尿6-酮-前列腺素F1α测定的简易放射免疫分析法。

Simple radioimmunological method for urinary 6-keto-PGF1 alpha measurement.

作者信息

Lahera V, Durán F, Cachofeiro V, del Cañizo F J, Rodríguez F J, Tresguerres J A

出版信息

Rev Esp Fisiol. 1986 Jun;42(2):233-8.

Abstract

A radioimmunoassay for the determination of 6-keto-PGF1 alpha (stable metabolite of Prostacyclin) in urine using a specific antiserum, is described. The antibody used, showed no significant cross-reactivity with other PGS. (3H)-6-keto-PGF1 alpha has been used as tracer and a 30% Hydroxyapatite suspension was utilized for the separation of bound and free fractions. Standard curve ranges between 2 and 1000 pg thus allowing measurements in urine. Aliquots of 0.2 to 1 ml of 24 hours urinary samples were submitted to a preextraction step with Hexane in order to eliminate neutral fats. After acidifying with 0.1 N HCl to pH = 3.5-4, an Ethyl Ether extraction step was performed showing a recovery of 66%. Repeated analysis of a rat urine pool, showed 7.7 intraassay and 9.8 interassay variation coefficients. Values obtained amounted to 38.3 +/- 4.6 ng/24 h in normal rats and 133 +/- 32 ng/24 h in 5% NaCl loaded rats (p less than 0.001). The values obtained in healthy men were 508 +/- 94 ng/24 h and in women 375 +/- 136 (p less than 0.05).

摘要

本文描述了一种使用特异性抗血清测定尿液中6-酮-前列腺素F1α（前列环素的稳定代谢产物）的放射免疫分析法。所用抗体与其他前列腺素无明显交叉反应。（3H）-6-酮-前列腺素F1α用作示踪剂，30%的羟基磷灰石悬浮液用于分离结合和游离部分。标准曲线范围为2至1000 pg，因此可用于尿液测量。将0.2至1 ml的24小时尿液样本等分试样用己烷进行预萃取步骤，以去除中性脂肪。用0.1 N盐酸酸化至pH = 3.5 - 4后，进行乙醚萃取步骤，回收率为66%。对大鼠尿液池进行重复分析，结果显示批内变异系数为7.7%，批间变异系数为9.8%。正常大鼠的测定值为38.3±4.6 ng/24 h，5%氯化钠负荷大鼠的测定值为133±32 ng/24 h（p<0.001）。健康男性的测定值为508±94 ng/24 h，女性为375±136（p<0.05）。