Pullen R H, Howell J A, Cox J W
Drug Metabolism Research, Upjohn Company, Kalamazoo, MI 49001.
Prostaglandins Leukot Med. 1987 Oct;29(2-3):205-19. doi: 10.1016/0262-1746(87)90010-2.
A high performance liquid chromatographic (HPLC) method was developed to measure thromboxane B2 (TxB2) levels in human serum. Serum samples (2 mL) were extracted using solid phase extraction columns in a C 18/silica mode sequencing approach. The internal standard, 6-ketoprostaglandin F1 alpha, was added to the serum extracts. The eicosanoids were doubly derivatized, first with panacyl bromide, then with methoxyamine to form methoxime-panacyl ester derivatives. The eicosanoid derivatives were chromatographed using a reverse phase HPLC system with UV detection (254 nm). Assay linearity was demonstrated with fortified TxB2 standards in 3% bovine serum albumin over a range of 25 to 500 ng/mL (r greater than or equal to 0.994). There was no significant interday difference or bias in assay results for pooled standards at 75, 226 and 376 ng/mL concentrations (p greater than 0.05). Pooled estimates of precision at these levels indicate an assay relative standard deviation 6-9%. The HPLC assay was used to quantitate TxB2 levels in human serum. Results were consistent with previously published values when drug-free serum was analyzed to assess ex vivo TxB2 formation.
开发了一种高效液相色谱（HPLC）法来测定人血清中血栓素B2（TxB2）的水平。血清样本（2 mL）采用C 18/硅胶模式测序方法的固相萃取柱进行提取。将内标物6-酮-前列腺素F1α添加到血清提取物中。类花生酸进行双重衍生化，首先用泛酰溴，然后用甲氧基胺形成甲氧肟-泛酰酯衍生物。使用带有紫外检测（254 nm）的反相HPLC系统对类花生酸衍生物进行色谱分析。在3%牛血清白蛋白中，添加的TxB2标准品在25至500 ng/mL范围内显示出分析线性（r大于或等于0.994）。在75、226和376 ng/mL浓度下，混合标准品的分析结果没有显著的日间差异或偏差（p大于0.05）。这些水平下的合并精密度估计表明分析相对标准偏差为6-9%。HPLC法用于定量人血清中TxB2的水平。当分析无药物血清以评估离体TxB2形成时，结果与先前发表的值一致。
