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Standardization of a Preparative Ultracentrifuge Method for Quantitative Determination of Protein Binding of Seven Antibiotics

Lance R. Peterson, Wendell H. Hall, Horace H. Zinneman and Dale N. Gerding
The Journal of Infectious Diseases
Vol. 136, No. 6 (Dec., 1977), pp. 778-783
Published by: Oxford University Press
Stable URL: http://www.jstor.org/stable/30107058
Page Count: 6
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Standardization of a Preparative Ultracentrifuge Method for Quantitative Determination of Protein Binding of Seven Antibiotics
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Abstract

A preparative ultracentrifuge method was standardized for determination of quantitative binding of cephalothin, cefamandole, cefazolin, cefaclor, erythromycin, gentamicin, and chloramphenicol to human serum proteins. At achievable in vivo concentrations, serum binding was 78.5% for cephalothin, 79.9% for cefamandole, 88.5% for cefazolin, 23.5% for cefaclor, 41.9% for erythromycin, 22.7% for gentamicin, and 59.5% for chloramphenicol. Techniques that use semipermeable cellophane or diaflow membranes, cross-linked dextran, inhibition of bacterial growth, protein precipitation, or liquid partitioning all have inherent problems with either the ligand or the antibiotic adversely interacting with the experimental apparatus. Ultracentrifugation provides a rapid, reproducible technique for protein-binding determinations of the classes of antibiotics described.

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