bims-metlip 2024-04-14 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2024–04–14
twenty-two papers selected by
Sofia Costa, Matterworks

Highly sensitive liquid chromatography-mass spectrometry method for the quantitative analysis of mometasone furoate in human plasma: Method validation and application to clinical pharmacokinetic studies.
Bayesian functional analysis for untargeted metabolomics data with matching uncertainty and small sample sizes.
Simultaneous mass spectrometric quantification of trace amines, their precursors and metabolites.
Method development and validation for the estimation of gedatolisib in mouse plasma by tandem mass spectrometry and its application to pharmacokinetics studies.
Systematic evaluation of benzoylation for liquid chromatography-mass spectrometry analysis of different analyte classes.
A rapid and sensitive LC-MS/MS method for quantifying oxycodone, noroxycodone, oxymorphone and noroxymorphone in human plasma to support pharmacokinetic drug interaction studies of oxycodone.
Quantitative determination of R/S-methadone in human serum using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry: A method for routine use.
MetaboAnalyst 6.0: towards a unified platform for metabolomics data processing, analysis and interpretation.
Isoporous Membrane Mediated Imprinting Mass Spectrometry Imaging for Spatially-Resolved Metabolomics and Rapid Histopathological Diagnosis.
Automated Extraction and LC-MS/MS Analysis of 11-Nor-9-carboxy-tetrahydrocannabinol Isomers and Prevalence in Authentic Urine Specimens.
Development of a two-dimensional LC-MS/MS system for the determination of proline and 4-hydroxyproline enantiomers in biological and food samples.
Determination of Pralsetinib in Human Plasma and Cerebrospinal Fluid for Therapeutic Drug Monitoring by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS).
An on-line SPE-LC-MS/MS method for quantification of nucleobases and nucleosides present in biological fluids.
[Determination of twelve halobenzoquinones in drinking water by solid phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry].
Anthocyanins: Quantification by Tandem Mass Spectrometry.
Metabolomics-based analysis of the diatom Cheatoceros tenuissimus combining NMR and GC-MS techniques.
Plasma drug screening using paper spray mass spectrometry with integrated solid phase extraction.
High-resolution ion mobility separations coupled to mass spectrometry: What's next?
Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples.
Determination of calcium, iron, and selenium in human serum by isotope dilution analysis using nitrogen microwave inductively coupled atmospheric pressure plasma mass spectrometry (MICAP-MS).
Screening of leaf extraction and storage conditions for eco-metabolomics studies.
Improving the Accuracy of Predictive 2D-LC Optimization Strategies: Incorporation of Simulated Elution Profiles to Account for Injection Band Broadening in Online Comprehensive Two-Dimensional Liquid Chromatography.

Biomed Chromatogr. 2024 Apr 10. e5871

Highly sensitive liquid chromatography-mass spectrometry method for the quantitative analysis of mometasone furoate in human plasma: Method validation and application to clinical pharmacokinetic studies.

Olivier Heudi, Monish Jain, Serge Winter.

  We report the development and the validation of a sensitive liquid chromatography-mass spectrometry (LC-MS/MS) method for mometasone furoate (MF) analysis in human plasma. Plasma samples were processed through liquid-liquid extraction and analyzed using LC-MS/MS operating in positive mode using multiple reaction monitoring of transitions m/z 520.9 → 355.0 and m/z 525.8 → 355.0 for MF and the internal standard (IS), respectively. Separation was achieved at 1.0 mL/min on a C18 column using a gradient elution of mobile phase of 0.05% ammonia in water (phase A) and acetonitrile (phase B). The assay range was 0.250-100 pg/mL and proved to be accurate and precise MF. Normalized recoveries were consistent and reproducible with a coefficient of variation (CV%) value of 6.0. The CV (%) of the IS normalized matrix factor was not observed in normal, lipemic, and hemolyzed plasmas. Dilutions of 1:10 were accurately quantified. A cycle of three freeze and thaw and stabilities at room temperature and on the autosampler were demonstrated. In addition, MF in the presence of indacaterol and glycopyrronium was proven to be stable at -70°C for at least 157 days. The present method was successfully applied to quantify MF in patients receiving MF, indacaterol, and glycopyrronium as a fixed-dose combination.

Keywords:  LC‐MS/MS; asthma; mometasone furoate; pharmacokinetic; validation and fixed‐dose combination

DOI:  https://doi.org/10.1002/bmc.5871
Brief Bioinform. 2024 Mar 27. pii: bbae141. [Epub ahead of print]25(3):

Bayesian functional analysis for untargeted metabolomics data with matching uncertainty and small sample sizes.

Guoxuan Ma, Jian Kang, Tianwei Yu.

