Quantification of homocysteine thiolactone in human saliva and urine by gas chromatography-mass spectrometry

Justyna Piechocka, Monika Wrońska, Grażyna Chwatko, Hieronim Jakubowski, Rafał Głowacki

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9 Scopus citations


Homocysteine thiolactone (HTL) is a chemically reactive thioester that has been implicated in cardiovascular disease. So far, its presence has been documented in human and mouse plasma and urine. Here, using a new method, we show that HTL is present in human saliva. The assay involves chloroform-methanol extraction of HTL, lyophilization, and derivatization with N-trimethylsilyl-N-methyl trifluoroacetamide (MSTFA) and trimethylchlorosilane (TMCS). The method is based on a gas chromatography coupled with mass spectrometry (GC–MS) and quantifies HTL in a linear range from 0.05 to 1 µmol L−1 saliva and urine. The limit of quantification (LOQ) was 0.05 µmol L−1. With respect to saliva specimen, the accuracy was 98.7–112.6%, and 90.2–100.5%, while the precision was 7.1–13.5% and 12.5–15.0% for the intra- and inter-day variation, respectively. In relation to urine samples, the accuracy was 91.9–110.9% and 91.2–103.3%, while the precision varied from 2.2% to 14.5% and 7.4% to 14.3% for intra- and inter-day measurements, respectively. Using this method, we show that in apparently healthy individuals (n = 18), HTL levels in saliva are not positively correlated with urinary HTL levels. Undoubtedly, larger population should be investigated to get more meaningful results.

Original languageEnglish (US)
Article number122155
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
StatePublished - Jul 15 2020

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology


  • Gas chromatography – mass spectrometry
  • Homocysteine thiolactone
  • Human saliva
  • Human urine
  • Liquid-liquid extraction
  • N-trimethylsilyl-N-methyl trifluoroacetamide


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