Document Detail


A monobromobimane-based assay to measure the pharmacokinetic profile of reactive sulphide species in blood.
MedLine Citation:
PMID:  20590590     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND AND PURPOSE: Hydrogen sulphide (H(2)S) is a labile, endogenous metabolite of cysteine, with multiple biological roles. The development of sulphide-based therapies for human diseases will benefit from a reliable method of quantifying H(2)S in blood and tissues.
EXPERIMENTAL APPROACH: Concentrations of reactive sulphide in saline and freshly drawn whole blood were quantified by reaction with the thio-specific derivatization agent monobromobimane, followed by reversed-phase fluorescence HPLC and/or mass spectrometry. In pharmacokinetic studies, male rats were exposed either to intravenous infusions of sodium sulphide or to H(2)S gas inhalation, and levels of available blood sulphide were measured. Levels of dissolved H(2)S/HS(-) were concomitantly measured using an amperometric sensor.
KEY RESULTS: Monobromobimane was found to rapidly and quantitatively derivatize sulphide in saline or whole blood to yield the stable small molecule sulphide dibimane. Extraction and quantification of this bis-bimane derivative were validated via reversed-phase HPLC separation coupled to fluorescence detection, and also by mass spectrometry. Baseline levels of sulphide in blood were in the range of 0.4-0.9 microM. Intravenous administration of sodium sulphide solution (2-20 mg x kg(-1) x h(-1)) or inhalation of H(2)S gas (50-400 ppm) elevated reactive sulphide in blood in a dose-dependent manner. Each 1 mg x kg(-1) x h(-1) of sodium sulphide infusion into rats was found to be pharmacokinetically equivalent to approximately 30 ppm of H(2)S gas inhalation.
CONCLUSIONS AND IMPLICATIONS: The monobromobimane derivatization method is a sensitive and reliable means to measure reactive sulphide species in whole blood. Using this method, we have established a bioequivalence between infused sodium sulphide and inhaled H(2)S gas.
Authors:
Edward A Wintner; Thomas L Deckwerth; William Langston; Asa Bengtsson; Dina Leviten; Paul Hill; Michael A Insko; Ronald Dumpit; Emily VandenEkart; Christopher F Toombs; Csaba Szabo
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  British journal of pharmacology     Volume:  160     ISSN:  1476-5381     ISO Abbreviation:  Br. J. Pharmacol.     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-07-01     Completed Date:  2010-11-03     Revised Date:  2011-07-28    
Medline Journal Info:
Nlm Unique ID:  7502536     Medline TA:  Br J Pharmacol     Country:  England    
Other Details:
Languages:  eng     Pagination:  941-57     Citation Subset:  IM    
Affiliation:
Ikaria, Seattle, WA, USA.
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MeSH Terms
Descriptor/Qualifier:
Analytic Sample Preparation Methods / methods*
Animals
Bicyclo Compounds / chemistry*
Chromatography, High Pressure Liquid
Fluorescent Dyes / chemistry
Free Radicals / analysis,  blood,  chemistry
Humans
Hydrogen Sulfide / administration & dosage,  blood,  chemistry,  pharmacokinetics
Kinetics
Limit of Detection
Male
Rats
Rats, Sprague-Dawley
Spectrometry, Fluorescence
Spectrometry, Mass, Electrospray Ionization
Sulfhydryl Compounds / analysis,  blood*,  chemistry
Sulfhydryl Reagents / chemistry*
Sulfides / administration & dosage,  blood*,  chemistry,  pharmacokinetics*,  therapeutic use
Tandem Mass Spectrometry
Chemical
Reg. No./Substance:
0/Bicyclo Compounds; 0/Fluorescent Dyes; 0/Free Radicals; 0/Sulfhydryl Compounds; 0/Sulfhydryl Reagents; 0/Sulfides; 1313-82-2/sodium sulfide; 71418-44-5/monobromobimane; 7783-06-4/Hydrogen Sulfide
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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