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目的确立一种快速、准确检测水溶性卟啉(TPPS4)的分析方法,从而进一步了解水溶性卟啉和牛血清白蛋白(BSA)之间相互作用的机制。方法采用电化学法对TPPS4的极谱伏安行为进行了研究,同时采用电化学法、荧光法和紫外法对TPPS4与BSA之间的相互作用分别进行了研究。3种方法相互辅证使得试验结果更加可靠。结果在底液NaH2PO4-Na2HPO4缓冲液(pH 7.18)中,TPPS4在-0.70 V(vsSCE)处有一个稳定而灵敏的还原峰,其峰电流与TPPS4浓度在1.0×10-7~1.0×10-5mol.L-1有良好的线形关系(r2=0.998 3,0.999 3),检测限LOD为3.0×10-8mol.L-1。平均标准回收率为99.59%,精密度较好,RSD为0.56%(n=5)。在NH4Cl-NH3.H2O缓冲液(pH9.05)中,实验结果表明BSA与TPPS4相互作用生成1∶1的TPPS4-BSA超分子体系。另外,加入环糊精体系后,磺丁醚-β-CD(SBE-β-CD)和羟丙基-β-CD(HP-β-CD)均能促进TPPS4与BSA发生反应。结论建立了一种简单、快速、准确的水溶性卟啉四-(4-磺基苯)卟啉(TPPS4)的电化学分析方法。卟啉类药物被环糊精包合后更容易与人体内的蛋白质进行作用,环糊精在卟啉类药物的控制、释放中具有重要的作用和意义。
OBJECTIVE: To establish a rapid and accurate method for the determination of water-soluble porphyrins (TPPS4) to further understand the mechanism of the interaction between water-soluble porphyrins and bovine serum albumin (BSA). Methods The polarographic voltammetric behavior of TPPS4 was studied by electrochemical method. The interaction between TPPS4 and BSA was studied by electrochemical method, fluorescence method and UV method. Three kinds of methods complement each other to make the test results more reliable. Results TPPS4 showed a stable and sensitive reduction peak at -0.70 V (vsSCE) in the buffer solution of NaH2PO4-Na2HPO4 (pH 7.18). The peak current of TPPS4 was in the range of 1.0 × 10-7 ~ 1.0 × 10- 5mol.L-1 has a good linear relationship (r2 = 0.998 3,0.999 3), the detection limit of LOD is 3.0 × 10-8mol.L-1. The average standard recovery was 99.59% with better precision with a RSD of 0.56% (n = 5). In NH4Cl-NH3.H2O buffer (pH9.05), the experimental results show that BSA interacts with TPPS4 to generate a 1: 1 TPPS4-BSA supramolecular system. In addition, both SBE-β-CD (SBE-β-CD) and HP-β-CD (HP-β-CD) promoted the reaction of TPPS4 with BSA. Conclusion A simple, rapid and accurate method for the electrochemical analysis of water-soluble porphyrin tetrakis (4-sulfophenyl) porphyrin (TPPS4) was established. Porphyrin drugs cyclodextrin inclusion easier with the human body after the role of protein, cyclodextrins in porphyrin drug control, release has an important role and significance.