Desenvolvimento de métodos baseados em espectrometria de massas e cromatografia líquida para análise de compostos químicos produzidos por bioconversão de glicerina
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2018-07-19
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Universidade Federal de Goiás
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Biodiesel production has been increasing every year, which has generated surpluses of
glycerin, the main co-product of the transesterification reaction. In the search for applications to
this surplus, microbial fermentation has been highlighted as an alternative to add value to this co-
product. In this process, microorganisms are used to convert this abundant carbon source into
compounds of high economic value, including organic acids and polyols. However, there are still
some technological challenges, such as low yield during the fermentation processes and high
dependence of new analytical methods, more sensitive and selective, able to identify and quantify
the compounds produced.
The objective of this work was to establish new analytical methodologies to identify and
quantify compounds from bioconversion processes involving the production of organic acids and
polyols. To guarantee the quantification of the target compounds, methods based on ultra-high-
performance liquid chromatography (UHPLC) were developed and the photodiode array (PDA)
and mass spectrometry (MS) detectors were evaluated as alternatives for the monitoring of these
two classes of chemical compounds. The analytes investigated in this study were short chain
organic acids: citric acid, fumaric acid, glyceric acid, kojic acid, lactic acid, muconic acid, oxalic
acid, propionic acid, succinic acid, xylonic acid; and polyols with spatial isomers, including
polyols with 4 carbons (C4: erythritol and threitol), 5 carbons (C5: arabitol, ribitol, xylitol), and 6
carbons (C6: dulcitol, iditol, mannitol, sorbitol) and 7 carbons (C7: volemitol).
The UHPLC-PDA method developed for the analysis of organic acids was validated by
parameters of linearity, limit of detection and quantification, precision and accuracy. The
analytical curves showed good linearity with determination coefficients (R²) higher than 0.999 for
all organic acids. Precision and accuracy were adequate with intra-day and inter-day variation
coefficients below 4.06% and recovery ranged from 94.90% to 109.63%. The developed UHPLC-
MS methods presented greater sensitivity and selectivity in the detection of organic acids and
polyols, being able to monitor these analytes even in low quantity.
The analytical methods developed in this work are crucial tools in the selection or screening
of promising microorganisms for low added value substrates conversion and important to optimize
biotechnological processes for production of these compounds of interest.
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VIEIRA, F. S. Desenvolvimento de métodos baseados em espectrometria de massas e cromatografia líquida para análise de compostos químicos produzidos por bioconversão de glicerina. 2018. 95 f. Dissertação (Mestrado em Química) - Universidade Federal de Goiás, Goiânia, 2018.