Abstract
Study on the stoichiometry and affinity of the arsenicals bound to HSA is an important step toward a better understanding of arsenic toxic effects. After incubation of AsIII or AsV with HSA at the physiological conditions (pH 7.43 and 37 °C), the free arsenicals and arsenic-HSA complexes were separated and detected by the combined techniques of microdialysis and liquid chromatography with hydride generation atomic fluorescence spectroscopy (MD–LC–HGAFS). The decrease of AsIII peak response rather than AsV indicated that HSA reacted with AsIII but not AsV. The binding plots indicated that the binding between HSA and AsIII was in Scatchard pattern when the concentration ratios of AsIII to HSA were ≤1:1. The strong binding sites (n 1) were 1.6 and the stability constant (K 1) was 1.54 × 106 M−1. When the concentration ratios of AsIII to HSA were >1:1, the binding was in Plasvento pattern with the stability constant K 2 ≅ 0 and no specific binding of AsIII with HSA. On the contrary, AsV did not show binding with HSA. The results showed that AsIII reacted with HSA more readily than AsV, which provides a chemical basis for arsenic toxicity.
References
Sun GF, Li X, Pi JB, Sun Y, Li B, Xu YY (2006) J Health Popul Nutr 24:176–181
Baastrup R, Sørensen M, Balstrøm T, Frederiksen K, Larsen CL, Tjønneland A, Overvad K, Raaschou-Nielsen O (2008) Environ Health Perspect 116:231–237
Smith AH, Smith MMH (2004) Toxicology 198:39–44
Gailer J, Madden S, Buttigieg GA, Denton MB, Younis HS (2002) Appl Organometal Chem 16:72–75
Ngu TT, Stillman MJ (2006) J Am Chem Soc 128:12473–12483
Wang ZW, Zhang HQ, Li XF, Le XC (2007) Rapid Commun Mass Spectrom 21:3658–3666
Suwalsky M, Rivera C, Villena F, Sotomayor CP, Malgorzato JR, Strzalka K (2007) Biophys Chem 127:28–35
Kitchin KT, Wallace K (2008) J Inorg Biochem 102:532–539
Jiang DF, Gong ZL, Li XF, Cullen WR, Le XC (2003) Chem Res Toxicol 16:873–880
Tsai TH (2003) J Chromatogr B 797:161–173
Wen XD, Qi LW, Chen J, Song Y, Yi L, Yang XW, Li P (2007) J Chromatogr B 852:598–604
Lange CME, Boer AG, Breimer DD (2000) Adv Drug Deliv Rev 45:125–148
Traunmuller F, Steiner I, Zeitlinger M, Joukhadar C (2006) J Chromatogr B 843:142–146
Elmquist FW, Sawchuk RJ (2000) Adv Drug Deliv Rev 45:123–124
Jiang H, Ding JH, Zhang YH, Gong HZ, Gao S, Sun GF (2008) Gaodeng Xuexiao Huaxue Xuebao 29:488–492
Jiang H, Ding JH, Gao S, Gong HZ, Sun GF (2008) Huaxue Yanjiu 19:91–93
Shen XC, Liang H, Guo JH, Song C, He XW, Yuan YZ (2003) J Inorg Biochem 95:124–130
Pesavento M, Profumo A (1991) Talanta 38:1099–1106
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This research was financially supported by the Key Fund (30530640) from the national Science Foundation of China.
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Jiang, H., Ding, J., Chang, P. et al. Determination of the Interaction of Arsenic and Human Serum Albumin by Online Microdialysis Coupled to LC with Hydride Generation Atomic Fluorescence Spectroscopy. Chroma 71, 1075–1079 (2010). https://doi.org/10.1365/s10337-010-1573-2
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DOI: https://doi.org/10.1365/s10337-010-1573-2