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<p>In this study, a straightforward, rapid, and cost-effective preliminary screening method is described for the estimation of melatonin, using xanthydrol as a chromogenic reagent in thin-layer chromatography (TLC). Melatonin is a neurohormone predominantly produced by the pineal gland, which is essential for maintaining circadian rhythm regulation. Even though melatonin is a commonly used sleep aid, the detection of this compound is increasingly important, as it is widely misused in criminal activities. Although melatonin is widely regarded as safe, analytical identification may be required in certain forensic scenarios where its presence in questioned samples needs evaluation. This approach involves the derivatization of melatonin with xanthydrol to generate a stable blue-colored product, facilitating enhanced detection and visualization, employing TLC as an analytical method. The visual detection limit was observed at approximately 0.5&#xa0;µg/mL. Selected potentially interfering compounds did not exhibit an identical <i>R</i><sub>F</sub>–color response under the optimized conditions. The method is intended as a rapid preliminary screening tool and does not replace confirmatory instrumental techniques such as liquid chromatography–tandem mass spectrometry (LC–MS/MS). Limitations regarding selectivity toward other amide-containing compounds are acknowledged.</p>

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Thin-layer chromatographic detection of melatonin using xanthydrol as a chromogenic reagent

  • 期刊论文

  • 发表日期：

    2026年5月13日

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  • Sachin R. Golhe,
  • Santosh R. Kote,
  • Dadasaheb M. Bansode,
  • Asif M. Pathan,
  • Vijay J. Thakare,
  • Anil B. Naik

摘要

In this study, a straightforward, rapid, and cost-effective preliminary screening method is described for the estimation of melatonin, using xanthydrol as a chromogenic reagent in thin-layer chromatography (TLC). Melatonin is a neurohormone predominantly produced by the pineal gland, which is essential for maintaining circadian rhythm regulation. Even though melatonin is a commonly used sleep aid, the detection of this compound is increasingly important, as it is widely misused in criminal activities. Although melatonin is widely regarded as safe, analytical identification may be required in certain forensic scenarios where its presence in questioned samples needs evaluation. This approach involves the derivatization of melatonin with xanthydrol to generate a stable blue-colored product, facilitating enhanced detection and visualization, employing TLC as an analytical method. The visual detection limit was observed at approximately 0.5 µg/mL. Selected potentially interfering compounds did not exhibit an identical RF–color response under the optimized conditions. The method is intended as a rapid preliminary screening tool and does not replace confirmatory instrumental techniques such as liquid chromatography–tandem mass spectrometry (LC–MS/MS). Limitations regarding selectivity toward other amide-containing compounds are acknowledged.