CJC‐1295 is a 30 amino acid peptide‐based drug that stimulates the release of growth hormone (GH) from the pituitary gland. It is unique among performance‐enhancing peptides due to the presence of a reactive maleimidopropionic acid group that covalently links the peptide to free thiols on the surface of plasma proteins. Once conjugated, CJC‐1295 remains active in the bloodstream for significantly longer than non‐conjugated peptide‐based drugs that are rapidly excreted. Conjugation of CJC‐1295 to plasma proteins prevents its detection by top‐down mass‐spectrometry‐based peptide screening protocols as it effectively becomes a macromolecular protein with an undefined molecular weight. Using a pair of monoclonal antibodies raised against the CJC‐1295 peptide, we present an immuno‐polymerase chain reaction (I‐PCR) assay that is capable of detecting the CJC‐1295‐protein conjugate at concentrations down to 0.8 pg/mL. Detection of endogenous equine GHRH necessitated a screening threshold for CJC‐1295 in equine plasma of 50 pg/mL. The effectiveness of the assay for controlling the illicit use of CJC‐1295 was confirmed in equine blood samples after administration in thoroughbred race horses.
The administration of alkalinising agents including bicarbonate is of concern to racing authorities because resultant alkalosis may enhance performance and interfere with the detection of drugs in post‐race urine. A threshold for total carbon dioxide (TCO2) of 36.0 mmol/L in plasma (with action limit of 37.0 mmol/L) has been set. Serial dosing of sodium bicarbonate has gained popularity in human athletes but has not been studied in horses previously. Sodium bicarbonate (200 g per horse) and 60 g of an electrolyte‐vitamin complex was administered in 2‐L water via nasogastric intubation to five Standardbred horses for three consecutive days (total dose bicarbonate 0.42 ± 0.02 g/kg). Serial blood samples were taken over Days 1–5, with the final day (5) intended to simulate a ‘clear day’, and TCO2 was analysed. Following the first bicarbonate administration, plasma TCO2 peaked at 6 h (34.8 ± 1.3 mmol/L), returning to baseline by 23 h. On Day 2, four out of the five horses showed a peak greater than 36.0 mmol/L (mean 37.0 ± 2.1 mmol/L). With daily repeated dosing, plasma TCO2 peaked progressively earlier, and by Day 3, the peak occurred at 2 h and concentrations declined more rapidly. On Days 4 and 5, TCO2 levels remained low (<32.1 mmol/L on Day 4 and between 27.0–31.2 mmol/L on Day 5). These studies demonstrate that serial dosing of a ‘split dose’ of sodium bicarbonate on three consecutive days does not result in the accumulation or carry‐over of plasma TCO2 levels beyond the levels observed following a single dose.
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