In this work a fast and simple multi-target gas chromatography–mass spectrometry (GC-MS) method for the simultaneous detection and absolute quantification of amino acids, fatty acids and sterols in marine organisms is proposed. The methodology was applied to the characterization of the echinoderm Marthasterias glacialis Linnaeus (sea star) extracts. The main factors influencing the extraction of target compounds were evaluated by using different extraction procedures, solvent systems and temperature conditions and a comparison with a reference technique was performed. The most suitable procedure, capable of successfully extract the three classes of target compounds, was ethanol as solvent at 40 ºC under magnetic stirring. Good analytical parameters were obtained since calibrations curves for the 40 compounds under analysis (15 amino acids, 16 fatty acids and 9 sterols) showed regression coefficients (r2) ranging from 0.9844 to 0.9978, with low RSD (from 0.00 to 9.45%), and detection limits varying from 0.03 to 15.40 mg/L. The RSD values for intra- and interday variations studies were also good (RSD < 13.5%, for both) and recoveries were higher than 92 %. Variation in samples from different harvests and origins and their chemical composition during the year is reported. The fact that no previous treatment of samples is required can make this a useful technique for metabolite profiling in marine organisms, among others, both in biomedical and nutritional studies. Moreover, due to the fast and robust character of the proposed method it seems to be suitable for the implementation as routine analysis.