Department of Medical Oncology, Erasmus MC-Daniel den Hoed Cancer Center, University Medical Center, Rotterdam, The Netherlands.
J Pharm Biomed Anal. 2009 Dec 5;50(5):977-82. doi: 10.1016/j.jpba.2009.06.048. Epub 2009 Jul 7.
A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantitative determination of RBG-286638, a novel multi-targeted protein kinase inhibitor, in 200 microl aliquots of human potassium EDTA plasma with deuterated RGB-286638 as internal standard. The sample extraction and cleaning-up involved a simple liquid-liquid extraction with 100 microl aliquots of acetonitrile and 1 ml aliquots of n-butylchloride. Urine was accurately 5- and 10-fold diluted in blank plasma prior to extraction. Chromatographic separations were achieved on a reversed phase C18 column eluted at a flow-rate of 0.250 ml/min on a gradient of 0.2 mM ammonium formate and acetonitrile both acidified with 0.1% formic acid. The overall cycle time of the method was 7 min, with RGB-286638 eluting at 1.9 min. The multiple reaction monitoring transitions were set at 546>402 (m/z), and 549>402 (m/z) for RGB-286638 and the internal standard, respectively. The calibration curves were linear over the range of 2.00 to 1000 ng/ml with the lower limit of quantitation validated at 2.00 ng/ml. The within-run and between-run precisions were within 7.90%, while the accuracy ranged from 92.2% to 99.7%. The method was successfully applied to samples derived from a clinical study.
一种快速灵敏的液相色谱/串联质谱（LC-MS/MS）方法已经建立并验证，用于定量测定新型多靶点蛋白激酶抑制剂 RBG-286638，在 200 微升人钾 EDTA 血浆中，以氘代 RBG-286638 为内标。样品提取和净化涉及简单的液-液萃取，用 100 微升乙腈和 1 毫升正丁基氯提取。尿液在提取前用空白血浆准确稀释 5 倍和 10 倍。色谱分离在反相 C18 柱上进行，以 0.2 mM 甲酸铵和乙腈的梯度洗脱，流速为 0.250 ml/min，均用 0.1%甲酸酸化。该方法的总循环时间为 7 分钟，RBG-286638 在 1.9 分钟洗脱。多重反应监测的转换分别设定为 546>402（m/z）和 549>402（m/z），用于 RBG-286638 和内标。校准曲线在 2.00 至 1000 ng/ml 范围内呈线性，验证的定量下限为 2.00 ng/ml。批内和批间精密度均在 7.90%以内，而准确度范围为 92.2%至 99.7%。该方法成功应用于临床研究样本。
