achieved with a mobile phase composed of acetonitrile and an aqueous solution of 0.1% formic acid, using a reversed-phase column with a binary gradient elution. Using multiple reaction monitoring (MRM) technology, the analytes were detected in the positive electrospray ionization mode. Favorable linearity was observed in the calibration range of 2.0-10000 ng/mL for nirmatrelvir and 1.0-5000 ng/mL for ritonavir, respectively, within plasma samples. The lower limits of quantification (LLOQ) attained were 2.0 ng/mL for nirmatrelvir and 1.0 ng/mL for ritonavir, respectively. Both drugs demonstrated inter-day and intra-day precision below 15%, with accuracies ranging from -7.6% to 13.2%. Analytes were extracted with recoveries higher than 90.7% and without significant matrix effects. Likewise, the stability was found to meet the requirements of the analytical method under different conditions. This UPLC-MS/MS method, characterized by enabling accurate and precise quantification of nirmatrelvir and ritonavir in plasma, was effectively utilized for in vivo pharmacokinetic studies in rats.

Keywords:  Nirmatrelvir; Pharmacokinetics; Ritonavir; UPLC-MS/MS; rat

DOI:  https://doi.org/10.1016/j.heliyon.2024.e32187
Anal Chem. 2024 Jun 13.

Pillar Array Mixer for Postcolumn Derivatization Integrated into Liquid Chromatography-Based Microfluidic Device.

Muneki Isokawa, Kanki Nakanishi, Takahiro Kanamori, Tetsushi Sekiguchi, Takashi Funatsu, Shuichi Shoji, Makoto Tsunoda.

The chemical derivatization of target analytes can enhance the sensitivity and selectivity of separation-based methods for metabolite analysis using microfluidic devices. However, the development of chromatography-based microfluidic devices with integrated derivatization units is challenging. In this study, a novel derivatization unit with a pillar array (PA)-based mixing channel was developed for postcolumn derivatization during on-chip liquid chromatography (LC). The PA mixer enhanced mixing between the derivatization reagents and analytes in the transverse direction, while preventing analyte dispersion in the flow direction. After the concept was confirmed using computational fluid dynamics analysis, microfluidic devices with a LC column and PA mixer were fabricated on a 20 × 20 mm silicon plate. Fluid experiments were performed using a PA mixer with a pillar size of 5 or 10 μm or a hollow-channel mixer, which revealed that the PA mixer enhanced transverse mixing without increasing the width of the analyte peak. Moreover, the developed device enabled the analysis of three amino acids within 40 s by separation via hydrophilic interaction chromatography followed by postcolumn fluorogenic derivatization with naphthalene-2,3-dicarboxaldehyde and fluorescence detection. Our results demonstrate the potential of integrated derivatization units for the development of micrototal analysis systems for use in bioanalysis.

DOI: https://doi.org/10.1021/acs.analchem.4c01669
J Sep Sci. 2024 Jun;47(11): e2400181

A sensitive bioanalytical ultra-high-performance liquid chromatography-tandem mass spectrometry method for the simultaneous quantitation of lactone and carboxylate forms of topotecan in plasma and vitreous.

Barbora Mudrova, Katerina Hrabakova, Petr Kozlik, Radka Hobzova, Jakub Sirc, Zuzana Bosakova.

  Topotecan (TPT) is used in the treatment of retinoblastoma, the most common malignant intraocular tumor in children. TPT undergoes pH-dependent hydrolysis of the lactone ring to the ring-opened carboxylate form, with the lactone form showing antitumor activity. A selective, and highly sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry method was developed for the determination of both forms of TPT in one mobile phase composition in plasma and vitreous humor matrices. The method showed an excellent linear range of 0.375-120 ng/mL for the lactone. For the carboxylate, the linear range was from 0.75 to 120 ng/mL. The matrix effect and the recovery for the lactone ranged from 98.5% to 106.0% in both matrices, for the carboxylate form, it ranged from 94.9% to 101.2%. The dynamics of the transition between TPT lactone and TPT carboxylate were evaluated at different pH environments. The stability of TPT forms was assessed in plasma and vitreous humor at 8 and 37°C and a very fast conversion of lactone to carboxylate form occurred at 37°C in both matrices. The method developed facilitates the investigation of TPT pharmacodynamics and the release kinetics in the development of the innovative local drug delivery systems.

