Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, School of Chemistry and Chemical Engineering, Southwest University, 400715, Chongqing, People's Republic of China.
Luminescence. 2013 Sep-Oct;28(5):648-55. doi: 10.1002/bio.2410. Epub 2012 Sep 12.
In pH 4.0 Britton-Robinson buffer medium, PdCl2 was able to react with enzymes (EZ) such as lysozyme (LYSO) and papain (PAP) to form a coordination complex (EZ-PdCl2 ), which further reacted with MoO4 (2-) to form a ternary complex (MoO4 (2-) -EZ-PdCl2 ). As a result, the absorption and fluorescence spectra changed; new spectra of resonance Rayleigh scattering (RRS), second-order scattering (SOS) and frequency-doubling scattering (FDS) appeared and their intensities were enhanced greatly. The maximum RRS, SOS and FDS wavelengths of two ternary complexes were located at 310, 560 and 350 nm, respectively. The increments of scattering intensity were directly proportional to the concentrations of EZ within certain ranges. The detection limits (3σ) of LYSO and PAP were 4.5 and 14.0 ng/mL (RRS method), 9.6 and 57.8 ng/mL (SOS method), and 5.2 and 106.0 ng/mL (FDS method). Taking the MoO4 (2-) -LYSO-PdCl2 system, which was more sensitive, as an example, the effects of coexisting substances were evaluated. The methods showed excellent selectivity. Accordingly, new rapid, convenient, sensitive and selective scattering methods for the determination of LYSO and PAP were proposed and applied to determine LYSO in egg white with satisfactory results. The reaction mechanism and basis of the enhancement of scattering were discussed.
在 pH 4.0 的 Britton-Robinson 缓冲介质中,PdCl2 能够与酶(如溶菌酶(LYSO)和木瓜蛋白酶(PAP))反应形成配位配合物(EZ-PdCl2),该配合物进一步与 MoO4(2-)反应形成三元配合物(MoO4(2-)-EZ-PdCl2)。结果,吸收和荧光光谱发生变化;出现了新的共振瑞利散射(RRS)、二级散射(SOS)和倍频散射(FDS)光谱,其强度大大增强。两个三元配合物的最大 RRS、SOS 和 FDS 波长分别位于 310、560 和 350nm。散射强度的增量与一定范围内 EZ 的浓度成正比。LYSO 和 PAP 的检测限(3σ)分别为 4.5 和 14.0ng/mL(RRS 法)、9.6 和 57.8ng/mL(SOS 法)和 5.2 和 106.0ng/mL(FDS 法)。以更灵敏的 MoO4(2-)-LYSO-PdCl2 体系为例,评估了共存物质的影响。这些方法表现出优异的选择性。因此,提出了新的快速、方便、灵敏和选择性的散射方法来测定 LYSO 和 PAP,并将其应用于蛋清中 LYSO 的测定,结果令人满意。讨论了反应机理和散射增强的基础。