Label-Free in Situ Quantification of Drug in Living Cells at Micromolar Levels Using Infrared Spectroscopy
Label-Free in Situ Quantification of Drug in Living Cells at Micromolar Levels Using Infrared Spectroscopy
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DOI：
10.1021/ac503915c
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发表时间：
2014-12-02
影响因子：
7.4
通讯作者：
Fale, Pedro L. V.
中科院分区：
文献类型：
--
作者：
Chan, K. L. Andrew;Fale, Pedro L. V.
定量药物进入活细胞的速度和数量是药物开发过程中必不可少的一部分。红外光谱是一种无标记的化学选择工具，用于分析培养活细胞的组成，有可能以无损的方式就地量化进入活细胞的药物量，尽管其灵敏度目前有限。本文首次展示了利用傅里叶变换红外光谱在治疗相关浓度的活细胞中对癌症药物氟尿嘧啶进行原位定量。为了提高红外测量的检出限和定量限，采用了两种策略。首先，采用多反弹衰减全反射采样方法优化信号；其次，采用长通滤波器与汞镉碲化探测器相结合来降低仪器噪声。利用这些新的适应性，可以使用标准FTIR仪器量化细胞培养基中20 μ M的氟尿嘧啶，同时可以量化和测量用80 μ M药物处理的活细胞中氟尿嘧啶的原位通量。
Quantifying the rate and the amount of drug entering live cells is an essential part of the medicine development process. Infrared spectroscopy is a label-free, chemically selective tool for analyzing the composition of live cells in culture that has the potential to quantify, in situ, the amount of drug entering living cells in a nondestructive manner, although its sensitivity is currently limited. This paper is the first to demonstrate in situ quantification of the cancer drug, fluorouracil, in live cells at a therapeutically relevant concentration using Fourier transform infrared spectroscopy. To achieve the required improvement in detection and quantitation limits of the IR measurement, two strategies were exploited. First, a sampling method called multibounce attenuated total reflection was used to optimize the signal while second, a long pass filter in combination with a mercury cadmium telluride detector was used to reduce the instrument noise. Using these novel adaptations, it was possible to quantify 20 mu M of fluorouracil in cell culture medium using a standard FTIR instrument, while it was possible to quantify and measure the flux of fluorouracil in situ in living cells treated with an 80 mu M drug.