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Clinical Chemistry 38: 2025-2032, 1992;
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Clinical Chemistry, Vol 38, 2025-2032, Copyright © 1992 by American Association for Clinical Chemistry

Specific and sensitive measurement of FK506 and its metabolites in blood and urine of liver-graft recipients

U Christians, F Braun, M Schmidt, N Kosian, HM Schiebel, L Ernst, M Winkler, C Kruse, A Linck and KF Sewing
Institute fur Allgemeine Pharmakologie, Medizinische Hochschule Hannover, FRG.

A specific and sensitive assay for quantifying the immunosuppressant FK506 and its metabolites in blood and urine was developed. 32-O-Acetyl FK506 was synthesized and used as internal standard. FK506 and its metabolites were purified from the samples by solid-liquid extraction and were injected into a high-performance liquid chromatographic (HPLC) system linked to a mass spectrometer (MS) by particle-beam interface. The FK506 derivatives were separated from interfering material by use of a 100 x 4 mm C8 analytical column and water/acetonitrile or water/methanol gradient elution; they were detected by negative chemical ionization with methane as reagent gas. The limit of detection was 25 pg in a standard solution, and the limit of quantification in blood was 250 pg (extracted from 1 mL of blood). The CV was 11.3% at 5 ng, and no interferences with other drugs were found.


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