Determination of 2,4-dihydroxy Benzoic AcidConcentration of Soil Using HPLC Method
The concentration of 2,4-dihydroxy benzoic acid was determined based on the established High Performance Liquid Chromatography (HPLC) method and the amount of 2,4-dihydroxy benzoic acid was analyzed in 4 different land uses in the vertical distribution.The 2,4-dihydroxy benzoic acid in sample was extracted by 24 hours-water-bath heating with ethyl acetate,and then was evaporated and dissolved in chromatographic grade methanol.The sample elution was taken by the mobile phase which was composed of 3% glacial acetic acid and methanol (75∶25) with the flowing rate of 1 mL/min.The chromatographic column was Agilent ZORBAX SB-C18(4.6 mm×150 mm,5 μm) while the detecting wavelength was 280 nm and the column temperature was 25℃.The result showed that a good linear relationship existed between the concentration of 2,4-dihydroxy benzoic acid and the peak area in the range of 80~800 μg/mL.The obtained linear regression equation was Y=13.315X-12.803 (r2=0.999 9).The relative standard deviation (RSD) of repeatability,precision and stability of 2,4-dihydroxy benzoic acid were 0.778%,1.623% and 1.918% respectively.The average recovery rate was 97.675% and the RSD was 0.771% (n=6),all of which met the requirements of qualitative and quantitative analysis.The determination of 2,4-dihydroxy benzoic acid content using HPLC method was simple,rapid and accurate.The amount of the 2,4-dihydroxy benzoic acid in soil of natural secondary forest was found the greatest among 4 kinds of land.A uses and the concentration of 2,4-dihydroxy benzoic acid decreased with the depth of the soil.
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