To probe into regulating functions of allicin, diallyldisulfid-S-oxide, on electron transport in the inner membrane of mitochondria isolated from beef heart, under the anaerobic, hypoxia, and oxygenating conditions in the presence of rotenone(complex I inhibitor) at 37 °C, we optimized the isolation method for isolating mitochondria from beef heart and used the technique for measuring dissolved oxygen content at the right moment so as to investigate the effects of allicin on the respiratory control of mitochondria in state 4 and state 3, P/O ratio, and respiratory control ratio(RCR). The results show that the optimized isolated requirements were that all the operations were always kept at 0―4 °C, the ratio between sample and isolation medium was 1:4, the filter was a four-layer gauze, two centrifugations in different sections. First section: 2500g, 10 min; second section 7000g, 20 min. The results indicate that the capacity of dissolved oxygen in state 3 was increased and that of dissolved oxygen in state 4 was almost stabilized in mitochondria from beef heart. To probe into regulating functions of allicin, diallyldisulfid-S-oxide, on electron transport in the inner membrane of mitochondria isolated from beef heart, under the anaerobic, hypoxia, and oxygenating conditions in the presence of rotenone (complex I inhibitor) at 37 ° C, we optimized the isolation method for isolating mitochondria from beef heart and used the technique for measuring dissolved oxygen content at the right moment so as to investigate the effects of allicin on the respiratory control of mitochondria in state 4 and state 3, P / O ratio, and respiratory control ratio (RCR). The results show that the optimized isolated requirements were that all the operations were always kept at 0-4 ° C, the ratio between sample and isolation medium was 1: 4, the filter was a four -layer gauze, two centrifugations in different sections. First section: 2500g, 10 min; second section 7000g, 20 min. The results indicate that the capacity of dissolved oxygen in state 3 was increased and that of d issuedolved oxygen in state 4 was almost stabilized in mitochondria from beef heart.