Hamilton C L, Kirkwood J A
Lilly Laboratory for Clinical Research, Indianapolis, IN 46202.
J Chromatogr B Biomed Appl. 1994 Apr 1;654(2):297-303. doi: 10.1016/0378-4347(93)e0447-x.
A reversed-phase, column-switching high-performance liquid chromatographic (HPLC) method is described for the determination of a new thymidylate synthase inhibitor in human plasma. The compound and an internal standard are extracted from plasma using a Certify II solid-phase cartridge. Extracts are evaporated to dryness and the residue is reconstituted with mobile phase buffer. The analytes are separated from polar interferences and buffer salts originating from the elution step on a 4-mm YMC Basic pre-column. The fraction containing the analytes is further separated on a 25-cm YMC Basic column. The analytes are detected by their absorbance at 250 nm. The limit of quantitation is 10 ng/ml. The method is linear from 10 ng/ml to 80 micrograms/ml using three standard curve ranges. Validation studies for all three ranges show the method to be reproducible. The method has been successfully used to support pharmacokinetic studies.
描述了一种反相柱切换高效液相色谱（HPLC）法，用于测定人血浆中一种新型胸苷酸合成酶抑制剂。使用Certify II固相萃取柱从血浆中提取该化合物和内标物。提取物蒸发至干，残留物用流动相缓冲液复溶。在4 mm YMC Basic预柱上，分析物与来自洗脱步骤的极性干扰物和缓冲盐分离。含有分析物的馏分在25 cm YMC Basic柱上进一步分离。通过在250 nm处的吸光度检测分析物。定量限为10 ng/ml。该方法在10 ng/ml至80 μg/ml范围内使用三个标准曲线范围呈线性。对所有三个范围的验证研究表明该方法具有可重复性。该方法已成功用于支持药代动力学研究。
