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Hydrogenated oil has been widely used for production of shortenings or margarine, however, the presence of trans fatty acids may be detrimental to human health. The objectives of this study were to develop an improved method for analysis of trans fatty acids and evaluate their formation in both unhydrogenated and hydrogenated soybean oil during heating at 160, 180 and 200°C for varied length of time. Results showed that among the four columns tested, an Agilent HP-88 column (100×0.25mm I.D.,0.2-μm film thickness) could resolve eight trans fatty acids and nine cis fatty acids simultaneously within 31min with injector temperature 240°C, detector temperature 250°C, and column temperature 170°C in the beginning, maintained for 24min, increased to 220°C at 7.5°C/min, 230°C at 10°C/min, and maintained for 5min. The contents of both cis and trans fatty acids showed a decreased trend for the increase of …