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在pH=2.8~4.0的BR缓冲溶液中,硫酸软骨素(CS)由于磺酸基离解而成为带多个负电荷的大阴离子,而低于其等电点(pI=4.7)的牛血清白蛋白(BSA)则以带正电荷的大阳离子存在,两者之间可结合形成复合物.此时将引起共振瑞利散射(RRS)显著增强,并产生新的RRS光谱,其最大散射波长位于304 nm处.与此同时,它的二级散射(SOS)和倍频散射(FDS)也明显增强,且最大SOS和FDS分别位于471 nm和292 nm.散射增强的强度(ΔIRRS、ΔISOS和ΔIFDS)与CS浓度在一定范围内成正比,可用于CS的定量测定.对于CS的检出限(3σ)分别为2.0 ng/mL(RRS),2.9 ng/mL(SOS)和13.2 ng/mL(FDS).研究了适宜的反应条件,考察了共存物质的影响,表明方法有较好的选择性,可用于市售滴眼液中硫酸软骨素含量的测定.
Chondroitin sulphate (CS) is a large anion with multiple negative charges due to dissociation of sulfonate groups in BR buffer solution at pH = 2.8-4.0, while bovine serum albumin (pI = 4.7) lower than its isoelectric point Proteins (BSA) exist as positively charged macrocations and combine to form a complex that causes a significant increase in Resonance Rayleigh Scattering (RRS) and a new RRS spectrum with a maximum scattering wavelength at 304 nm at the same time, its SOS and FDS also increased significantly, and the maximum SOS and FDS were located at 471 nm and 292 nm, respectively.The intensities of scattering enhancement (ΔIRRS, ΔISOS and ΔIFDS ) Is proportional to the concentration of CS in a certain range and can be used for the quantitative determination of CS.The detection limits (3σ) for CS are 2.0 ng / mL (RRS), 2.9 ng / mL (SOS) and 13.2 ng / mL FDS) .The suitable reaction conditions were studied, and the influence of coexisting substances was investigated, indicating that the method has good selectivity and can be used for the determination of chondroitin sulfate in the commercially available eye drops.