Determination of trace amounts of sulphide in human red blood cells by high-performance liquid chromatography with fluorimetric detection after derivatization with p-phenylenediamine and iron(III).
A high-performance liquid chromatographic method based on pre-column fluorescence derivatization has been developed for the determination of trace amounts of sulphide. After the sulphide had been converted into a fluorescent derivative, thionine, by the reaction with p-phenylenediamine and Fe3+, it was separated on a reversed-phase column and detected fluorimetrically (excitation, 600 nm; emission, 623 nm). Sulphide ion can be determined in the range from 0.01 to 3.0 mumol dm-3 with a relative standard deviation (n = 5) of 2.54% at 0.02 mumol dm-3 and 1.74% at 1.0 mumol dm-3. The proposed method was applied to the determination of sulphide in human red blood cells from ten healthy subjects, by generating sulphide in a microdiffusion apparatus; the concentrations found ranged from 0.123 to 0.189 mumol dm-3.
Ogasawara
Y
Y
Department of Hygienic Chemistry, Meiji College of Pharmacy, Tokyo, Japan.
Ishii
K
K
Togawa
T
T
Tanabe
S
S
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004912:Erythrocytes / Q000737:chemistry*; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D007501:Iron; D008297:Male; D008751:Methylene Blue; D008832:Microchemistry; D010655:Phenylenediamines; D012016:Reference Values; D013050:Spectrometry, Fluorescence / Q000379:methods; D013440:Sulfides / Q000097:blood*
D007202:Indicators and Reagents; D010655:Phenylenediamines; D013440:Sulfides; D007501:Iron; D008751:Methylene Blue; C029728:4-phenylenediamine