Keywords:  UHPLC‐MS/MS; anti‐cancer drug topotecan; carboxylate form; lactone form; topotecan stability

DOI:  https://doi.org/10.1002/jssc.202400181
ACS Omega. 2024 Jun 04. 9(22): 23476-23484

Advanced Method for the Detection of Saturated Monoglycerides in Biodiesel Using GC-EI-MS/MS.

Jessica Pichler, Marcella Frauscher, Martina Marchetti-Deschmann.

Common B7 biodiesels consist of mixtures of mineral oil-based diesel and 7% fatty acid methyl ester (FAME). While biocontent increase can be achieved with these blends at high-quality levels during cold temperature periods, fuel filter blocking events are reported from time to time. Based on a preliminary study on fuel filters, the selection of compounds responsible for filter blocking could be narrowed down to saturated monoglycerides (SMGs). The most abundant SMGs in Europe are 1- and 2-monopalmitin (1-C16:0, 2-C16:0) and 1-monostearin (C18:0), based on the FAME origin. Until now, there has been no simple, precise, and accurate method to quantitatively detect those SMGs in the B7 matrix, which was the aim of the following work. An improved gas chromatography electron ionization tandem mass spectrometry method was developed for the quantitative detection of 1-C16:0, 2-C16:0, C18:0, and C20:0 SMGs. During the method improvement, (a) the sample preparation and (b) the calibration were optimized for low concentrations. (c) The samples were analyzed by multiple reaction monitoring focusing on specific qualifier and quantifier ions with optimized collision energies, (d) time segments and improved scan time were implemented, and (e) limits of detection and limits of quantification were determined. The time-stability of SMG standards in CHCl3 with 4% neat biodiesel and the discrimination effects of the standard components were evaluated to assess method reliability. Overall, a highly sensitive and precise method for the improved detection of SMGs in biodiesel is presented.

DOI: https://doi.org/10.1021/acsomega.4c00513
Methods Mol Biol. 2024 ;2792 195-208

Combining Gas Exchange and Rapid Quenching of Leaf Tissue for Mass Spectrometry and NMR Analysis Using an External Chamber.

Cyril Abadie, Julie Lalande, Guillaume Tcherkez.

  We describe here a method to study and manipulate photorespiration in intact illuminated leaves. When the CO2/O2 mole fraction ratio changes, instant sampling is critical, to quench leaf metabolism and thus trace rapid metabolic modification due to gaseous conditions. To do so, we combine 13CO2 labeling and gas exchange, using a large custom leaf chamber to facilitate fast sampling by direct liquid nitrogen spraying. Moreover, the use of a high chamber surface area (about 130 cm2) allows one to sample a large amount of leaf material to carry out 13C-nuclear magnetic resonance (NMR) analysis and complementary analyses, such as isotopic analyses by high-resolution mass spectrometry (by both GC and LC-MS). 13C-NMR gives access to absolute 13C amounts at the specific carbon atom position in the labeled molecules and thereby provides an estimate of 13C-flux of photorespiratory intermediates. Since NMR analysis is not very sensitive and can miss minor metabolites, GC or LC-MS analyses are useful to monitor metabolites at low concentrations. Furthermore, 13C-NMR and high-resolution LC-MS allow to estimate isotopologue distribution in response to 13CO2 labeling while modifying photorespiration activity.

Keywords:   13C labeling; Custom leaf chamber; High-resolution mass spectrometry; Nuclear Magnetic Resonance; Photorespiration

DOI:  https://doi.org/10.1007/978-1-0716-3802-6_16
J Chromatogr A. 2024 Jun 08. pii: S0021-9673(24)00434-5. [Epub ahead of print]1730 465060

Solving the retention time repeatability problem of hydrophilic interaction liquid chromatography.

Kristian Serafimov, Cornelius Knappe, Feiyang Li, Adrian Sievers-Engler, Michael Lämmerhofer.

