Ramezani Amir M, Manzoori Jamshid L, Amjadi Mohammad, Jouyban Abolghasem
Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran.
ScientificWorldJournal. 2012;2012:940541. doi: 10.1100/2012/940541. Epub 2012 May 2.
A spectrofluorimetric method is proposed for the determination of human serum albumin (HSA) and bovine serum albumin (BSA) using terbium-danofloxacin (Tb(3+)-Dano) as a fluorescent probe. These proteins remarkably enhance the fluorescence intensity of the Tb(3+)-Dano complex at 545 nm, and the enhanced fluorescence intensity of Tb(3+)-Dano is proportional to the concentration of proteins (HSA and BSA). Optimum conditions for the determination of HSA were investigated and found that the maximum response was observed at: pH = 7.8, [Tb(3+)] = 8.5 × 10(-5) mol L(-1), [Dano] = 1.5 × 10(-4) mol L(-1). The calibration graphs for standard solutions of BSA, HSA, and plasma samples of HSA were linear in the range of 0.2 × 10(-6) - 1.3 × 10(-6) mol L(-1), 0.2 × 10(-6) - 1.4 × 10(-6) mol L(-1), and 0.2 × 10(-6) - 1 × 10(-6) mol L(-1), respectively. The detection limits (S/N = 3) for BSA, HSA, and plasma sample of HSA were 8.7 × 10(-8) mol L(-1), 6.2 × 10(-8) mol L(-1), and 8.1 × 10(-8) mol L(-1), respectively. The applicability of the method was checked using a number of real biological plasma samples and was compared with the UV spectrometric reference method. The results was showed that the method could be regarded as a simple, practical, and sensitive alternative method for determination of albumin in biological samples.
提出了一种以铽-达氟沙星（Tb(3+)-Dano）作为荧光探针测定人血清白蛋白（HSA）和牛血清白蛋白（BSA）的荧光光谱法。这些蛋白质显著增强了Tb(3+)-Dano络合物在545nm处的荧光强度，且Tb(3+)-Dano增强的荧光强度与蛋白质（HSA和BSA）的浓度成正比。研究了测定HSA的最佳条件，发现在pH = 7.8、[Tb(3+)] = 8.5×10(-5) mol L(-1)、[Dano] = 1.5×10(-4) mol L(-1)时观察到最大响应。BSA标准溶液、HSA标准溶液以及HSA血浆样品的校准曲线分别在0.2×10(-6) - 1.3×10(-6) mol L(-1)、0.2×10(-6) - 1.4×10(-6) mol L(-1)和0.2×10(-6) - 1×10(-6) mol L(-1)范围内呈线性。BSA、HSA以及HSA血浆样品的检测限（S/N = 3）分别为8.7×10(-8) mol L(-1)、6.2×10(-8) mol L(-1)和8.1×10(-8) mol L(-1)。使用多个实际生物血浆样品检验了该方法的适用性，并与紫外光谱参考方法进行了比较。结果表明，该方法可被视为一种测定生物样品中白蛋白的简单、实用且灵敏的替代方法。
