Key Laboratory of Combinatorial Biosynthesis and Drug Discovery, Ministry of Education, Wuhan, China.
School of Pharmaceutical Sciences, Wuhan University, Wuhan, China.
J Sep Sci. 2018 Jul;41(13):2767-2773. doi: 10.1002/jssc.201701417. Epub 2018 May 28.
A poly(vinylphenylboronic acid-ethylene glycol dimethacrylate) monolithic material incorporated with graphene oxide was synthesized inside a poly(ether ether ketone) tube. This tube with boronate affinity monolith was coupled with a high-performance liquid chromatography system through a six-port valve to construct an online solid-phase microextraction with high-performance liquid chromatography system. The performance of this solid-phase microextraction with high-performance liquid chromatography system was demonstrated by standard glycoprotein in aqueous samples, namely, horseradish peroxidase. Some parameters that affect the extraction performance were investigated, including sampling rate, pH of sample solution, and sampling volume. Under the optimized conditions, the developed method showed high extraction efficiency toward horseradish peroxidase. The addition of graphene oxide greatly increased the extraction efficiency of boronate affinity monolith for horseradish peroxidase. The limit of detection of the proposed method was as low as 0.01 μg/mL by using ultraviolet detection. The recognition specificity was also evaluated by analyzing the mixture of bovine serum albumin (nonglycoprotein) and horseradish peroxidase. The results showed that this material could selectively extract horseradish peroxidase from the mixture, indicating its good specificity toward glycoproteins. The proposed method was further applied for analyzing rat plasma samples spiked with horseradish peroxidase. Good recovery and repeatability were obtained.
一种聚（苯乙烯基硼酸-乙二醇二甲基丙烯酸酯）整体材料与氧化石墨烯结合，在聚醚醚酮管内合成。该带有硼酸亲和性整体的管通过六通阀与高效液相色谱系统相连，构建了带有高效液相色谱系统的在线固相微萃取。通过在水性样品中标准糖蛋白（即辣根过氧化物酶）对该高效液相色谱系统的固相微萃取进行了性能验证。考察了一些影响萃取性能的参数，包括采样速率、样品溶液 pH 值和采样体积。在优化条件下，该方法对辣根过氧化物酶表现出高萃取效率。氧化石墨烯的加入大大提高了硼酸亲和整体对辣根过氧化物酶的萃取效率。通过紫外检测，该方法的检出限低至 0.01μg/mL。通过分析牛血清白蛋白（非糖蛋白）和辣根过氧化物酶的混合物来评估该方法的识别特异性。结果表明，该材料可从混合物中选择性地提取辣根过氧化物酶，表明其对糖蛋白具有良好的特异性。该方法进一步应用于分析加标辣根过氧化物酶的大鼠血浆样品，获得了良好的回收率和重现性。
