Carcinogenesis vol.13 no.8 pp.1397-1401, 1992
Study of interaction of styrene oxide with Angiotensin by mass spectrometry
Pasquale Ferranti, Virginia Carbone1, Nicola Sannolo1, Immacolata Fiume1, Alfredo Milone, Margherita Ruoppolo, Monica Gallo and Antonio Malorni ICMIB and Servizio di Spettromctria di Massa del CNR, U° Facolta di Medicina, Via Pansini 5, 80131 Napoli and 'Sezione di Medicina Occupazionale e Igiene Industriale, Dipartimento di Biochimica e Biofisica, Universita di Napoli, Piazza Miraglia 3, 80138 Napoli, Italy 2
To whom correspondence should be addressed
Introduction Most chemical carcinogens are electrophilic reagents which form covalent adducts with nucleophilic centres of biomolecules present in living cells, either directly or after metabolic activation (1,2). Electrophiles bind covalently to nucleophilic sites present in DNA, the target for the mutagenic effect, and in proteins, as demonstrated for hemoglobin (3,4). It has been demonstrated that a constant ratio exists between DNA and protein adduct formation over a wide range of doses for some compounds (4—6). The amount of protein adducts is therefore closely related to the tissue dose, i.e. the actual dose responsible for genetic damage. Thus, biomonitoring the protein adducts represents a specific and sufficiently sensitive approach to allow the determination of the tissue dose absorbed in working places and in the environment. Styrene oxide is the major metabolite of styrene (7), widely used in industry. Styrene is mainly absorbed via inhalation and an uptake of ~ 60-70% of the amount inhaled has been reported •Abbreviations: GC/MS, gas chromatography-mass spectrometry; FAB/MS, fast atom bombardment mass spectrometry; MS-MS, tandem mass spectrometry; TFA, tnfluoroacetic acid; MTBSTFA, W-methyl-iV-^rt-butyl-dimethylsilyl)trifiuoroacetamide; RP-HPLC, reverse-phase HPLC; El, electron ionization; TIC, total ion chromatogram; TBDMS, terr-butyldimethylsilyl. © Oxford University Press
Materials and methods Trifluoroacetic acid (TFA) and sodium dihydrogen phosphate (reagent grade), acetonitrile, diethyl ether and methylene chloride (HPLC grade) were obtained from Carlo Erba. Angiotensin I was purchased from Novabiochem. Chymotrypsin and Pronase E were obtained from Sigma and aminopeptidase M from BoehringerMannheim. W-Methyl-AKr
Study of interaction of styrene oxide with Angiotensin by mass spectrometry
Pasquale Ferranti, Virginia Carbone1, Nicola Sannolo1, Immacolata Fiume1, Alfredo Milone, Margherita Ruoppolo, Monica Gallo and Antonio Malorni ICMIB and Servizio di Spettromctria di Massa del CNR, U° Facolta di Medicina, Via Pansini 5, 80131 Napoli and 'Sezione di Medicina Occupazionale e Igiene Industriale, Dipartimento di Biochimica e Biofisica, Universita di Napoli, Piazza Miraglia 3, 80138 Napoli, Italy 2
To whom correspondence should be addressed
Introduction Most chemical carcinogens are electrophilic reagents which form covalent adducts with nucleophilic centres of biomolecules present in living cells, either directly or after metabolic activation (1,2). Electrophiles bind covalently to nucleophilic sites present in DNA, the target for the mutagenic effect, and in proteins, as demonstrated for hemoglobin (3,4). It has been demonstrated that a constant ratio exists between DNA and protein adduct formation over a wide range of doses for some compounds (4—6). The amount of protein adducts is therefore closely related to the tissue dose, i.e. the actual dose responsible for genetic damage. Thus, biomonitoring the protein adducts represents a specific and sufficiently sensitive approach to allow the determination of the tissue dose absorbed in working places and in the environment. Styrene oxide is the major metabolite of styrene (7), widely used in industry. Styrene is mainly absorbed via inhalation and an uptake of ~ 60-70% of the amount inhaled has been reported •Abbreviations: GC/MS, gas chromatography-mass spectrometry; FAB/MS, fast atom bombardment mass spectrometry; MS-MS, tandem mass spectrometry; TFA, tnfluoroacetic acid; MTBSTFA, W-methyl-iV-^rt-butyl-dimethylsilyl)trifiuoroacetamide; RP-HPLC, reverse-phase HPLC; El, electron ionization; TIC, total ion chromatogram; TBDMS, terr-butyldimethylsilyl. © Oxford University Press
Materials and methods Trifluoroacetic acid (TFA) and sodium dihydrogen phosphate (reagent grade), acetonitrile, diethyl ether and methylene chloride (HPLC grade) were obtained from Carlo Erba. Angiotensin I was purchased from Novabiochem. Chymotrypsin and Pronase E were obtained from Sigma and aminopeptidase M from BoehringerMannheim. W-Methyl-AKr
