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Validation of microwave-assisted sample extraction-high performance liquid chromatography (HPLC) method for quantitative analysis of hydrolysed amino acids in aquatic biological samples

A sensitive and widely applicable High-Performance Liquid Chromatography (HPLC) with fluorescence detector (FLD) method for determination hydrolysed amino acids in fish and aquatic products was developed and validated through study. The method involved a microwave-assisted sample extraction and protein hydrolysed with the hydrochloric acid and propionic acid. The separation was performed by Shim-pack HR-ODS column with a gradient mobile phase. The total separation time was 25 min per run. The method showed satisfactory linearity, repeatability, accuracy, and limit of detection for 21 amino acids with the 1-100 mg/L calibration range based on 10 µL injection. The fish, fish feed, canned fish and seaweeds samples were analysed successfully by using this developed method.