Yue Hongfei, Strauss Kenneth I, Borenstein Michael R, Barbe Mary F, Rossi Luella J, Jansen Susan A
Chemistry Department, Analytical Chemistry, Temple University, 1901 N. 13th Street, Philadelphia, PA 19122, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Apr 25;803(2):267-77. doi: 10.1016/j.jchromb.2003.12.027.
Arachidonic acid (AA) is metabolized to prostaglandins (PGs) via cyclooxygenases (COX) catalysis, and to epoxyeicosatrienoic acids (EETs), dihydroxyeicosatrienoic acids (DiHETrEs), and hydroxyeicosatetraenoic acids (HETEs) via cytochrome P450 (CYP450) enzymes. A reliable and robust fluorescence based HPLC method for these eicosanoids was developed. A new selective reverse-phase solid phase extraction (SPE) procedure was developed for PG, DiHETrEs, HETE, and EETs of interest from rat cortical brain tissue. The eicosanoids were derivatized with 2-(2,3-naphthalimino)ethyl-trifluoromethanesulphonate (NE-OTf), followed by separation and quantification at high sensitivity using reverse-phase HPLC with fluorescent detection, and further identified via LC/MS. The derivatization was studied and optimized to obtain reproducible reactions. Various PGs, DiHETrEs, HETEs, EETs, and AA were sensitively detected and baseline resolved simultaneously. LC/MS under positive electrospray ionization selected ion monitoring (SIM) mode was developed to further identify the peaks of these eicosanoids in cortical brain tissue. The method was applied in the traumatic brain injured rat brain.
花生四烯酸（AA）通过环氧化酶（COX）催化代谢为前列腺素（PGs），并通过细胞色素P450（CYP450）酶代谢为环氧二十碳三烯酸（EETs）、二羟基二十碳三烯酸（DiHETrEs）和羟基二十碳四烯酸（HETEs）。我们开发了一种可靠且强大的基于荧光的高效液相色谱法来检测这些类花生酸。我们开发了一种新的选择性反相固相萃取（SPE）程序，用于从大鼠大脑皮质组织中提取感兴趣的PG、DiHETrEs、HETE和EETs。这些类花生酸用2-（2,3-萘二甲酰亚胺基）乙基三氟甲磺酸酯（NE-OTf）进行衍生化，然后使用带荧光检测的反相高效液相色谱法进行高灵敏度的分离和定量，并通过液相色谱/质谱进一步鉴定。对衍生化反应进行了研究和优化，以获得可重复的反应。各种PGs、DiHETrEs、HETEs、EETs和AA均能被灵敏地检测到，并同时实现基线分离。开发了正电喷雾电离选择离子监测（SIM）模式下的液相色谱/质谱法，以进一步鉴定大脑皮质组织中这些类花生酸的峰。该方法应用于创伤性脑损伤大鼠的大脑。
