HPLC-ECD测定人血及尿中儿茶酚胺浓度

Determination of catecolamies in plasma and urine by HPLC-ECD

目的建立简便快速准确测定体内儿茶酚胺的方法，为临床检测和研究提供技术支持。方法血浆和尿样经Al2O3吸附，净化，0．2mol·L－1HCl洗脱，以C8为固定相，磷酸缓冲液（pH2．7）、200mg·L－1SDS、10mg·L－1EDTA、6％乙晴为流动相，分离去甲肾上腺素（NE）肾上腺素（E）、多巴胺（DA），电化学检测，工作电压＋0．75V，灵敏度4nA，以外标法定量。结果NE，E，DA分离良好，且不受体内其他内源性物质及代谢物干扰。绝对回收率血浆为（71．58±5．57）％，尿样为（75．65±6．22）％。血浆相对回收率为（95．83±4．98）％，尿样为（101．5±7．17）％，血样线性范围0．1～20μg·L-1，尿样0．5～80μg·L-1，最低检测浓度0．05μg·L－1。结论操作、分析简便快速，专一性强，无干扰，重复性较好，适合于儿茶酚胺的临床测定与研究。

Purpose To establish a rapid,simple and specific HPLC method with electrochedrical detection for the determination of free catecholamines (norepinephnine NE,epinephrine E,dopamine DA)in plasma and urine. Methods The catecholamines were extrated from plasma and urine by alumina absorption,sample was separated on shimpack CKC-C8 cloumn with a moblie phase of water (containing 0.08mol· L-1 Na2H2PO4 pH 2. 7, 200mg· L-1 SDS and 10mg· L-1 EDTA) -acetonitrile(94: 6). The detection potentials were setat+0. 75V,an Ag/AgCl reference electrode and glass calbon working elecfrode at 4n A full scale. Results The separation of NE,E,DA was highly satisfactory. The plasma recoveries of added CSs ranged from 89. 9 ~ 105. 7 % (mean 95. 83 ± 4. 98%), and urine ranged from 91. 7 ~ 112. 8% (mean 101. 5±7. 71%). The linearrty was from 0. 1~20μg. L-1 for plasma from 0. 5~80μg· L-1 for urine. The detection Limits are 0. 05μg· L-1,0.1μg· L-1,0. 1μg· L-1 for NE,E,DA. Conclusions This method simple and good reappear,The method could be in the deterrmination of plasma and urine concentration in clinical and study.