  Hydrophilic interaction (liquid) chromatography (HILIC) has become the first choice LC mode for the separation of hydrophilic analytes. Numerous studies reported the poor retention time repeatability of HILIC. The problem was often ascribed to slow equilibration and insufficient re-equilibration time to establish the sensitive semi-immobilized water layer at the interface of the polar stationary phase and the bulk mobile phase. In this study, we compare retention time repeatability in HILIC for borosilicate glass and PFA (co-polymer of tetrafluoroethylene and perfluoroalkoxyethylene) solvent bottles. During this study, we observed peak patterns shifting towards higher retention times (for metabolites and peptides) and lower retention times (oligonucleotide sample) with ongoing analysis time when standard borosilicate glass bottles were used as solvent reservoirs. It was hypothesized that release of ions (sodium, potassium, borate, etc.) from the borosilicate glass bottles leads to alterations (thickness and electrostatic screening effects) in the semi-immobilized water layer which is adsorbed to the polar stationary phase surface under acetonitrile-rich eluents in HILIC with concomitant shifts in retention. When PFA solvent bottles were employed instead of borosilicate glass, retention time repeatability was greatly improved and changed from average 8.4 % RSD for the tested metabolites with borosilicate glass bottles to 0.14 % RSD for the PFA solvent bottles (30 injections over 12 h). Similar improvements were observed for peptides and oligonucleotides. This simple solution to the retention time repeatability problem in HILIC might contribute to a better acceptance of HILIC, especially in fields like targeted and untargeted metabolomics, peptide and oligonucleotide analysis.

Keywords:  HILIC; Liquid chromatography-mass spectrometry; Metabolomics; Oligonucleotide; Peptide

DOI:  https://doi.org/10.1016/j.chroma.2024.465060
Biomed Chromatogr. 2024 Jun 11. e5933

Metabolite profiling of liquiritin in acute myocardial infarction model rat after intragastric administration using an information-dependent acquisition-mediated ultra-high-performance liquid chromatography-tandem mass spectrometry method.

Jian Chen, Jing Li, Feng Wang, Ruirui Ge, Liang Wang, Jinling Huang.

  Liquiritin (LQ), a kind of flavonoid isolated from licorice, was proven to have great potential in treating heart failure. Pharmacokinetic evaluation is important for demonstrating clinical efficacy and mechanisms, and the prototype drug and its metabolite profiling are important for drug discovery and development. However, the metabolism of LQ in acute myocardial infarction (AMI) model rats still needs to be studied in depth. An information-dependent acquisition (IDA)-ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was applied to profile LQ metabolites in AMI model rat plasma. Protein precipitation and extraction were used for sample preparation. Chromatographic separation was achieved using an XSelect BEH C18 column (2.1 × 150 mm, 2.5 μm) using gradient elution method combining 0.1% formic acid and acetonitrile with a flow rate of 0.3 mL/min. Twelve metabolites were identified in IDA mode, sulfation, glucuronidation, methylation, methyl esterification, glutamine conjugation, and valine conjugation, and their composite reactions were presumed as the primary pathways of LQ metabolism. The variation in the peak areas showed that the time to reach the peak drug concentration of LQ and 12 metabolites was within 5 h. In summary, IDA-bridged UHPLC-MS/MS from characteristic fragment ions toward confidence-enhanced identification could effectively screen and profile metabolites.

Keywords:  UHPLC‐MS/MS; acute myocardial infarction; information‐dependent acquisition; liquiritin; metabolites

DOI:  https://doi.org/10.1002/bmc.5933
Anal Chem. 2024 Jun 11.

Ultrafast, Selective, and Highly Sensitive Nonchromatographic Analysis of Fourteen Cannabinoids in Cannabis Extracts, Δ8-Tetrahydrocannabinol Synthetic Mixtures, and Edibles by Cyclic Ion Mobility Spectrometry-Mass Spectrometry.

Si Huang, Laura Righetti, Frank W Claassen, Akash Krishna, Ming Ma, Teris A van Beek, Bo Chen, Han Zuilhof, Gert Ij Salentijn.

