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Emerging Trends in Microextraction Techniques for Bioanalytical Applications
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Emerging Trends in Microextraction Techniques for Bioanalytical Applications

A special issue of Molecules (ISSN 1420-3049). This special issue belongs to the section "Analytical Chemistry".

Deadline for manuscript submissions: 31 August 2026 | Viewed by 4571

Special Issue Editor

Dr. Constantinos K. Zacharis

E-Mail Website
Guest Editor
Laboratory of Pharmaceutical Analysis, Department of Pharmaceutical Technology, School of Pharmacy, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece
Interests: pharmaceutical analytical chemistry; method development and validation; sample preparation (derivatization, microextraction, etc.); liquid and gas chromatography; capillary electrophoresis; mass spectrometry
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Special Issue Information

Dear Colleagues,

Microextraction techniques have gained significant attention from researchers working in bioanalysis due to their ability to efficiently isolate and preconcentrate analytes from complex biological matrices (e.g., blood, urine, plasma). These techniques are crucial for enhancing sensitivity and selectivity in detecting biomarkers and drugs in biological systems.

Recent trends in this field have involved the development of faster and greener and automated approaches. These aim to reduce solvent consumption, decrease sample size, and streamline the extraction process. Advances in technology have also led to the development of automated sample preparation systems, improving both efficiency and consistency. Sorbent-based or liquid-based microextraction techniques are the most promising for meeting these requirements and are increasingly replacing traditional liquid extraction approaches. Typical examples of these techniques include solid phase microextraction, stir-bar sorptive extraction, ionic-liquid-based or deep eutectic solvent-based liquid phase microextraction, microextraction by packed sorbent, micro solid phase extraction, etc.

This Special Issue focuses on the latest research trends and applications of sorbent-based and liquid-phase microextraction strategies. Researchers across all areas of basic research and biomedical sciences are cordially invited to submit a research or review article for this Special Issue.

Dr. Constantinos K. Zacharis
Guest Editor

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Keywords

  • sorbent-based microextraction (i.e., solid phase microextraction (SPME), matrix solid phase dispersion (MSPD), pipette-tip micro solid phase extraction, fabric sorptive phase extraction (FPSE), capsule phase microextraction (CPME), stir bar sorptive extraction (SBSE), etc.)
  • liquid-phase microextraction (dispersive liquid–liquid microextraction, single-drop microextraction, electromembrane extraction, etc.)
  • automation
  • new materials/sorbents (MOF, COFs, nanomaterials, conductive polymers, magnetic materials, etc.)
  • miniaturization in sample preparation

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Published Papers (4 papers)

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Research

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14 pages, 2178 KB  
Article
Composition Study of Polyphyllin in Paris polyphylla by Ultrasound-Assisted Deep Eutectic Solvent Extraction Combined with UHPLC-MS/MS
by Jinyu Guo, Jiajia Liu, Minlong Li, Zhenlin Tan, Huayin Lu and Yuting Zhou
Molecules 2026, 31(3), 473; https://doi.org/10.3390/molecules31030473 - 29 Jan 2026
Viewed by 556
Abstract
Paris polyphylla (Chonglou), a medicinal herb documented in Shennong’s Classic of Materia Medica and a key component of formulas such as Yunnan Baiyao, is a rare and endangered plant prized for its bioactive steroidal saponins, notably polyphyllin I (PPI) and II (PPII). However, [...] Read more.
Paris polyphylla (Chonglou), a medicinal herb documented in Shennong’s Classic of Materia Medica and a key component of formulas such as Yunnan Baiyao, is a rare and endangered plant prized for its bioactive steroidal saponins, notably polyphyllin I (PPI) and II (PPII). However, its pharmacological potential is hampered by inefficient extraction and unreliable compound identification. Herein, we developed a sustainable and efficient extraction strategy using ultrasound-assisted deep eutectic solvents (DES), optimized via an L9(34) orthogonal experimental design. Extraction efficiencies across the seven Paris species ranged from 2.04% to 16.51%, achieved by systematically optimizing key parameters such as the choline chloride-to-ethanol molar ratio (1:1.8), material-to-liquid ratio (1:20 g mL−1), and extraction time (100 min). By ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) analysis, PPI and PPII were quantified using specific retention times and characteristic fragment ions, revealing content ranges of 3.282–21.452 mg g−1 and 4.201–17.975 mg g−1, respectively. This methodology provides a robust platform for quality control and standardization of Paris-derived medicines, while paving the way for sustainable utilization and in-depth study of its steroidal saponins. Full article
(This article belongs to the Special Issue Emerging Trends in Microextraction Techniques for Bioanalytical Applications)
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14 pages, 1487 KB  
Article
Sexual Hormones Determination in Biofluids by In-Vial Polycaprolactone Thin-Film Microextraction Coupled with HPLC-MS/MS
by Francesca Merlo, Silvia Anselmi, Andrea Speltini, Clàudia Fontàs, Enriqueta Anticó and Antonella Profumo
Molecules 2026, 31(2), 255; https://doi.org/10.3390/molecules31020255 - 12 Jan 2026
Viewed by 615
Abstract
The in-vial microextraction technique is emerging as an alternative sample treatment, as it integrates sorbent preparation, adsorption, and desorption of analytes in a single device before instrumental analysis. In this work, the applicability of polycaprolactone polymeric film, recently used for the in-vial microextraction [...] Read more.
The in-vial microextraction technique is emerging as an alternative sample treatment, as it integrates sorbent preparation, adsorption, and desorption of analytes in a single device before instrumental analysis. In this work, the applicability of polycaprolactone polymeric film, recently used for the in-vial microextraction of sex hormones from environmental waters, is studied in a low-capacity format for unconjugated sex hormones determination in biological samples by HPLC-MS/MS. Its performance was evaluated in urine and serum, achieving extraction in a short time (10 and 30 min, in turn) and satisfactory elution with ethanol, with recovery in the range of 65–111% in urine, 55–122% in bovine serum albumin (BSA) solution, and 66–121% in fetal bovine serum (FBS). In the case of protein matrices, a dilution to 20 g L−1 protein content and washing step (3 × 1 mL ultrapure water) afore the elution are required to achieve clean extract, as verified by a Bradford assay. Matrix-matched calibration was used for quantification, obtaining correlation coefficients greater than 0.9929; limits of detection and quantification were in the range of 0.01–0.65 and 0.03–1.96 ng mL−1 in urine, 0.02–0.8 and 0.05–2.5 ng mL−1 in BSA, and 0.02–1.0 and 0.06–3.0 g mL−1 in FBS, respectively. The in-vial polycaprolactone film proved to be reusable for several cycles (up to ten), and the greenness assessment revealed a good adhesion to green sample preparation principles. All these achievements further strengthen its feasibility for efficient extraction/clean-up of trace sex hormones in complex biological samples. Full article
(This article belongs to the Special Issue Emerging Trends in Microextraction Techniques for Bioanalytical Applications)
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Review

