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Identification and verification of the anabolic steroid boldenone in equine blood and urine by HPLC/ELIS
Author(s) -
Hagedorn HeinzWerner,
Schulz Rüdiger,
Jaeschke Gerhard
Publication year - 1994
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130080204
Subject(s) - chemistry , chromatography , anabolic steroid , urine , anabolism , steroid , high performance liquid chromatography , biochemistry , hormone
An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone‐17‐hemisuccinate–bovine serum albumin as antigen. Boldenone‐17‐hemisuccinate–horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 ± 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross‐reactivities, 3.4 and 2.5%, respectively. These crossreactivities are of no importance for the boldenone assay. For the reduction of background levels, srceening for boldenone of equine serum was performed after extraction. Urine samples were determined directly after dilution, omitting hydrolysis of boldenone conjugates. Positive screening results were confirmed by means of two independent HPLC systems combined with off‐line detection, employing the boldenone ELISA. Methandienone served as internal standard to ascertain retention factors. In horses treated with boldenone‐17‐undecylenate the presence of boldenone in serum was confirmed up to 28 days and in unhydrolysed urine up to 56 days post applicationem .