The DPRA is an in chemico method which quantifies the remaining concentration of Cysteine- or Lysine-containing peptide following 24 hours incubation with the test chemical at room temperature. The peptide is a custom material containing phenylalanine to aid in the detection. Relative peptide concentration is measured by high-performance liquid chromatography (HPLC) with gradient elution and UV detection at 220 nm. Cysteine- and Lysine peptide Percent Depletion Values are then calculated and used in a prediction model (see paragraph 27) which allows assigning the substance to one of four reactivity classes used to discriminate between sensitisers and non-sensitisers.
The peptide depletion (sign of protein reactivity) is measured by HPLC UV. The induction of allergic contact dermatitis typically involves an initial exposure of the chemical or hapten to the skin and then binding of the chemical to a protein carrier in a process known as haptenisation.

DPRA results generated during ECVAM validation and in other published studies suggest accuracy of DPRA in discriminating between non-sensitisers and sensitisers is 80% with a sensitivity of 80% and a specificity of 77% when compared with the local lymph node assay (LLNA).

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