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目的:建立高效液相色谱荧光法检测尿液中儿茶酚胺的浓度。方法:通过自制氧化铝固相萃取小柱对尿液样本预处理,以3,4-二羟基苄胺(DHBA)为内标,色谱柱为Agilent Zorbax SB-C_(18)(250mm×4.6mm,5μm);流动相为甲醇:0.02mol·L~(-1)磷酸二氢钾=2:98(含有0.2 mmol·L~(-1)乙二胺四乙酸二钠);流速0.9 ml·min~(-1);进样量:20μl,柱温:23℃;荧光检测器,λEx为286nm,λEm为318nm。结果:去甲肾上腺素、肾上腺素和多巴胺的线性范围分别为3.6～115.2,14.85～475.2和28.65～916.8 ng·ml~(-1),提取回收率约为80%左右,RSD<7%,且在避光条件下较稳定。结论:本法灵敏准确可靠,可用于测定尿液中儿茶酚胺浓度。
Objective: To establish a HPLC method for the determination of catecholamine in urine. Methods: Urine samples were pretreated with self-made alumina solid-phase extraction cartridges using 3,4-dihydroxybenzylamine (DHBA) as internal standard. The column was Agilent Zorbax SB-C 18 (250 mm × 4.6 mm , 5μm). The mobile phase consisted of methanol: 0.02mol·L -1 potassium dihydrogen phosphate = 2: 98 (containing 0.2 mmol·L -1 disodium ethylenediaminetetraacetate); the flow rate was 0.9 ml · min -1; injection volume: 20 μl, column temperature: 23 ° C .; fluorescence detector, λEx 286 nm, λ Em 318 nm. Results: The linear ranges of norepinephrine, epinephrine and dopamine ranged from 3.6 to 115.2, 14.85 to 475.2 and 28.65 to 916.8 ng · ml -1, respectively. The extraction recovery was about 80% with RSD <7% And in the dark conditions more stable. Conclusion: This method is sensitive and reliable, and can be used to determine the concentration of catecholamines in urine.