Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
Institute for Occupational and Social Medicine, Medical Faculty, RWTH Aachen University, Pauwelsstrasse 30, D-52074 Aachen, Germany.
Talanta. 2015 Mar;134:310-316. doi: 10.1016/j.talanta.2014.11.034. Epub 2014 Nov 28.
Hair analysis has been increasingly used to establish long-term biomarkers of exposure to both endogenous and exogenous substances, with a special emphasis on steroidal hormones. Hair cortisol and cortisone have been associated to physiological and psychological strains, anxiety and depression. Hair is a very complex matrix, which might jeopardize analyte detection at low concentrations. A new, highly selective and sensitive method based on fragments of second order, MS(3) (MS/MS/MS), was developed and validated for the analysis of hair cortisol and cortisone. An online solid phase extraction was performed on a C8 restricted access material (RAM) phase following by separation on a reversed-phase C18 column using methanol and 0.02% ammonium hydroxide as mobile phase. The developed method required minimal sample preparation and the injection of only 50 µL of sample leading to a LOQ of 2 pg mg(-1). Good linear responses were observed in the range 2-200 pg mg(-1) (R(2)>0.99) and extraction recoveries ranged between 77-125% and 70-123% for cortisol and cortisone, respectively. Intra- and inter-assay coefficients of variation were between 1.4 and 14%. In order to evaluate the applicability of the method, preliminary tests (N=33) were conducted in 3 cm hair samples (close to scalp) of healthy volunteers with an age range of 4-63. Average concentrations in hair were 12.7±14 pg mg(-1) and 41.6±42 pg mg(-1) for cortisol and cortisone, respectively. Further investigations on cortisol and cortisone as biomarkers for chronic psychological strain will be assessed as a next step.
头发分析越来越多地被用于建立内源性和外源性物质的长期生物标志物，特别强调甾体激素。头发皮质醇和皮质酮与生理和心理压力、焦虑和抑郁有关。头发是一种非常复杂的基质，这可能会危及低浓度分析物的检测。本研究建立了一种基于二级碎片（MS(3) (MS/MS/MS)）的新的、高度选择性和灵敏的方法，用于分析头发皮质醇和皮质酮。采用 C8 受限访问材料（RAM）进行在线固相萃取，然后在反相 C18 柱上用甲醇和 0.02%氨水溶液作为流动相进行分离。该方法需要的样品制备最少，只需注入 50 μL 样品，LOQ 为 2 pg mg(-1)。在 2-200 pg mg(-1)范围内观察到良好的线性响应（R(2)>0.99），皮质醇和皮质酮的提取回收率分别在 77-125%和 70-123%之间。内标和日间精密度的变异系数在 1.4-14%之间。为了评估该方法的适用性，在 33 名年龄在 4-63 岁的健康志愿者的 3 cm 头发样本（接近头皮）中进行了初步测试。头发中皮质醇和皮质酮的平均浓度分别为 12.7±14 pg mg(-1)和 41.6±42 pg mg(-1)。下一步将进一步研究皮质醇和皮质酮作为慢性心理压力的生物标志物。