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39 pages, 2305 KB  
Review
Green and Emerging Microextraction Strategies for Bioanalytical Determination of Hormones: Trends, Challenges, and Applications
by David Vicente-Zurdo, Sonia Morante-Zarcero and Isabel Sierra
Molecules 2025, 30(22), 4471; https://doi.org/10.3390/molecules30224471 - 19 Nov 2025
Cited by 1 | Viewed by 1471
Abstract
Accurate and sensitive determination of hormones in biological matrices is essential for clinical diagnostics, therapeutic monitoring, and endocrine research. However, hormone determination presents significant challenges due to their typically low concentrations, complex sample matrices, and structural diversity. In recent years, microextraction techniques have [...] Read more.
Accurate and sensitive determination of hormones in biological matrices is essential for clinical diagnostics, therapeutic monitoring, and endocrine research. However, hormone determination presents significant challenges due to their typically low concentrations, complex sample matrices, and structural diversity. In recent years, microextraction techniques have emerged as strategic tools in bioanalytical chemistry, offering advantages in terms of miniaturization, enhanced selectivity, and compatibility with the principles of green analytical chemistry (GAC). This review provides a comprehensive overview of green and emerging microextraction approaches for the determination of steroidal, thyroid, peptide, and other hormones in biological samples. Key techniques such as solid-phase microextraction (SPME) and dispersive liquid–liquid microextraction (DLLME), followed by high-performance liquid chromatography (HPLC) coupled to diode array detectors (DADs) or mass spectrometry (MS), are critically discussed. Special emphasis is placed on the use of environmentally friendly solvents, such as deep eutectic solvents (DESs), supramolecular solvents (SUPRASs), and advanced sorbents including molecularly imprinted polymers (MIPs) and nanostructured magnetic phases. Applications across various bioanalytical matrices (urine, plasma, serum, saliva, tissues…) are examined in terms of sensitivity, selectivity, and validation parameters. Finally, current challenges, method development gaps, and future directions are highlighted to support the continued advancement of sustainable hormone determination in complex biological systems. Full article
(This article belongs to the Special Issue Emerging Trends in Microextraction Techniques for Bioanalytical Applications)
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41 pages, 1247 KB  
Review
Miniaturised Extraction Techniques in Personalised Medicine: Analytical Opportunities and Translational Perspectives
by Luana M. Rosendo, Tiago Rosado, Mário Barroso and Eugenia Gallardo
Molecules 2025, 30(21), 4263; https://doi.org/10.3390/molecules30214263 - 31 Oct 2025
Cited by 2 | Viewed by 1343
Abstract
Miniaturised sampling and extraction are redefining therapeutic drug monitoring (TDM) by enabling low-volume sampling, simplifying collection, and improving patient acceptability, while also promoting decentralised workflows and more sustainable laboratory practices. This review critically appraises the current landscape, with emphasis on analytical performance, matrix [...] Read more.
Miniaturised sampling and extraction are redefining therapeutic drug monitoring (TDM) by enabling low-volume sampling, simplifying collection, and improving patient acceptability, while also promoting decentralised workflows and more sustainable laboratory practices. This review critically appraises the current landscape, with emphasis on analytical performance, matrix compatibility, and readiness for clinical implementation. It examines validation requirements, the extent of alignment and existing gaps across major regulatory guidelines, and recurrent challenges such as haematocrit bias, real-world stability and transport, incurred sample reanalysis, device variability, commutability with conventional matrices, and inter-laboratory reproducibility. To make the evidence actionable, operational recommendations are distilled into a practical ten-point checklist designed to support validation and translation of miniaturised approaches into routine laboratory practice. Looking ahead, priorities include automation and portable platforms, advanced functional materials, and integration with digital tools and biosensors, alongside the development of harmonised frameworks tailored to miniaturised methods and prospective clinical studies that demonstrate impact on dosing decisions, adherence, and clinical outcomes. Overall, this review aims to equip researchers, laboratory professionals, and regulators with the knowledge to implement miniaturised bioanalysis and advance personalised medicine through TDM. Full article
(This article belongs to the Special Issue Emerging Trends in Microextraction Techniques for Bioanalytical Applications)
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