Determination of cortisol in guinea pig urine by high performance thin-layer chromatography, high performance liquid chromatography, and thin-layer chromatography/radioimmunoassay

Fenske, M.

doi:10.1007/bf02466515

Cited by 10 publications

(4 citation statements)

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“…Similarly, the excretion of cortisol was augmented in ACTH-or cortisol-treated animals (saline compared with ACTH, P < 0.02; ACTH compared with cortisol, P < 0.01). When the samples were measured by TLC-radioimmunoassay [9], similar results were obtained ( Figure 7). Comparison of cortisol concentrations measured by the INH method and by the TLC-radioimmunoassay technique resulted in correlation coefficients, r, of 0.98 and 0.97 for plasma and urine, respectively.…”

Section: Figurementioning

confidence: 51%

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Determination of cortisol in plasma and urine by thin-layer chromatography and fluorescence derivatization with isonicotinic acid hydrazide

Fenske

1998

Chromatographia

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Key WordsThin-layer chromatography Cortisol Isonicotinic acid hydrazide Plasma and urine SummaryA simple, inexpensive and reliable method is described for the measurement of cortisol in guinea-pig plasma and urine by thin-layer chromatography (TLC) and densitometry. Plasma and urine were extracted with dichloromethane and the extracts chromatographed on aluminium-backed silica gel TLC plates with concentrating zones. The plates were then dipped into the isonicotinic acid hydrazide (INH) reagent (3 g INH, 5 g trichloroacetic acid, 300 mL ethanol). The fluorescence of the cortisol hydrazone was further increased by dipping the plates into chloroform-liquid paraffin (9:1, v/v). The fluorescence was determined densitometrically (excitation: 366 nm, cut-off filter: >460 nm). The fluorescence intensity was linearly dependent on the amount of cortisol between i ng and 200 ng; the coefficients of variation ranged between 6.3% (1 ng) and 1.4% (200 ng).The sensitivity of this method (< 1 ng) enables the measurement of cortisol in the plasma or urine of saline-, ACTH-or cortisol-treated guinea-pigs. Comparison of the cortisol concentration measured by this method or by a TLCradioimmunoassay technique gave correlation coefficients of r = 0.98 (plasma) and r = 0.97 (urine). Thus, in conjunction with the isolation of cortisol by TLC, the densitometric measurement of the fluorescent hydrazone of cortisol is a sensitive and rapid routine means of determination of the concentration of cortisol in biological materials such as plasma or urine.

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Section: Figurementioning

confidence: 51%

“…Aliquots (0.5 mL, duplicates) of each wash or cortisol standards (20-200 ng/0.5 mL of acid-washed shavings) were placed on kieselguhr-fllled minicolumns [9]. Cortisol was extracted with dichloromethane (3 mL); after evaporation of the solvent the dry residue was dissolved in ethanol (0.2 mL).…”

Section: Extraction Of Plasma or Urine Samples Cortisol Measurementmentioning

confidence: 99%