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Direct determination of ephedrine intermediate in a biotransformation reaction using infrared spectroscopy and PLS.


PMID 18585178

Abstract

Fourier transform mid-infrared spectroscopy (FT-MIR) coupled with a homemade attenuated total reflectance (ATR) flow-cell was used for on-line monitoring of a biotransformation reaction. The reaction was also monitored off-line by gas chromatography-mass spectrometry (GC-MS) enabling to establish a multivariate model for the infrared data based on partial least squares (PLS) regression. The method developed allowed the simultaneous determination of the substrate, two intermediates and the final product involved in the reduction reaction of 1-phenyl 1,2-propanedione at an initial concentration of 0.5% (v/v). The reaction was accomplished with a whole-cell suspension of Saccharomyces cerevisiae in a phosphate buffer of pH 3.0 at 32+/-1 degrees C. The ATR infrared monitoring was performed directly on the suspension cell without any separation process or extraction over 3h, totaling 188 spectra. The data were split into two subsets, with 158 times for calibration and 30 times for validation. The results showed that the proposed method may be used for on-line monitoring of the biotransformation reactions when the initial concentration is very low.

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