bims-metlip 2023-12-31 papers

bims-metlip

Biomed News

on Methods and protocols in metabolomics and lipidomics

Issue of 2023–12–31
ten papers selected by
Sofia Costa, Matterworks

Determination of bile acids in serum of pigs exposed to polychlorinated biphenyls by liquid chromatography-mass spectrometry.
Dilute and shoot approach for toxicology testing.
Improved quantitative determination of (R)- and (S)-rabeprazole sodium and its metabolites in rat plasma by LC-MS/MS and its application to a toxicokinetic study.
Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of the acetyl-coenzyme A competitive p300/CBP catalytic inhibitor A-485 in biological samples.
A highly efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for etomidate and etomidate acid in urine, liver and kidney.
Development of a simultaneous quantification method of imatinib and sunitinib and their main metabolites and its application in patients with gastrointestinal stromal tumor.
Mobilized Electrospray Device for On-Tissue Chemical Derivatization in MALDI-MS Imaging.
Construction and application of a QSRR approach for identifying flavonoids.
Arc-Induced Electrospray Ionization Mass Spectrometry.
Integrating Post-Ionization Separation via Differential Mobility Spectrometry into Direct Analysis in Real Time Mass Spectrometry for Toy Safety Screening.

J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Dec 18. pii: S1570-0232(23)00386-0. [Epub ahead of print]1232 123976

Determination of bile acids in serum of pigs exposed to polychlorinated biphenyls by liquid chromatography-mass spectrometry.

Luca Narduzzi, Maykel Hernández-Mesa, Bruno Le Bizec, Ana M García-Campaña, Gaud Dervilly.

  Exposure to polychlorinated biphenyls (PCBs) has been linked to dyslipidemia. Under acute exposure to PCBs, it has been observed that the secretion of bile acids (BAs) can be impacted, limiting (indirectly) lipid absorption in the gut. In this context, two non-targeted metabolomics studies on pig serum have recently suggested that BA concentrations may fluctuate under exposure to current non-dioxin-like (NDL)-PCB levels in food, reflecting the acute effects of such chronic exposure. The objective of this research is to implement a targeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for BA analysis in order to validate the findings of previous metabolomics studies, in which BA levels in serum samples from pigs exposed to environmental doses of NDL-PCBs were highlighted to be affected. The proposed LC-MS method involves the use of a C18-pentafluorophenyl LC column, which is not usually selected for the separation of BAs, but shows better performance for the separation of isomers than typical C18 columns. This LC-MS method shows excellent analytical performance such as low limits of detection (LODs) (≤1 ng/mL for most BAs) and good linearity (R2 > 0.994), while no matrix effect was observed. A total of 13 BAs have been quantified, while further BA isomers could be detected and semi-quantified. The application of this targeted LC-MS method confirmed previous findings, suggesting that exposure to low doses of NDL-PCBs decreases the concentration of BAs (i.e., glycochenodeoxycholic acid, hyodeoxycholic acid and taurochenodeoxycholic acid) while the effect on the precursors (cholic acid and chenodeoxycholic acid) is less pronounced.

Keywords:  Bile acids; Biomarkers of effect; LC-MS/MS; PCBs; Serum

DOI:  https://doi.org/10.1016/j.jchromb.2023.123976
Front Chem. 2023 ;11 1278313

Dilute and shoot approach for toxicology testing.

Kenichi Tamama.

  Toxicology testing is performed in clinical settings, forensic settings, and for controlling doping. Drug screening is a toxicology test to determine if drugs are present in biological samples. The most common specimen type for drug testing is urine, as drugs and/or their metabolites are often more concentrated in the urine, extending the detection window of drugs. The dilute-and-shoot method is a simple procedure used in toxicology testing, where a sample is diluted before being directly injected into the liquid chromatography-mass spectrometry (LC-MS) system. This method is easy, quick, and cost-saving, and can be used for protein-poor liquid specimens such as urine. Thus, it is reasonable and attractive for busy toxicology laboratories to combine the dilute-and-shoot method with high-resolution hyphenated-MS for urine drug screening. This method has several disadvantages, including a suboptimal detection capability for certain analytes, as well as interference from co-eluting matrix components called matrix effects, in which co-eluting matrix molecules alter the ionization efficiency of the analyte molecules at the ionization source in LC-MS, altering (mostly reducing) the analyte detection capability. The matrix effect testing is essential for the validation of LC-MS-based assays. A reasonable approach to addressing these undesirable effects would be to minimize these components. The most straightforward approach is to reduce the amounts of matrix components by using a higher dilution of the specimen and a lower volume for specimen injection. Optimization of the chromatographic separation is another reasonable approach for reducing co-eluting matrix components with the analyte.

