Institute of Legal Medicine, University of Strasbourg, Strasbourg, France.
Anal Bioanal Chem. 2011 Sep;401(4):1153-62. doi: 10.1007/s00216-011-5172-3. Epub 2011 Jul 1.
Steroid hormone concentrations are mostly determined by using different body fluids as matrices and applying immunoassay techniques. However, usability of these approaches may be restricted for several reasons, including ethical barriers to invasive sampling. Therefore, we developed an ultra-performance LC-MS-MS method for high-throughput determination of concentrations of cortisol, cortisone, dehydroepiandrosterone (DHEA), and DHEA sulfate (DHEAS) in small quantities of human nails. The method was validated for linearity, limits of detection and quantification, recovery, intra and interassay precision, accuracy, and matrix effect. Samples from 10 adult women were analyzed to provide proof-of-principle for the method's applicability. Calibration curves were linear (r(2) > 0.999) in the ranges 10-5000 pg mg(-1) for cortisol, cortisone, and DHEAS, and 50-5000 pg mg(-1) for DHEA. Limits of quantification were 10 pg mg(-1) for cortisol, cortisone, and DHEAS, and 50 pg mg(-1) for DHEA. The sensitivity and specificity of the method were good, and there was no interference with the analytes. Mean recovery of cortisol, cortisone, DHEA, and DHEAS was 90.5%, 94.1%, 84.9%, and 95.9%, respectively, with good precision (coefficient of variation <14% for all analytes) and accuracy (relative error (%) -8.3% to 12.2% for all analytes). The median (pg mg(-1), range) hormone concentrations were 69.5 (36-158), 65 (32-133), 212 (50-1077), and 246 (115-547) for cortisol, cortisone, DHEA, and DHEAS, respectively. This method enables measurement of cortisol, cortisone, DHEA, and DHEAS in small quantities of human nails, leading to the development of applications in endocrinology and beyond.
类固醇激素浓度主要通过使用不同的体液作为基质并应用免疫测定技术来确定。然而，由于侵入性采样的伦理障碍等原因，这些方法的可用性可能会受到限制。因此，我们开发了一种超高效 LC-MS-MS 方法，用于高通量测定小量人指甲中皮质醇、皮质酮、脱氢表雄酮 (DHEA) 和硫酸脱氢表雄酮 (DHEAS) 的浓度。该方法经过线性、检测限和定量限、回收率、日内和日间精密度、准确性和基质效应的验证。分析了 10 名成年女性的样本，为该方法的适用性提供了原理证明。皮质醇、皮质酮和 DHEAS 的校准曲线在 10-5000 pg mg(-1) 范围内呈线性 (r(2) > 0.999)，DHEA 的校准曲线在 50-5000 pg mg(-1) 范围内呈线性。皮质醇、皮质酮和 DHEAS 的定量限为 10 pg mg(-1)，DHEA 的定量限为 50 pg mg(-1)。该方法的灵敏度和特异性良好，无分析物干扰。皮质醇、皮质酮、DHEA 和 DHEAS 的平均回收率分别为 90.5%、94.1%、84.9%和 95.9%，所有分析物的精密度均良好（变异系数 <14%），准确性（相对误差（%）-8.3%至 12.2%）良好。皮质醇、皮质酮、DHEA 和 DHEAS 的中位数 (pg mg(-1)，范围) 浓度分别为 69.5 (36-158)、65 (32-133)、212 (50-1077) 和 246 (115-547)。该方法能够测量小量人指甲中的皮质醇、皮质酮、DHEA 和 DHEAS，从而开发了内分泌学等领域的应用。
