Study of deuterium isotope effects on protein binding by gas chromatography/mass spectrometry. Caffeine and deuterated isotopomers.
A study of the binding to human serum albumin (HSA) of caffeine and its deuterated isotopomers, 1-C2H3-,3-C2H3-, 1,7-(C2H3)2-, 3,7-(C2H3)2- and 1,3,7-(C2H3)3-caffeine, was performed by equilibrium dialysis. Free and bound fractions were measured by gas chromatography/mass spectrometry. Important and significant (Fischer and Student tests) isotope effects were observed on binding parameters: sites total concentration (N = 1732 microM for 1,3,7-(C2H3)3-caffeine versus 822 microM for caffeine; number of sites (n = 3 for 1,3,7-(C2H3)3-caffeine v. 1 for caffeine); and extent of binding (46% for 1,3,7-(C2H3)3-caffeine v. 27% for caffeine). A study of competition for HSA binding between caffeine and its 1,3,7-(C2H3)3- and 3,7-(C2H3)2-isotopomers confirmed the results obtained in direct binding studies. These isotope effects are discussed in terms of (a) tools for molecular pharmacology, (b) precautions to be taken when such labelled drugs are used in clinical pharmacology.
D002110:Caffeine / Q000097:blood*; D003903:Deuterium; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D007700:Kinetics; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*
D012709:Serum Albumin; D002110:Caffeine; D003903:Deuterium