Validation of a Single-Extraction Procedure for Sequential Analysis of Vitamin E, Cholesterol, Fatty Acids, and Total Fa
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Validation of a Single-Extraction Procedure for Sequential Analysis of Vitamin E, Cholesterol, Fatty Acids, and Total Fat in Seafood Rebeca Cruz & Susana Casal & Eulália Mendes & Ana Costa & Catarina Santos & Simone Morais
Received: 10 August 2012 / Accepted: 4 November 2012 # Springer Science+Business Media New York 2012
Abstract Food lipid major components are usually analyzed by individual methodologies using diverse extractive procedures for each class. A simple and fast extractive procedure was devised for the sequential analysis of vitamin E, cholesterol, fatty acids, and total fat estimation in seafood, reducing analyses time and organic solvent consumption. Several liquid/liquid-based extractive methodologies using chlorinated and non-chlorinated organic solvents were tested. The extract obtained is used for vitamin E quantification (normal-phase HPLC with fluorescence detection), total cholesterol (normal-phase HPLC with UV detection), fatty acid profile, and total fat estimation (GC-FID), all accomplished in 0.99), even when standards were extracted, demonstrate the accuracy of the methodology. Concerning Table 2 Validation parameters achieved for the global method after the two extractive methods tested (Folch/Smedes)
Folch/Smedes results, when different DW dry weight, FW fresh weight
Linearity Working range r2 after extraction LOD Intra-day precision (RSD) Inter-day precision (RSD) Recovery
vitamin E, validation was accomplished only for αtocopherol, once it was the only detected tocochromanol in the samples analyzed. As for cholesterol, a good linearity was observed (r2 >0.99). For sensitivity evaluation, the limits of detection (LOD) and quantitation (LOQ) were calculated. The LOD was defined as the lowest concentration of each compound that gave an average signal-to-noise ratio >3 over five replicate injections (Table 2). The LOQ was defined as the lowest concentration of each compound that gave peak area counts or peak-to-IS area ratios with a relative standard deviation (RSD)
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