Keywords:  dilute-and-shoot; liquid chromatography-mass spectrometry; matrix effect; sample preparation; toxicology testing; urine

DOI:  https://doi.org/10.3389/fchem.2023.1278313
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Dec 13. pii: S1570-0232(23)00379-3. [Epub ahead of print]1232 123969

Improved quantitative determination of (R)- and (S)-rabeprazole sodium and its metabolites in rat plasma by LC-MS/MS and its application to a toxicokinetic study.

Jianman Guo, Xiaoyuan Ma, Fanlong Bu, Jian Wang, Jie Sun, Rui Zhang.

  There exist two enantiomers: (R)- and (S)-rabeprazole. (R)-rabeprazole offers specific pharmacokinetic advantages and enhanced therapeutic efficacy, warranting further investigation and development. Here, we developed a simple and rapid chiral liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to simultaneously quantify rabeprazole enantiomers and their metabolites (rabeprazole sulfoxide and desmethyl rabeprazole enantiomers) and a LC-MS to quantify rabeprazole thioether. As for the chiral LC-MS/MS method, Chiral-AGP column (150 × 4 mm, 5 μm) was used and its mobile phase was acetonitrile (mobile phase A) and 10 mmol/L ammonium acetate (mobile phase B) (linear gradient profile: 0 min, 10 % B; 5 min, 15 % B; 9 min, 15 % B; 9.01 min, 10 % B; 13 min, 10 % B). The multiple reactions monitoring transitions of m/z 360.3 → 242.1, 376.2 → 240.1, 346.2 → 228.2 and 368.2 → 190.2 were opted for quantifying rabeprazole enantiomers, rabeprazole sulfoxide, desmethyl rabeprazole enantiomers and internal standard omeprazole. The analyte samples were prepared by a simple liquid-liquid extraction method. As for the LC-MS method, analytes were separated on a Inertsil® ODS-3 column (4.6 × 150 mm, 5 μm). The mobile phase was acetonitrile-5 mmol/L ammonium acetate water solution (65:35, v/v). ESI+ was used and ion peaks with m/z 344.2 (rabeprazole thioether) and 285.1 (internal standard diazepam) were monitored. Both these 2 methods were validated for specificity, linearity, precision, accuracy, matrix effect and extraction recovery, and, particularly, the stability of analytes under various conditions. We successfully applied these methods to a 13-week toxicokinetic study of rabeprazole in rats after intravenous administration of (R)- (80, 20, 5 mg/kg/d) and racemic (80 mg/kg/d) rabeprazole sodium. The results showed that rabeprazole and its metabolites did not accumulate in rats. However, desmethyl rabeprazole and rabeprazole thioether showed higher exposure and lower clearance rate in the last administration than in the first one. (R)-rabeprazole showed a higher exposure and a slower elimination rate than (S)-rabeprazole in rats. These findings offer experimental evidence and a theoretical foundation for further preclinical investigations and clinical applications of (R)-rabeprazole.

Keywords:  Desmethyl rabeprazole; LC-MS/MS; Rabeprazole; Rabeprazole sulfoxide; Rabeprazole thioether; Toxicokinetic

DOI:  https://doi.org/10.1016/j.jchromb.2023.123969
Biomed Chromatogr. 2023 Dec 26. e5819

Development and validation of a liquid chromatography-tandem mass spectrometry method for the quantification of the acetyl-coenzyme A competitive p300/CBP catalytic inhibitor A-485 in biological samples.

Reinhard Oertel, Nikolai Jaschke, Annemarie Lippert, Bertold Renner.

