阴离子固相萃取结合高效液相色谱技术检测尿儿茶酚胺

Extraction of Urinary Catecholamines Using Anion Solid Phase Extraction Column and High Performance Liquid Chromatography

目的对阴离子固相萃取技术在尿儿茶酚胺（包括NE，E，DA）提取过程中的应用进行了研究，并在临床标本中应用。方法收集14例嗜铬细胞瘤患者和15例正常人24h尿液，利用强阴离子目相萃取柱萃取样本中的儿茶酚胺，用高效液相色谱与电化学检测仪联用测定其含量，对两组结果进行对比分析。结果NE，E和DA的回收率分别为93％～105％，94％～104％和89％～112％；批内变异系数分别为8．9％，8．4％和9．3％；批间变异系数分别为9．3％，9．6％和10．7％。在0．01-100mg／L的浓度范围内，线性良好（r=0．9999）。正常对照组的结果为NE28．67±11．98ug／24h，E4．08±2．34ug／24h，DA225．76±104．83ug／24h。嗜铬细胞瘤组的结果为NE300．29±212．79ug／24h，E11．76±7．58ug／24h，DA235．49±79．85ug／24h。嗜铬细胞瘤组的NE高于正常对照组（t=5．402，P〈0．01），嗜铬细胞瘤纽的E高于正常对照组（t=3．998，P〈0．01），嗜铬细胞瘸组的DA与正常对照组差异无统计学显著性意义（t=0．283，P=0．779）。结论该方法操作简便，结果可靠，优于酸性氧化铝提取儿茶酚胺方法，适合临床应用。

Objective The study has established a method that extraction urine catecholamine with anion solid phase extraction column,and used for clinical samples. Methods 24-hour urine of 14 patients with pheochromocytoma and 15 healthy subjects were collected. Extracted catecholamine from urine by anion solid phase extraction column and tested content by HPLC-ECD. Two groups of results were compared and analysed. Results The intra-assay CV,inter-assay CV and recovery of norepinephrine （NE） were 8.9%, 9.3% and 93%- 105% ;the intra-assay CV,inter-assay CV and recovery of epinephrine （E） were 8. 4% ,9.6% and 94%-104% ; the intra-assay CV,inter-assay CV and recovery of dopamine （DA） were 9.3% ,10.7 % and 89%-112%. Good linearity in the range of 0. 01-100 mg/L（r=0. 999 9）. The contents of group that patients with pheochromocytoma were （NE） 300. 29±212. 79 ug/24h, （E） 11.76±7.58 ug/24h and （DA） 235.49±79. 85 ug/24h. The contents of group that healthy subjects were （NE） 28.67±11.98 ug/24h, （E） 4. 08±2.34 ug/24h and （DA） 225.76±104.83 ug/24h. （NE） of pheochromoeytoma was high than healthy subjects （t = 5. 402,P〈 0.01）. （E） of pheochromocytoma was high than healthy subjects （t= 3. 998, P〈0.01）. （DA） of pheochromoeytoma and healthy subiects with no significant different （t=0. 283,P=0. 779）. Conclusion This method is simple and result is reliable,better than the method that using acid alumina extraction of catecholamines from urine. It can be used for clinical applications.