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 Nicotiana tabacum 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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We determine the pH dependency of the mid-infrared spectra in aqueous solution of the organic dissociative materials in the metabolic pathway: saccharide phosphates (G6P, F6P), adenosine, and its phosphates (ATP, ADP, AMP). The series of molar absorbance spectra for these reagents were obtained in a pH range of about 2 to 11 with a Fourier transform infrared (FT-IR) spectrometer equipped with a horizontal diamond attenuated total reflection (ATR) sampling accessory. We also provide a method of infrared spectral extraction of ionic dissociative materials by performing a linear least-square fitting utilizing the formulas of ionic dissociation equilibrium shift, and we obtain the infrared spectrum of each ionic species of the dissociative materials: G6P-, G6P2-; F6P-, F6P2-; ATP2-, ATP3-, ATP4-; ADP-, ADP2-, ADP3-; AMP, AMP-, AMP2-; and adenosine+, adenosine0. The infrared spectral structure of each ionic species of the dissociative materials in the metabolic pathway are discussed. Additionally, the possibility for a quantification system of the concentrations of the organic dissociative materials in varying pH is suggested.

A simple, rapid and accurate mid-infrared (MIR) spectroscopic method for simultaneously determining the product (ethanol) content and the nutrient (sugar) content in plant-cell culture media was developed using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We assessed the potential of this method by comparing it to a high-performance liquid chromatography (HPLC) method, and using the developed method to measure the ethanol and sugar contents simultaneously in liquid culture media with rice and tabacum cell suspensions, respectively. The experimental results suggest that the sugar consumption and ethanol production behaviors of the plant cell suspensions can be non-destructively and simultaneously monitored using the developed method. Furthermore, the spectroscopic method provided in this study could be developed into a technique that could be used to analyze the overall kinetics of the metabolism of the plant cell suspensions.

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