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Identification and Quantitation of Reaction Intermediates and Residuals in Lipase-Catalyzed Transesterified Oils by HPLC

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Abstract

A high-performance liquid chromatography (HPLC) unit equipped with size exclusion column and a refractive index detector was used for simultaneous monitoring, identification, and quantitation of the reaction components from lipase-catalyzed transesterification of three oils. The procedure simultaneously separated and detected the unreacted triacylglycerols (TAG), diacyl-, and monoacyl-glycerol (DAG and MAG) co-products, residual alcohol as well as free fatty acid (FFA) based on retention times. The chromatograms showed well separated and resolved peaks. The elution of the components from the transesterification reaction in increasing order was: TAG < DAG < FFA < MAG. Generally, higher alcohol ratios decreased the conversion of TAG in all the oils studied with between 14% and 94% of TAG remaining at all the treatment combinations. Higher amount of salmon skin oil (SSO) TAG was generally converted to DAG than Rothsay composite (RC) and olive oil (OO) TAG. Relatively higher amount of OO DAG was converted to MAG than SSO and RC with only 5–14% DAG remaining in OO. RC and OO generally accumulated less MAG, and this was reflected as lower MAG levels in RC (<6%) and OO (<14%) compared with SSO (<27%). For the various treatment combinations and the three oils used in this study, the least amount of FFA was recorded in transesterified OO with a maximum of approximately 4%. This HPLC method can be used as a simple and fast technique to analyze the reaction components and products of transesterification reactions without the need for additional derivatization steps.

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Acknowledgments

The authors acknowledge the financial support from the Natural Sciences and Engineering Research Council (NSERC-Strategic Program) of Canada, and the provision of research samples by Atkins and Frères Inc. and Rothsay® Biodiesel, as well as the technical assistance with the HPLC from Dr. Veronique Fournier.

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Authors and Affiliations

  1. Department of Food Science and Agricultural Chemistry, McGill University (Macdonald Campus), 21,111 Lakeshore Rd., Ste. Anne de Bellevue, QC, Canada, H9X 3V9

    Alberta N. A. Aryee & Benjamin K. Simpson

  2. Department of Animal Science, Faculty of Agricultural and Environmental Sciences, McGill University (Macdonald Campus), 21,111 Lakeshore Rd., Ste. Anne de Bellevue, QC, Canada, H9X 3V9

    Leroy E. Phillip & Roger I. Cue

Authors
  1. Alberta N. A. Aryee
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  2. Leroy E. Phillip
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  3. Roger I. Cue
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  4. Benjamin K. Simpson
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Correspondence to Benjamin K. Simpson.

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Aryee, A.N.A., Phillip, L.E., Cue, R.I. et al. Identification and Quantitation of Reaction Intermediates and Residuals in Lipase-Catalyzed Transesterified Oils by HPLC. Appl Biochem Biotechnol 165, 155–177 (2011). https://doi.org/10.1007/s12010-011-9241-z

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  • DOI: https://doi.org/10.1007/s12010-011-9241-z

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