Institute of Pharmaceutical Sciences, King's College London, Franklin-Wilkins Building, 150 Stamford Street, London SE1 9NH, United Kingdom; Waters Corporation, 34 Maple Street, Milford, MA 01757, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2013 Nov 1;938:96-104. doi: 10.1016/j.jchromb.2013.08.027. Epub 2013 Sep 3.
Teriparatide, the 1-34 fragment of human parathyroid hormone, is used to treat osteoporosis patients with a high risk of fracture by stimulating new bone formation. Routinely teriparatide is quantified using radioimmunoassay however the LC-MS/MS described here has the potential to achieve greater accuracy and precision, higher specificity, and is readily implemented in routine bioanalytical laboratories. Hence a complete method combining effective sample prep with appropriate LC separation and selected reaction monitoring (SRM) MS detection was developed to selectively separate teriparatide from closely related endogenous peptides and to reduce interferences. Samples were concentrated without evaporation, minimizing the risk of adsorptive losses. Chromatography was performed on a sub 2μm particle charged surface hybrid column, which provided significantly higher peak capacity than a traditional C18 column when formic acid was used as the mobile phase modifier. Total LC cycle time was 6min. An LOD of 15pg/mL (3.6fmol/mL) from 200μL of human plasma was readily achieved and standard curves were accurate and precise from 15pg/mL to 500pg/mL. Mean QC accuracies ranged from 90% to 106%. Mean QC precision was better than 7%. The CV of matrix factors across 6 sources of human plasma was 5%. The assay presented here is the first LC-MS method which reaches clinically relevant detection limits for teriparatide.
特立帕肽，甲状旁腺激素 1-34 片段，通过刺激新骨形成用于治疗骨折风险高的骨质疏松症患者。常规使用放射免疫法来定量特立帕肽，但本文所述的 LC-MS/MS 具有更高的准确性和精密度、更高的特异性，并易于在常规生物分析实验室中实施。因此，开发了一种完整的方法，将有效的样品制备与适当的 LC 分离和选择反应监测 (SRM) MS 检测相结合，以选择性地将特立帕肽与密切相关的内源性肽分离，并减少干扰。样品在不蒸发的情况下浓缩，最大限度地减少了吸附损失的风险。色谱在亚 2μm 颗粒带电表面混合柱上进行，当使用甲酸作为流动相改性剂时，与传统的 C18 柱相比，提供了更高的峰容量。从 200μL 人血浆中可轻松实现 15pg/mL（3.6fmol/mL）的检测限 (LOD)，并且标准曲线从 15pg/mL 到 500pg/mL 都准确且精密。QC 平均准确度范围在 90%到 106%之间。QC 平均精密度优于 7%。来自 6 个人源血浆来源的基质因子的 CV 为 5%。本文介绍的方法是第一个达到特立帕肽临床相关检测限的 LC-MS 方法。