The diversity of cannabinoid isomers and complexity of Cannabis products pose significant challenges for analytical methodologies. In this study, we developed a method to analyze 14 different cannabinoid isomers in diverse samples within milliseconds by leveraging the unique adduct-forming behavior of silver ions in advanced cyclic ion mobility spectrometry-mass spectrometry. The developed method achieved the separation of isomers from four groups of cannabinoids: Δ3-tetrahydrocannabinol (THC) (1), Δ8-THC (2), Δ9-THC (3), cannabidiol (CBD) (4), Δ8-iso-THC (5), and Δ(4)8-iso-THC (6) (all MW = 314); 9α-hydroxyhexahydrocannabinol (7), 9β-hydroxyhexahydrocannabinol (8), and 8-hydroxy-iso-THC (9) (all MW = 332); tetrahydrocannabinolic acid (THCA) (10) and cannabidiolic acid (CBDA) (11) (both MW = 358); Δ8-tetrahydrocannabivarin (THCV) (12), Δ8-iso-THCV (13), and Δ9-THCV (14) (all MW = 286). Moreover, experimental and theoretical traveling wave collision cross section values in nitrogen (TWCCSN2) of cannabinoid-Ag(I) species were obtained for the first time with an average error between experimental and theoretical values of 2.6%. Furthermore, a workflow for the identification of cannabinoid isomers in Cannabis and Cannabis-derived samples was established based on three identification steps (m/z and isotope pattern of Ag(I) adducts, TWCCSN2, and MS/MS fragments). Afterward, calibration curves of three major cannabinoids were established with a linear range of 1-250 ng·ml-1 for Δ8-THC (2) (R2 = 0.9999), 0.1-25 ng·ml-1 for Δ9-THC (3) (R2 = 0.9987), and 0.04-10 ng·ml-1 for CBD (4) (R2 = 0.9986) as well as very low limits of detection (0.008-0.2 ng·ml-1). Finally, relative quantification of Δ8-THC (2), Δ9-THC (3), and CBD (4) in eight complex acid-treated CBD mixtures was achieved without chromatographic separation. The results showed good correspondence (R2 = 0.999) with those obtained by gas chromatography-flame ionization detection/mass spectrometry.

DOI: https://doi.org/10.1021/acs.analchem.3c05879
J Pharm Biomed Anal. 2024 May 28. pii: S0731-7085(24)00303-0. [Epub ahead of print]248 116263

Untargeted metabolomics of blood plasma samples of patients with hepatocellular carcinoma.

Adéla Böhmová, Miloš Mikoška, Kamila Syslová, Dominika Šindelářová, Petr Hříbek, Petr Urbánek, Vladimír Setnička.

  Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths in the world. HCC is often diagnosed late because patients with early-stage cancer have no apparent symptoms. Therefore, it is desirable to find a reliable method for an early diagnosis based on the detection of metabolites - biomarkers, that can be detected in the early stages of the disease. Untargeted metabolomics is often used as a tool to find a suitable biomarker for several diseases. In this work, untargeted metabolomics was performed on blood plasma samples of HCC patients and compared with healthy individuals and patients with liver cirrhosis. A combination of liquid chromatography and high-resolution mass spectrometry was used as an analytical method. More than a thousand peaks were detected in the blood plasma samples, from which mainly amino acids, carboxylic acids, lipids, and their derivatives were evaluated as potential biomarkers. The data obtained were statistically processed using the analysis of variance, correlation analysis, and principal component analysis.

Keywords:  Biomarkers; Cirrhosis; Hepatocellular carcinoma; LC-MS; Metabolomics

DOI:  https://doi.org/10.1016/j.jpba.2024.116263
PLoS One. 2024 ;19(6): e0304405

Impact of the delay in cryopreservation timing during biobanking procedures on human liver tissue metabolomics.

Corentine Goossens, Vincent Tambay, Valérie-Ann Raymond, Louise Rousseau, Simon Turcotte, Marc Bilodeau.

The liver is a highly specialized organ involved in regulating systemic metabolism. Understanding metabolic reprogramming of liver disease is key in discovering clinical biomarkers, which relies on robust tissue biobanks. However, sample collection and storage procedures pose a threat to obtaining reliable results, as metabolic alterations may occur during sample handling. This study aimed to elucidate the impact of pre-analytical delay during liver resection surgery on liver tissue metabolomics. Patients were enrolled for liver resection during which normal tissue was collected and snap-frozen at three timepoints: before transection, after transection, and after analysis in Pathology. Metabolomics analyses were performed using 1H Nuclear Magnetic Resonance (NMR) and Liquid Chromatography-Mass Spectrometry (LC-MS). Time at cryopreservation was the principal variable contributing to differences between liver specimen metabolomes, which superseded even interindividual variability. NMR revealed global changes in the abundance of an array of metabolites, namely a decrease in most metabolites and an increase in β-glucose and lactate. LC-MS revealed that succinate, alanine, glutamine, arginine, leucine, glycerol-3-phosphate, lactate, AMP, glutathione, and NADP were enhanced during cryopreservation delay (all p<0.05), whereas aspartate, iso(citrate), ADP, and ATP, decreased (all p<0.05). Cryopreservation delays occurring during liver tissue biobanking significantly alter an array of metabolites. Indeed, such alterations compromise the integrity of metabolomic data from liver specimens, underlining the importance of standardized protocols for tissue biobanking in hepatology.

