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A method was established to measure the concentrations of dissolved reactive phosphate (DRP) and dissolved ferrous iron (Fe) in micro-volume solution samples through colorimetric determination in large batch using a 384-well Microplate Spectrophotometer. Concentrations of DRP and dissolved Fe were determined by the molybdenum blue and phenanthroline colorimetric methods, respectively. The results showed that the sample consumption used for each parameter was between 20 and 50 microL after dilution, and the detection limits for DRP and dissolved Fe were 0.006 mg x L(-1) and 0.010 mg x L(-1) respectively, while the analytical precision varied between 1% and 5%. The established method was applied to measure DRP and dissolved Fe in pore waters of sediment profiles in Lake Taihu, which were collected by a high-resolution Peeper (HR-Peeper) device with a vertical resolution of 2 mm. The results showed a simultaneous increase of DRP and dissolved Fe in their concentrations with the depth of two sediment profiles investigated.