  The small molecule A-485 competitively inhibits the histone acetyltransferase domain of CBP (cyclic-adenosine monophosphate response element-binding protein)/p300. Apart from its antineoplastic activity, researchers are exploring its potential benefits in treating osteoporosis and its impact on energy metabolism. However, so far, only limited pharmacokinetic data are available, and the crucial determination of A-485 concentration in various biological materials with small sample volumes remains unpublished. A rapid and sensitive LC-tandem mass spectrometry method has been developed and validated to quantify A-485 in mouse serum and tissue. In this method, serum samples underwent precipitation with acetonitrile, while cell lysates were appropriately diluted. The determination of A-485 utilized a reversed-phase column with a mobile phase gradient, and detection was carried out in multiple reaction monitoring mode. The lower standard sample, with a concentration of 7.8 ng/mL, served as the lower limit of quantification, while the upper standard was established at 1000 ng/mL. A-485 concentrations were assessed in both serum samples and the lysate of all examined tissues, revealing swift metabolic clearance. The analytical method outlined here is deemed appropriate for subsequent studies. The ability to measure the active ingredient in various compartments facilitates the determination of accurate pharmacokinetic parameters. In the event of human use of A-485, the analysis method can be seamlessly transferred to human samples.

Keywords:  A-485; LC-MS/MS; protein precipitation; serum and tissue samples

DOI:  https://doi.org/10.1002/bmc.5819
J Pharmacol Toxicol Methods. 2023 Dec 21. pii: S1056-8719(23)00241-1. [Epub ahead of print]125 107490

A highly efficient liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for etomidate and etomidate acid in urine, liver and kidney.

Tian-Fu He, Huan-Hui Zhu, Xian-Wen Lin, Yuan-Yuan Tian, Li-Min Sun, Xu Guan, Hai-Yan Zhang, Li Tan, Song-Cai Wang.

  Etomidate (ETO) is a highly-efficient drug that can induce anesthesia with increasing doses, thus subject to strict regulation. However, an accurate and efficient method for ETO intake detection is currently lacking. Therefore, this study developed a straightforward sample preparation method using LC-MS/MS to analyze ETO and its primary metabolite, etomidate acid (ETA), in urine, liver, and kidney samples. Snap frozen pig liver and kidney samples were ground into a fine powder. Then, all the biological samples, including human urine, pig liver and kidney tissues, were deproteinized using acetonitrile and filtered for analysis. The separation was achieved in 9.01 min with gradient elution. The calibration curves ranged from 0.5 to 50 ng/mL for ETO in urine and 0.5 to 50 ng/g in liver and kidney, while the curves ranged from 1 to 100 ng/mL for ETA in urine and 1 to 100 ng/g in liver and kidney. The correlation coefficients (R2) were greater than 0.9957. The Limit of detection (LOD) and limit of quantitation (LOQ) for ETO were 0.2 and 0.5 ng/mL in urine samples and 0.2 and 0.5 ng/g in liver and kidney samples, respectively. For ETA, the LOD and LOQ were 0.5 and 1 ng/mL in urine samples and 0.5 and 1 ng/g in liver and kidney samples. This method was assessed by validation parameters, including selectivity, intra- and inter-day precision and accuracy, recovery, matrix effect, dilution integrity and stability. It was successfully applied to a practical case, revealing ETO and ETA concentrations in urine of 1.01 and 5.58 μg/mL, in liver samples of 12.30 and 1.13 μg/g, and in kidney samples of 6.95 and 4.23 μg/g. This suggests that the method is suitable for routine forensic detection of illicit ETO abuse.

Keywords:  Etomidate; Etomidate acid; Forensic toxicology; Kidney; Liquid chromatography–tandem mass spectrometry (LC-MS/MS); Liver; Urine

DOI:  https://doi.org/10.1016/j.vascn.2023.107490
Biomed Chromatogr. 2023 Dec 28. e5804

Development of a simultaneous quantification method of imatinib and sunitinib and their main metabolites and its application in patients with gastrointestinal stromal tumor.

Hefen Chen, Jianbo Lyu, Zhujun Gong, Yong Han, Kaixiong Tao, Hong Zhou.

