基于Lab-on-a-Colloidosome快速检测方法的开发及其在生命早期抗抑郁药物暴露与不良出生结局之间关联性研究中的应用

Evaluation of the internal exposure of antidepressant drugs plays a significant role in health risk assessment, especially for the sensitive populations such as pregnant women and infants. The prospective birth cohort study about the effect of the internal exposure of antidepressant drugs on adverse birth outcomes always has to analyze a lot of samples with complex matrices, so it’s necessary to develop a rapid analytical method integrated with efficient sample preparation techniques. Surface enhanced Raman scattering (SERS) has shown high potential for rapid detection in analytical field. However, the selectivity of SERS substrate nanoparticles is limited, and the application of SERS in complex samples always suffers matrix effects. It's difficult to solve these problems by a single sample preparation technique. Among sample prepration techniques, electromembrane extraction (EME) has displayed excellent sample clean-up, and molecular imprinting provides high specificity. Utilizing the advantages of EME, molecular imprinting and SERS, as well as the synergistic effects between them, we hereby propose an intergrated EME/Lab-on-a-Colloidosome (LoCo) system for rapid determination of trace antidepressant drugs in complex samples. After the construction of LoCo sensor with SERS substrate nanoparticles and monodispersed molecularly imprinted particles (MIPs), the feasibility of EME/LoCo system will be explored by investigating the suitability of solvent for both EME and MIPs. Subsequently, we will focus on the study of synergistic effects between EME, MIPs and SERS. After the validation, this EME/LoCo system will be used for accurate, sensitive and rapid determination of trace antidepressant drugs in blood and urinary for the study of birth cohort conducted by Tongji Medical School to evaluate the association between the exposure of antidepressant drug at early life and adverse birth outcomes. We believe this LoCo-based approach will open new direction for development of rapid analytical methods and can provide necessary information in relatively shorter time for accurate health risk assessment.

在前瞻性出生队列研究中，样本数量庞大，样品基质复杂，环境与健康领域亟需开发可替代色谱法的抗抑郁药快检方法。SERS技术在快检方面显示了良好的应用前景，但单一样品前处理技术难以同时解决SERS技术选择性不高、受基质干扰的问题。根据申请人前期的研究基础，本项目利用具有特异性富集能力的分子印迹微球和纳米SERS基底为结构模块通过界面自组装构建自分离式Lab-on-a-Colloidosome(LoCo)敏感元件，结合电膜萃取(EME)技术良好的样品净化能力，建立一种适用于复杂样品分析的EME/LoCo一体化快检体系。项目将验证该快检方法的有效性，进而将其用于同济医学院出生队列研究，评估孕妇抗抑郁药物内暴露水平，分析生命早期抗抑郁药物暴露与新生儿不良出生结局之间的关联性。EME/LoCo体系的建立与研究，既为开发新快检方法提供思路又为我国掌握敏感人群抗抑郁药的暴露水平和分布特征快速提供基础数据。

抗抑郁药所引起的环境与健康问题已不容忽视，在前瞻性出生队列研究中，存在样本数量庞大和样品基质复杂的问题。为实现人群中抗抑郁药内暴露水平的快速检测分析，本项目将样品净化能力突出的电膜萃取（EME）技术、特异性分子识别能力强的分子印迹技术（MIP）与灵敏度高的表面增强拉曼（SERS）技术联用，开发一种新型的一体化快检体系。本项目首先利用稳定的支撑液膜（SLM）组建了平板薄膜EME装置，然后研究了该EME体系在不同条件下的样品净化能力及萃取效率。在最优条件下，该EME体系可以达到生物检材中八种典型抗抑郁药的高效萃取。血液和尿液中八种典型抗抑郁药的萃取回收率分别为78%-101%和95-105%。由于EME难以分离理化性质相似的物质，从而难以实现抗抑郁药的选择性分离。因此，本项目使用MIP对抗抑郁药进行选择性富集，以舍曲林和文拉法辛（摩尔比1：2）为混合模版，合成了对目标抗抑郁药具有选择性吸附能力的MIP材料，并对该MIP材料进行表征和吸附性能的研究。在此基础上，对EME体系与MIP材料进行匹配性研究，建立了EME与MIP联用的新体系。为了实现高灵敏度和快速SERS检测，本项目明确了多种抗抑郁药的SERS特征谱图，并针对目标抗抑郁药合成了具有较高灵敏度的SERS基底，研究证明了拉曼检测方法对抗抑郁药的适用性。随后，尝试将单分散MIP微球、SERS基底和EME结合构建Lo-Co检测体系。最后，我们对人群队列中的抗抑郁药内暴露水平进行检测，抗抑郁药内暴露与不良健康结局的分析结果表明健康人群抗抑郁药检出率为28.3%，环境水平抗抑郁药暴露可能会使普通人群谷丙转氨酶、谷草转氨酶、总胆红素、总胆固醇、高密度脂蛋白胆固醇、血糖、血红蛋白水平更高，使BMI、腰臀比、白细胞、血小板水平更低，增加内分泌和泌尿生殖系统疾病的患病风险。该项目为深入开展抗抑郁药的毒性机制研究和健康风险评价、制定抗抑郁药的环境健康基准提供科学依据。