A novel approach for identification and measurement of hemoglobin adducts with 1,2,3,4-diepoxybutane by liquid chromatography/electrospray ionisation mass spectrometry and matrix-assisted laser desorption/ionisation tandem mass spectrometry.
The structural characterisation of the adducts formed by in vitro interaction of hemoglobin (Hb) with 1,2,3,4-diepoxybutane (DEB), the most reactive 1,3-butadiene (BD) metabolite, was obtained by liquid chromatography/electrospray ionisation mass spectrometry (LC/ES-MS) analysis of modified tryptic peptides of human hemoglobin chains. The reactive sites of human hemoglobin towards DEB and its hydroxylated derivatives (trihydroxybutyl (THB)-derivatives) were identified through the characterisation of alkylated tryptic peptides by matrix-assisted laser desorption/ionisation tandem mass spectrometry (MALDI-MS/MS). Based on this characterisation, a procedure was set up to measure the Hb-adducts of THB-derivatives by isotope dilution mass spectrometry with the use of a deuterated peptide standard. The results obtained here could permit optimisation of molecular dosimetry of BD-adducts, and extension of the analysis to the biological monitoring of occupational exposure to butadiene.
Basile
A
A
Centro Internazionale di Servizi di Spettrometria di Massa, Istituto di Scienze dell'Alimentazione del C.N.R., Via Roma 52, I-83100 Avellino, Italy.
Ferranti
P
P
Pocsfalvi
G
G
Mamone
G
G
Miraglia
N
N
Caira
S
S
Ambrosi
L
L
Soleo
L
L
Sannolo
N
N
Malorni
A
A
D000478:Alkylation; D002851:Chromatography, High Pressure Liquid; D004852:Epoxy Compounds / Q000737:chemistry*; D005914:Globins / Q000032:analysis; D006454:Hemoglobins / Q000032:analysis*; D007202:Indicators and Reagents; D010455:Peptides / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry
D004852:Epoxy Compounds; D006454:Hemoglobins; D007202:Indicators and Reagents; D010455:Peptides; C007366:diepoxybutane; D005914:Globins; D014357:Trypsin