A high sensitivity, low volume HPLC method to determine soluble reactive phosphate in freshwater and saltwater

A reverse phase High Performance Liquid Chromatography (HPLC) method was developed to detect soluble reactive phosphate (SRP) in marine and freshwater samples. This technique is scalable over a range of sample volumes from 10 μl to 10 ml, requires minimal sample preparation, and responds linearly from 1 nM to 100 μM (R2>0.964). Standard Murphy–Riley molybdenum blue reagents were used to react with phosphate in samples for analysis. Solid phase extraction tubes were used to clean the mixed complexing reagent prior to combining the reagent with samples. The method was applied to freshwater samples collected in July 2001 along an Andean mountain stream located in the Eastern Cordillera ( 2400 m) of Peru and marine samples collected in January 2002 within the low marsh of the estuarine Nueces Delta System, Texas Coastal Bend. For method comparison, field samples were analyzed using a standard manual spectrophotometric method. Field sample phosphate concentrations produced from each method were nearly identical. This method offers the advantages of low sample volume (1 ml or less) with high sensitivity, precision, and potential automation.