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

1.描述了人体尿液中11-脱氢-血栓烷B3（11-脱氢- txb3）的微量测定。我们制备了[^<18>]11-脱氢血栓烷B3作为内标（IS）。加入[^<18>]类似物的样品，采用Sep Pak tC18和硅胶柱进行色谱制样。将提取的11-脱氢血栓烷B3转化为1-甲酯(ME) -11-正丙酰胺(PA) -甲氧肟-9,11,15-三-二甲基异丙基硅基（DMIPS）醚衍生物，采用气相色谱/高分辨率选择离子监测（GC/HR-SIM）。在GC/HR-SIM分析中，使用MP-65HT柱消除尿基质中的干扰物质。在10 pg-100 ng/管范围内具有良好的线性响应。我们能够在人类尿液的26至375 pg/ml范围内检测到11-脱氢txb3。本方法可用于人尿中11-脱氢- txb3的测定。研究了人类尿液中2,3-dinor-6-keto-pros - More taglandin F1alpha (I)的微量分析。加入[^2H4] -类似物作为内标的样品，用Bond Elut C18和硅胶进行色谱样品制备。通过气相色谱/高分辨率选择离子监测（GC/HR-SIM），将提取的I转化为1-甲酯-6-甲氧基肟-9,11,15-三-二甲基-异丙基硅醚衍生物。在GC/HR-SIM分析中消除了尿基质中的干扰物质。建立了一种微量测定PGI3尿液代谢物2,3-dinor- delta ^<17>-6-keto- pgf1 α的方法。将DELTA^<17>-6-keto- pgf1 α与大鼠肝脏均质液孵育，制备了正宗的2,3-dinor-DELTA^<17>-6-keto- pgf1 α。在[^<18>O]水中通过重复碱催化水解甲酯衍生物合成了DELTA^<17>-6-keto- pgf1 α，并获得DELTA^<17>-6-keto- pgf1 α的气相色谱/选择离子监测（GC/SIM）内标。在10pg -10ng范围内具有良好的线性响应。使用MP-65HT色谱柱的色谱条件显示DELTA^<17>-6-keto- pgf1 α和6-keto- pgf1 α的分离接近基线。我们能够在6到26 pg/ml的人血浆中检测到DELTA^<17>-6-keto-PGF1alpha。本方法可用于测定人尿液和血浆中DELTA^<17>-6-keto- pgf1 α。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