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。

• 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

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.

循环多巴胺主要以结合形式存在，其硫酸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是未知的。为了检查这个问题，需要一种灵敏和特异的方法来测定它们。因此，我们建立了一种灵敏和特异的HPLC荧光法测定生物样品中多巴胺、多巴胺-3-O-（3-O）和多巴胺-4-O-硫酸盐（4-O），并将该方法应用于药理学和临床研究，结果如下：杂种狗肾上腺、肾脏、肝脏、小肠和心脏中多巴胺-3-O-硫酸盐的水平为1.17 ± 0.55，0.13 ± 0.02、0.05 ± 0.02、0.03 ± 0.01和0.01 ± 0.002纳摩尔/g（平均值± S.D）。在狗的所有检查组织中发现了酚磺基转移酶（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还原的酶）的活性。当这两种异构体注入狗静脉时，它们在约60分钟的半衰期内从血液中消失，并且只有一小部分给药的4-O被还原形成多巴胺。在输注过程中，这两种异构体对血压、心率或尿量都没有影响。人体血浆中的3-O水平被发现受食物摄入的影响，餐后增加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（pmole/ml）。原发性高血压患者血浆中的β-淀粉样蛋白水平显著高于正常血压对照组。当口服多巴胺或左旋多巴给健康男性时，观察到多巴胺硫酸盐水平迅速显著增加。

项目成果

T.Yamamoto: Jpn.J.Pharmacol.40Supple. 116 (1986)

T.Yamamoto：Jpn.J.Pharmacol.40Suple。

A.Yamatodani, T.Yamamoto and H.Wada: "A direct determination method for dopamine-3-O- and -4-O-sulfate in plasma, urine and tissue by HPLC." Jpn. J. Pharmacology. 41 suppl.35P (1987)

A.Yamatodani、T.Yamamoto 和 H.Wada：“通过 HPLC 直接测定血浆、尿液和组织中多巴胺-3-O-和-4-O-硫酸盐的方法。”

T.Yamamoto, A.Yamatodani, K.Hashizume, H.Fukuda and H.Wada: "Metabolic fate of dopamine-3-O- and -4-O-sulfate in the dog." Jpn. J. Pharmacology. 40 suppl.116P (1986)

T.Yamamoto、A.Yamatodani、K.Hashizume、H.Fukuda 和 H.Wada：“狗体内多巴胺-3-O-和-4-O-硫酸盐的代谢命运。”

A.Yamatodani, T.Yamamoto, K.Hashizume and H.Wada: "Determination method and physiological significance of sulfo-conjugated dopamines." Byoutaiseiri (Medicina Philosophica). 6. (1987)

A.Yamatodani、T.Yamamoto、K.Hashizume 和 H.Wada：“磺基共轭多巴胺的测定方法及其生理意义”。

A.Yamatodani: Jpn.J.Pharmacol.41Supple. 35 (1987)

A.Yamatodani：Jpn.J.Pharmacol.41Suple。