Olthof Anouk, de Kleijne Vera H, Boelen Anita, Heijboer Annemieke C
Endocrine Laboratory, Department of Laboratory Medicine, Amsterdam UMC, Location University of Amsterdam, Amsterdam Gastroenterology Endocrinology & Metabolism, Meibergdreef 9, Amsterdam 1105 AZ, the Netherlands; Endocrine Laboratory, Department of Laboratory Medicine, Amsterdam UMC, Location Vrije Universiteit Amsterdam, Amsterdam Gastroenterology Endocrinology & Metabolism, De Boelelaan 1117, Amsterdam 1081 HV, the Netherlands.
Endocrine Laboratory, Department of Laboratory Medicine, Amsterdam UMC, Location University of Amsterdam, Amsterdam Gastroenterology Endocrinology & Metabolism, Meibergdreef 9, Amsterdam 1105 AZ, the Netherlands; Endocrine Laboratory, Department of Laboratory Medicine, Amsterdam UMC, Location University of Amsterdam, Amsterdam Reproduction & Development, Meibergdreef 9, Amsterdam 1105 AZ, the Netherlands.
Clin Chim Acta. 2025 Feb 1;567:120099. doi: 10.1016/j.cca.2024.120099. Epub 2024 Dec 17.
An important aspect of the shift towards dried blood spots (DBS) as a sample matrix for laboratory measurements, is the availability of robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) methods that can reliably quantify analyte concentrations in DBS. The development and validation of these LC-MS/MS methods, however, concerns an extensive process, for which large amounts of DBS samples are required. DBS are usually obtained from capillary blood samples, but they can also be prepared from venous (residual) blood samples, which are widely available in clinical laboratories. Therefore, we aimed to determine whether DBS prepared from (residual) venous blood samples, collected in EDTA blood tubes, can be used for future development and validation of LC-MS/MS methods to quantify steroid hormones in DBS.
Capillary DBS and venous blood samples (EDTA tube and tube without additives) were collected from twenty healthy volunteers (12F/8M). From both venous blood samples, DBS were prepared volumetrically. Samples were analyzed using in-house developed LC-MS/MS methods for testosterone, androstenedione, 17-hydroxyprogesterone (17-OHP), cortisol, cortisone, corticosterone, and for dehydroepiandrosterone sulfate (DHEA-S).
DBS made from venous blood collected in EDTA tubes compared with capillary blood showed a correlation coefficient of ≥ 0.89 for all steroid hormones except corticosterone (0.67). DBS made from venous blood collected in tubes without additives showed a strong correlation with both DBS made from venous blood collected in EDTA tubes (≥0.97 for all steroid hormones) and capillary DBS (>0.90) except corticosterone (0.64).
DBS prepared from (residual) venous blood collected in EDTA blood tubes can be used for future development and validation of LC-MS/MS methods to quantify steroid hormones, except for corticosterone, in capillary DBS.
向将干血斑（DBS）用作实验室检测样本基质转变的一个重要方面，是要有可靠的液相色谱 - 串联质谱（LC - MS/MS）方法，该方法能够可靠地定量测定DBS中的分析物浓度。然而，这些LC - MS/MS方法的开发和验证涉及一个广泛的过程，为此需要大量的DBS样本。DBS通常从毛细血管血样中获取，但也可以从静脉（残余）血样中制备，而静脉（残余）血样在临床实验室中广泛可得。因此，我们旨在确定从收集于乙二胺四乙酸（EDTA）血样管中的（残余）静脉血样制备的DBS，是否可用于未来开发和验证LC - MS/MS方法以定量测定DBS中的甾体激素。
从20名健康志愿者（12名女性/8名男性）采集毛细血管DBS和静脉血样（EDTA管和无添加剂管）。从两份静脉血样中，按体积制备DBS。使用内部开发的LC - MS/MS方法分析样本中的睾酮、雄烯二酮、17 - 羟孕酮（17 - OHP）、皮质醇、可的松、皮质酮以及硫酸脱氢表雄酮（DHEA - S）。
与毛细血管血相比，由收集于EDTA管中的静脉血制备的DBS，除皮质酮（0.67）外，所有甾体激素的相关系数均≥0.89。由收集于无添加剂管中的静脉血制备的DBS，与由收集于EDTA管中的静脉血制备的DBS（所有甾体激素≥0.97）以及毛细血管DBS（>0.90）均显示出强相关性，但皮质酮除外（0.64）。
从收集于EDTA血样管中的（残余）静脉血制备的DBS，可用于未来开发和验证LC - MS/MS方法以定量测定毛细血管DBS中的甾体激素，但皮质酮除外。
