COMPARISON AND VALIDATION OF a FAST METHOD FOR THE EXTRACTION AND QUANTIFICATION OF Α-Linolenic, EICOSAPENTAENOIC, AND DOCOSAHEXAENOIC FATTY ACIDS

Abstract

Background: Omega 3 family fatty acids are currently widely studied for the diverse effects they have on cardiovascular disease risk factors. Seafood is the main natural source of these compounds. Objetives: In this paper the accuracy is verified on the fat extraction in fish, between two methods, using one of the most employees as it is the Bligh & Dyer since 1959, compared with SOXTEC, one more updated method with more modern equipment and validated quantification method for some omega 3 fatty acids by gas chromatography. Methods: Fat extraction was performed by SOXTEC and Bligh & Dyer with the use of a Certified Reference Material (SRM 1946). After having assessed the accuracy of both systems, a quantification method of α-Linolenic, Eicosapentaenoic, and Docosahexaenoic fatty acids was validated through gas chromatography. Results: It was found that the method proposed by Bligh & Dyer in 1959 is the most accurate because it showed better recovery percentages. Also, it was found that the quantification validation method of the most important omega 3 fatty acids complies with every parameter assessed throughout the validation. Conclusions: In conclusion SOXTEC being a more modern method which uses the latest technology, the Bligh & Dyer is the most effective method for removing fat in fish and gas chromatography allows quantification of fatty acids with precision and accuracy as meets the evaluated parameters.

Method Development and Validation for Omega-3 Fatty Acids (DHA and EPA) in Fish Using Gas Chromatography with Flame Ionization Detection (GC-FID)

“…According to the FDA Reviewer Guidance for the Validation of Chromatographic Methods (1994), the acceptable value for each concentration should be less than 1% [27]. The good precision value indicated no variability in precision at different concentrations measured on the same and different days The study result was in line with a study by Oviedo Castrillon et al ( 2016) that found repeatability (RSD%) of less than 1%, in which n-3 PUFA (C18:3n-3, C20:5n-3 and C22:6n-3) were 0.6%, 0.5% and 0.1%, respectively, in a commercial tuna sample that was prepared in sextuplet and injected in triplicate [28]. In a study by Trbovic et al ( 2018), the reproducibility (RSD%) for majority n-3 PUFA (C18:3n-3, C20:5n-3 and C22:6n-3) in fish tissues were found to be less than 3%, a bit higher compared to this study [29].…”

“…According to C. Truzzi et al (2017), the mean recovery FA fortified with low and high concentrations of FAME standard were 96 ± 9% (min-max, 81-115%) and 96 ± 7% (min-max, 81-111%), respectively, in an Antarctic fish muscle [26]. Meanwhile, the recovery (R%) of n-3 PUFA (C18:3n-3, C20:5n-3 and C22:6n-3) were 93.5%, 116.2% and 105.3%, respectively, as conducted by Oviedo Castrillon et al (2016) using a commercial tuna oil sample extracted using the Bligh and Dyer method [28]. Most of the R% in this study was comparable to our previous study that indicated satisfactory R% at about 90-120% in extracted fishes using the Bligh and Dyer method spiked with FA standard [33].…”