  Untargeted metabolomics based on liquid chromatography-mass spectrometry technology is quickly gaining widespread application, given its ability to depict the global metabolic pattern in biological samples. However, the data are noisy and plagued by the lack of clear identity of data features measured from samples. Multiple potential matchings exist between data features and known metabolites, while the truth can only be one-to-one matches. Some existing methods attempt to reduce the matching uncertainty, but are far from being able to remove the uncertainty for most features. The existence of the uncertainty causes major difficulty in downstream functional analysis. To address these issues, we develop a novel approach for Bayesian Analysis of Untargeted Metabolomics data (BAUM) to integrate previously separate tasks into a single framework, including matching uncertainty inference, metabolite selection and functional analysis. By incorporating the knowledge graph between variables and using relatively simple assumptions, BAUM can analyze datasets with small sample sizes. By allowing different confidence levels of feature-metabolite matching, the method is applicable to datasets in which feature identities are partially known. Simulation studies demonstrate that, compared with other existing methods, BAUM achieves better accuracy in selecting important metabolites that tend to be functionally consistent and assigning confidence scores to feature-metabolite matches. We analyze a COVID-19 metabolomics dataset and a mouse brain metabolomics dataset using BAUM. Even with a very small sample size of 16 mice per group, BAUM is robust and stable. It finds pathways that conform to existing knowledge, as well as novel pathways that are biologically plausible.

Keywords:  Bayesian latent factor model; matching uncertainty; metabolite network analysis

DOI:  https://doi.org/10.1093/bib/bbae141
J Chromatogr B Analyt Technol Biomed Life Sci. 2024 Mar 18. pii: S1570-0232(24)00106-5. [Epub ahead of print]1238 124098

Simultaneous mass spectrometric quantification of trace amines, their precursors and metabolites.

Krisztina de Bruyn, Eugene F Diekman, Claude P van der Ley, Martijn van Faassen, Ido P Kema.

   OBJECTIVES: Trace amines are powerful neuromodulators influencing the release and reuptake of catecholamines. These low concentrated endogenous amines impact mood, cognition, and hormone regulation. Dysregulation of trace amines have been associated with a variety of diseases, such as schizophrenia, Parkinson's disease, migraine, depression and more. Succesfull simultaneous quantification of trace amines, their precursors and metabolites would benefit both research and patient care. Since these compounds have various functional groups and are present in biological matrices with large concentration difference, their simultaneous quantification is an analytical challenge. Our goal was to develop a highly sensitive LC-MS/MS assay to simultaneously quantify trace amines, their precursors and metabolites in plasma.
METHODS: Our method is based on a simple two-step in-matrix derivatization protocol: propionic anhydride (PA) and 3-Ethyl-1-[3-(dimethylamino)propyl]carbodiimide (EDC) in combination with 2,2,2-trifluoroethylamine (TFEA) followed by online solid phase extraction combined with LC-MS/MS. Fifteen metabolites can be measured simultaneously, three precursors, eight trace amines and four metabolites. Validation of this method was performed according to international validation guidelines. The pre-analytical stability of trace amines was assessed.
RESULTS: This novel method was successful in quantifying trace amines, their precursors, and metabolites in plasma. Using just 50 µl human plasma, we were able to accomplish limit of quantification for 2-phenylethylamine and N-methyl-phenylethylamine of 0.2 nmol/L and 0.1 nmol/L for tyramine and n-methyltyramine. Inter-and intra-assay imprecision was < 15 % for all analytes. Stability assessment showed susceptibility of certain trace amines e.g. 2-phenylethylamine and N-methyl-phenylethylamine to enzymatic degradation in plasma. The addition of the monoamine oxidase inhibitor pargyline to plasma prevented this enzymatic degradation.
CONCLUSIONS: We developed a novel LC-MS/MS method that1) uses a new double derivatization technique, 2) is automated with online SPE, 3) uses far less sample volume then previous methods and 4) detects more components in the same sample (eight trace amines, three precursors, and four metabolites) with high specificity and selectivity. Furthermore, addition of MAO A/B inhibitor prevents degradation and guarantees more accurate quantification of trace amines.

Keywords:  Derivatization; Monoamine oxidase inhibitor; Online solid phase extraction; Quantification; Schizophrenia; Trace amine

DOI:  https://doi.org/10.1016/j.jchromb.2024.124098
Biomed Chromatogr. 2024 Apr 10. e5878

Method development and validation for the estimation of gedatolisib in mouse plasma by tandem mass spectrometry and its application to pharmacokinetics studies.

Vinod A B, Arunava Das.

  A simple and a sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for the quantification of gedatolisib in mouse plasma. The extraction technique involved a simple precipitation method to extract gedatolisib and idelalisib (internal standard) from mouse plasma. A clean chromatographic separation of gedatolisib and the internal standard was achieved on an Atlantis dC18 column using an isocratic mobile phase (10 mm ammonium formate and acetonitrile; 30:70% v/v, both supplemented with 0.1% formic acid) delivered at a flow rate of 0.7 ml/min. The total run time was 2.0 min, and gedatolisib and idelalisib were eluted at 0.80 and 0.95 min, respectively. Gedatolisib was monitored at m/z 616.40 → 488.20 and idelalisib at 416.05 → 176.10. All the required parameters for the method validation were performed as per US Food and Drug Administration guidelines, and the results were within the acceptance criteria. The method was accurate and proved to be precise at a linearity range of 1.33-2667 ng/ml. The accuracy for gedatolisib in mouse plasma was in the ranges 0.99-1.06% (intra-day) and 0.96-1.04% (inter-day). Gedatolisib appeared to be stable in a series of stability conditions. Gedatolisib showed a good intravenous profile when administered through a solution formulation.

