Influence of Assay Methodology on the Measurement of Free Serum Ceftriaxone Concentrations

Author:

Kohlhepp Sue J.1,Gilbert David N.12,Leggett James E.12

Affiliation:

1. Earle A. Chiles Research Institute, Providence Portland Medical Center,1 and

2. Oregon Health Sciences University,2Portland, Oregon

Abstract

ABSTRACT The influence of assay methodology on the measurement of the active free fraction of ceftriaxone in plasma was determined. The free fraction was measured by three methods: agar diffusion bioassay, precipitation of plasma protein with methanol followed by high-performance liquid chromatography (HPLC) of the supernatant, and ultrafiltration of plasma followed by HPLC of the filtrate. In human serum, the free ceftriaxone levels were significantly lower ( P = 0.03) when measured on ultrafiltrates compared to the other two methods. This difference disappeared when dolphin serum was studied. After ultrafiltration, human serum was shown, by Scatchard plot analysis, to have two ceftriaxone binding sites. Species differences were also demonstrated. Hence, in humans, determination of free plasma ceftriaxone varies with the assay method employed.

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Pharmacology (medical),Pharmacology

Reference14 articles.

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4. Craig W. A. Suh B. Protein binding and the antimicrobial effects: methods for the determination of protein binding Antibiotics in laboratory medicine. Lorian V. 1986 477 517 Williams & Wilkins Baltimore Md

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