Measurement of H2S in vivo and in vitro by the monobromobimane method.
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.
Department of Pathology, Louisiana State University Health Sciences Center-Shreveport, Shreveport, Louisiana, USA.
Xinggui
X
Shen
Department of Pathology, Louisiana State University Health Sciences Center-Shreveport, Shreveport, Louisiana, USA.
Gopi K
GK
Kolluru
Department of Pathology, Louisiana State University Health Sciences Center-Shreveport, Shreveport, Louisiana, USA.
Shuai
S
Yuan
Department of Pathology, Louisiana State University Health Sciences Center-Shreveport, Shreveport, Louisiana, USA. Electronic address: ckevil@LSUHSC.edu.
Christopher G
CG
Kevil
D000818:Animals; D001643:Bridged Bicyclo Compounds / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006862:Hydrogen Sulfide / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D012015:Reference Standards; D013050:Spectrometry, Fluorescence; D021241:Spectrometry, Mass, Electrospray Ionization
D001643:Bridged Bicyclo Compounds; D005456:Fluorescent Dyes; C028381:monobromobimane; D006862:Hydrogen Sulfide
Fluorescence; High-performance liquid chromatography; Mass spectrometry; Oxygen; Sulfide; pH