DOI: https://doi.org/10.1371/journal.pone.0304405
Anal Bioanal Chem. 2024 Jun 11.

Deep profiling of plasmalogens by coupling the Paternò-Büchi derivatization with tandem mass spectrometry.

Yichun Wang, Yu Xia.

  Plasmalogens are a special class of glycerophospholipids characterized by a vinyl ether bond (-C = C-O-) at the sn-1 position of the glycerol backbone. Altered plasmalogen profiles have been observed in neurodegenerative diseases and cancers. Profiling of plasmalogens requires specifying the vinyl ether bond and differentiating them from various types of isobars and isomers. Herein, by coupling C = C derivatization via offline Paternò-Büchi reaction with liquid chromatography-tandem mass spectrometry, we have developed a sensitive workflow for analysis of plasmalogens from biological samples. Using bovine heart lipid extract as a model system, we profiled more than 100 distinct structures of plasmenylethanolamines (PE-Ps) and plasmenylcholines (PC-Ps) at the C = C location level, far exceeding previous reports. Analysis of human glioma and normal brain tissue samples revealed elevated n-10 C = C isomers of PE-Ps in the glioma tissue samples. These findings suggest that the developed workflow holds potential in aiding the study of altered metabolism of plasmalogens in clinical samples.

Keywords:  Double bond position; Lipidomics; Liquid chromatography; Plasmalogen; Tandem mass spectrometry; The Paternò–Büchi reaction

DOI:  https://doi.org/10.1007/s00216-024-05376-9
Ther Drug Monit. 2024 Jun 11.

Quantification of Efavirenz Hydroxymetabolites in Human Plasma Using LC-HRMS/MS.

Madeleine Pettersson Bergstrand, Sandra Soeria-Atmadja, Victoria Barclay, Jelena Tolic, Lars Navér, Lars L Gustafsson, Anton Pohanka.

BACKGROUND: Efavirenz (EFV) is a drug used to treat HIV. Low plasma concentrations of EFV result in suboptimal viral suppression, whereas high concentrations can cause adverse neuropsychiatric side reactions. Some studies have identified a correlation between the plasma concentrations of EFV metabolites and neurotoxicity. To our knowledge, no studies have investigated the metabolism of EFV in young children and its effect on treatment outcomes. Therefore, the aim of this study was to develop and validate a method for quantifying EFV and its metabolites in human plasma derived from children.METHODS: Sample preparation was performed using protein precipitation of 100 µL plasma. Thereafter, an aliquot of the supernatant was used to quantify EFV, 7-hydroxyefavirenz (7-OH-EFV), 8-hydroxyefavirenz (8-OH-EFV), and a newly discovered metabolite ("EFAdeg") associated with 8-OH-EFV. A second aliquot of the supernatant was hydrolyzed using β-glucuronidase/arylsulfatase and used with the first aliquot to quantify phase II metabolites. The analyses were performed using a Dionex Ultimate 3000RS LC-system coupled with a Q Exactive Orbitrap mass spectrometer.
RESULTS: The method has a measuring range of 100-50,000 ng/mL (EFV, 8-OH-EFV), 125-25,000 ng/mL (7-OH-EFV), and 200-10,000 ng/mL ("EFAdeg"). All criteria of the European Medicines Agency guidelines regarding precision, accuracy, and selectivity were met. Of note, carryover must be considered for 8-OH-EFV. Overall, the validated method was successfully applied to plasma samples obtained from children and confirmed the presence of the newly discovered metabolite, "EFAdeg."
CONCLUSIONS: An LC-HRMS/MS method for the quantification of EFV and its phase I and II metabolites was developed and validated. This method is suitable for analyzing plasma samples from children. Furthermore, studies using this method identified an additional metabolite that may influence the concentration of 8-OH-EFV in patient samples.

