LAQV, REQUIMTE, University of Porto, Department of Chemical Sciences, Faculty of Pharmacy, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal.
Centre of Biological Engineering (CEB), University of Minho, 4710-057 Braga, Portugal.
Anal Chem. 2023 Mar 14;95(10):4619-4626. doi: 10.1021/acs.analchem.2c04631. Epub 2023 Feb 21.
Nanoparticles (NPs) concentration directly impacts the dose delivered to target tissues by nanocarriers. The evaluation of this parameter is required during NPs developmental and quality control stages, for setting dose-response correlations and for evaluating the reproducibility of the manufacturing process. Still, faster and simpler procedures, dismissing skilled operators and post-analysis conversions are needed to quantify NPs for research and quality control operations, and to support result validation. Herein, a miniaturized automated ensemble method to measure NPs concentration was established under the lab-on-valve (LOV) mesofluidic platform. Automatic NPs sampling and delivery to the LOV detection unit were set by flow programming. NPs concentration measurements were based on the decrease in the light transmitted to the detector due to the light scattered by NPs when passing through the optical path. Each analysis was accomplished in 2 min, rendering a determination throughput of 30 h (6 samples h for = 5) and only requiring 30 μL (≈0.03 g) of NPs suspension. Measurements were performed on polymeric NPs, as these represent one of the major classes of NPs under development for drug-delivery aims. Determinations for polystyrene NPs (of 100, 200, and 500 nm) and for NPs made of PEGylated poly-d,l-lactide--glycolide (PEG-PLGA, a biocompatible FDA-approved polymer) were accomplished within 10-10 particles mL range, depending on the NPs size and composition. NPs size and concentration were maintained during analysis, as verified for NPs eluted from the LOV by particle tracking analysis (PTA). Moreover, concentration measurements for PEG-PLGA NPs loaded with an anti-inflammatory drug, methotrexate (MTX), after their incubation in simulated gastric and intestinal fluids were successfully achieved (recovery values of 102-115%, as confirmed by PTA), showing the suitability of the proposed method to support the development of polymeric NPs targeting intestinal delivery.
纳米粒子（NPs）的浓度直接影响纳米载体向靶组织输送的剂量。在 NPs 的开发和质量控制阶段，需要评估这个参数，以建立剂量-反应相关性，并评估制造过程的重现性。然而，为了在研究和质量控制操作中定量 NPs，以及支持结果验证，需要更快、更简单的程序，摒弃熟练操作人员和分析后转换。在此，在阀上实验室（LOV）中观流控平台下建立了一种用于测量 NPs 浓度的小型化自动化集合方法。通过流程序设计实现 NPs 的自动采样和输送到 LOV 检测单元。NPs 浓度的测量基于光穿过光路时被 NPs 散射而导致传输到检测器的光减少。每次分析在 2 分钟内完成，这使得测定通量达到 30 h（6 个样品 h， = 5），仅需要 30 μL（≈0.03 g）的 NPs 悬浮液。测量是在聚合物 NPs 上进行的，因为它们是为药物输送目的开发的主要 NPs 类别之一。完成了对聚苯乙烯 NPs（100、200 和 500 nm）和由聚乙二醇化聚-d,l-乳酸-乙醇酸（PEG-PLGA，一种生物相容性的 FDA 批准的聚合物）制成的 NPs 的测定，测定范围在 10-10 个粒子 mL 之间，具体取决于 NPs 的大小和组成。通过粒子跟踪分析（PTA）验证了在 LOV 中洗脱的 NPs 分析过程中 NPs 的大小和浓度保持不变。此外，成功地实现了负载抗炎药物甲氨蝶呤（MTX）的 PEG-PLGA NPs 在模拟胃液和肠液孵育后的浓度测量（通过 PTA 确认回收率为 102-115%），表明该方法适合支持针对肠道输送的聚合物 NPs 的开发。