  Correlations between plasma concentrations of imatinib and sunitinib with efficacy and toxicity have been established. It is crucial to develop a sensitive and precise method for determining the plasma concentrations of imatinib and sunitinib, along with their active metabolites, to facilitate therapeutic drug monitoring and individualized therapy. Plasma samples were separated on an Agilent ZORBAX SB-C18 chromatographic column using gradient elution. Quantification was performed using a mass spectrometer equipped with electrospray ionization in multiple reaction monitoring. The analysis time was 18 min per run, with all analytes and internal standards eluting within 8 min. The calibration range was 25-4000 ng/mL for imatinib, 5-800 ng/mL for N-desmethyl imatinib (CGP74588), and 2.5-400 ng/mL for sunitinib and N-desethyl sunitinib (SU12662). Intra- and inter-assay precision were both below 15%, and accuracy ranged between 90.0% and 101.9%. The method was successfully applied to determine blood samples from 120 patients with gastrointestinal stromal tumors who received imatinib (n = 115) and sunitinib (n = 5). It has been validated as linear, accurate, precise, and robust, making it suitable for therapeutic drug monitoring of imatinib and sunitinib in routine clinical practice.

Keywords:  LC-MS/MS; gastrointestinal stromal tumor; imatinib; sunitinib

DOI:  https://doi.org/10.1002/bmc.5804
J Am Soc Mass Spectrom. 2023 Dec 26.

Mobilized Electrospray Device for On-Tissue Chemical Derivatization in MALDI-MS Imaging.

Andrew E Paulson, Evan A Larson, Young Jin Lee.

  Applying solutions of a matrix or derivatization agent via microdroplets is a common sample preparation technique for matrix-assisted laser desorption/ionization-mass spectrometry imaging (MALDI-MSI) experiments. Mobilized nebulizer sprayers are commonly used to create a homogeneous matrix or reagent layer across large surfaces. Electrospray devices have also been used to produce microdroplets for the same purpose but are rarely used for large tissues due to their immobility. Herein, we present a movable electrospray device that can be used for large tissue sample preparation through a simple modification to an automatic commercial nebulizer device. As demonstrated for on-tissue chemical derivatization (OTCD) with Girard's reagent T using a mimetic tissue model, the sprayer has the additional benefit of being able to investigate reaction acceleration in OTCD when comparing electrostatically charged spray to electrostatically neutral spray. Finally, MALDI-MSI of fatty aldehydes is successfully demonstrated in rat brain tissues using this device for both OTCD and matrix application.

Keywords:  electrospray; mass spectrometry imaging; on-tissue chemical derivatization; reaction acceleration

DOI:  https://doi.org/10.1021/jasms.3c00290
J Pharm Biomed Anal. 2023 Dec 21. pii: S0731-7085(23)00698-2. [Epub ahead of print]240 115929

Construction and application of a QSRR approach for identifying flavonoids.

Shiyuan Sun, Biyue Cui, Fanyu Kong, Zitong Zhang, Youfu Qiao, Shuting Zhang, Xinran Zhang, Changhai Sun.

  A quantitative structure retention relationship (QSRR) method was developed to identify flavonoid isomers auxiliary using an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method based on the linear relationships between the Ln(k') values of flavonoids and their hydrogen bonding energy (XAH) and dissolution energy (ES). Chromatographic separation was achieved with a Hypersil GOLD C18 (100 mm × 2.1 mm, 1.9 µm) column and Agilent SB-C18 (2.1 ×50 mm, 1.8 µm) column on a Dionex Ultimate 3000 RSLC chromatograph. Compounds were eluted isocratically using a mobile phase containing 0.1% formic acid/water solution and methanol at a ratio of 55:45 (v/v). Mass spectrometry was performed in the negative and positive ionization modes on a Thermo Fisher Q Exactive Orbitrap mass spectrometer equipped with an electrospray ionization interface. The established QSRR model was Ln(k') = 5.6163 + 0.0469ES - 0.0984XAH, with a determination coefficient (R2) of 0.9981, adjusted determination coefficient (adjR2) of 0.9976, and corrected root mean square error of 0.0682. The determination coefficient of the leave-one-out (LOO) cross-validation (Q2LOO) was 0.9976, and the cross-verification root mean square error was 0.0754. Simulated samples containing 7 flavonoids were used to validate the feasibility of the method. The classical method (UHPLC-MS/MS combined the CD software and the mzCloud, mzVault and Chemspider databases) was used to identify the seven flavonoids in the simulated samples. This classic identification strategy cannot provide accurate identification results, which provided multiple identification results for each compound in the simulated samples. On the basis of the results, the 7 flavonoids were accurately identified by the established QSRR model, and the reference standards were used to validate it. The relative error of retention time(RE(tR)) between the model calculation and experimental results was less than 10%. This method effectively complements and improves the classical methods, that UHPLC-MS/MS combined the CD software and the mass spectra databases were used to identify flavonoids identification.