DOI: https://doi.org/10.1097/FTD.0000000000001173
Anal Bioanal Chem. 2024 Jun 14.

Biomonitoring urinary organophosphorus flame retardant metabolites by liquid-liquid extraction and ultra-high performance liquid chromatography-tandem mass spectrometry and their association with oxidative stress.

Yongxian Li, Yingyi Dai, Xinni Luo, Lin Zhang, Jun Yuan, Lei Tan.

  Organophosphate flame retardants (OPFRs) are widely used as substitutes for traditional brominated flame retardants, necessitating a reliable and sensitive method for biomonitoring their urinary metabolites to assess human exposure. This study conducted biomonitoring of 10 metabolites of OPFRs in 152 adults and assessed their association with oxidative stress biomarkers 8-hydroxydeoxyguanosine and 8-hydroxyguanosine. Urinary metabolites of OPFRs were released via enzymatic deconjugation. The addition of sodium chloride to the urine samples increases the ionic strength, inducing a salting-out effect that reduces the solubility of these compounds, thereby facilitating their extraction with a mixture of ethyl acetate and acetonitrile. Then, the metabolites of OPFRs were quantified by ultra-high performance liquid chromatography-tandem mass spectrometry, and we validated the method for linear range, precision, matrix effect, and method detection limit. The detection limit of the metabolites of OPFRs ranged from 0.01 to 0.2 μg/L, and these metabolites were detected with high frequencies ranging from 25.0 to 98.68% in the urine samples. The concentration of bis (2-chloroethyl) phosphate was significantly higher in males than in females, with the geometric mean concentration of 0.88 μg/L for males and 0.53 μg/L for females, respectively. Spearman correlation analysis revealed weak but statistically significant positive correlations among the urinary metabolites. Bayesian kernel machine regression analysis showed a significant positive association between elevated urinary concentrations of metabolites of OPFRs and increased oxidative stress levels. Di-n-butyl phosphate was identified as the metabolite that significantly contributed to the elevated level of 8-hydroxyguanosine.

Keywords:  Human biomonitoring; Liquid–liquid extraction; Organophosphate flame retardants; Oxidative stress; Ultra-high performance liquid chromatography-tandem mass spectrometry

DOI:  https://doi.org/10.1007/s00216-024-05393-8
Anal Chem. 2024 Jun 12.

Wideband PRM: Highly Accurate and Sensitive Method for High-Throughput Targeted Proteomics.

Dowoon Nam, Minyoung Ji, Chaewon Kang, Hokeun Kim, Hyunju Yang, Kwon Hee Bok, Jingi Bae, Jiwon Hong, Sang-Won Lee.

Targeted mass spectrometry (MS) approaches, which are powerful methods for uniquely and confidently quantifying a specific panel of proteins in complex biological samples, play a crucial role in validating and clinically translating protein biomarkers discovered through global proteomic profiling. Common targeted MS methods, such as multiple reaction monitoring (MRM) and parallel-reaction monitoring (PRM), employ specific mass spectrometric technologies to quantify protein levels by comparing the transitions of surrogate endogenous (ENDO) peptides with those of stable isotope-labeled (SIL) peptide counterparts. These methods utilizing amino acid analyzed (AAA) SIL peptides warrant sensitive and precise measurements required for targeted MS assays. Compared with MRM, PRM provides higher experimental throughput by simultaneously acquiring all transitions of the target peptides and thereby compensates for different ion suppressions among transitions of a target peptide. However, PRM still suffers different ion suppressions between ENDO and SIL peptides due to spray instability, as the ENDO and SIL peptides were monitored at different liquid chromatography (LC) retention times. Here we introduce a new targeted MS method, termed wideband PRM (WBPRM), that is designed for high-throughput targeted MS analysis. WBPRM employs a wide isolation window for simultaneous fragmentation of both ENDO and SIL peptides along with multiplexed single ion monitoring (SIM) scans for enhanced MS sensitivity of the target peptides. Compared with PRM, WBPRM was demonstrated to provide increased sensitivity, precision, and reproducibility of quantitative measurements of target peptides with increased throughput, allowing more target peptide measurements in a shortened experiment time. WBPRM is a straightforward adaptation to a manufacturer-provided MS method, making it an easily implementable technique, particularly in complex biological samples where the demand for higher precision, sensitivity, and efficiency is paramount.

