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Contribution to the quantification of humic acids by thin layer chromatography
Authors:Ahmed H Khairy
Abstract:Increasing amounts from 2.5 to 45 μg of one synthetic and seven natural humic acids were chromatographed on silica gel thin layers using ammonia 25% and propanol in the ratio 70:30 v/v as mobile phase. The fragments of humic acids produced under the influence of the alkaline mobile phase were separated in three band-shaped chromatographic fractions, whose lengths were transverse to the direction of the mobile phase. Since the fragments were not formed in constant ratios from different quantities of the same humic acid, the length of a fraction could only reflect its content of humic substance, whereas the sum of lengths of the three fractions (F) reflected quantities of the humic acid (x) at starting points. Similar F-x curves were obtained from humic acids extracted from similar origins, even when the locations of the samples, particle weights of the humic acids or the pH values at which they were extracted were different. The mathematical relation between F and x for quantities of humic acids up to 15–20 μg possessed high correlation coefficients and may be thus applied for the estimation of unknown concentrations of humic acids extracted from similar origins.
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