Radioimmunoassay for dexamethasone 17,21-dipropionate and its metabolites in plasma and urine after topical application

Add time:08/19/2019    Source:sciencedirect.com

A sensitive radioimmunoassay for dexamethasone 17,21-dipropionate and its four metabolites in human plasma and urine has been developed using single anti-dexamethasone antiserum. The antiserum was obtained by immunizing rabbits with dexamethasone-3-oxime-bovine serum albumin conjugate. All of the endogenous steroids tested cross-reacted less than 0.07%. Before radioimmunoassay, dexamethasone 17,21-dipropionate and dexamethasone 17-propionate were hydrolyzed to dexamethasone, and 6β-OH-dexamethasone 17-propionate was hydrolyzed to 6β-OH-dexamethasone in 3% ammoniamethanol at 5 C for 16 h. A standard curve was established with a useful range between 0.005 and 2 ng in the case of dexamethasone, between 0.05 and 5 ng in the case of 6β-OH-dexamethasone. Measurement of plasma concentrations and percent urinary excretion of the metabolites in healthy men was performed following occlusive dressing of dexamethasone 17,21-dipropionate cream and ointment. The main metabolites in plasma were dexamethasone 17-propionate and dexamethasone, which increased gradually and reached maximum levels (160–200 pg/mL) at 24–32 h after application. The major metabolites observed in urine were 6β-OH-dexamethasone 17-propionate and 6β-OH-dexamethasone. Total percentage of their urinary excretions within 72 h after application amounted to 0.28–0.50% of the dose administered.

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