The invention discloses a method for determining compositions of fatty acids and acylglycerols at alpha and beta positions of structured lipid by combining solid-phase extraction and gas chromatography. The method comprises utilizing pure arachidonic acid-triacylglycerol (ARA-TAG), docosahexaenoic acid-diacylglycerol (DAH-DAG), linolenic acid- monoacylglycerol (Ln-MAG) and an oleic acid-free fatty acid (O-FFA) standard sample, and optimizing eluent elution conditions with different polarity ratios, so as to effectively separate TAG, DAG, MAG and FFA in mixtures in a same SPE (Solid Phase Extraction) microcolumn; and the method is characterized by also comprising the following steps: 1) separation of a mixed standard sample and analysis on compositions of fatty acids; and 2) analysis on compositions of fatty acids and acylglycerols at alpha and beta positions of the structured lipid. The method has the advantages of simple operation, low sample demanded amount, less consumption amount of a needed solvent, accurate qualitative quantitative analysis results, low detection limit and short analysis time, and is capable of realizing high-flux detection on TAG, DAG, MAG and FFA compositions and content in structured lipid and compositions of fatty acids at alpha and beta positions of structured lipid.