Keywords:  Flavonoids; Quantitative structure-retention relationships; Reverse phase chromatography; UHPLC-MS/MS

DOI:  https://doi.org/10.1016/j.jpba.2023.115929
Anal Chem. 2023 Dec 28.

Arc-Induced Electrospray Ionization Mass Spectrometry.

Kaineng Huang, Hui Zeng, Xingyue Li, Xiaoting Li, Yuanjiang Pan, Yuanji Gao.

Arc-induced electrospray ionization mass spectrometry (AESI-MS) was developed during which alternating current electrospray is simply achieved through the arc plasma. The AESI source exploits the arc's temperature and charge properties to generate aerosols consisting of charged microdroplets. The electrospray region, in which organic molecules are contained within microdroplets, partially overlaps with the arc plasma region. Guided by the electric field, these molecules undergo ionization, yielding ionic target analytes. AESI represents a soft ionization method that combines the mechanisms of atmospheric pressure chemical ionization and electrospray ionization, facilitating the ionization of analytes with wide ranging polarities. The precisely targeted spraying area enhances ion entry into the mass analyzer, thereby enabling excellent ionization efficiency. The AESI source exhibits several notable advantages over the electrospray ionization source, including an elevated but comparable level of active species concentrations and types, simplified mass spectra for direct amino acid analysis, high salt tolerance, versatile analysis of compounds with varying polarities, and reliable quantitative analysis of amino acids in complex matrices. Overall, AESI broadens the methodologies employed to generate microdroplets, providing a technological and scientific framework for creating distinctive electrospray ionization techniques.

DOI: https://doi.org/10.1021/acs.analchem.3c04125
Anal Chem. 2023 Dec 28.

Integrating Post-Ionization Separation via Differential Mobility Spectrometry into Direct Analysis in Real Time Mass Spectrometry for Toy Safety Screening.

Mengmeng Yan, Nan Zhang, Xiaoxu Li, Jianqiang Xu, Haimin Lei, Qiang Ma.

Direct analysis in real time (DART) enables direct desorption and ionization of analytes, bypassing the time-consuming chromatographic separation traditionally required for mass spectrometry (MS) analysis. However, DART-MS suffers from matrix interference of complex samples, resulting in compromised detection sensitivity and quantitation accuracy. In this study, DART-MS was combined with differential mobility spectrometry (DMS) to provide an additional dimension of post-ionization ion mobility separation within a millisecond time scale, compensating for the lack of separation in DART-MS analysis. As proof-of-concept, primary aromatic amines (PAAs), a class of potentially hazardous chemicals, were analyzed in various toy products, including bubble solutions, finger paints, and plush toys. In addition to commercial Dip-it glass rod and metal mesh sampling tools, a customized rapid extractive evaporation device was designed for the accelerated extraction and sensitive analysis of solid toy samples. The incorporation of DMS in DART-MS analysis enabled the rapid separation and differentiation of isomeric analytes, leading to improved accuracy and reliability. The developed protocols were optimized and validated, achieving good linearity with correlation coefficients greater than 0.99 and acceptable repeatability with relative standard deviations less than 10%. Moreover, satisfactory sensitivity was realized with limits of detection and quantitation ranges of 0.2-5 and 1-20 μg/kg (μg/L) for the 11 PAA analytes. The established methodology was applied for the analysis of real toy samples (n = 18), which confirmed its appealing potential for toy safety screening and consumer health protection.

DOI: https://doi.org/10.1021/acs.analchem.3c03915