Mid-Infrared Spectroscopic Determination of Sugar Contents in Plant-Cell Culture Media Using an ATR Method

Abstract

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.

Simple and rapid determination of metabolite content in plant cell culture medium using an FT-IR/ATR method

“…In our previous study [20], we developed a method of determining the sugar content in MS media using FT-IR/ATR, that focused on sugar uptake by TBY-2 cells [21]. In this study, we investigated the simultaneous quantification of both the nutrient sugars and the product ethanol.…”

“…We have also quantitatively analyzed sugars in aqueous solutions using the FT-IR/ATR method [18,19], and applied this technique to quantifying actual liquid medium suspended-plant cells [20]. As a result, the content of each sugar in the culture medium in which the plant cells were suspended was simultaneously and accurately measured by MIR spectroscopy.…”

Discrimination and classification of adulterants in maple syrup with the use of infrared spectroscopic techniques

“…16,17 Past applications include establishing the authenticity of foods such as vegetable oils, 18 olive oil, [19][20][21][22] coffee, 23 honey, 24,25 raspberry purées, 26 orange juice 27 and meat. [28][29][30] Attempts were also made to determine the presence and quantity of sugars in aqueous mixtures 31 and in plant cell culture media 32 using Fourier transform infrared attenuated total reflectance (FTIR-ATR). Infrared spectroscopic methods are simple, costeffective, rapid and non-destructive and could serve as a potential tool for detecting economic adulteration or for routine analysis if proper calibration and validation procedures with data acquisition protocols are established.…”

Development of a quantification system of ionic dissociative metabolites using an FT-IR/ATR method

“…The accuracy of the developed method was almost comparable to that of a high-performance liquid chromatography method [5]. The kinetic sugar uptake phenomena by suspending cells were overall well confirmed using the developed method [7].…”

“…Using the FT-IR/ATR method, we studied the quantitative analysis of sugars in aqueous solution [3,4] and developed the infrared spectroscopic determination of the sugar content in culture media suspending cells [5,6]. The accuracy of the developed method was almost comparable to that of a high-performance liquid chromatography method [5].…”