Non-radioactive immunoassays in toxicology: Instrumentation and analysis of hormones
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We have designed and executed comparative studies with the objective of selecting instrumentation for the non-isotope immunological determination of free and total thyroxine (fT4 and tT4), and total triiodothyronine (tT3) and progesterone in rat plasma for general toxicity studies. In addition, this instrumentation has been used for the determination of progesterone in rabbits for early pregnancy diagnosis in reprotoxicity studies.
During instrument selection, special emphasis has been given to automation and on-line coupling capabilities, maximal flexibility in method development, walkaway capability, manufacturers' computer validation and GLP performance, linearity and intra-assay CV.
For tT3 and tT4, seven instruments have been compared with each other whereas for progesterone and fT4, three instruments were compared.
Normal rat plasma values have been subjected to variance analysis, followed by Duncan testing.
The instrument selection process finally indicated the ES300 Enzymun ELISA of Boehringer Mannheim as the best candidate on the aspects defined above. Spiking recovery values on the ES300 are presented.
The ES300 provides a reliable pregnancy diagnosis for rabbits as early as day +4 after mating in the predosing period.
KeywordsT3 T4 Progesterone Rabbit Rat ELISA Automation
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