DEVELOPMENT OF A RAPID IN SITU TRANSESTERIFICATION METHOD FOR FATTY ACID ANALYSIS IN MICROALGAE
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The FAME yield from microalgae of two in situ transesterification methods were compared to a typical Folch et al. (1957) extraction followed by transesterification using the Hilditch et al. (1964) procedure. A method based on Park & Goins (1994), utilizing 0.5 N NaOH in methanol, then 14 % BCl3 in methanol, was found to be superior to a method based on Lepage & Roy (1986), utilizing acetyl chloride in methanol. The Park & Goins (1994) method was equivalent to the traditional method and was, therefore, selected for further study. In establishing the parameters of the method, water contents up to 0.55 mL were not found to inhibit the reaction within the maximum lipid load, conservatively assessed at ~1 mg. The reaction time and temperature required to produce a maximum FAME yield was 10 min at 90 °C for the BCl3-catalyzed reaction, while the NaOH-catalyzed reaction happened instantaneously at ambient temperature.