Paired Ion Reversed-Phase HPLC Assay for the Simultaneous Determination of Iothalamic Acid and Para Aminohippuric Acid in Plasma

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Abstract
An ion paired reversed—phase high performance liquid chromatographic assay for iothalamic acid and para aminohippuric acid in the same sample of plasma is described. The analysis uses one internal standard for both drugs. Sample preparation consists of precipitating plasma proteins with methanol and centrifuging to settle the proteins. The supernatant is evaporated and the residue reconstituted with mobile phase for injection. For HPLC a C8 column and a mobile phase consisting of potassium phosphate buffer with dodecyl triethyl ammonium phosphate IP reagent, 22.5% methanol and 2.5% acetonitrile with UV detection at 254 nm was used. Coefficients of variation for the assay were in the range of 1.6–12.1% for iothalamic acid and 4.3–17.7% for para aminohippuric acid for four levels of concentration. Limits of quantitation were 3.0 μg/mL for iothalamic acid and 5.0 μg/mL for para aminohippuric acid. This isocratic HPLC assay is simple, rapid and relatively inexpensive.

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