Label-free and de-conjugation-free workflow to simultaneously quantify trace amount of free/conjugated and protein-bound estrogen metabolites in human serum

Kafeenah, Husam; Kuo, Chin-Ming; Chang, Ting-Yao; Jen, Hung-Hsiang; Yang, Jou-Hsin; Shen, Yan-Shan; Wu, Chih‐Hsing; Chen, Shuhui

doi:10.1016/j.aca.2022.340457

Cited by 5 publications

(2 citation statements)

References 33 publications

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“…The applications also demonstrated that the overall procedure, with sample extraction and analysis on the same platform, was fast and effective for the analysis of serum samples. The developed platform performed similarly to chemiluminescent immunoassays (E2 concentrations >50–100 pmol L –1 ) but needs further improvement to match the sensitivity of LC–MS/MS methods with traditional offline sample preparation, where E2 LOD and LOQ values reached 0.8 pg mL –1 and 3 pg mL –1 , respectively …”

Section: Resultsmentioning

confidence: 99%

“…Liquid chromatography–tandem mass spectrometry (LC–MS/MS) has emerged as a popular alternative for estrogen analysis, especially when high sensitivity and throughput are required. − In a recent study, acetonitrile was employed for the dual purpose of eliminating proteins and DNA, followed by salting out in a QuEChERS-type extraction. Microflow LC-multiple reaction monitoring (MRM) analysis was utilized for the sensitive quantitation of estrogens at subpicogram levels and for hundreds of sample runs, demonstrating a significant improvement over previously reported methods . While these methods proved to be efficient, they all necessitated a dedicated sample preparation protocol.…”

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confidence: 99%

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Preparation of Monolith for Online Extraction and LC–MS Analysis of β-Estradiol in Serum Via a Simple Multicomponent Reaction

Cerrato,

Aita,

Cavaliere

et al. 2024

Anal. Chem.

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Multicomponent reactions offer efficient and environmentally friendly strategies for preparing monoliths suitable for applications in analytical chemistry. In the described study, a multicomponent reaction was utilized for the one-pot miniaturized preparation of a poly(propargyl amine) polymer inside commercial silica-lined PEEK tubing. The reaction involved only small amounts of reagents and was characterized by atom economy. The resulting monolithic column was incorporated into an autosampler system for the online extraction and cleanup of β-estradiol from human serum. Sample pretreatment was simplified to a simple dilution with methanol and centrifugation to remove proteins. The resulting platform included LC−MS analysis in multiple reaction monitoring for quantitative analysis of β-estradiol. The method was validated in serum, demonstrating practical applicability for the monitoring of fertile women. Recoveries were above 94%, and LOD and LOQ values at 0.008 and 0.18 ng mL −1 , respectively. The developed platform proved to be competitive with previous methods for solidphase microextraction of β-estradiol in serum, with comparable recovery and sensitivity but with the advantage of nearly complete automation. The environmental impact of the process was evaluated as acceptable due to the miniaturization of the monolith synthesis and the automation of extraction. The drawback associated with the LC−MS technique can be reduced by the inclusion of additional analytes in a single investigation. The work demonstrates that multicomponent reactions are versatile, economical, and possibly a green methodology for producing reversed-phase and mixed-mode sorbents, enabling miniaturization of the entire analytical procedure from the preparation of extraction sorbents to analysis.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Preparation of Monolith for Online Extraction and LC–MS Analysis of β-Estradiol in Serum Via a Simple Multicomponent Reaction

Cerrato,

Aita,

Cavaliere

et al. 2024

Anal. Chem.

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Hyphenation of microflow chromatography with electrospray ionization mass spectrometry for bioanalytical applications focusing on low molecular weight compounds: A tutorial review

Girel,

Meister,

Glauser

et al. 2024

Mass Spectrometry Reviews

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Benefits of miniaturized chromatography with various detection modes, such as increased sensitivity, chromatographic efficiency, and speed, were recognized nearly 50 years ago. Over the past two decades, this approach has experienced rapid growth, driven by the emergence of mass spectrometry applications serving ‐omics sciences and the need for analyzing minute volumes of precious samples with ever higher sensitivity. While nanoscale liquid chromatography (flow rates <1 μL/min) has gained widespread recognition in proteomics, the adoption of microscale setups (flow rates ranging from 1 to 100 μL/min) for low molecular weight compound applications, including metabolomics, has been surprisingly slow, despite the inherent advantages of the approach. Highly heterogeneous matrices and chemical structures accompanied by a relative lack of options for both selective sample preparation and user‐friendly equipment are usually reported as major hindrances. To facilitate the wider implementation of microscale analyses, we present here a comprehensive tutorial encompassing important theoretical and practical considerations. We provide fundamental principles in micro‐chromatography and guide the reader through the main elements of a microflow workflow, from LC pumps to ionization devices. Finally, based on both our literature overview and experience, illustrated by some in‐house data, we highlight the critical importance of the ionization source design and its careful optimization to achieve significant sensitivity improvement.

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High-performance peptide and disulfide mapping by direct injection of intact proteins using on-line coupled UV-liquid chromatography microdroplet mass spectrometry (UVLC-MMS)

Kuo,

Jen,

Chen

et al. 2023

Analytica Chimica Acta

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Label-free and de-conjugation-free workflow to simultaneously quantify trace amount of free/conjugated and protein-bound estrogen metabolites in human serum

Cited by 5 publications

References 33 publications

Preparation of Monolith for Online Extraction and LC–MS Analysis of β-Estradiol in Serum Via a Simple Multicomponent Reaction

Preparation of Monolith for Online Extraction and LC–MS Analysis of β-Estradiol in Serum Via a Simple Multicomponent Reaction

Hyphenation of microflow chromatography with electrospray ionization mass spectrometry for bioanalytical applications focusing on low molecular weight compounds: A tutorial review

High-performance peptide and disulfide mapping by direct injection of intact proteins using on-line coupled UV-liquid chromatography microdroplet mass spectrometry (UVLC-MMS)

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