Nichols J H, Charlson J R, Lawson G M
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN 55905.
Clin Chem. 1994 Jul;40(7 Pt 1):1312-6.
This automated assay determines the concentration of the antidepressant fluoxetine (Prozac) and its active metabolite norfluoxetine in serum by reversed-phase HPLC with spectrophotometric detection. Extraction, injection, and quantification are performed by the Gilson Aspec automated sample handler. The patient's specimen, with added protriptyline as internal standard, is extracted with solid-phase and liquid-liquid methods. Separation is conducted isocratically on a 5-microns (particle size) Supelcosil LC-8-DB reversed-phase column with a triethylamine acetate:acetonitrile mobile phase. The detection limit is 10 micrograms/L and absorbance varies linearly with concentration between 20 and 1,000 micrograms/L for both compounds. Mean recovery was 62% for fluoxetine and 70% for norfluoxetine over linear limits. Within-run and day-to-day imprecision (CV), evaluated at three concentrations (50, 200, and 500 micrograms/L), ranged from 2% to 7%. An extensive interference study of 108 drugs was conducted. Results (n = 58) by the automated method (y) correlated well with those by a manual HPLC method (x): y = 0.96x + 10.20 (r = 0.951, Sylx = 42.9) for fluoxetine, and y = 0.95x - 1.37 (r = 0.917, Sylx = 47.2) for norfluoxetine.
这种自动化检测方法通过反相高效液相色谱法（HPLC）和分光光度检测法来测定血清中抗抑郁药氟西汀（百忧解）及其活性代谢物去甲氟西汀的浓度。提取、进样和定量分析由吉尔森Aspec自动进样器完成。患者样本加入丙咪嗪作为内标，采用固相和液液萃取方法进行提取。在一根5微米（粒径）的Supelcosil LC - 8 - DB反相柱上，以三乙胺醋酸盐：乙腈为流动相进行等度分离。两种化合物的检测限均为10微克/升，在20至1000微克/升浓度范围内吸光度与浓度呈线性关系。在该线性范围内，氟西汀的平均回收率为62%，去甲氟西汀为70%。在三个浓度（50、200和500微克/升）下评估的批内和日间不精密度（CV）范围为2%至7%。对108种药物进行了广泛的干扰研究。自动化方法（y）的结果（n = 58）与手动HPLC方法（x）的结果相关性良好：氟西汀的相关性为y = 0.96x + 10.20（r = 0.951，Sylx = 42.9），去甲氟西汀的相关性为y = 0.95x - 1.37（r = 0.917，Sylx = 47.2）。
