Combined high-performance liquid chromatography and radioimmunoassay for ceruletide and its metabolites in dog plasma and urine

A combined high-performance liquid chromatographic (HPLC) and competitive radioimmunoassay (RIA) method for ceruletide (CLT). an analogue of cholecystokinin-8, was developed to investigate the behaviour of CLT in dogs. Dog plasma samples after administration of CLT were deproteinized and separated b...

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Bibliographic Details
Published in:Journal of pharmaceutical and biomedical analysis 1994-03, Vol.12 (3), p.413-418
Main Authors: Kominami, Goro, Okabe, Hiroshi, Imoda, Keiko, Mizojiri, Kenji
Format: Article
Language:English
Subjects:
Animals
ceruletide
Ceruletide - administration & dosage
Ceruletide - blood
Ceruletide - pharmacokinetics
Ceruletide - urine
Chromatography, High Pressure Liquid
combined method
competitive radioimmunoassay
Dogs
Injections, Intramuscular
metabolite
peptide
Peptide Fragments - blood
Peptide Fragments - urine
Radioimmunoassay
Reproducibility of Results
Reversed-phase high-performance liquid chromatography
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Summary:A combined high-performance liquid chromatographic (HPLC) and competitive radioimmunoassay (RIA) method for ceruletide (CLT). an analogue of cholecystokinin-8, was developed to investigate the behaviour of CLT in dogs. Dog plasma samples after administration of CLT were deproteinized and separated by reversed-phase HPLC. Fractions for the HPLC eluate were measured by a RIA and two immunoreactive components were found in dog plasma. One fraction was assumed to be unchanged CET and the other was the (1–6) fragment peptide of CLT [CLT(1–6)] in accordance with the retention times. For the simultaneous determination by the combined method in dog plasma, the assay recoveries of both compounds were 100% and the values for assay precision (RSD intra-assay) were 8–19% for the CLT assay and 2–17% for the CLT(1–6) peptide assay. The lower limit of quantitation in dog plasma was estimated as 18 pg ml −1 for CLT and 14–20 pg ml −1 for the CLT(1–6) peptide. Diluted urine samples from dogs were directly injected into the HPLC and the immunoreactivities of the fractions were measured. More than 98% immunoreactivity was found at the retention time of CLT(1–6) peptide. Thus only the CLT(1–6) peptide was measured in dog urine by the RIA without HPLC separation. Urine samples could be assayed directly only after dilution 1:20. The assay recovery for urine was 100% and the precision (RSD) was estimated to be
ISSN:0731-7085
1873-264X
DOI:10.1016/0731-7085(94)90019-1