Oliw Ernst H, Wennman Anneli
Division of Biochemical Pharmacology, Department of Pharmaceutical Biosciences, Uppsala Biomedical Center, Uppsala University, 591, 75124, Uppsala, Sweden,
Methods Mol Biol. 2015;1208:85-95. doi: 10.1007/978-1-4939-1441-8_7.
Fatty acid oxygenases are often characterized by steric analysis of their hydroxy or hydroperoxy metabolites. Chiral phase-HPLC (CP-HPLC) can be used to separate enantiomeric hydroperoxyoctadecenoic acids. This method is based on analysis of seven octadecenoic fatty acids with double bonds at positions 6Z to 13Z, which were oxidized to hydroperoxides by photooxidation. A stationary phase, Reprosil Chiral NR, was found to resolve these hydroperoxy fatty acids with 1-hydroperoxy-2-propene and with 3-hydroperoxy-1-propene elements so that the S hydroperoxy fatty acids consistently eluted before the R stereoisomers. The chiral selector has not been disclosed, but it is described as an aromatic chiral phase with π-donor and π-acceptor groups of Pirkle type. The MS(3) spectra of the hydroperoxides showed characteristic fragments, which were influenced by the distance between the hydroperoxy and the carboxyl groups and the relative position of the double bond. Octadecenoic fatty acids can be oxidized by fungal and bacterial dioxygenases to hydroperoxides with cis or trans double bond configuration. Steric analysis of the hydroperoxy metabolites can be performed by this method, and it can also be used for preparative purposes.
脂肪酸氧化酶通常通过对其羟基或氢过氧化物代谢产物进行空间分析来表征。手性相高效液相色谱法（CP-HPLC）可用于分离对映体氢过氧化十八碳烯酸。该方法基于对6Z至13Z位置带有双键的七种十八碳烯酸脂肪酸的分析，这些脂肪酸通过光氧化被氧化为氢过氧化物。发现一种固定相Reprosil Chiral NR能够分离这些带有1-氢过氧基-2-丙烯和3-氢过氧基-1-丙烯元素的氢过氧基脂肪酸，使得S型氢过氧基脂肪酸始终比R型立体异构体先洗脱。手性选择剂尚未公开，但被描述为具有Pirkle型π供体和π受体基团的芳香手性相。氢过氧化物的MS(3)光谱显示出特征性碎片，这些碎片受氢过氧基与羧基之间的距离以及双键的相对位置影响。十八碳烯酸脂肪酸可被真菌和细菌双加氧酶氧化为具有顺式或反式双键构型的氢过氧化物。通过该方法可对氢过氧基代谢产物进行空间分析，并且它也可用于制备目的。
