Covance Laboratories, Inc., 8211 SciCor Drive, Indianapolis, IN, 46214, USA.
Covance Laboratories, Inc., 3301 Kinsman Blvd, Madison, WI, 53704, USA.
J Pharm Biomed Anal. 2021 May 10;198:114028. doi: 10.1016/j.jpba.2021.114028. Epub 2021 Mar 16.
Cortisol is a steroid hormone that is frequently measured as a marker of stress, inflammation, and immune function. While commonly analyzed in saliva, hair, blood plasma and urine, a recent trend towards whole blood-based at-home collection devices has emerged, which necessitates development of more sensitive assays for cortisol in whole blood. To support the implementation of a patient-centric sampling approach in a drug development program, a fit-for-purpose surrogate analyte-based liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for cortisol in whole blood was developed using C-cortisol as a surrogate analyte and cortisol-d6 as the internal standard. The surrogate analyte approach was chosen due to a lack of available cortisol-free whole blood and the absence of appropriately representative surrogate matrices. Samples were prepared using supported liquid extraction, and the LC-MS/MS analysis consisted of a 4.00 min analytical run. The method demonstrated linearity between 0.500 and 500 ng/mL of C-cortisol, and accuracy, precision and robustness were all acceptable per current regulatory guidance for bioanalytical method validation of chromatographic assays for cortisol- and C-cortisol-based quality control (QC) samples when quantified against a C-cortisol calibration curve. The acceptable robustness of cortisol-based QCs when quantified against a C-cortisol-based calibration curve also suggests parallelism between the analytes. These results indicate a viable surrogate analyte method, that is fit-for-purpose to analyze whole blood cortisol levels using a surrogate analyte LC-MS/MS approach. Evaluation of patient samples showed very promising comparability between whole blood and plasma cortisol concentrations, suggesting that whole blood could be used in place of or in addition to a plasma-based sampling protocol in clinical trials analyzing cortisol. Overall, this method presents a novel tool that is a first step in supporting the trend towards sample miniaturization and at-home sample collection, and may be readily used in clinical and diagnostic settings.
皮质醇是一种类固醇激素，常被作为应激、炎症和免疫功能的标志物进行测量。虽然通常在唾液、头发、血浆和尿液中进行分析，但最近出现了一种基于全血的家庭采集设备的趋势，这就需要开发更灵敏的全血皮质醇检测方法。为了支持药物开发项目中以患者为中心的采样方法的实施，我们开发了一种基于合适的替代分析物的适用于全血的液相色谱-串联质谱（LC-MS/MS）皮质醇检测方法，该方法以 C-皮质醇作为替代分析物，皮质醇-d6 作为内标。由于缺乏可用的无皮质醇全血以及缺乏具有代表性的替代基质，因此选择了替代分析物方法。样品采用支持液萃取法进行预处理，LC-MS/MS 分析包括 4.00 分钟的分析运行。该方法在 0.500-500ng/mL 的 C-皮质醇范围内显示出线性关系，并且根据当前用于基于皮质醇和 C-皮质醇的质量控制（QC）样品的色谱测定的生物分析方法验证的监管指南，准确性、精密度和稳健性均可接受。当使用 C-皮质醇校准曲线定量时，基于皮质醇的 QC 的可接受稳健性也表明了分析物之间的平行性。这些结果表明，该方法是一种可行的替代分析物方法，适用于使用替代分析物 LC-MS/MS 方法分析全血皮质醇水平。对患者样本的评估表明，全血和血浆皮质醇浓度之间具有非常有前景的可比性，这表明在分析皮质醇的临床试验中，全血可以替代或补充基于血浆的采样方案。总的来说，这种方法提供了一种新颖的工具，是支持样品微型化和家庭采集趋势的第一步，并且可能在临床和诊断环境中得到广泛应用。
