【摘要】 目的 评估动力学直接多肽反应试验(kinetic direct peptide reactivity assay,k DPRA)对化学物皮肤致敏性的鉴别能力。方法 选用已知皮肤致敏强度的31种化学物，将化学物用磷酸盐缓冲液或乙腈配制成1.25、2.5、5.0、10.0和20.0mmol/L溶液，分别取40μl上述溶液与120μl的0.667 mmol/L半胱氨酸肽混合，同时设空白对照、溶剂对照、测试物对照和阳性对照（肉桂醛），在25℃条件下孵育10、30、90、150、210和1 440 min，作用相应时间后，每孔加入40μl 3mmol/L的单溴二胺作用5 min，在酶标仪上以390 nm的激发光和480 nm的发射光测定荧光强度；计算半胱氨酸肽的消耗率和速率常数，取6个时间点中最大对数速率常数(log kmax)作为评价指标，评价各种化学物的皮肤致敏性。结果应用2,4-二硝基氯苯的3次测试获得log kmax分别为-0.39、-0.28、-0.31，结果均判定为1A类致敏物；对10种化学物的3次重复测试均能获得一致的致敏性分类；扩展对20种化学物测试的结果，6种化学物判为1A类，14种判为非1A类，所测31种化学物皮肤致敏性分类结果均与欧洲化学品管理局(European Chemicals Agency,ECHA)数据库一致。结论该方法对化学物皮肤致敏性具有较好的鉴别能力。更多还原

【Abstract】 Objective To evaluate the discriminative ability of the Kinetic Direct Peptide Reactivity Assay(kDPRA) for identifying the skin sensitization potential of chemicals. Methods 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 0.667mmol/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.Results 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.Conclusion This method demonstrates good discriminative ability for assessing the skin sensitization potential of chemicals.更多还原