Measurement of AM19 and other cyclosporine metabolites in the blood of liver transplant patients with stable liver function.

Abstract

Chronic renal impairment represents on of the major side effects of cyclosporine (CsA) immunotherapy for organ transplantation. The clinical relevance of selective measurement of CsA metabolites to correlating nephrotoxic activities remains inconclusive. A relatively simple and reliable method was developed to measure AM19 and four other CsA metabolites in whole blood by sequential solid-phase extraction and reversed phase high-pressure liquid chromatography (HPLC). The procedures were modified from a well-established method developed to quantitate CsA, the most important difference being in the elution step. The use of acetonitrile/methanol instead of ethylacetate/isopropanol in the elution procedure enhanced the absolute recoveries of metabolites. The washing steps were also slightly modified to recover AM19 quantitatively and specifically. Isocratic chromatographic conditions allowed very good separation of AM19, AM1, AM9, AM1c, AM4N, CsA, and the internal standard, cyclosporin C (CsC). Analytical recoveries for CsA and five of its metabolites ranged from 82 to 92%. No interfering substance from the matrix was found. The detection limit was 10 micrograms/l. The objectives of this study were to measure trough concentration of AM19 in whole blood, expressed as percentage of total metabolites plus parent CsA, and to determine if the blood level of AM19 could be used to predict subsequent changes in serum creatinine level in liver transplant patients. Twenty-three patients, who underwent liver transplantation between January and June 1993 were studied. Trough concentration of CsA in whole blood, serum creatinine and liver enzymes were constantly monitored. AM19 level was determined 5-8 months after transplantation. Preliminary results suggest an inverse relationship between AM19 and serum creatine levels.(ABSTRACT TRUNCATED AT 250 WORDS)