How to use the internal standard method to determine the fatty acids in fish, determine the absolute content, there are detailed steps?
Prama Hartami
Lipid Extraction and Methanolysis: The standard procedure involves lipid extraction using organic solvents (such as chloroform/methanol) to isolate lipids from fish tissues. Next, methanolysis is performed to convert the extracted lipids into fatty acid methyl esters (FAMEs). These FAMEs are then analyzed using gas chromatography (GC) or GC-mass spectrometry (GC/MS) to determine the fatty acid composition. Gas Chromatography (GC): GC is widely used for fatty acid analysis. Separation of FAMEs occurs on a GC column, and quantification is usually done using internal standards. This method provides detailed information about individual fatty acids present in the sample. Quantification and Reporting: Fatty acids can be expressed as absolute content (e.g., mg/g of lipids or tissue) or relative proportions (% of total FA mass or molar concentration). Concentrations are commonly used in nutrition studies, while proportions are useful for understanding relative contributions of different fatty acids.
Some methods commonly used for fatty acid analysis in fish:
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