An HPLC method for determination of dopamine, dopamine-3-O- and dopamine-4-O-sulfate in plasma, urine and tissues.

高效液相色谱法测定血浆、尿液和组织中的多巴胺、3-O-多巴胺和4-O-硫酸多巴胺。

基本信息

批准号：
60870015
负责人：
YAMATODANI Atsushi
金额：
$ 4.42万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-60870015/
关键词：
DOPAMINE SULFATES CONJUGATED DOPAMINE DOPAMINE HPLC FLUOROMETRY TISSUE DISTRIBUTION 生体内動態イヌヒト

项目摘要

Circulating dopamine is mainly present in conjugated forms, and its sulfate esters are the predominant conjugates in human plasma. The physiological significance of these dopamine conjugates is unknown. For examination of this problem, a sensitive and specific method is required for their determination. Thus, we developed a sensitive and specific HPLC-fluorometric method for determination of dopamine, dopamine-3-O- (3-O) and dopamine-4-O-sulfate (4-O) in biological samples, and we applied the method to pharmacological and clinical studies as follows.Levels of dopamine-3-O-sulfate in the adrenal, kidneys, liver, small intestine and heart of the mongrel dogs were 1.17+0.55, 0.13+0.02, 0.05+0.02, 0.03+0.01 and 0.01+0.002 nmoles/g (Mean+S.D), respectively. The activities of phenolsulfotransferase, an enzyme responsible for dopamine-sulfate synthesis, and arylsulfatase, an enzyme responsible for desulfation of the dopamine sulfates, were found all the tissues examined in the dog. When the two isomers were infused into the dog vein, they disappeared from the blood with the half time of about 60 min, and only a small portion of the administered 4-O was desulfated to form dopamine. During the infusion, neither isomer had effects on blood pressure, heart rate or urine output.The level of 3-O in human plasma was found to be influenced by food intake, and to increase 10- to 20-fold after meals. The plasma levels of the two isomers on early morning were determined to be 16.8+1.9 for 3-O and 3.2+0.5 for 4-O (pmoles/ml). The levels in patients with essential hypertension was significantly higher than the normotensive controls. When dopamine or L-DOPA was administered orally to healthy men, a rapid and marked increase of the levels of dopamine sulfates were observed.

循环多巴胺主要以缀合物形式存在，其硫酸酯是人血浆中的主要缀合物。这些多巴胺缀合物的生理意义尚不清楚。为了检查这个问题，需要一种灵敏且特异的方法来确定它们。因此，我们开发了一种灵敏且特异的高效液相色谱荧光法来测定生物样品中的多巴胺、多巴胺-3-O- (3-O) 和多巴胺-4-O-硫酸盐 (4-O)，并应用该方法药理和临床研究如下。杂种犬肾上腺、肾脏、肝脏、小肠和心脏中3-O-硫酸多巴胺的水平为1.17±0.55，分别为 0.13+0.02、0.05+0.02、0.03+0.01 和 0.01+0.002 纳摩尔/克（平均值+标准差）。在狗的所有组织中都发现了苯酚磺基转移酶（一种负责多巴胺硫酸盐合成的酶）和芳基硫酸酯酶（一种负责多巴胺硫酸盐脱硫的酶）的活性。当这两种异构体被输注到狗静脉中时，它们在约60分钟的半衰期内从血液中消失，并且只有一小部分所施用的4-O脱硫形成多巴胺。在输注过程中，两种异构体均不会对血压、心率或尿量产生影响。人血浆中3-O的水平被发现受食物摄入的影响，饭后会增加10至20倍。清晨两种异构体的血浆水平测定为：3-O 为 16.8+1.9，4-O 为 3.2+0.5（皮摩尔/毫升）。原发性高血压患者的水平显着高于血压正常对照者。当健康男性口服多巴胺或左旋多巴时，观察到硫酸多巴胺水平迅速而显着增加。