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麦角甾-4,6,8,22-四烯-3-酮(ergone)为猪苓中主要甾体之一,具有多种药理和生理活性.研究了不同极性溶剂中ergone的光学性质,考察了人血清蛋白(HSA),牛血清蛋白(BSA)对ergone荧光和紫外光谱的影响,结果表明血清蛋白加入后ergone的光谱信号强度显著增强,并发生蓝移.根据Benesi-Hildebrand方程求得了结合常数和自由能变.基于血清蛋白对ergone具有良好的荧光增强作用,在模拟生理条件下以ergone为荧光探针,建立了一种灵敏的蛋白质定量分析方法,HSA和BSA线性响应浓度范围分别为(0.38～16.67)×10-7 mol·L-1和(0.42～15.25)×10-7 mol·L-1,检测限(3σ)分别为1.01×10-10和1.22×10-10mol·L-1.考察共存物质对测定结果的影响中发现Fe3+会显著淬灭ergone荧光.该方法用于人血清中总蛋白含量测定结果与考马斯亮蓝法基本一致.
Ergone, one of the major steroids in Polyporus umbellatus, has a variety of pharmacological and physiological activities.The optical properties of ergone in different polar solvents, The effects of human serum albumin (HSA) and bovine serum albumin (BSA) on the fluorescence and UV spectra of ergone were investigated. The results showed that the signal intensity of ergone increased significantly and the blue shift occurred. According to the Benesi-Hildebrand equation, Binding constant and free energy.Based on the good fluorescence enhancement of ergone by serum protein and ergone as fluorescent probe under the simulated physiological conditions, a sensitive protein quantitative analysis method was established. The linear response concentration range of HSA and BSA was (0.38-16.67) × 10-7 mol·L-1 and (0.42-15.25) × 10-7 mol·L-1, the detection limits (3σ) were 1.01 × 10-10 and 1.22 × 10-10 mol · L-1. Examining the effect of coexisting substances on the determination results, it was found that Fe3 + significantly quenched ergone fluorescence. The method for the determination of total protein content in human serum is basically consistent with Coomassie Brilliant Blue method.