逆相C18シリカカラムを用いた高感度Radioimmunoassayによる血漿8-arginine vasopressinの測定法
逆相C18シリカカラムを用いた高感度Radioimmunoassayによる血漿8-arginine vasopressinの測定法
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使用反相C18硅胶柱的高灵敏度放射免疫测定法测定血浆8-精氨酸加压素的方法
DOI：
10.1507/endocrine1927.61.7_724
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
K. Shizume
中科院分区：
文献类型：
--
作者：
Hyoichiro Sakurai;Fumihiko Kurimoto;Hideto Ohono;Akira Kanai;Kaoru Nomura;Hiroshi Demura;K. Shizume
建立了一种测定人血浆中8-精氨酸抗利尿素（AVP）的高灵敏度、简便的放射免疫测定方法。剂量响应曲线范围为0.025 ~ 8pg /支。采用ODS C18色谱柱提取AVP，加入0.5 ml血浆中，AVP回收率为87.1 +/- 10.4（平均+/- SD）%，范围为1- 10pg /ml。通过Sephadex G25 （1 X 25 cm）凝胶过滤，测定血浆中AVP的各个部分，发现血浆中AVP的部分与真实的AVP重叠，非特异性物质的干扰被ODS C18柱完全消除。16例尿囊症（DI）患者中有13例血浆AVP浓度在0.03 ~ 0.21 pg/ml之间，另外3例低于0.03 pg/ml。DI的AVP浓度与正常人明显不同（0.30 ~ 4.20 pg/ml, n = 65）。测定内部和测定之间的变异性各约为10%。正常受试者（n = 6）站着、坐着和仰卧在剥夺液体一整晚后的血浆AVP浓度（平均+/- SD）分别为2.41 +/- 1.15、1.95 +/- 0.85和0.97 +/- 0.48 pg/ml （30 min后）。正常受试者（n = 6）在水负荷（20 ml/kg wt）后血浆AVP浓度明显从1.89 +/- 1.00（前）降至0.42 +/- 0.21（站立60分钟），也从0.89 +/- 0.41降至0.40 +/- 0.22 pg/ml（仰卧60分钟）。
: A highly sensitive and simple radioimmunoassay for the measurement of 8-arginine vasopressin (AVP) in human plasma has been developed. The dose response curve ranges from 0.025 to 8 pg/tube. This simple extraction technique employing an ODS C18 column recovered 87.1 +/- 10.4 (mean +/- SD)% of AVP in the range of 1-10 pg/ml added to 0.5 ml plasma. Determination of AVP in each fraction of plasma, which was gel-filtrated through a Sephadex G25 (1 X 25 cm), revealed that the fraction of plasma AVP was superimposed on that of authentic AVP, and interference of non-specific substances was completely eliminated by an ODS C18 column. Using the assay, 13 of 16 patients with diabetes insipidus (DI) showed plasma AVP concentrations ranging from 0.03 to 0.21 pg/ml, and the other 3 patients had less than 0.03 pg/ml. The AVP concentrations of DI were clearly distinguished from those obtained in normal subjects (0.30-4.20 pg/ml, n = 65). The within and between assay variability was about 10% each. Plasma AVP concentrations (mean +/- SD) of normal subjects (n = 6) standing, sitting, and supine after an overnight of fluid deprivation were 2.41 +/- 1.15, 1.95 +/- 0.85 and 0.97 +/- 0.48 pg/ml (30 min after) respectively. Plasma AVP concentrations of normal subjects (n = 6) after water load (20 ml/kg wt) were clearly reduced from 1.89 +/- 1.00 (before) to 0.42 +/- 0.21 (standing for 60 min) and also from 0.89 +/- 0.41 to 0.40 +/- 0.22 pg/ml (supine for 60 min).