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An improved direct transesterification method for fatty acid determination of Phaeodactylum tricornutum

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Abstract

Development of microalgae biotechnology demands a rapid and accurate method of fatty acid determination. In this study, an improved two-step direct transesterification (TSDT) using a sequential combination of alkaline catalyst (NaOH) and acid catalyst (AcCl) was found to enhance the recovery yield of fatty acids in Phaeodactylum tricornutum compared with the methods of traditional solvent extraction (the Folch, the Bligh and Dyer, or the soxhlet methods) followed by transesterification. There was no significant difference (p > 0.05) in the transesterification efficiency between TSDT and AOAC 991.39. TSDT converted intracellular lipid to saponifiable lipids, which were then in situ transesterified to fatty acid methyl esters. All the reactions occurred in one tube, so the extraction step was eliminated and the small samples were enough to assay. Higher levels of fatty acid content with recovery yield above 96 % were obtained with TSDT compared with the traditional extraction-transesterification methods. In the presence of water, TSDT was more effective than each catalyst used alone. A water content up to 10 % of total reaction volume was allowed in the TSDT determination with no effect on results. TSDT is a simple and more accurate method suitable for wet and small samples assay in P. tricornutum cultures.

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Acknowledgments

The study was supported by the Research Award Fund for Outstanding Middle-aged and Young Scientists of Shandong Province of China (BS2010SW025 and BS2013HZ018), the Taishan Scholars Station of Aquatic Animal Nutrition and Feed (HYK201004), the National Natural Science Foundation of China (31201973), the National Marine Public Welfare Research Project of China (201205025), the Marine Biological Industry of China: Aquatic Animal Nutrition and Feed Research and Innovation Demonstration Platform (201301003); and the Modern Agricultural Industry System of Shandong Province of China: Industrial Innovation Team of Sea Cucumber (2012–2014).

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Authors and Affiliations

  1. Shandong Provincial Key Laboratory of Restoration for Marine Ecology, Shandong Marine Resource and Environment Research Institute, 264006, Yantai, People’s Republic of China

    Hongjin Qiao, Jiying Wang, Limin Zhang, Chunxiao Sun, Jingjing Ma, Zhidong Song & Baoshan Li

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  1. Hongjin Qiao
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  2. Jiying Wang
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  3. Limin Zhang
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  4. Chunxiao Sun
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  5. Jingjing Ma
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  6. Zhidong Song
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  7. Baoshan Li
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Corresponding authors

Correspondence to Jiying Wang or Limin Zhang.

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Fig. S1

The wet algal pellet was resuspended in NaOH/MeOH. a, 500 μL of toluene was added. b, algal pellet was sucked in after pipetting. c, algal pellet mixed in toluene was transferred into the tube. d, algal pellet was resuspended in solution without mixing after 1 mL NaOH/MeOH addition. (DOC 399 kb)

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Qiao, H., Wang, J., Zhang, L. et al. An improved direct transesterification method for fatty acid determination of Phaeodactylum tricornutum . J Appl Phycol 27, 697–701 (2015). https://doi.org/10.1007/s10811-014-0376-5

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