Simões Nathália Soares, de Oliveira Hanna Leijoto, da Silva Ricky Cássio Santos, Teixeira Leila Suleimara, Sales Thaís Lorenna Souza, de Castro Whocely Victor, de Paiva Maria José Nunes, Sanches Cristina, Borges Keyller Bastos
Departamento de Ciências Naturais, Universidade Federal de São João del-Rei, Campus Dom Bosco, São João del-Rei, Minas Gerais, Brazil.
Universidade Federal de São João del-Rei, Campus Centro Oeste, Divinópolis, Minas Gerais, Brazil.
Electrophoresis. 2018 Oct;39(20):2581-2589. doi: 10.1002/elps.201800165. Epub 2018 Jun 13.
In this work a hollow mesoporous structured molecularly imprinted polymer was synthetized and used as adsorbent in pipette-tip solid-phase extraction for the determination of lamivudine (3TC), zidovudine (AZT) and efavirenz (EFZ) from plasma of human immunodeficiency virus (HIV) infected patients by high-performance liquid chromatography (HPLC). All parameters that influence the recovery of the pipette tip based on hollow mesoporous molecularly imprinted polymer solid-phase extraction (PT-HM-MIP-SPE) method were systematically studied and discussed in detail. The adsorbent material was prepared using methacrylic acid and 4-vinylpyridine as functional monomers, ethylene glycol dimethacrylate as crosslinker, acetonitrile as solvent, 4,4'-azobis(4-cyanovaleric acid) as radical initiator, benzalkonium chloride as surfactant, 3TC, and AZT as templates. The simultaneous separation of 3TC, AZT and EFZ by HPLC-UV was performed using a Gemini C18 Phenomenex® column (250 mm × 4.6 mm, 5 μm) and mobile phase consisting of acetonitrile: water pH 3.2 (68:32, v/v), flow rate of 1.0 mL/min and λ = 260 nm. The method was linear over the concentration range from 0.25 to 10 μg/mL for 3TC and EFZ, and 0.05 to 2.0 μg mL for AZT, with correlation coefficients larger than 0.99 for all analytes. Recovery ± relative standard deviations (RSDs %) were 41.99 ± 2.38%, 82.29 ± 1.63%, and 83.72 ± 7.52% for 3TC, AZT, and EFZ, respectively. The RSDs and relative errors (REs) were lower than 15% for intra and interday assays. The method has been successfully applied for monitoring HIV-infected patients outside the therapeutic dosage.
在本研究中，合成了一种中空介孔结构的分子印迹聚合物，并将其用作移液器吸头固相萃取的吸附剂，用于通过高效液相色谱法（HPLC）测定人类免疫缺陷病毒（HIV）感染患者血浆中的拉米夫定（3TC）、齐多夫定（AZT）和依非韦伦（EFZ）。系统研究并详细讨论了影响基于中空介孔分子印迹聚合物固相萃取（PT-HM-MIP-SPE）方法的移液器吸头回收率的所有参数。以甲基丙烯酸和4-乙烯基吡啶为功能单体，乙二醇二甲基丙烯酸酯为交联剂，乙腈为溶剂，4,4'-偶氮双（4-氰基戊酸）为自由基引发剂，苯扎氯铵为表面活性剂，3TC和AZT为模板制备吸附材料。使用Gemini C18 Phenomenex®柱（250 mm×4.6 mm，5μm）和由乙腈：pH 3.2的水（68:32，v/v）组成的流动相、流速为1.0 mL/min以及λ=260 nm，通过HPLC-UV对3TC、AZT和EFZ进行同时分离。该方法在3TC和EFZ的浓度范围为0.25至10μg/mL、AZT的浓度范围为0.05至2.0μg/mL时呈线性，所有分析物的相关系数均大于0.99。3TC、AZT和EFZ的回收率±相对标准偏差（RSDs%）分别为41.99±2.38%、82.29±1.63%和83.72±7.52%。日内和日间测定的RSDs和相对误差（REs）均低于15%。该方法已成功应用于监测治疗剂量以外的HIV感染患者。
