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Related Experiment Video

Updated: Jan 29, 2026

Assessment of Sexual Behavior of Male Mice
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Sexual Hormones Determination in Biofluids by In-Vial Polycaprolactone Thin-Film Microextraction Coupled with

Francesca Merlo1, Silvia Anselmi1, Andrea Speltini1

  • 1Department of Chemistry, University of Pavia, Via Taramelli 12, 27100 Pavia, Italy.

Molecules (Basel, Switzerland)
|January 28, 2026
PubMed
Summary

Polycaprolactone films offer efficient in-vial microextraction for sex hormones in biological samples. This green method provides rapid extraction and reusable films for accurate HPLC-MS/MS analysis.

Keywords:
bioanalysisestrogensgreen sample preparationin-vial microextractionpolymeric filmprogestinsserumurine

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Area of Science:

  • Analytical Chemistry
  • Environmental Chemistry
  • Biochemistry

Background:

  • In-vial microextraction (IVME) integrates sample preparation steps into a single device.
  • Polycaprolactone (PCL) films are explored for IVME applications.
  • Determination of unconjugated sex hormones in biological matrices requires efficient sample preparation.

Purpose of the Study:

  • To evaluate the applicability of PCL polymeric films for low-capacity IVME of unconjugated sex hormones.
  • To assess the performance of PCL films in biological samples like urine and serum using HPLC-MS/MS.
  • To determine the efficiency, reusability, and greenness of the PCL-based IVME method.

Main Methods:

  • Utilized PCL polymeric films for in-vial microextraction of sex hormones.
  • Analyzed urine, bovine serum albumin (BSA) solution, and fetal bovine serum (FBS) samples.
  • Employed High-Performance Liquid Chromatography-Tandem Mass Spectrometry (HPLC-MS/MS) for quantification.
  • Implemented matrix-matched calibration and Bradford assay for protein quantification.

Main Results:

  • Achieved rapid extraction times (10-30 min) with satisfactory elution using ethanol.
  • Obtained good recovery rates (55-122%) across different biological matrices.
  • Demonstrated the need for protein matrix dilution and washing steps for clean extracts.
  • Reported low limits of detection and quantification (0.01-3.0 ng mL⁻¹).
  • Confirmed PCL film reusability for up to ten cycles.

Conclusions:

  • PCL films are effective for the efficient extraction and clean-up of trace sex hormones in complex biological samples.
  • The IVME method using PCL films adheres to green sample preparation principles.
  • This technique offers a feasible alternative for routine analysis of sex hormones in biological fluids.