对硝基酚荷移分光光度法测定组氨酸含量的研究

Determination of histidine based on charge transfer reaction with p-nitrophenol by spectrophotometry

目的建立测定组氨酸含量的对硝基酚荷移分光光度法。方法组氨酸与对硝基酚在室温下可生成稳定1∶1型的荷移络合物。荷移络合物的最大吸收波长λmax=400 nm,表观摩尔吸光系数ε=1.0×104 L/(mol.cm)。结果组氨酸浓度在1-15μg/ml的范围内遵循比尔定律,回收率在93.3%-99.9%之间,相对标准偏差在2.5%以内。结论本方法简便、稳定性好,其测定组氨酸样品的含量结果与文献方法一致。

Objective To develop a spectrophotometric method for determining the content of histidine based on a charge transfer reaction with p-nitrophenol. Methods A stable charge transfer complex of histidine with p-nitrophenol was formed at room temperature.The maximum absorption was at 400 nm.The apparent molar absorptivity was determined to be 1.0×104 L/（mol·cm）. Results The calibration curve was linear in the range of 1-15 μg/ml.The recovery was 93.3%-99.9%.The relative standard deviation was less than 2.5%. Conclusion The method is simple and repeatable,and the result is consistent with the literature for determining the content of histidine.