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A novel aptamer-based online magnetic solid phase extraction method for the selective determination of 8-hydroxy-2′-deoxyguanosine in human urine
Analytica Chimica Acta ( IF 5.7 ) Pub Date : 2018-05-01 , DOI: 10.1016/j.aca.2017.12.032
Haijiao Gan , Hui Xu

In this work, an innovative magnetic aptamer adsorbent (Fe3O4-aptamer MNPs) was synthesized for the selective extraction of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Amino-functionalized-Fe3O4 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-1, S/N = 3), limit of quantity (0.03 ng mL-1, S/N = 10) and wide linear range with a satisfactory correlation coefficient (R2 ≥ 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.

中文翻译:

一种用于选择性测定人尿中 8-羟基-2'-脱氧鸟苷的基于适配体的在线磁性固相萃取新方法

在这项工作中,合成了一种创新的磁性适体吸附剂 (Fe3O4-适体 MNPs),用于选择性提取 8-羟基-2'-脱氧鸟苷 (8-OHdG)。氨基官能化的 Fe3O4 通过戊二醛与 8-OHdG 适配体交联,并固定在不锈钢管中作为磁性固相萃取 (MSPE) 的吸附剂。通过适体吸附剂选择性提取后,吸附的 8-OHdG 被动态解吸并通过高效液相色谱-质谱 (HPLC-MS) 进行在线分析。合成的吸附剂具有突出的特点,包括特异性选择性、高富集能力、稳定性和生物相容性。而且,这种提出的MSPE-HPLC-MS可以实现吸附和解吸操作一体化,大大简化了分析过程,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。对吸附剂用量、样品流速、样品体积等实验参数进行了系统优化。在最佳条件下,检测下限(0.01 ng mL-1,S/N = 3)、数量限(0.03 ng mL-1,S/N = 10)和宽线性范围,具有令人满意的相关系数(R2 ≥ 0.9992) 获得。尿样中 8-OHdG 的回收率从 82% 到 116% 不等。所有这些结果表明,该方法简单、快速、选择性、灵敏和自动化,有望成为选择性测定复杂尿样中痕量 8-OHdG 的潜在方法。对吸附剂用量、样品流速、样品体积等实验参数进行了系统优化。在最佳条件下,检测下限(0.01 ng mL-1,S/N = 3)、数量限(0.03 ng mL-1,S/N = 10)和宽线性范围,具有令人满意的相关系数(R2 ≥ 0.9992) 获得。尿样中 8-OHdG 的回收率从 82% 到 116% 不等。所有这些结果表明,该方法简单、快速、选择性、灵敏和自动化,有望成为选择性测定复杂尿样中痕量 8-OHdG 的潜在方法。对吸附剂用量、样品流速、样品体积等实验参数进行了系统优化。在最佳条件下,检测下限(0.01 ng mL-1,S/N = 3)、数量限(0.03 ng mL-1,S/N = 10)和宽线性范围,具有令人满意的相关系数(R2 ≥ 0.9992) 获得。尿样中 8-OHdG 的回收率从 82% 到 116% 不等。所有这些结果表明,该方法简单、快速、选择性、灵敏和自动化,有望成为选择性测定复杂尿样中痕量 8-OHdG 的潜在方法。获得了定量限 (0.03 ng mL-1, S/N = 10) 和宽线性范围,具有令人满意的相关系数 (R2 ≥ 0.9992)。尿样中 8-OHdG 的回收率从 82% 到 116% 不等。所有这些结果表明,该方法简单、快速、选择性、灵敏和自动化,有望成为选择性测定复杂尿样中痕量 8-OHdG 的潜在方法。获得了定量限 (0.03 ng mL-1, S/N = 10) 和宽线性范围,具有令人满意的相关系数 (R2 ≥ 0.9992)。尿样中 8-OHdG 的回收率从 82% 到 116% 不等。所有这些结果表明,该方法简单、快速、选择性、灵敏和自动化,有望成为选择性测定复杂尿样中痕量 8-OHdG 的潜在方法。
更新日期:2018-05-01
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