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Investigation of Complex Formation PDTC with Ni(II) in Physiological Conditions 336 The absorbance maxima at 324 nm reflect nickel binding at the two binding sites of PDTC. The spectrum reflects that the maximum absorbance at about 324 nm increased rapidly, until the mol ratio 1 : 2 metal/ligand reached. Figure 3: Spectrum of PDTC at 10 -6 M. In titrations of PDTC with Ni 2+ alone, there was an isosbestic point at 296 nm (fig.4). The binding constant of metal to ligand was determined by using the continuous variation method (Job's method) at 324 nm (the obtained maximum absorbance from fig.4). The corrected absorbance was plotted against the volume of fraction of PDTC, that is, VM/(VM + VL), where VM and VL are the volumes of Ni 2+ and PDTC solutions, respectively (Fig. 5). The maximum values of VM/(VM + VL) = 0.33 and VL/(VM + VL) = 0.66 obtained from figure 5, thus VL/VM is 2, which suggested that the complex had the formula Ni(PDTC)2. It is clear that nickel is binding to both specific metal-binding sites of PDTC. The binding of Ni to PDTC can be quite fast. If the nickel is added as a NiCl2, then the binding takes many seconds to reach completion. The effective Ni-PDTC binding constant is logk=11.1±0.1, this is clearly higher than effective Ni-Tf binding constants (logk1=4.1±0.15, logk2=3.23±0.31) [22].
337 S. J. Fatemi, A. Badiei and A. Hooshmand Figure 4: A series of difference spectra for Ni 2+ binding to PDTC in 10mM hepes buffer, 5mM sodium bicarbonate, pH 7.4, and 25°C [PDTC] ~ 10 -6 M. Figure 5: Plot of corrected absorbance against VM/(VM + VL) for determination of molar ratio of Ni 2+ to PDTC. [PDTC] = 10 -5 M, [Ni 2+ ] = 10 -5 M, pH 7.4, and 25°C. Conclusion Complex formation of nickel(II) ion with PDTC, was studied using UV-vis spectroscopy. PDTC can be an oral chelator with pKa= 3.300±0.002. It behaves as a bidentate ligand at serum pH. The effect of pH on Ni 2+ -PDTC indicated that the maximum adsorption was at pH=7.4. The effective Ni-PDTC binding constant was determined (logk=11.1±0.1) for the 1:2 Ni(PDTC)2 complex by job's method using UV-vis spectra.
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