Doss: Thin-layer chromatography of porphyrin methyl esters
نویسنده
چکیده
Thin-layer chromatography methods on the preparative and analytical scale are presented for the separation of porphyrin methyl esters according to the number of ester groups in each porphyrin. Complete and sharp separations are obtained on silica gel glass plates or aluminium cards run in benzene-ethyl acetate-methanol solvent systems. Dicarboxylic porphyrin esters from proto-, deuteroand hematoporphyrin IX have also been separated. Modifications of the analytical methods are described for the rapid separation of porphyrins bearing eight to two methyl ester groups within 10 min on small chromatocards in chloroform-ethyl acetate solvent mixtures. The porphyrins are determined quantitatively by spectrophotometric analysis of the eluted substances, or by fluorescence scanning of the bands of thin-layer chromatograms, using the Camag-Turner TLC-scanner. The fluorometric method is highly sensitive and permits a simultaneous relative measurement of all bands in the chromatogram. Examples of thin-layer chromatograms in combination with fluorescence scanning are presented, demonstrating the application of this new analytical method.
منابع مشابه
InhibitorMethodfor the Determination of Pancreatic Isoenzymes
1. Gray CH, Urn CK, Nicholson DC. The differentiation of the porphyrins by means of high pressure liquid chromatography. Clin Chim Acta 77, 167-178 (1977). 2. Doss M. Analytical and preparative thinlayer chromatography of porphyrin methyl esters. Z Kiln Chem Kiln Biochem 8, 197207 (1970). 3. Straka JG, Kushner, JP, Burnham BF. High-performance liquid chromatography of porphyrin esters. Identifi...
متن کاملRapid procedure for fecal porphyrin assay.
Hydrochloric acid extraction of feces in the presence of ether yields an extract suitable for spectrophotometric estimation of total porphyrin and for further separation by "high-performance" liquid chromatography (HPLC) or thin-layer chromatography. A total porphyrin reference interval of less than 200 nmol/g dry weight of feces was established from data on 106 normal subjects on an unrestrict...
متن کاملThe pattern of porphyrin isomer accumulation and excretion in symptomatic porphyria.
1 . Thin-layer chromatography of porphyrin methyl esters provides a useful and rapid technique for resolving mixtures of porphyrins from biological samples. 2. Application of this technique to urinary, faecal and hepatic porphyrins from patients with symptomatic porphyria revealed five porphyrins of interest which could be identified by mass spectrometry as the methyl esters of porphyrins with ...
متن کاملRapid semiquantitative measurement of total porphyrins in urine and feces by magnetic circular dichroism.
Magnetic circular dichroism is a suitable technique for semiquantitative determination of total porphyrins in urine and feces, being rapid, reliable, and involving little sample preparation. The sensitivity of the assay suffices to distinguish normal from above-normal concentrations of total porphyrins in urine and feces. Results for urine compare well with those obtained by thin-layer chromato...
متن کاملRapid determination of urinary total porphyrins by ion exchange chromatography.
Total urinary porphyrins were eluted from a commercially available anion exchange Chromatographie column with hydrochloric acid and determined spectrophotometrically. For 1 ml samples the optimum concentration range for measurement was found to lie between 0.1 and 1.5 mg of total porphyrin/1 urine. Assay of a single sample takes 12 min. The specificity (measurement at three wave lengths), sensi...
متن کامل