Determination of urinary cortisol:creatinine ratios by sequential thin layer chromatography and ELISA in dogs

Sohn, D.H.; Oh, T.H.; Han, H.R. . College of Veterinary Medicine; Na, K.J. . College of Veterinary Medicine; Lee, H.S. .

Determination of urinary cortisol:creatinine ratios by sequential thin layer chromatography and ELISA in dogs

1999

Sohn, D.H. | Oh, T.H. | Han, H.R. (Seoul National University, Suwon (Korea Republic). College of Veterinary Medicine) | Na, K.J. (Chung National University, Cheongju (Korea Republic). College of Veterinary Medicine) | Lee, H.S. (National Veterinary Research & Quarantine Service, Anyang (Korea Republic).)

This study was conducted to evaluate the ELISA kit for measuring the level of cortisol in the urine. The CV of within-run variation and day to day variation were 0.4-2.8 and 1.8-5.7, respectively. the minimum limitation of measurement was 1ng/ml. the cross reaction was high (CR50(%)=11.4-43.2) in prednisolone, 11-deoxycortisol, 21-deoxycortisol and predinosone. There was low and no cross reaction in other steroid. To develop the ELISA kit we measured the cortisol level in diluted urine with PBS (procedure I), extracted urine with methylene chloride (procedure II) and extracted methylene chloride-extracted urine from thin-layer chromatography (procedure III). The CV value of procedure I, II, III was 9.4-28.3%, 7.2-8.9% and 2.5-5.7%, respectively. There was significant difference between procedure I with II, and pro-cedure I with III(p0.01), but no difference between procedure II with III significantly(p0.01). The mean UCCR of urine collected through am 8 to 10 ws 9.5+_7.6(0.14-28.0) in 12-month-old dog(n

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