Woldegiorgis G, Spennetta T, Corkey B E, Williamson J R, Shrago E
Anal Biochem. 1985 Oct;150(1):8-12. doi: 10.1016/0003-2697(85)90434-8.
Long-chain acyl-CoA esters were extracted from freeze-clamped livers of fed and fasted rats according to the method of Mancha et al. [M. Mancha, G. B. Stokes, and P. K. Stumpf (1975) Anal. Biochem. 68, 600-608] and analyzed on a radially compressed C18, 5 microns, reverse-phase column using a gradient system consisting of acetonitrile and 25 mM KH2PO4, pH 5.3, at 254 nm. Total analysis time was 25 min. Eight peaks in the extract with carbon chain lengths of 12 to 18, which subsequently disappeared on alkaline hydrolysis, were identified. The major acyl-CoA peaks in the extract in order of increasing retention times were 14:0, 16:1, 18:2, 16:0, 18:1, and 18:0. Total liver long-chain acyl-CoA esters were 108 +/- 11 and 248 +/- 19 nmol/g protein for fed and fasted rats, respectively. On fasting (48 h) the levels of 18:2, 16:0, and 18:1 increased two-to threefold and that of 18:0 sixfold. The advantages of this method are that it not only provides a more direct determination of total tissue long-chain acyl-CoA esters, in that no decomposition of the CoA ester is involved, but it also detects the constituent molecular species.
按照曼查等人的方法[M. 曼查、G. B. 斯托克斯和P. K. 斯顿夫（1975年）《分析生物化学》第68卷，第600 - 608页]，从喂食和禁食大鼠的冷冻钳夹肝脏中提取长链酰基辅酶A酯，并在一个5微米的径向压缩C18反相柱上进行分析，使用由乙腈和25 mM磷酸二氢钾（pH 5.3）组成的梯度系统，检测波长为254 nm。总分析时间为25分钟。提取物中鉴定出8个碳链长度为12至18的峰，这些峰在碱性水解后消失。提取物中按保留时间增加顺序排列的主要酰基辅酶A峰为14:0、16:1、18:2、16:0、18:1和18:0。喂食和禁食大鼠肝脏中的总长链酰基辅酶A酯分别为108±11和248±19 nmol/g蛋白质。禁食（48小时）后，18:2、16:0和18:1的水平增加了两到三倍，18:0的水平增加了六倍。该方法的优点是，它不仅能更直接地测定组织中的总长链酰基辅酶A酯，因为不涉及辅酶A酯的分解，而且还能检测出组成分子种类。
