Heavey D J, Soberman R J, Lewis R A, Spur B, Austen K F
Prostaglandins. 1987 May;33(5):693-708. doi: 10.1016/0090-6980(87)90035-9.
A sensitive and specific assay has been developed for measurement of total sulfidopeptide leukotrienes (LT) in plasma. LTC4 and LTD4 in plasma are converted to LTE4 which is then extracted by C18 Sep-Pak binding and elution. Total LTE4 is resolved by reverse phase high performance liquid chromatography (RP-HPLC) and quantitated by radioimmunoassay (RIA). A [3H]LTE4 internal standard is added to the starting plasma sample to allow overall recovery to be calculated and to define the fractions from RP-HPLC to be assayed for LTE4-like immunoreactivity. The correlation between the measured increase in LTE4 concentration after addition of incremental amounts of LTC4 and LTE4 to plasma was 0.989 and 0.978, respectively, with slopes of 1.05 and 1.11. Addition of 51 pg/ml LTE4 to 5 ml plasma was detectable; the measured increase was 48 +/- 12 pg/ml (mean +/- SE, n = 7). The intra-assay coefficient of variation for 341 pg/ml of added LTC4 was 3.2% (n = 6). Sulfidopeptide leukotrienes could not be detected in blood samples taken from 12 normal volunteers in whom the theoretical detection limit, calculated from the sensitivity of the RIA, the overall recovery of LTE4, and the volume of plasma extracted, was 83 +/- 4 pg LTE4/ml plasma (0.19 +/- 0.01 pmol sulfidopeptide leukotriene/ml plasma; mean +/- SE).
已开发出一种灵敏且特异的检测方法，用于测定血浆中总硫化肽白三烯（LT）。血浆中的LTC4和LTD4转化为LTE4，然后通过C18 Sep-Pak结合和洗脱进行提取。总LTE4通过反相高效液相色谱（RP-HPLC）分离，并通过放射免疫分析（RIA）进行定量。向起始血浆样品中加入[3H]LTE4内标，以计算总体回收率，并确定RP-HPLC中用于检测LTE4样免疫反应性的馏分。向血浆中添加递增剂量的LTC4和LTE4后，测得的LTE4浓度增加之间的相关性分别为0.989和0.978，斜率分别为1.05和1.11。向5 ml血浆中添加51 pg/ml LTE4可被检测到；测得的增加量为48±12 pg/ml（平均值±标准误，n = 7）。添加341 pg/ml LTC4的批内变异系数为3.2%（n = 6）。在12名正常志愿者采集的血样中未检测到硫化肽白三烯，根据RIA的灵敏度、LTE4的总体回收率和提取的血浆体积计算，理论检测限为83±4 pg LTE4/ml血浆（0.19±0.01 pmol硫化肽白三烯/ml血浆；平均值±标准误）。
