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Purpose <p>Ethylene glycol (EG) is a typical antifreeze compound and a significant analyte in forensic toxicology. Current EG analytical method for biological samples in forensic toxicology employ liquid chromatography–tandem mass spectrometry (LC–MS/MS), however, they exhibit low sensitivity and reliability. Therefore, in this study, we aimed to establish a highly sensitive, selective, and reliable EG assay system for human serum analysis using a hydroxyl derivatization-aided LC–MS/MS technique. </p> Methods <p><i>p</i>-Toluenesulfonyl isocyanate (PTSI) was applied for precolumn derivatization of EG in human serum, to enhance the sensitivity of LC–MS/MS for EG detection.</p> Results <p>The optimal derivatization conditions were 200 µL/mL PTSI in acetonitrile at 25&#xa0;°C for 10&#xa0;min. A highly sensitive and reliable LC–MS/MS detection of EG in human serum was achieved, with the calibration curve exhibiting a good linearity (<i>r</i> &gt; 0.999, 10–1000 µg/mL of EG). The proposed PTSI-derivatization-LC–MS/MS method exhibited high reliability (1.4–1.8%) for the intra-day and inter-day repeatability (%RSD), and accuracy (96.7–102.4%), with the limits of detection and quantification in human serum being 0.023 µg/mL (<i>S/N</i> = 3) and 0.077 µg/mL (<i>S/N</i> = 10), respectively. </p> Conclusions <p>A novel PTSI derivatization-aided LC–MS/MS method was developed, offering a highly sensitive, selective, and reliable analytical tool for EG quantification in human serum for forensic toxicology applications.</p>

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Quantitative analysis of ethylene glycol in human serum by liquid chromatography–tandem mass spectrometry with p-toluenesulfonyl isocyanate derivatization

  • 期刊论文

  • 发表日期：

    2025年6月24日

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  • Shin Ogawa,
  • Ryosuke Shiraki,
  • Kengo Wakigawa,
  • Hidehiko Okazaki,
  • Akira Tsujita,
  • Akinaga Gohda,
  • Toshiro Matsui

摘要

Purpose

Ethylene glycol (EG) is a typical antifreeze compound and a significant analyte in forensic toxicology. Current EG analytical method for biological samples in forensic toxicology employ liquid chromatography–tandem mass spectrometry (LC–MS/MS), however, they exhibit low sensitivity and reliability. Therefore, in this study, we aimed to establish a highly sensitive, selective, and reliable EG assay system for human serum analysis using a hydroxyl derivatization-aided LC–MS/MS technique.

Methods

p-Toluenesulfonyl isocyanate (PTSI) was applied for precolumn derivatization of EG in human serum, to enhance the sensitivity of LC–MS/MS for EG detection.

Results

The optimal derivatization conditions were 200 µL/mL PTSI in acetonitrile at 25 °C for 10 min. A highly sensitive and reliable LC–MS/MS detection of EG in human serum was achieved, with the calibration curve exhibiting a good linearity (r > 0.999, 10–1000 µg/mL of EG). The proposed PTSI-derivatization-LC–MS/MS method exhibited high reliability (1.4–1.8%) for the intra-day and inter-day repeatability (%RSD), and accuracy (96.7–102.4%), with the limits of detection and quantification in human serum being 0.023 µg/mL (S/N = 3) and 0.077 µg/mL (S/N = 10), respectively.

Conclusions

A novel PTSI derivatization-aided LC–MS/MS method was developed, offering a highly sensitive, selective, and reliable analytical tool for EG quantification in human serum for forensic toxicology applications.