Keywords:  LC–MS/MS; gedatolisib; idelalisib; method validation; mouse plasma; pharmacokinetics

DOI:  https://doi.org/10.1002/bmc.5878
J Chromatogr A. 2024 Apr 04. pii: S0021-9673(24)00246-2. [Epub ahead of print]1722 464872

Systematic evaluation of benzoylation for liquid chromatography-mass spectrometry analysis of different analyte classes.

Ashley E Lenhart, Peter-Philip M Booth, Kaley M Simcox, Brianna A Ramos, Robert T Kennedy.

  LC-MS is an indispensable tool for small molecule analysis in many fields; however, many small molecules require chemical derivatization to improve retention on commonly used reversed-phase columns and increase ionization. Benzoyl chloride (BzCl) derivatization is commonly used for derivatization of primary and secondary amines and phenolic alcohols, though evidence exists that with proper reaction conditions (i.e., specific bases), other hydroxyl groups may be derivatized too. Previous studies have examined BzCl concentration, reaction times, and reaction temperatures for derivatization of amines and phenols for LC-MS analysis; however, use of different bases, base concentration, and extending to conditions to hydroxyl groups for LC-MS analysis has not been well-studied. To address this understudied area and identify reaction conditions for both amino and hydroxyl groups, we performed a systematic study of reaction conditions on multiple classes of potential targets. For selected derivatization methods, detection limits and performance in a variety of biological matrices were assessed. Results highlight the importance of tailoring derivatization methods for a given application as they varied by molecule and/or molecule class. Compared to the standard BzCl method commonly used, alternative methods were identified to better derivatize challenging analytes (glucosamine, choline, cortisol, uridine, cytidine) with detection limits reaching 1100, 9, 38, 170, and 67 nM compared to undetectable, 170, 86, 1000, and 86 nM respectively. Sub-nanomolar detection limits were achieved for norepinephrine with alternative derivatization approaches. Improved derivatization methods for several classes and molecules including nucleosides, steroids, and molecules containing hydroxyl groups were also identified.

Keywords:  Benzoyl chloride; Chemical derivatization; Liquid chromatography-mass spectrometry; Small molecules

DOI:  https://doi.org/10.1016/j.chroma.2024.464872
Biomed Chromatogr. 2024 Apr 08. e5874

A rapid and sensitive LC-MS/MS method for quantifying oxycodone, noroxycodone, oxymorphone and noroxymorphone in human plasma to support pharmacokinetic drug interaction studies of oxycodone.

Lotte M G Hulskotte, Inge Wilbrink-Pijffers, Maurits E L Arbouw, Guillemette E Benoist, Frank G A Jansman, Inge R F van Berlo-van de Laar.

  A sensitive and reliable LC-MS/MS method was developed and validated for the quantification of oxycodone and metabolites in human plasma. The method has a runtime of 6 min and a sensitivity of 0.1 μg/L for all analytes. Sample preparation consisted of protein precipitation. Separation was performed on a Kinetix biphenyl column (2.1 × 100 mm, 1.7 μm), using ammonium formate 5 mm in 0.1% aqueous formic acid and methanol LC-MS grade 100% in gradient elution at a flow rate of 0.4 ml/min. Detection was performed in multiple reaction monitoring mode using positive electrospray ionization. The method was linear over the calibration range of 0.1-25.0 μg/L for oxycodone, noroxycodone and noroxymorphone and 0.1-5.0 μg/L for oxymorphone. The method demonstrated good performance in terms of intra- and interday accuracy (86.5-110.3%) and precision (CV 1.7-9.3%). The criteria for the matrix effect were met (CV < 15%) except for noroxymorphone, for which an additional method was applied to compensate for the matrix effect. Whole blood samples were stable for 4 h at room temperature. Plasma samples were stable for 24 h at room temperature and 3 months at -20°C. Furthermore, the method was successfully applied in a pharmacokinetic drug interaction study of oxycodone and enzalutamide in patients with prostate cancer.

Keywords:  chromatography; drug monitoring; oxycodone; pharmacokinetics; tandem mass spectrometry

DOI:  https://doi.org/10.1002/bmc.5874
Drug Test Anal. 2024 Apr 10.

Quantitative determination of R/S-methadone in human serum using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry: A method for routine use.

Miriam Hansen, Hilde Havnen, Trine Naalsund Andreassen, Olav Spigset, Solfrid Hegstad.

  Methadone has two enantiomers, which exhibit differences in pharmacological effects, with R-methadone being the active and S-methadone the inactive enantiomer. A robust, simple and rapid method for chiral separation of the two enantiomers in serum samples using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry (UHPSFC-MSMS) has been developed and validated. Enantiomeric separation was achieved using a Chiralpak IH-3 column with a mobile phase consisting of CO2 and 30mM ammonium acetate in methanol/water (98/2, v/v). Runtime was 4 minutes. Sample preparation was semi-automated using a Hamilton ML Star robot with protein precipitation, and phospholipid removal was carried out using a Waters OSTRO™ 96-well plate. The calibration range was 50.0-1,500 nM for each enantiomer. The between-assay relative standard deviations were in the range of 1.2-3.6%. Matrix effects ranged from 99% to 115% corrected with internal standard. The method has been implemented in our laboratory and has proven to be a robust and reliable method for determining the ratio of R/S-methadone in authentic patient samples.

