Andre J C, Funk M O
Anal Biochem. 1986 Nov 1;158(2):316-21. doi: 10.1016/0003-2697(86)90556-7.
High-performance liquid chromatography has been found to be an effective method for the determination of absolute configuration in the products of the lipoxygenase-catalyzed oxygenation of polyunsaturated fatty acids. Methyl esters of fatty acid hydroperoxides that had been reduced to the corresponding alcohols were converted into (+)-alpha-methoxy-alpha-trifluoromethylphenylacetic acid esters. Enantiomeric alcohols were converted into diastereomeric esters that were readily resolved by normal-phase HPLC. The instrumental requirements for the technique are an isocratic HPLC and a uv absorbance monitor. The method was found to be effective in the determination of stereochemistry in the products derived from the action of plant lipoxygenases on linoleic acid. In addition, the chromatography of the derivatives obtained from lipoxygenase catalysis on arachidonic acid was found to be effective for the assignment of stereochemistry in those products. A comparison of the chromatography of these derivatives with that for the corresponding menthyloxycarbonyl derivatives demonstrated the superiority of this approach for the resolution of the diastereomeric pairs. The technique was applied to the determination of stereochemistry in the products derived from soybean lipoxygenase isoenzymes under a variety of experimental conditions.
高效液相色谱法已被证明是一种测定多不饱和脂肪酸经脂氧合酶催化氧化产物绝对构型的有效方法。已还原为相应醇的脂肪酸氢过氧化物甲酯被转化为(+)-α-甲氧基-α-三氟甲基苯乙酸酯。对映体醇被转化为非对映体酯，这些酯很容易通过正相高效液相色谱法分离。该技术的仪器要求是等度高效液相色谱仪和紫外吸光度监测器。该方法被发现对于确定植物脂氧合酶作用于亚油酸产生的产物的立体化学是有效的。此外，发现从脂氧合酶催化花生四烯酸得到的衍生物的色谱法对于确定这些产物的立体化学是有效的。将这些衍生物的色谱法与相应的薄荷氧基羰基衍生物的色谱法进行比较，证明了该方法在分离非对映体对方面的优越性。该技术被应用于在各种实验条件下测定大豆脂氧合酶同工酶产生的产物的立体化学。
