In recent years, there has been an increasing worldwide interest in the use of alternative sample preparation methods that are proceeded by separation techniques. Fused deposition modeling (FDM) is a 3D printing technique that is based the consecutive layering of softened/melted thermoplastic materials.

In this study, a group of natural steroids and sexual hormones – namely, aldosterone, cortisol, β-estradiol, testosterone, dihydrotestosterone, and synthetic methyltestosterone and betamethasone – were separated and determined using an optimized high-performance liquid chromatography coupled to mass spectrometry (LC–MS) method in positive ionization mode. 3D-printed sorbents were selected as the pre-concentration technique because they are generally low cost, fast, and simple to make and automate. Furthermore, the use of 3D-printed sorbents helps to minimize potential errors due to their repeatability and reproducibility, and their ability to eliminate carry over by using one printed sorbent for a single extraction of steroids from biological matrices.

The extraction procedure was optimized and the parameters influencing 3D-printed Layfomm 60^® based sorbent and LC–MS were studied, including the type of extraction solvent used, sorption and desorption times, temperature, and the salting-out effect. To demonstrate this method’s applicability for biological sample analysis, the SPME-LC–MS method was validated for its ability to simultaneously quantify endogenous steroids. This evaluation confirmed good linearity and an R² that was between 0.9970 and 0.9990. The recovery rates for human plasma samples were 86.34–93.6% for the studied steroids with intra- and inter-day RSDs of 1.44–7.42% and 1.44–9.46%, respectively. To our knowledge, this study is the first time that 3D-printed sorbents have been used to extract trace amounts of endogenous low-molecular-weight compounds, such as steroids, from biological samples, such as plasma.

近年来，全世界对通过分离技术进行的其他样品制备方法的使用越来越感兴趣。熔融沉积建模（FDM）是一种3D打印技术，该技术基于软化/熔融热塑性材料的连续分层。

在这项研究中，一组自然类固醇和性激素-醛固酮，皮质醇，β-雌二醇，睾丸激素，二氢睾丸激素以及合成的甲基睾丸激素和倍他米松被分离并使用优化的高效液相色谱-质谱联用测定（ LC-MS）方法在正电离模式下。选择3D打印的吸附剂作为预浓缩技术，因为它们通常成本低，速度快，易于制造和自动化。此外，使用3D打印的吸附剂有助于降低潜在的误差，这是由于它们的可重复性和可重复性，以及通过使用一种印刷的吸附剂从生物基质中一次提取类固醇来消除残留的能力。

提取过程进行优化，所述参数影响3D印刷Layfomm 60 ^®基于吸附剂和LC-MS进行了研究，包括所使用的提取溶剂的种类，吸附和解吸时间，温度和盐析的效果。为了证明该方法在生物样品分析中的适用性，对SPME-LC-MS方法同时定量内源性类固醇的能力进行了验证。该评估证实了良好的线性和R ^2。在0.9970和0.9990之间。研究的类固醇的人血浆样品的回收率为86.34–93.6％，日内和日间RSD分别为1.44–7.42％和1.44–9.46％。就我们所知，这项研究是首次使用3D打印的吸附剂从生物样品（例如血浆）中提取痕量内源性低分子量化合物（例如类固醇）。