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To evaluate the discriminative ability of the Kinetic Direct Peptide Reactivity Assay (kDPRA) for identifying the skin sensitization potential of chemicals.
A total of 31 chemicals with known skin sensitization strengths were selected.These chemicals were prepared in phosphate-buffered saline or acetonitrile to create solutions at concentrations of 1.25, 2.5, 5.0,10.0, and 20.0 mmol/L. A mixture of 40 μl of the above solutions with 120 μl of a 0.667 mmol/L cysteine peptide was prepared, alongside blank controls, solvent controls, test substance controls, and a positive control (cinnamaldehyde). The mixtures were incubated at 25°C for 10, 30, 90, 150, 210, and 1 440 minutes. After the respective incubation times, 40 μl of 3 mmol/L monobromination was added for 5 minutes, and fluorescence intensity was measured using an enzyme-linked immunosorbent assay (ELISA) reader with excitation at 390 nm and emission at 480 nm. The consumption rate and rate constant of the cysteine peptide were calculated, with the maximum logarithmic rate constant (log kmax) from the six time points serving as the evaluation metric for the skin sensitization potential of various chemicals.
Testing 2,4-dinitrochlorobenzene yielded log kmax values of -0.39,-0.28, and -0.31 across three tests, all classified as Category 1A sensitizers. Consistent sensitization classifications were obtained from three repeated tests on 10 chemicals. Expanding to 20 additional chemicals, 6 were classified as Category 1A and 14 as non-1A. The skin sensitization classifications for all 31 tested chemicals were consistent with the European Chemicals Agency (ECHA) database.
This method demonstrates good discriminative ability for assessing the skin sensitization potential of chemicals.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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