Key Laboratory of Pesticide & Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan, China.
Key Laboratory of Pesticide & Chemical Biology, Ministry of Education, College of Chemistry, Central China Normal University, Wuhan, China.
Anal Chim Acta. 2018 May 30;1008:48-56. doi: 10.1016/j.aca.2017.12.032. Epub 2018 Jan 8.
In this work, an innovative magnetic aptamer adsorbent (FeO-aptamer MNPs) was synthesized for the selective extraction of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Amino-functionalized-FeO was crosslinked with 8-OHdG aptamer by glutaraldehyde and fixed into a steel stainless tube as the sorbent of magnetic solid phase extraction (MSPE). After selective extraction by the aptamer adsorbent, the adsorbed 8-OHdG was desorbed dynamically and online analyzed by high performance liquid chromatography-mass spectrometry (HPLC-MS). The synthesized sorbent presented outstanding features, including specific selectivity, high enrichment capacity, stability and biocompatibility. Moreover, this proposed MSPE-HPLC-MS can achieve adsorption and desorption operation integration, greatly simplify the analysis process and reduce human errors. When compared with offline MSPE, a sensitivity enhancement of 800 times was obtained for the online method. Some experimental parameters such as the amount of the sorbent, sample flow rate and sample volume, were optimized systematically. Under the optimal conditions, low limit of detection (0.01 ng mL, S/N = 3), limit of quantity (0.03 ng mL, S/N = 10) and wide linear range with a satisfactory correlation coefficient (R ≥ 0.9992) were obtained. And the recoveries of 8-OHdG in the urine samples varied from 82% to 116%. All these results revealed that the method is simple, rapid, selective, sensitive and automated, and it could be expected to become a potential approach for the selective determination of trace 8-OHdG in complex urinary samples.
在这项工作中，合成了一种创新的磁性适体吸附剂（FeO-适体 MNPs），用于选择性提取 8-羟基-2'-脱氧鸟苷（8-OHdG）。通过戊二醛将氨基功能化的-FeO 与 8-OHdG 适体交联，并固定在不锈钢管中作为磁性固相萃取（MSPE）的吸附剂。通过适体吸附剂进行选择性萃取后，动态解吸吸附的 8-OHdG，并通过高效液相色谱-质谱联用（HPLC-MS）在线分析。合成的吸附剂具有突出的特点，包括特异性选择性、高富集容量、稳定性和生物相容性。此外，该提出的 MSPE-HPLC-MS 可以实现吸附和解吸操作的集成，大大简化了分析过程并减少了人为错误。与离线 MSPE 相比，在线方法的灵敏度提高了 800 倍。系统优化了一些实验参数，如吸附剂的用量、样品流速和样品体积。在最佳条件下，获得了低检测限（0.01ng/mL，S/N=3）、定量限（0.03ng/mL，S/N=10）和宽线性范围，相关系数令人满意（R≥0.9992）。并且，尿液样品中 8-OHdG 的回收率在 82%至 116%之间变化。所有这些结果表明，该方法简单、快速、选择性好、灵敏度高且自动化，有望成为复杂尿液样品中痕量 8-OHdG 选择性测定的潜在方法。