DOI: https://doi.org/10.1021/acs.analchem.4c00518
Talanta. 2024 Jun 11. pii: S0039-9140(24)00789-6. [Epub ahead of print]277 126410

Chain electrospray ionization mass spectrometry for ultra-low volume sample analysis.

Yu Wang, Lu Chen, Xia Huang, Bing Xia, Yan Zhou.

  In this work, chain electrospray ionization (chain-ESI) was developed to efficiently ionize trace samples for mass spectrometry analysis. The primary ion source was found to have the ability to induce secondary electrospray ionization with an extraordinarily low sample consumption rate in the picoliters per minute (pLs/min). This allows low volume sample to generate substantial tandem mass spectrum (MS2) data for metabolite annotations. Notably, chain-ESI can effectively prevent the electro-redox reaction in the process of electrospray, so as to reflect the native state of the analytes. Furthermore, from a single Broussonetia papyrifera (B. papyrifera) trichome and a single A549 cancer cell, 1426 and 617 metabolites were detected respectively. All of those observations demonstrated that chain-ESI offers the advantages of direct, rapid analysis with extreme-low volumes and high coverage, enabling the measurement of bio-information in low volume samples.

Keywords:  Ambient mass spectrometry; Low flowrate; Low volume sample analysis; Untargeted metabolic analysis

DOI:  https://doi.org/10.1016/j.talanta.2024.126410
Anal Bioanal Chem. 2024 Jun 12.

Development and validation of a method for analysis of 25 cannabinoids in oral fluid and exhaled breath condensate.

Meena Swaminathan, Anamary Tarifa, Anthony P DeCaprio.

  Currently, there is a significant demand in forensic toxicology for biomarkers of cannabis exposure that, unlike ∆9-tetrahydrocannabinol, can reliably indicate time and frequency of use, be sampled with relative ease, and correlate with impairment. Oral fluid (OF) and exhaled breath condensate (EBC) are alternative, non-invasive sample matrices that hold promise for identifying cannabis exposure biomarkers. OF, produced by salivary glands, is increasingly utilized in drug screening due to its non-invasive collection and is being explored as an alternative matrix for cannabinoid analysis. EBC is an aqueous specimen consisting of condensed water vapor containing water-soluble volatile and non-volatile components present in exhaled breath. Despite potential advantages, there are no reports on the use of EBC for cannabinoid detection. This study developed a supported liquid extraction approach and LC-QqQ-MS dMRM analytical method for quantification of 25 major and minor cannabinoids and metabolites in OF and EBC. The method was validated according to the ANSI/ASB 036 standard and other published guidelines. LOQ ranged from 0.5 to 6.0 ng/mL for all cannabinoids in both matrices. Recoveries for most analytes were 60-90%, with generally higher values for EBC compared to OF. Matrix effects were observed with some cannabinoids, with effects mitigated by use of matrix-matched calibration. Bias and precision were within ± 25%. Method applicability was demonstrated by analyzing ten authentic OF and EBC samples, with positive detections of multiple analytes in both matrices. The method will facilitate comprehensive analysis of cannabinoids in non-invasive sample matrices for the development of reliable cannabis exposure biomarkers.

Keywords:  Biomarkers of cannabis exposure; Cannabinoids; Exhaled breath condensate; LC-QqQ-MS/MS analysis; Oral fluid

DOI:  https://doi.org/10.1007/s00216-024-05369-8
J Chromatogr A. 2024 May 17. pii: S0021-9673(24)00386-8. [Epub ahead of print]1729 465012

Development and validation of LC-MS/MS method for trace analysis of acrylamide, acrylic acid and N, N-methylene bis acrylamide in sandy loam soil.

Harshang Talaviya, Anupama Singh, Neera Singh, Suman Manna, Tirthankar Banerjee.

