摘要
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 simple and accuracy method for determine the concentration of catecholamines in urine by HPLC-FLD. Method： Samples were cleaned- up by alumina solid-phase extraction cartridge homemade,and DHBA was used as interhal standard. The solid phase was Agilent Zorbax SB-C18 （250 mm ×4.6mm,5μm） ,and the mobile phase was a mixture of methanol - 0.02mol. L- 1 Kalium Hydrogen Phospharate （contain 0.2 mmol·L^-1 EDTA-2Na）with a flow rate at 0.9 ml· min^- 1 and the temperature was 23℃. Result： The linear concentration range for norepinephrine was 3.6 - 115.2 ng· ml^-1, for epinephrine was 14.85 -475.2 ng· ml^-1 and for dopamine was 28.65 -916.8 ng· ml^-1. The average recovery was about 80% with both the intra-day RSD and the interday RSD at less than 7%. And all of catecholamines were very stable when protected from light. Conclusion： The method is high sensibility and accuracy, which can be used for concentration detecting of catecholamines and correlative diseases study at clinic.
出处
《中国药师》
CAS
2009年第9期1205-1207,共3页
China Pharmacist
