Li Nan, Hai Xiaoman, Yu Xiaoling, Dang Fuquan
School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710119, China.
School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an 710119, China; Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, Xi'an 710119, China.
J Chromatogr A. 2017 Jan 20;1481:152-157. doi: 10.1016/j.chroma.2016.12.049. Epub 2016 Dec 19.
A facile and efficient dynamic coating method using an ionic complementary peptide was established for high-performance separation of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled carbohydrates in a hybrid poly(dimethylsiloxane) (PDMS)/glass microfluidic channel. EAK16-II with a sequence of [(Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys)] can readily self-organize into a complete coating layer tightly adsorbed on both hydrophobic PDMS and hydrophilic glass surfaces, which efficiently suppressed nonspecific analyte adsorption and minimized electroosmotic flow (EOF). Separation conditions were systematically investigated with respect to EAK16-II concentration, running buffer, buffer pH, and field strength (E). Under the optimal conditions, rapid and reproducible separations of maltodextrin ladder, glycans from glucosamine capsules, tablets, and pomegranate peel extracts were achieved with over 450000 theoretical plates per meter in the hybrid PDMS/glass microchannels dynamically coated with 1.0mg/mL EAK16-II-0.05% n-dodecyl β-d-maltoside (DDM), and the relative standard deviation (RSD) values were less than 3.2% (n=4) for the migration times. The present work provides a facile and efficient means to minimize EOF and nonspecific analyte adsorption in microfluidic chips fabricated in various substrates, thereby broadening the applications of microfluidic chips in complicated biological assays.
建立了一种简便高效的动态涂层方法,该方法使用离子互补肽,用于在混合聚二甲基硅氧烷(PDMS)/玻璃微流控通道中对8-氨基芘-1,3,6-三磺酸(APTS)标记的碳水化合物进行高效分离。序列为[(Ala-Glu-Ala-Glu-Ala-Lys-Ala-Lys)]的EAK16-II能够轻松自组装成完整的涂层,紧密吸附在疏水的PDMS和亲水的玻璃表面上,有效抑制了非特异性分析物吸附,并使电渗流(EOF)最小化。系统研究了EAK16-II浓度、运行缓冲液、缓冲液pH值和场强(E)等分离条件。在最佳条件下,在动态涂覆有1.0mg/mL EAK16-II-0.05%正十二烷基-β-D-麦芽糖苷(DDM)的混合PDMS/玻璃微通道中,实现了麦芽糖糊精梯、氨基葡萄糖胶囊、片剂和石榴皮提取物中聚糖的快速且可重复的分离,每米理论塔板数超过450000,迁移时间的相对标准偏差(RSD)值小于3.2%(n = 4)。本工作提供了一种简便高效的方法,可最大限度地减少在各种基材上制造的微流控芯片中的EOF和非特异性分析物吸附,从而拓宽了微流控芯片在复杂生物分析中的应用。