Development of microdetermination of 3-series prostanoid and the study of the physiological significance in diseases

1.建立了人尿中11-脱氢血栓素B_3(11-dedro-TXB_3)的微量测定方法。我们制备了[^&lt；18；O]11-脱氢血栓素B3作为内标。加入类似物的样品用Sep Pak tC18和硅胶柱进行层析制样。用气相色谱/高分辨选择离子监测(GC/HR-SIM)技术将11-脱氢血栓素B3转化为1-甲酯(ME)-11-正丙酰胺(PA)-methoxime-9，11，15-tris-dimethylisopropylsilyl(DMIPS)醚衍生物。在使用MP-65HT色谱柱进行GC/HR-SIM分析时，消除了尿基质中的干扰物质。在10 pg-100 ng/管范围内线性关系良好。我们能够在人体尿液中检测到26到375 pg/ml范围内的11-脱氢TXB3。本方法可用于人尿中11-脱氢-血栓素B_3的测定。2，3-二氢-6-酮-丙酮…的微量分析人尿中更多的Taglandin F1α(I)被开发出来。加入[^2H4]-类似物作为内标的样品通过Bond Elut C18和硅胶制备的层析样品被提取。提取的I转化为1-甲基ester-6-methoxime-9，11，15-tris-dimethy-lisopropylsilyl醚衍生物后，用气相色谱/高分辨选择离子监测(GC/HR-SIM)进行检测。在GC/HR-SIM分析过程中，消除了尿基质中的干扰物质，建立了尿液中PGI3代谢产物2，3-dior-Delta；17-gt；-6-keto-PGF1α的微量测定方法。用大鼠肝匀浆孵育，制备了正宗的2，3-二甲基-17-酮-前列腺素F1α。在[^&lt；18&gt；O]水中，通过重复碱催化的甲酯类化合物的水解反应，合成了Delta^；lt；17&gt；-6-keto-PGF1pha，得到了Delta^&lt；17&gt；-6-keto-PGF1pha的气相色谱/选择离子监测内标。在10 pg-10 ng范围内线性关系良好。使用MP-65HT色谱柱的条件下，Delta^&lt；17&gt；-6-keto-PGF1pha和6-keto-PGF1pha几乎达到基线分离。我们能够在6到26 pg/ml的人血浆中检测到Delta^&lt；17&gt；-6-keto-PGF1α。本方法可用于人尿和血浆中6-酮-前列腺素F1α的测定。较少

1.The microdetermination of 11-dehydro-thromboxane B3 (11-dehydro-TXB3) in human urine is described. We prepared [^<18>O] 11-dehydro-thromboxane B3 for use as an internal standard (IS). Samples to which was added [^<18>O] analogue were extracted by chromatographic sample preparation using Sep Pak tC18 and silica gel column. Conversion of the extracted11-dehydro-thromboxane B3 to 1-methyl ester (ME) -11-n-propylamide (PA) -methoxime-9,11,15-tris-dimethylisopropylsilyl (DMIPS) ether derivative was followed by gas chromatography/high resolution-selected ion monitoring (GC/HR-SIM). Interfering substances from the urine matrix were eliminated during GC/HR-SIM analysis using a MP-65HT column. Good linear response over the range of 10 pg-100 ng/tube was demonstrated. We were able to detect 11-dehydro-TXB3 in the range from 26 to 375 pg/ml of the human urine. The present method can be applied to the determination of 11-dehydro-TXB3 in the human urine2.The microanalysis of 2,3-dinor-6-keto-pros … More taglandin F1alpha (I) in human urine was developped. Samples to which was added [^2H4] -analogue as an internal standard were extracted by chromatographic sample preparation using Bond Elut C18, and silica gel. Conversion of the extracted I to 1-methyl ester-6-methoxime-9,11,15-tris-dimethy-lisopropylsilyl ether derivative was followed by gas chromatography/high resolution-selected ion monitoring (GC/HR-SIM). Inrerfering substances from the urine matrix were eliminated during GC/HR-SIM analysis.A method of the microdetermination of 2,3-dinor-DELTA^<17>-6-keto-PGF1alpha, a urinary metabolite of PGI3, is developped. An authentic 2,3-dinor-DELTA^<17>-6-keto-PGF1alpha was preparedfrom DELTA^<17>-6-keto-PGF1alpha incubated with homogenate of rat liver. [^<18>O] DELTA^<17>-6-keto-PGF1alpha was synthesized by repeating base-catalyzed hydrolysis of methyl ester derivatives in [^<18>O] water, to obtain an internal standard in gas chromatography/selected ion monitoring (GC/SIM) of DELTA^<17>-6-keto-PGF1alpha. Good linear response over the range of 10 pg-10ng was demonstrated. Chromatographic conditions using a MP-65HT column presented nearly baseline separation of DELTA^<17>-6-keto-PGF1alpha and 6-keto-PGF1alpha. We were able to detect DELTA^<17>-6-keto-PGF1alpha in the range from 6 to 26 pg/ml of the human plasma. The present method can be applied to the determination of DELTA^<17>-6-keto-PGF1alpha in the human urine and plasma. Less