Rapid and Direct Compositional Analysis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in Whole Bacterial Cells by Thermally Assisted Hydrolysis and Methylation–Gas Chromatography

Siti BAIDURAH; Yasuko KUBO; Mitsuhiro KUNO; Kazuho KODERA; Yasuyuki ISHIDA; Tsuneo YAMANE; Hajime OHTANI

doi:10.2116/analsci.31.79

Original Papers

Rapid and Direct Compositional Analysis of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) in Whole Bacterial Cells by Thermally Assisted Hydrolysis and Methylation–Gas Chromatography

Siti BAIDURAH, Yasuko KUBO, Mitsuhiro KUNO, Kazuho KODERA, Yasuyuki ISHIDA, Tsuneo YAMANE, Hajime OHTANI

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Keywords: Poly(3-hydroxybutyrate-co-3-hydroxyvalerate), thermally assisted hydrolysis and methylation–GC, tetramethylammonium hydroxide, copolymer composition

JOURNAL FREE ACCESS

2015 Volume 31 Issue 2 Pages 79-83

DOI https://doi.org/10.2116/analsci.31.79

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Abstract

Thermally assisted hydrolysis and methylation–gas chromatography (THM-GC) in the presence of an organic alkali was applied to the direct analysis of copolymer composition for poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] accumulated in whole bacterial cells. Cupriavidus necator was grown on a liquid medium with different molar ratios of valeric acid (V) to glucose (G) in order to control the compositions of P(3HB-co-3HV) produced in the cells. Trace amounts (0.03 mg) of dried Cupriavidus necator cells were directly subjected to THM-GC in the presence of tetramethylammonium hydroxide (TMAH) at 400°C. The obtained chromatograms clearly showed nine characteristic peaks, attributed to the THM products from 3HB and 3HV units in the polymer chains, without any appreciable interference by the bacterial matrix components. Based on these peak intensities, the copolymer compositions were determined rapidly without using any cumbersome sample pretreatment. Moreover, the compositions thus obtained were in good agreement with those obtained by the conventional technique.

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