Gruenstein Yehudah
Department of Biochemistry and Molecular Biology University of Miami Coral Gables Florida USA.
Anal Sci Adv. 2024 Nov 2;5(11-12):e202400035. doi: 10.1002/ansa.202400035. eCollection 2024 Dec.
Testosterone is a critical hormone involved in regulating various physiological processes in both men and women. Accurate testosterone measurement is essential for diagnosing endocrine disorders such as hypogonadism and polycystic ovary syndrome and for routine testing. Traditionally, testosterone levels are measured using serum or plasma samples, which present challenges in sample collection, storage, and transport, particularly in resource-limited settings. Dried blood spot (DBS) sampling has emerged as an effective alternative for hormone analysis, offering significant advantages in terms of sample stability, ease of collection, and simplified logistics. This study aimed to validate a DBS-based testosterone assay using liquid chromatography-tandem mass spectrometry (LC-MS/MS) to ensure accuracy and precision comparable to conventional serum-based methods. Drops of whole blood samples were collected from adult volunteers using a single-use safety lancet for finger pricks, with blood applied onto DBS cards (PerkinElmer 226 Spot Saver RUO card) for further analysis. The testosterone was extracted from DBS using a liquid-liquid method and analyzed with LC-MS/MS. The assay demonstrated excellent linearity across a wide concentration range (0.1-100 ng/mL) with a correlation coefficient (r) of 0.999 and achieved a lower limit of detection of 0.058 ng/mL and a lower limit of quantification of 0.086 ng/mL. The method showed high precision, with intra- and inter-day coefficients of variation below 10%, and satisfactory recovery rates. Hematocrit correction and matrix effect evaluations confirmed the robustness of the assay for clinical and research applications. Additionally, the assay displayed a strong clinical correlation between testosterone levels in DBS and venous serum samples, supporting its reliability for testosterone monitoring. This validation study supports that the DBS-based LC-MS/MS testosterone assay is a reliable tool for testosterone quantification for routine testing.
睾酮是一种关键激素，参与调节男性和女性的各种生理过程。准确测量睾酮对于诊断性腺功能减退和多囊卵巢综合征等内分泌疾病以及进行常规检测至关重要。传统上，睾酮水平是使用血清或血浆样本进行测量的，这在样本采集、储存和运输方面存在挑战，尤其是在资源有限的环境中。干血斑（DBS）采样已成为激素分析的一种有效替代方法，在样本稳定性、采集便利性和物流简化方面具有显著优势。本研究旨在验证一种基于DBS的睾酮检测方法，该方法使用液相色谱-串联质谱（LC-MS/MS），以确保其准确性和精密度与传统的基于血清的方法相当。使用一次性安全采血针从成年志愿者手指采血，将血液滴在DBS卡（珀金埃尔默226 Spot Saver RUO卡）上进行进一步分析。采用液-液法从DBS中提取睾酮，并使用LC-MS/MS进行分析。该检测方法在较宽的浓度范围（0.1 - 100 ng/mL）内显示出良好的线性，相关系数（r）为0.999，检测下限为0.058 ng/mL，定量下限为0.086 ng/mL。该方法具有很高的精密度，日内和日间变异系数均低于10%，回收率令人满意。血细胞比容校正和基质效应评估证实了该检测方法在临床和研究应用中的稳健性。此外，该检测方法显示DBS中的睾酮水平与静脉血清样本之间具有很强的临床相关性，支持其在睾酮监测方面的可靠性。这项验证研究支持基于DBS的LC-MS/MS睾酮检测方法是一种用于常规检测中睾酮定量的可靠工具。
