Derivatisierungsreaktionen im Bereich der quantitativen Dünnschicht-Chromatographie

Funk, W.

doi:10.1007/bf00528583

Cited by 22 publications

(7 citation statements)

References 24 publications

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“…The linear relationship of increasing amounts of cortisol and cortisone and INH-derived fluorescence is summarised in Figure 6. In agreement with previous work [9,10] fluorescence intensity of isonicotinyl hydrazones was significantly increased by dipping the chromatograms into paraffin/chloroform (1:2, v/v) and scanning them 10 min after the last dipping. The sensitivity was 12ng spot 1 for cortisol and 16ngspot 1 for cortisone (INH, without dipping into paraffin/chloroform) and 3 ng spot 1 for cortisol and 6 ng spot 1 for cortisone (INH, with dipping into paraffin/chloroform).…”

Section: Resultssupporting

confidence: 91%

Determination of cortisol and cortisone in human morning and overnight urine by thin-layer chromatography and fluorescence derivatisation with isonicotinic acid hydrazide

Fenske

2000

Chromatographia

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Key WardsThin-layer chromatography Cortisol Cortisone Isonicotinic acid hydrazide Human urine SummaryA routine method for the measurement of cortisol and cortisone in human morning and overnight urine has been developed. The method includes liquid-liquid extraction, alkaline wash of the solvent, thin-layer chromatography (TLC), formation of isonicotinyl hydrazones of A4-3 -keto-corticosteroids and their quantification by fluorescence densitometry. Cortisol excretion (mean values, n = 22) was 3918 ng h 1 (07.00-08.00), 1743 ng h 1 (08.00-09.00) and 24300 ng night 1; corresponding cortisone values were 2356 ng h 1, 2144 ng h 1 and 22780 ng night 1. The advantage of the method is its sensitivib,, being comparable with that of HPLC methods, its specifici b, (fluorescence of isonicotinyl hydrazones) and simplici b, of separation (TLC) and quantification (densitometry).

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Section: Resultssupporting

confidence: 91%

Determination of cortisol and cortisone in human morning and overnight urine by thin-layer chromatography and fluorescence derivatisation with isonicotinic acid hydrazide

Fenske

2000

Chromatographia

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“…Figure 3 shows that a time of 30 min was necessary for maximum fluorescence. This is approximately ten times longer than that used by others [7,8]. This difference cannot be explained for certain, but might arise because of the use of different acids -Funk and coworkers used trifluoroacetic acid whereas we used trichloroacetic acid.…”

Section: Discussionmentioning

confidence: 47%

“…Because of these limitations, other reagents have been used for detection of steroids, e.g. 2,4-dinitrophenylhydrazine [5,6,14], zirconyl chloride [15], dansyl hydrazine [7,16] or INH [4][5][6][7][8]. During initial tests we found that although the INH reagent described by Funk et al [8] gave the most encouraging results there were three obvious limitations: 1.…”

Section: Discussionmentioning

confidence: 97%

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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“…In most instances, subsequent heating is necessary. Postchromatographic derivatizations have been extensively reviewed (90)(91)(92). In 1990, a new method, overpressured derivatization, was reported (93); this can be done by pressing an absorbent polymeric pad (prewetted with derivatization reagent) on the TLC plate.…”

Section: A Conventional Detection Modesmentioning

confidence: 99%

Pharmaceuticals and Drugs

Nyiredy¹,

V√©gh²,

Szepesi³

et al. 2003

Handbook of Thin-Layer Chromatography

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Derivatisierungsreaktionen im Bereich der quantitativen Dünnschicht-Chromatographie

Cited by 22 publications

References 24 publications

Determination of cortisol and cortisone in human morning and overnight urine by thin-layer chromatography and fluorescence derivatisation with isonicotinic acid hydrazide

Determination of cortisol and cortisone in human morning and overnight urine by thin-layer chromatography and fluorescence derivatisation with isonicotinic acid hydrazide

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

Pharmaceuticals and Drugs

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