Improved spray reagent for thin layer chromatographic method for detecting uric acid: collaborative study

A collaborative study was conducted to validate the substitution of an improved single spray in the official AOAC thin layer chromatographic method for identifying uric acid (UA) from bird and insect excreta. The proposed reagent, which is a dilute aqueous solution of ferric chloride and potassium ferricyanide, requires neither a heating step nor a pH indicator. Its preparation time, specificity, and sensitivity to low levels (5-50 ng) of UA were compared with those of the official sprays. The improved spray took 1/5 as long to prepare as the official sprays. Neither the proposed spray nor the official sprays gave false positive reactions with compounds similar to UA. For bird and insect excreta samples, at the 95% confidence limits, the false negative rate was between 0 and 9.7% for the proposed spray and between 0.7 and 18.7% for the official sprays. Sensitivity results showed that the proportion positive for the proposed spray was significantly higher (P less than 0.05) than for the official sprays at the 15 ng UA level. The proposed changes have been adopted official first action.