Keywords:  chiral methadone quantitation; human serum; supercritical fluid chromatography; therapeutic drug monitoring

DOI:  https://doi.org/10.1002/dta.3693
Nucleic Acids Res. 2024 Apr 08. pii: gkae253. [Epub ahead of print]

MetaboAnalyst 6.0: towards a unified platform for metabolomics data processing, analysis and interpretation.

Zhiqiang Pang, Yao Lu, Guangyan Zhou, Fiona Hui, Lei Xu, Charles Viau, Aliya F Spigelman, Patrick E MacDonald, David S Wishart, Shuzhao Li, Jianguo Xia.

We introduce MetaboAnalyst version 6.0 as a unified platform for processing, analyzing, and interpreting data from targeted as well as untargeted metabolomics studies using liquid chromatography - mass spectrometry (LC-MS). The two main objectives in developing version 6.0 are to support tandem MS (MS2) data processing and annotation, as well as to support the analysis of data from exposomics studies and related experiments. Key features of MetaboAnalyst 6.0 include: (i) a significantly enhanced Spectra Processing module with support for MS2 data and the asari algorithm; (ii) a MS2 Peak Annotation module based on comprehensive MS2 reference databases with fragment-level annotation; (iii) a new Statistical Analysis module dedicated for handling complex study design with multiple factors or phenotypic descriptors; (iv) a Causal Analysis module for estimating metabolite - phenotype causal relations based on two-sample Mendelian randomization, and (v) a Dose-Response Analysis module for benchmark dose calculations. In addition, we have also improved MetaboAnalyst's visualization functions, updated its compound database and metabolite sets, and significantly expanded its pathway analysis support to around 130 species. MetaboAnalyst 6.0 is freely available at https://www.metaboanalyst.ca.

DOI: https://doi.org/10.1093/nar/gkae253
Small Methods. 2024 Apr 09. e2301644

Isoporous Membrane Mediated Imprinting Mass Spectrometry Imaging for Spatially-Resolved Metabolomics and Rapid Histopathological Diagnosis.

Xingyue Liu, Liye Tao, Xinrong Jiang, Xuetong Qu, Wei Duan, Jiekai Yu, Xiao Liang, Jianmin Wu.

  Surface-assisted laser desorption/ionization (SALDI) mass spectrometry imaging (MSI) holds great value in spatial metabolomics and tumor diagnosis. Tissue imprinting on the SALDI target can avoid laser-induced tissue ablation and simplifies the sample preparation. However, the tissue imprinting process always causes lateral diffusion of biomolecules, thereby losing the fidelity of metabolite distribution on tissue. Herein, a membrane-mediated imprinting mass spectrometry imaging (MMI-MSI) strategy is proposed using isoporous nuclepore track-etched membrane as a mediating imprinting layer to selectively transport metabolites through uniform and vertical pores onto silicon nanowires (SiNWs) array. Compared with conventional direct imprinting technique, MMI-MSI can not only exclude the adsorption of large biomolecules but also avoid the lateral diffusion of metabolites. The whole time for MMI-based sample preparation can be reduced to 2 min, and the lipid peak number can increase from 46 to 113 in kidney tissue detection. Meanwhile, higher resolution of MSI can be achieved due to the confinement effect of the pore channel in the diffusion of metabolites. Based on MMI-MSI, the tumor margins of liver cancer can be clearly discriminated and their different subtypes can be precisely classified. This work demonstrates MMI-MSI is a rapid, highly sensitive, robust and high-resolution technique for spatially-resolved metabolomics and pathological diagnosis.

Keywords:  mass spectrometry imaging; membrane‐mediated imprinting; rapid histopathological diagnosis; silicon nanowires; spatially resolved metabolomics

DOI:  https://doi.org/10.1002/smtd.202301644
J Anal Toxicol. 2024 Apr 06. pii: bkae031. [Epub ahead of print]

Automated Extraction and LC-MS/MS Analysis of 11-Nor-9-carboxy-tetrahydrocannabinol Isomers and Prevalence in Authentic Urine Specimens.

Larissa K Karas, Courtney Patterson, Zachary J Fuller, Erin L Karschner.

  11-Nor-9-carboxy-Δ9-tetrahydrocannabinol (Δ9-THCCOOH) is the most frequently detected illicit drug metabolite in the military drug testing program. An increasing number of specimens containing unresolved Δ8-THCCOOH prompted the addition of this analyte to the Department of Defense (DoD) drug testing panel. A method was developed and validated for the quantitative confirmation of the carboxylated metabolites of Δ8- and Δ9-THC in urine samples utilizing automated pipette tip dispersive solid phase extraction and analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS). Analytes were separated isocratically over an 8.5 min runtime and detected on an MS/MS system equipped with an electrospray ionization source operating in negative mode. A single point calibrator (15 ng/mL) forced through zero demonstrated linearity from 3 to 1,000 ng/mL. Intra- and inter-day precision were ≤9.1% CV, and bias was within ±14.1% for Δ8-THCCOOH and Δ9-THCCOOH. No interferences were found after challenging the method with different over-the-counter drugs, prescription pharmaceuticals, drugs of abuse, and several cannabinoids and cannabinoid metabolites, including Δ1°-THCCOOH. Urine specimens presumptively positive by immunoassay (n=2939; 50 ng/mL Δ9-THCCOOH cutoff) were analyzed with this confirmation method. Specimens that contained Δ8-THCCOOH often had Δ9-THCCOOH above the 15 ng/mL cutoff. However, nearly one-third of the specimens analyzed were positive for Δ8-THCCOOH only. This manuscript describes the first validated automated extraction and confirmation method for Δ8- and Δ9-THCCOOH in urine that provides adequate analyte separation in urine specimens with extreme isomer abundance ratios.

