A comparison of soil adenine nucleotide measurements by hplc and enzymatic analysis

  • Martens R
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A high pressure liquid chromatography (HPLC) method was developed to measure the soil adenine nucleotides ATP, ADP and AMP. This method was compared with conventional measurement of these nucleotides by enzymatic conversion of ADP and AMP to ATP and final measurement of ATP by the luciferin-luciferase enzyme system. Sufficiently sensitive measurement of these nucleotides with the HPLC system was achieved by chemical derivatization with chloracetaldehyde, which formed highly fluorescent 1-N6-ethenoadenosine nucleotides. The efficiency of this reaction was 100% for ATP and AMP and 93% for ADP. The nucleotides can be quantified in the low picomole range using this derivatization. The concentrations of adenine nucleotides in four arable soils were measured following extraction with two different reagents: an acid H2SO4-based reagent and an alkaline NaHCO3-based reagent. The difference between amounts of ATP, ADP measured by the HPLC and the enzymatic conversion technique was mostly

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  • R. Martens

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