茜素红-镧-牛血清白蛋白体系的共振瑞利散射及共振非线性散射光谱研究及应用

Investigation and application of alizarin red S-La3^+-BSA system by resonance rayleigh scattering and resonance non-linear scattering

在pH 4.5的HAc-NaAc缓冲液中,茜素红(ARS)-镧与牛血清白蛋白(BSA)形成三元离子缔合物,导致共振瑞利散射(RRS)、二级散射(SOS)和倍频散射(FDS)显著增强,光谱最大散射波长分别位于373 nm、620 nm和310 nm。体系的光散射强度与BSA浓度在一定范围内呈线性增强,线性范围分别是RRS在0.034～13.40μg/mL、SOS在0.201～13.40μg/mL和FDS在0.201～10.05μg/mL范围内,对于BSA的检出限分别为0.010μg/mL(RRS法)0、.052μg/mL(SOS法)和0.089μg/mL(FDS法),据此建立了灵敏的测定BSA的共振线性(RRS)和共振非线性光散射(RNLS)分析法。以RRS法考察了茜素红-镧与白蛋白形成三元离子缔合物的适宜条件、影响因素等,此法可用于血清、卵清和尿样中蛋白含量的测定。

In buffer solution of HAc-NaAc （ pH 4.5 ）, alizarin red S （ARS） and La3 ＋ was combined with bovine serum albumin （BSA） to form ternary ion-complexes, which resulted in significant enhancement of resonance Rayleigh scattering （RRS）, Second-Order scattering （SOS） and Frequency-Double scattering （FDS）. The maximum scattering wavelength is located at 373 nm for RRS, 620 nm for SOS and 310 nm for FDS, respectively. The light scattering intensity of the system was proportional to the concentration of BSA in certain ranges. The linear ranges were 0. 034 - 13.4μg/mL for RRS method, 0. 09-13.4μg/mL for SOS method and 0. 13-13.4μg/mL for FDS method, and the detection limits of BSA were 0. 031μg/mL （for RRS method）, 0. 083μg/mL （for SOS method） and 0. 117μg/mL （for FDS method）, respectively. Based on the enhancement of light scattering, the determination of BSA by the proposed methods （resonance linear and nonlinear scattering analysis methods） were established, sensitively. The reaction conditions and influence factors of the ARS-La-BSA system were investigated by RRS method. The proposed methods can be applied to the determination of protein contents in serum, ovalbumin and urine samples, and the results were satisfactory.