摘要
建立一种基于分光光度法和快速图像比色法双模式测定碱性磷酸酶的分析方法。以四甲基联苯胺(TMB)和邻菲啰啉(Phen)为显色探针,在650 nm处测定样品溶液的吸光度,以碱性磷酸酶浓度和吸光度绘制标准曲线。同时以氧化型TMB蓝色的褪去及Fe3+和Phen配合物橙红色的增强两种颜色变化对应的图像RGB变化值对碱性磷酸酶浓度绘制标准曲线。在最佳条件下,吸光度和颜色强度均与碱性磷酸酶浓度在5~100 U/L范围内呈现良好的线性关系,相关系数分别为0.97、0.992,检出限分别为0.35 U/L和5.5 U/L。吸收光谱法样品加标回收率为99.19%~102.72%,测定结果的相对标准偏差为1.31%~4.01%(n=5);图像快速比色法样品加标回收率为96.41%~105.2%,测定结果的相对标准偏差为1.81%~3.09%(n=5)。该方法满足碱性磷酸酶现场快速检测需要。
An analytical method for the determination of alkaline phosphatase(ALP)activity by absorbance and color intensity RGB values was established.Tetramethylbenzidine(TMB)and phenanthroline(Phen)were used as chromogenic probes,the absorbance of the sample solution was determined at 650 nm,and standard curves were drawn based on ALP concentration and absorbance.Simultaneously,the standard curve of ALP concentration was drawn based on the change of RGB values corresponding to the fading of oxTMB blue and the enhancement of orange red of Fe3+-Phen complexes.Under optimal conditions,both absorbance and color intensity had a good linear relationship with ALP concentration in the range of 5-100 U/L,with the correlation coefficients of 0.97 and 0.992,respectively.The detection limits were 0.35 U/L and 5.5 U/L,respectively.The recoveries of sample spiked by absorption spectroscopy were 99.19%-102.72%,and the relative standard deviation of the measurement results was 1.31%-4.01%(n=5).The recoveries of sample spiked by image rapid colorimetric method were 96.41%-105.2%,and the relative standard deviation of the measured results was 1.81%-3.09%(n=5).This method meets the needs of rapid field detection of ALP.
作者
赵灵芝
侯茹雪
刘永杰
杨国静
寇张文
赵柳
ZHAO Lingzhi;HOU Ruxue;LIU Yongjie;YANG Guojing;KOU Zhangwen;ZHAO Liu(Department of Pharmacy,Xi'an Medical University,Xi'an 710021,China;Shaanxi Agricultural Inspection and Testing Center,Xi'an 710003,China)
出处
《化学分析计量》
2025年第5期49-54,共6页
Chemical Analysis And Meterage
基金
2024年陕西省大学生创新项目(202411840003)
2023年陕西省大学生创新项目(S202311840059)
陕西省科技厅项目(2022JM-094,2024JC-YBMS-640,2023-JC-QN-0629)
2022年科研能力提升计划项目(2022NLTS072)
西安医学院2024年度大学生开发实验科研基金项目(2024DXS37,2024DXS38)。
