Results for Practical 2 A.G.

Hodges 10170757

Analysis of Hydrocortisone in Horse Urine

Figure.1 Calibration graph for horse urine with known concentrations of hydrocortisone

Sample Urine A Urine A Urine A Urine A Average Urine B Urine B Urine B Urine B Average

Retention Time (min) 5.848 5.837 5.831 5.810 5.832 5.820 5.818 5.830 5.831 5.825

Peak Area 10747 14331 23300 31271 19912.250 206994 213282 179524 189222 197255.500

ctions from the Table 1 . Retention times and peak areas obtained from 8 samples of two types of horse %. urine

Sample A B

Retention Time (min) 5.832 5.825

Peak Area 19912.3 197255

[Cortisol] in MeOH (mg/l) 0.861 8.528

[Cortisol] in 1ml of urine (mg/ml) 0.10mg/ml 0.967 mg/ml

Table 2 . Average retention times, peak areas and cortisol concentrations obtained from Samples A and B.

Results for Practical 2 A.G.Hodges 10170757

The threshold for cortisol levels in race horses according to the ARCI ( Association of Racing Commisioners International) currently stands at1mg/ml. In the case of this investigation, the sample taken for Horse A falls well beneath this threshold (0.10mg/ml). In the case of Horse B, urine levels of cortisol are measured at 0.967 mg/ml, although strictly speaking this is below the threshold of 1mg/ml, Horse B will be classified as the threshold limit for urine cortisol levels and would be disallowed to compete in any events. However, a degree of error may slightly hinder the accuracy of the results, as measurements were not calculated in a 1:1 ratio with keratenin levels which are at constant based on muscle mass, as this was not measured and the ratio applied accordingly, it cannot be disregarded that other physiological stressors may have induced such high cortisol levels.