Keywords:  Isomers; LC-MS/MS; automation; Δ8-THCCOOH

DOI:  https://doi.org/10.1093/jat/bkae031
Anal Sci. 2024 Apr 10.

Development of a two-dimensional LC-MS/MS system for the determination of proline and 4-hydroxyproline enantiomers in biological and food samples.

Chiharu Ishii, Yosuke Tojo, Komei Iwasaki, Akira Fujii, Takeyuki Akita, Masanobu Nagano, Masashi Mita, Tomomi Ide, Kenji Hamase.

  A two-dimensional LC-MS/MS system has been developed for the enantioselective determination of proline (Pro), cis-4-hydroxyproline (cis-4-Hyp) and trans-4-hydroxyproline (trans-4-Hyp) in a variety of biological samples. The amino acids were pre-column derivatized with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and the NBD-derivatives were separated by a reversed-phase column (Singularity RP18) as their D plus L mixtures in the first dimension. The collected target fractions were then introduced into the second dimension where the enantiomers were separated by a Pirkle-type enantioselective column (Singularity CSP-001S) and determined by a tandem mass spectrometer (Triple Quad™ 5500). The method was validated by the standard amino acids and also by human plasma, and sufficient results were obtained for the calibration, precision and accuracy. The method was applied to human plasma and urine, bivalve tissues and fermented food/beverages. D-Pro was widely found in the human physiological fluids, bivalves and several fermented products. Although trans-4-D-Hyp was not found in all the tested samples, cis-4-D-Hyp was present in human urine and tissues of the ark shell, and further studies focusing on the origin and physiological significance of these D-enantiomers are expected.

Keywords:  4-Hydroxyproline; Chiral separation; MS/MS; Proline; Two-dimensional LC

DOI:  https://doi.org/10.1007/s44211-024-00530-w
Anticancer Agents Med Chem. 2024 Apr 04.

Determination of Pralsetinib in Human Plasma and Cerebrospinal Fluid for Therapeutic Drug Monitoring by Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry (UPLC-MS/MS).

Zichen Zhao, Qianlun Pu, Tonglin Sun, Qian Huang, Liping Tong, Ting Fan, Jingyue Kang, Yuhong Chen, Yan Zhang.

   BACKGROUND: Ultra-performance Liquid Chromatography-tandem Mass Spectrometry (UPLC-MS/MS) is widely used for concentration detection of many Tyrosine Kinase Inhibitors (TKIs), including afatinib, crizotinib, and osimertinib. In order to analyze whether pralsetinib takes effect in Rearranged during Transfection (RET)-positive patients with central nervous system metastasis, we aimed to develop a method for the detection of pralsetinib concentrations in human plasma and Cerebrospinal Fluid (CSF) by UPLC-MS/MS.
METHODS: The method was developed using the external standard method, and method validation included precision, accuracy, stability, extraction recovery, and matrix effect. Working solutions were all obtained based on stock solutions of pralsetinib of 1mg/mL. The plasma/CSF samples were precipitated by acetonitrile for protein precipitation and then separated on an ACQUITY UPLC HSS T3 column (2.1×100 mm, 1.8 µm) with a gradient elution using 0.1% formic acid (solution A) and acetonitrile (solution B) as mobile phases at a flow rate of 0.4 mL/min. The tandem mass spectrometry was performed by a triple quadrupole linear ion trap mass spectrometry system (QTRAPTM 6500+) with an electrospray ion (ESI) source and Analyst 1.7.2 data acquisition system. Data were collected in Multiple Reaction Monitoring (MRM) and positive ionization mode.
RESULTS: A good linear relationship of pralsetinib in both plasma and CSF was successfully established, and the calibration ranges were found to be 1.0-64.0 µg/mL and 50.0ng/mL-12.8 µg/mL for pralsetinib in the plasma and CSF, respectively. Validation was performed, including calibration assessment, selectivity, precision, accuracy, matrix effect, extraction recovery, and stability, and all results have been found to be acceptable. The method has been successfully applied to pralsetinib concentration detection in a clinical sample, and the concentrations have been found to be 475ng/mL and 61.55 µg/mL in the CSF and plasma, respectively.
CONCLUSION: We have developed a quick and effective method for concentration detection in both plasma and CSF, and it can be applied for drug monitoring in clinical practice. The method can also provide a reference for further optimization.

Keywords:  Cerebrospinal Fluid; Plasma; Pralsetinib; UPLC-MS/MS; Ultra-performance Liquid Chromatography-Tandem Mass Spectrometry

DOI:  https://doi.org/10.2174/0118715206290110240326071909
Anal Methods. 2024 Apr 08.

