4.8 Article

Label-Free in Situ Quantification of Drug in Living Cells at Micromolar Levels Using Infrared Spectroscopy

期刊

ANALYTICAL CHEMISTRY
卷 86, 期 23, 页码 11673-11679

出版社

AMER CHEMICAL SOC
DOI: 10.1021/ac503915c

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资金

  1. Royal Society
  2. EPSRC [EP/L013045/1]
  3. Engineering and Physical Sciences Research Council [EP/L013045/1] Funding Source: researchfish
  4. EPSRC [EP/L013045/1] Funding Source: UKRI

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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.

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