高效液相色谱法测定单氨基四苯基卟啉衍生物的含量

Content determination of mono amino tetraphenylporphyrin derivative by high performance liquid chromatography

目的建立单氨基四苯基卟啉衍生物(DTP)的含量测定方法,为阴离子卟啉光敏剂的质量控制提供依据。方法采用高效液相色谱法(HPLC)测定DTP样品的含量,色谱柱为Waters Symmetry~C_(18)(4. 6 mm×250 mm,5μm),流动相为乙腈-0. 2%三氟乙酸水溶液(50∶50,V/V),检测波长414 nm,柱温30℃,流速1. 0 ml/min,进样体积10μl。从专属性、精密度、重复性、稳定性、检测限、定量限、线性范围、系统适用性和加样回收率方面对该方法的可行性和科学性进行考察。结果 DTP在1. 95-250μg/ml范围内峰面积与浓度呈良好的线性关系(r=0. 999 9);精密度、重复性、稳定性实验的RSD分别为0. 28%,0. 64%,0. 63%;检测限和定量限分别为1. 5 ng和4. 5 ng;低、中、高浓度的平均加样回收率分别为98. 6%,99. 1%,98. 2%,RSD分别为0. 26%,0. 32%,0. 36%。结论本文建立的高效液相色谱方法具有良好的精密度、准确度、专属性和重复性,可用于DTP的含量测定及其有关物质检查。

Objective To establish a method for content determination of mono amino tetraphenylporphyrin derivative(DTP),and to provide evidence for quality control of anion porphyrin photosensitizers.Methods The HPLC method for content determination of DTP was performed using a Waters Symmetry C 18 column(4.6 mm×250 mm,5μm)with a mobile phase of acetonitrile-water(containing 0.2%trifluoroacetic acid,50∶50).The detection wavelength was 414 nm.The column temperature was 30℃.The flow rate was 1.0 ml/min,and the injection volume was 10μl.The feasibility and scientificity of this method were examined by the terms of specificity,precision,reproducibility,stability,detection limit,quantitative limit,linear range,system suitability and sample recovery.Results There was a good linear relationship between peak area and DTP concentration in the range of 1.95-250μg/ml(r=0.999 9).RSDs of precision,reproducibility and stability tests were 0.28%,0.64%and 0.63%,respectively.The detection limit and quantitation limit were 1.5 ng and 4.5 ng.The average recoveries were 98.6%,99.1%and 98.2%,respectively,and RSDs were 0.26%,0.32%and 0.36%,respectively.Conclusion This HPLC method can be used for the content determination and related substances inspection of DTP,with good precision,accuracy,specificity and reproducibility.