An on-line SPE-LC-MS/MS method for quantification of nucleobases and nucleosides present in biological fluids.

Yi-Ming Liu, Shuguan Wang, Amani Dickenson, Jinghe Mao, Xiaolin Bai, Xun Liao.

Solid phase extraction (SPE) and liquid chromatographic (LC) separation of nucleobases and nucleosides are challenging due to the high hydrophilicity of these compounds. Herein we report a novel on-line SPE-LC-MS/MS method for their quantification after pre-column derivatization with chloroacetaldehyde (CAA). The method proposed is selective and sensitive with limits of detection at the nano-molar level. Analysis of urine and saliva samples by using this method is demonstrated. Adenine, guanine, cytosine, adenosine, guanosine, and cytidine were found in the range from 0.19 (guanosine) to 1.83 μM (cytidine) in urine and from 0.015 (guanosine) to 0.79 μM (adenine) in saliva. Interestingly, methylation of cytidine was found to be significantly different in urine from that in saliva. While 5-hydroxymethylcytidine was detected at a very low level (<0.05 μM) in saliva, it was found to be the most prominent methylated cytidine in urine at a high level of 3.33 μM. Since on-line SPE is deployed, the proposed LC-MS/MS quantitative assay is convenient to carry out and offers good assay accuracy and repeatability.

DOI: https://doi.org/10.1039/d4ay00100a
Wei Sheng Yan Jiu. 2024 Mar;53(2): 310-315

[Determination of twelve halobenzoquinones in drinking water by solid phase extraction-ultra-performance liquid chromatography-tandem mass spectrometry].

Danyang Yin, Li Tian, Minjuan Wang, Jiawei Hu.

   OBJECTIVE: To establish a method for twelve halobenzoquinones(HBQs) in drinking water by solid phase extraction-ultra-performance liquid chromatography coupled with electrospray-tandem mass spectrometry(SPE-UPLC-MS/MS).
METHODS: The drinking water was acidified with formic acid and concentrated by Bond Elut Plexa solid phase extraction column. The sample solution was separated using Waters ACQUITY HSS T3 column(100 mm×2.1 mm, 1.8 μm) with gradient elution using methanol-water containing 0.1% formic acid as mobile phase. The target compouds were detected in negtive electrospray ionization(ESI~-) and multiple reaction monitoring.
RESULTS: The concentration of twelve HBQs showed good linearity in the range 5.0-150.0 ng/mL, respectively, with the correlation coefficients greater than 0.999. The limits of detection(LOD) of twelve HBQs were lower than 2.0 ng/mL, and the limits of quantification(LOQ) for twelve HBQs were lower than 5.0 ng/mL, respectively. The recoveries of three levels in the matrix were 70.0%-84.0%. The matrix effffect was 0.08-0.64.
CONCLUSION: The SPE-UPLC-MS/MS method has high sensitivity, good accuracy and fast analysis speed for the detection of halobenzoquinones in drinking water.

Keywords:  disinfection byproducts; drinking water; halobenzoquinones; solid phase extraction-ultra-performance liquid chromatography coupled with electrospray-tandem mass spectrometry

DOI:  https://doi.org/10.19813/j.cnki.weishengyanjiu.2024.02.020
Methods Mol Biol. 2024 ;2798 153-159

Anthocyanins: Quantification by Tandem Mass Spectrometry.

Cindy Dias, Manuela Pintado, Clara Sousa.

  Mass spectrometry is a high throughput technique widely used for metabolic fingerprinting of plant material. Among the diverse plant metabolites, pigments such as anthocyanins play a determinant role in plant defence mechanisms, protecting them from biotic and abiotic stresses. Anthocyanins are phenolic water-soluble glycosides or acyl-glycosides of anthocyanidins which could be accurately detected and quantified through mass spectrometry. This chapter describes how to extract anthocyanins from higher plant materials and quantify them through a liquid chromatography-mass spectrometry (LC-MS) based method.

Keywords:  Anthocyanins; HPLC; Mass spectrometry (MS); Metabolites; Pigments; Plant; Solid phase extraction (SPE)

DOI:  https://doi.org/10.1007/978-1-0716-3826-2_10
MethodsX. 2024 Jun;12 102695

Metabolomics-based analysis of the diatom Cheatoceros tenuissimus combining NMR and GC-MS techniques.

Afrah Alothman, Abdul-Hamid Emwas, Upendra Singh, Mariusz Jaremko, Susana Agusti.

  Metabolomics, a recent addition to omics sciences, studies small molecules across plants, animals, humans, and marine organisms. Nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS) are widely used in those studies, including microalgae metabolomics. NMR is non-destructive and highly reproducible but has limited sensitivity, which could be supplemented by joining GC-MS analysis. Extracting metabolites from macromolecules requires optimization for trustworthy results. Different extraction methods yield distinct profiles, emphasizing the need for optimization. The results indicated that the optimized extraction procedure successfully identified NMR and GC-MS-based metabolites in MeOH, CHCl3, and H2O extraction solvents. The findings represented the spectral information related to carbohydrates, organic molecules, and amino acids from the water-soluble metabolites fraction and a series of fatty acid chains, lipids, and sterols from the lipid fraction. Our study underscores the benefit of combining NMR and GC-MS techniques to comprehensively understand microalgae metabolomes, including high and low metabolite concentrations and abundances.•In this study, we focused on optimizing the extraction procedure and combining NMR and GC-MS techniques to overcome the low NMR sensitivity and the different detected range limits of NMR and GC-MS.•We explored metabolome diversity in a tropical strain of the small cells' diatom Cheatoceros tenuissimus.

