Ha H R, Follath F
Department of Internal Medicine, University Hospital, Zurich, Switzerland.
Ther Drug Monit. 1994 Oct;16(5):499-503. doi: 10.1097/00007691-199410000-00010.
A rapid and selective high-performance liquid chromatographic (HPLC) method using solid-phase extraction (SPE) for measuring piroximone in plasma samples has been developed. Human plasma and internal standard were pipetted onto a Bond Elut C18 extraction cartridge conditioned with methanol and water. Impurities and proteins were washed out with water. Piroximone and internal standard were eluted with methanol. After evaporation, the residue was dissolved in the mobile phase and the aliquot injected into the HPLC system. Piroximone and its related compounds were separated on a reversed phase C18 HPLC column maintained at 50 degrees C using a mobile phase consisting of phosphate buffer and methanol. Piroximone-N-oxide, piroximone, and internal standard were detected spectrophotometrically at 340 nm. The extraction recovery for piroximone was 94%. The within- and between-run coefficients of variation were < 3% in the concentration range of 320-5,000 ng/ml and < 17.5% at lower concentrations, e.g., 20 ng/ml. The limit of detection was 10 ng/ml.
已开发出一种使用固相萃取（SPE）的快速、选择性高效液相色谱（HPLC）方法，用于测定血浆样品中的吡罗昔酮。将人血浆和内标移液至用甲醇和水预处理过的Bond Elut C18萃取柱上。用水洗去杂质和蛋白质。用甲醇洗脱吡罗昔酮和内标。蒸发后，将残渣溶解于流动相中，并取部分溶液注入HPLC系统。吡罗昔酮及其相关化合物在反相C18 HPLC柱上分离，该柱保持在50℃，使用由磷酸盐缓冲液和甲醇组成的流动相。吡罗昔酮 - N - 氧化物、吡罗昔酮和内标在340nm处用分光光度法检测。吡罗昔酮的萃取回收率为94%。在320 - 5000 ng/ml浓度范围内，批内和批间变异系数<3%，在较低浓度（如20 ng/ml）时<17.5%。检测限为10 ng/ml。
