A simple, rapid, and accurate method for determining sugar contents in plant-cell culture media by mid-infrared (MIR) spectroscopy was developed by using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We examined the potential of this method by comparison with a high-performance liquid chromatography (HPLC) method and applied it to the determination of sugar consumption by plant cells. The cultivation was carried out with <i>Nicotiana tabacum</i> cv. Bright Yellow No. 2 cells. We determined that MIR spectroscopy is at least as useful as the HPLC method for the sugar content determination in the culture medium and may be available to monitor the overall cell metabolisms. This study represents an important step in the development of nonintensive on-line monitoring devices for bioreactors and of simple, rapid, and accurate analysis of the cellular metabolism through the composition changes in media.

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