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.

循环多巴胺主要以共轭形式存在，其硫酸盐酯是人血浆中的主要结合物。这些多巴胺结合物的生理意义尚不清楚。为了检查此问题，确定其确定需要一种敏感和特定的方法。因此，我们开发了一种灵敏的HPLC-氟化方法来确定多巴胺，多巴胺-3-O-（3-O）（3-O）和多巴胺-4-O-硫酸盐（4-O）（4-O）（4-O），并且我们将这种方法应用于药理学和临床研究中，如下所示，请参见多巴胺液和肾上腺素的水平。杂种狗的中分别为1.17+0.55、0.13+0.02、0.05+0.02、0.03+0.01和0.01+0.002 nmoles/g（平均+s.d）。苯酚硫代转移酶是一种负责多巴胺 - 硫酸盐合成的酶，而芳基硫酸酯酶（一种负责脱硫硫酸盐硫酸盐的酶）的活性被发现了狗中检查的所有组织。当将两个异构体注入狗静脉中时，它们从血液中消失了大约60分钟，只有一小部分的4-O溶液硫化以形成多巴胺。在输注过程中，异构体对血压，心率或尿液输出都没有影响。发现人血浆中的3-O水平受到食物摄入的影响，并在进餐后增加10至20倍。确定3-O的两个异构体的血浆水平为16.8+1.9，4-O（PMOLES/mL）为3.2+0.5。基本高血压患者的水平明显高于正常血压对照。当口服多巴胺或L-DOPA到健康男性时，观察到多巴胺硫酸盐水平的迅速增加。