  Acrylamide and N, N-methylene bis acrylamide are most commonly used monomer and crosslinker compounds employed in synthesis of super absorbent hydrogels. When applied as soil conditioners, there are apprehensions that these hydrogels degrade over time and thus may release the toxic monomers in the soil. A method was thus developed using Liquid Chromatography tandem mass spectrometry (LC-MS/MS) for the trace level quantification of acrylamide (AD), acrylic acid (AA) and N,N-methylene-bis-acrylamide (MBA) in sandy loam soil amended by two test hydrogels the Pusa Hydrogel and SPG 1118 hydrogel prepared using AD and MBA. The MRM (multiple reaction monitoring) transitions were optimized for both the compounds. Soil samples were extracted using dispersive solid-phase extraction (dSPE) with a modified QuEChERS (quick, easy, cheap, effective, rugged, and safe) technique, employing acetonitrile. All analytes were quantified at trace levels within a five-minute run using UHPLC equipped with a C-18 column. Single laboratory validation of the developed method in soil matrix was conducted based on specificity, linearity, sensitivity, accuracy, precision, matrix effect and measurement of uncertainty. LC-MS/MS exhibited a linear response in the concentration range of 0.001 to 1 µg mL-1, with correlation coefficient >+0.99. Acceptable recovery (within 70-120 %) with repeatability (%RSD ≤20 %) was obtained at 0.01 to 1 µg g-1 fortification levels. LOQ (Limit of quantification) of the method for AD, AA and MBA in soil matrix were 0.05, 1 and 0.01 µg g-1, respectively. Both intra-laboratory repeatability and intermediate precision at LOQ suggested well acceptable precise (HorRat≈ 0.3) method for quantification. Matrix enhancement effect was observed in the order: AA>AD>MBA. The Expanded Uncertainty (EU) in soil matrix at LOQ was 21.64 %, 28 % and 19 % for AD, AA and MBA respectively. Groundnut and wheat grown with application of the hydrogels showed no detectable residues of monomers in soil samples (total n = 60) near the root zone at the time of crop harvesting.

Keywords:  Acrylamide; Acrylic acid; LC-MS/MS, soil; N,N-methylene-bis-acrylamide; QuEChERS

DOI:  https://doi.org/10.1016/j.chroma.2024.465012
Anal Chem. 2024 Jun 14.

Low-Cost Alternative for Online Analysis of Volatile Organic Compounds.

Benjamin Rico, Jon S Sauer, Kimberly A Prather, Robert S Pomeroy.

The use of online mass spectrometry for detecting volatile organic compounds (VOCs) has proven to be a powerful technique, allowing for real-time analysis of many chemical and biochemical processes. Unfortunately, online mass spectrometry has had limited application due to high instrument costs and limited availability. Here, we detail the design, construction, and performance characteristics of a custom ion-molecule reactor retrofitted to a commonly used single quadrupole mass spectrometer to operate as an online chemical ionization mass spectrometer (CIMS). This low-cost modified CIMS is capable of limits of detection below 10 parts per trillion for select VOCs including dimethyl sulfide, dimethylamine, and trimethylamine.

DOI: https://doi.org/10.1021/acs.analchem.4c00916
Chem Biodivers. 2024 Jun 10. e202400749

The Pre- and Post-column Derivatization on Monosaccharide Composition Analysis, a Review.

Xuexia Chen, Yinuo Wang, Yongjun Ye, Huali Yu, Bin Wu.

  Polysaccharides, as common metabolic products in organisms, play a crucial role in the growth and development of living organisms. For humans, polysaccharides represent a class of compounds with diverse applications, particularly in the medical field. Therefore, the exploration of the monosaccharide composition and structural characteristics of polysaccharides holds significant importance in understanding their biological functions. This review provides a comprehensive overview of extraction methods and hydrolysis strategies for polysaccharides. It systematically analyzes strategies and technologies for determining polysaccharide composition and discusses common derivatization reagents employed in further polysaccharide studies. Derivatization is considered a fundamental strategy for determining monosaccharides, as it not only enhances the detectability of analytes but also increases detection sensitivity, especially in liquid chromatography (LC), capillary electrophoresis (CE), and gas chromatography (GC) techniques. The critical comparison of pre- and post-column derivatization is elaborated in this paper, aiming to serve as a reference for selecting appropriate modes based on the structural characteristics, biological activities of polysaccharides, and reaction systems.

Keywords:  derivatization; monosaccharide; polysaccharides; post-column; pre-column

DOI:  https://doi.org/10.1002/cbdv.202400749