Qian M, O'Dwyer P J, Gallo J M
Department of Medical Oncology, Fox Chase Cancer Center, Philadelphia, PA 19111, USA.
J Chromatogr B Biomed Appl. 1995 Apr 21;666(2):307-14. doi: 10.1016/0378-4347(94)00589-w.
A reversed-phase isocratic high-performance liquid chromatographic method has been developed for the determination of AG-331, a novel thymidylate synthase inhibitor, in human serum and urine. The method involves a solid-phase extraction from C18 cartridges without addition of an internal standard. The methanol eluent is evaporated under nitrogen at 40 degrees C, and reconstituted in mobile phase, acetonitrile-water (35:65, v/v) containing 25 mM ammonium phosphate. Separation of AG-331 was obtained on a C18 column at a flow-rate of 1 ml/min. Chromatographic signals were monitored by a photodiode-array detector at a primary wavelength of 457 nm with a bandwidth of 4.8 nm. Standard curves are linear in the range of 22-2175 ng/ml in plasma and 44-2175 ng/ml in urine, respectively. The extraction recovery ranged from 92.9-102.4%. Intra-day coefficient of variation was less than 9.5%, and inter-day coefficient of variation was less than 14.3% for an AG-331 concentration of 44 ng/ml. This method has been used to characterize the pharmacokinetics of AG-331 in cancer patients as part of ongoing Phase I trials.
已开发出一种反相等度高效液相色谱法，用于测定人血清和尿液中新型胸苷酸合成酶抑制剂AG - 331。该方法包括从C18柱进行固相萃取，无需添加内标。甲醇洗脱液在40℃下用氮气蒸发，然后用流动相（含25 mM磷酸铵的乙腈 - 水，35:65，v/v）复溶。在C18柱上以1 ml/min的流速分离AG - 331。通过光电二极管阵列检测器在457 nm的主波长和4.8 nm的带宽下监测色谱信号。标准曲线在血浆中22 - 2175 ng/ml和尿液中44 - 2175 ng/ml范围内呈线性。萃取回收率在92.9% - 102.4%之间。对于44 ng/ml的AG - 331浓度，日内变异系数小于9.5%，日间变异系数小于14.3%。作为正在进行的I期试验的一部分，该方法已用于表征AG - 331在癌症患者中的药代动力学。