Keywords:  Diatoms; Extraction; GC–MS; Metabolites profiling; Metabolomics; Microalgae; NMR; Optimizing extraction procedure and combining NMR with GC–MS techniques for comprehensive metabolome analysis of microalgae

DOI:  https://doi.org/10.1016/j.mex.2024.102695
Drug Test Anal. 2024 Apr 07.

Plasma drug screening using paper spray mass spectrometry with integrated solid phase extraction.

Hannah Zimmerman-Federle, Greta Ren, Sarah Dowling, Cassandra Warren, Daniel Rusyniak, Robert Avera, Nicholas E Manicke.

  Drug overdoses have risen dramatically in recent years. We developed a simple nontargeted method using a disposable paper spray cartridge with an integrated solid phase extraction column. This method was used to screen for ~160 fentanyl analogs, synthetic cannabinoids, other synthetic drugs, and traditional drugs of abuse in over 300 authentic overdose samples collected at emergency departments in Indianapolis. A solid phase extraction step was implemented on the paper spray cartridge to enable subnanograms per milliliter synthetic drugs screening in plasma. Analysis was performed on a quadrupole orbitrap mass spectrometer using the sequential window acquisition of all theoretical fragment ion spectra approach in which tandem mass spectrometry was performed using 7 m/z isolation windows in the quadrupole. Calibration curves with isotopically labeled internal standards were constructed for 35 of the most frequently encountered synthetic and traditional illicit drugs by US toxicology labs. Additional qualitative-only drugs in a suspect screening list were also included. Limits of detection in plasma for synthetic cannabinoids ranged from 0.1 to 0.5 and 0.1 to 0.3 ng/mL for fentanyl and its analogs and between 1 and 5 ng/mL for most other drugs. Relative matrix effects were evaluated by determining the variation of the calibration slope in 10 different lots of biofluid and found to be between 3% and 20%. The method was validated on authentic overdose samples collected from two emergency departments in Indianapolis, Indiana, from suspected or known overdoses. Commonly detected synthetic drugs included fentanyl related substances, designer benzodiazepines such as flubromazolam, and the synthetic cannabinoid 5F-PB-22.

Keywords:  3D printing; ambient ionization; high resolution; methamphetamine; opiates

DOI:  https://doi.org/10.1002/dta.3687
J Mass Spectrom. 2024 May;59(5): e5014

High-resolution ion mobility separations coupled to mass spectrometry: What's next?

Gabe Nagy.

  Herein, I provide a personal perspective on high-resolution multipass ion mobility spectrometry-mass spectrometry (IMS-MS), with a specific emphasis on cyclic (cIMS) and structures for lossless ion manipulations (SLIM IMS)-based separations. My overarching goal for this perspective was to detail what I believe will be the key important areas in which IMS-MS will help shape the bioanalytical community and especially omics-based research.

Keywords:  high resolution; instrumentation; ion mobility spectrometry; mass spectrometry; separation science

DOI:  https://doi.org/10.1002/jms.5014
Forensic Sci Med Pathol. 2024 Apr 10.

Determination of lamotrigine in human plasma by HPLC-PDA. Application to forensic samples.

Inés Sánchez-Sellero, Iván Álvarez-Freire, Pamela Cabarcos-Fernández, Lidia Janza-Candal, María Jesús Tabernero-Duque, Ana María Bermejo-Barrera.

   PURPOSE: Therapeutic drug monitoring of plasma lamotrigine (LTG) has customarily been carried out in order to prevent some its adverse effects. For forensic purposes, determination of LTG in plasma is an useful tool in cases of accidental overdose or suicidal attempts. Currently, there are several analytical methods available including some based on LC tandem mass spectrometry techniques, but simple and accessible LC-UV methods still can be useful for the purpose. Here we report on a new high-performance liquid chromatography method for the determination of lamotrigine in human plasma which has been developed and validated including selectivity, sensitivity, accuracy, precision and recovery studies.
METHODS: Lamotrigine and the internal standard chloramphenicol were extracted from plasma using liquid-liquid extraction using small volumes of buffer and ethylacetate. Detection was monitored at 305.7 and 276.0 nm for lamotrigine and chloramphenicol, respectively.
RESULTS: The method was linear concentration dependence within the range of 0.1-10 µg/ml, with a mean coefficient of correlation r = 0.993. The limit of detection (LOD) was 0.04 µg/ml and the limit of quantification (LOQ) was 0.1 µg/ml. Intra and interday precision values were lower than 9.0% at all concentrations studied. The intra and interday accuracy values ranged from - 7.6 to 10.1%. Recovery was found to be 98.9% or higher. The method here described was successfully applied to 11 postmortem blood samples received at the Forensic Sciences Institute of Santiago de Compostela (Spain).
CONCLUSION: A new HPLC method for the determination of lamotrigine in human plasma was developed and validated. A liquid-liquid extraction using small volumes of buffer and ethylacetate was optimized. The proposed method is suitable for forensic toxicological analysis.

