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The present study investigated the effect of processing parameters comprising rigor temperature, ageing and display time on conjugated linoleic acid (CLA) concentrations, stability and the development of cholesterol oxidation products in hot boned beef <i>semimembranosus</i> muscles. Meat samples, having attained rigor mortis at 5 &#176;C and 25 &#176;C, were vacuum packed and aged for 7 and 14 days and then displayed under aerobic conditions for 7 days at 4 &#176;C. Lipid was extracted at each time interval then ¹H NMR and GC-FID were used for CLA quantification. The cholesterol oxidation products (COPs) were separated from lipids via column chromatography and derivatized for GC-FID. The CLA content was not affected by the rigor temperature, ageing and display time (<i>p</i> &gt; 0.05). The cholesterol oxidative stability was not affected by rigor temperature (<i>p</i> &gt; 0.05) but was affected by ageing and display time (<i>p</i> &lt; 0.05). The COPs, 7&#945;- and 7&#946;-hydroxycholesterol, and 7-ketocholesterol were positively identified and their quantities increased with ageing and display time (<i>p</i> &lt; 0.05). These results demonstrate that the production of COPs in semimembranosus muscle was significantly altered by the ageing and display time parameters but not by the rigor temperature used in this study.