Measurement of H2S in vivo and in vitro by the monobromobimane method

Xinggui Shen, Gopi K Kolluru, Shuai Yuan, Christopher G. Kevil

Research output: Chapter in Book/Published conference outputChapter (peer-reviewed)peer-review


The gasotransmitter hydrogen sulfide (H2S) is known as an important regulator in several physiological and pathological responses. Among the challenges facing the field is the accurate and reliable measurement of hydrogen sulfide bioavailability. We have reported an approach to discretely measure sulfide and sulfide pools using the monobromobimane (MBB) method coupled with reversed phase high-performance liquid chromatography (RP-HPLC). The method involves the derivatization of sulfide with excess MBB under precise reaction conditions at room temperature to form sulfide dibimane (SDB). The resultant fluorescent SDB is analyzed by RP-HPLC using fluorescence detection with the limit of detection for SDB (2 nM). Care must be taken to avoid conditions that may confound H2S measurement with this method. Overall, RP-HPLC with fluorescence detection of SDB is a useful and powerful tool to measure biological sulfide levels.

Original languageEnglish
Title of host publicationHydrogen sulfide in redox biology, part A
EditorsEnrique Cadenas, Lester Packer
Place of PublicationLondon (UK)
PublisherAcademic Press
Number of pages15
ISBN (Print)978-0-12-801512-4
Publication statusPublished - 2015

Publication series

NameMethods in Enzymology
PublisherAcademics Press
ISSN (Print)0076-6879


  • high pressure liquid chromatography
  • bicyclo compounds
  • fluorescence spectrometry
  • fluorescent dyes
  • electrospray ionization
  • hydrogen sulfide
  • reference standards
  • spectrometry
  • mass


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  • H2S regulation of nitric oxide metabolism

    Kolluru, G. K., Yuan, S., Shen, X. & Kevil, C. G., 9 Sept 2015, Hydrogen sulfide in redox biology, part A. Cadenas, E. & Packer, L. (eds.). London (UK): Academic Press, p. 271-297 27 p. (Methods in Enzymology; vol. 554).

    Research output: Chapter in Book/Published conference outputChapter (peer-reviewed)peer-review

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