Keywords:  Forensic; HPLC–PDA; Lamotrigine; Plasma

DOI:  https://doi.org/10.1007/s12024-024-00812-9
Anal Bioanal Chem. 2024 Apr 08.

Determination of calcium, iron, and selenium in human serum by isotope dilution analysis using nitrogen microwave inductively coupled atmospheric pressure plasma mass spectrometry (MICAP-MS).

Zengchao You, Alexander Winckelmann, Jochen Vogl, Sebastian Recknagel, Carlos Abad.

  In this study, we demonstrate the applicability of nitrogen microwave inductively coupled atmospheric pressure mass spectrometry (MICAP-MS) for Ca, Fe, and Se quantification in human serum using isotope dilution (ID) analysis. The matrix tolerance of MICAP-MS in Na matrix was investigated, revealing that high Na levels can suppress the signal intensity. This suppression is likely due to the plasma loading and the space charge effect. Moreover, 40Ca and 44Ca isotopic fractionation was noted at elevated Na concentration. Nine certified serum samples were analyzed using both external calibration and ID analysis. Overestimation of Cr, Zn, As, and Se was found in the results of external calibration, which might result from C-induced polyatomic interference and signal enhancement, respectively. Further investigations performed with methanol showed a similar enhancement effect for Zn, As, and Se, potentially supporting this assumption. The mass concentrations determined with ID analysis show metrological compatibility with the reference values, indicating that MICAP-MS combined with ID analysis can be a promising method for precise Ca, Fe, and Se determination. Moreover, this combination reduces the influence of matrix effects, broadening the applicability of MICAP-MS for samples with complex matrixes.

Keywords:  Calcium; Human serum; Iron; Isotope dilution; Nitrogen microwave inductively coupled atmospheric pressure mass spectrometry; Selenium

DOI:  https://doi.org/10.1007/s00216-024-05274-0
Plant Direct. 2024 Apr;8(4): e578

Screening of leaf extraction and storage conditions for eco-metabolomics studies.

Jakob Lang, Sergio E Ramos, Marharyta Smohunova, Laurent Bigler, Meredith C Schuman.

  Mass spectrometry-based plant metabolomics is frequently used to identify novel natural products or study the effect of specific treatments on a plant's metabolism. Reliable sample handling is required to avoid artifacts, which is why most protocols mandate shock freezing of plant tissue in liquid nitrogen and an uninterrupted cooling chain. However, the logistical challenges of this approach make it infeasible for many ecological studies. Especially for research in the tropics, permanent cooling poses a challenge, which is why many of those studies use dried leaf tissue instead. We screened a total of 10 extraction and storage approaches for plant metabolites extracted from maize leaf tissue across two cropping seasons to develop a methodology for agroecological studies in logistically challenging tropical locations. All methods were evaluated based on changes in the metabolite profile across a 2-month storage period at different temperatures with the goal of reproducing the metabolite profile of the living plant as closely as possible. We show that our newly developed on-site liquid-liquid extraction protocol provides a good compromise between sample replicability, extraction efficiency, material logistics, and metabolite profile stability. We further discuss alternative methods which showed promising results and feasibility of on-site sample handling for field studies.

Keywords:  UHPLC–MS; agroecology; chemical ecology; extract stability; maize (zea mays)

DOI:  https://doi.org/10.1002/pld3.578
Anal Chem. 2024 Apr 09.

Improving the Accuracy of Predictive 2D-LC Optimization Strategies: Incorporation of Simulated Elution Profiles to Account for Injection Band Broadening in Online Comprehensive Two-Dimensional Liquid Chromatography.

Magriet Muller, Tyler Brau, Thomas Lauer, Dwight Stoll, André de Villiers.

Method development in online comprehensive two-dimensional liquid chromatography (LC × LC) requires the selection of a large number of experimental parameters. The complexity of this process has led to several computer-based LC × LC optimization algorithms being developed to facilitate LC × LC method development. One particularly relevant challenge for predictive optimization software is to accurately model the effect of second dimension (2D) injection band broadening under sample solvent mismatch and/or sample volume overload conditions. We report a novel methodology that combines a chromatographic numerical simulation model capable of predicting elution profiles of analytes under conditions where peak distortion occurs with a predictive multiparameter Pareto optimization approach for online LC × LC. Preliminary method optimization is performed using a theoretical model to predict 2D injection profiles, and optimal experimental configurations obtained from the Pareto fronts are then subjected to further optimization using the simulation model. This approach drastically reduces the number of simulations and therefore the computational demand. We show that the optimal experimental conditions obtained in this manner are similar to those obtained using a complete optimization using only the simulation model. Online HILIC × RP-LC separation of phenolic compounds was used to compare experimental data to simulated two- and three-dimensional contour plots. The main advantage of the proposed approach is the ability to predict the formation of split or deformed peaks in the 2D, a significant benefit in online LC × LC method optimization, especially for separation combinations with mismatched mobile phases. A further benefit is that simulated elution profiles can be used for the visualization of predicted two-dimensional chromatograms for method selection.

DOI: https://doi.org/10.1021/acs.analchem.4c00491