Heinig Katja, Wirz Thomas
F. Hoffmann-La Roche Ltd., Pharma Research, Non-clinical Drug Safety, Drug Metabolism and Pharmacokinetics Department, Bioanalytical Section, Grenzacherstrasse 124, CH-4070 Basel, Switzerland.
Anal Chem. 2009 May 15;81(10):3705-13. doi: 10.1021/ac900422e.
A highly sensitive liquid chromatographic method with online solid-phase extraction (SPE) and tandem mass spectrometric detection was developed for the quantification of the synthetic peptide drug taspoglutide in human and animal plasma. Sample preparation consisted of simple protein precipitation or automated off-line SPE using mixed mode cation exchange material, optionally preceded by cleavage of potential antidrug antibody complexes. Excellent selectivity was obtained by a novel orthogonal column switching approach, employing hydrophilic interaction liquid chromatography (HILIC) for online SPE followed by chromatographic separation on a 300 A pore size C18 column. The use of a stable isotope labeled drug (C-terminal arginine containing six (13)C and four (15)N atoms) as internal standard improved the quality and ruggedness of the method. The lower limits of quantitation (LLOQ) in human and animal plasma were 10.0 and 50.0 pg/mL, respectively, extracting only 250 or 100 microL sample aliquots. Precisions and accuracies were below 15% CV and between 89% and 114%, respectively. The method was successfully employed to analyze samples from pharmacokinetic studies.
建立了一种采用在线固相萃取（SPE）和串联质谱检测的高灵敏度液相色谱法，用于定量测定人和动物血浆中的合成肽药物塔司谷氨酸。样品制备包括简单的蛋白质沉淀或使用混合模式阳离子交换材料的自动离线SPE，可选择在潜在的抗药抗体复合物裂解之前进行。通过一种新颖的正交柱切换方法获得了出色的选择性，该方法采用亲水作用液相色谱（HILIC）进行在线SPE，然后在孔径为300 Å的C18柱上进行色谱分离。使用稳定同位素标记药物（含有六个（13）C和四个（15）N原子的C末端精氨酸）作为内标提高了方法的质量和耐用性。人和动物血浆中的定量下限（LLOQ）分别为10.0和50.0 pg/mL，仅提取250或100 μL样品等分试样。精密度和准确度分别低于15% CV和在89%至114%之间。该方法成功用于分析药代动力学研究的样品。
