Albiach-Delgado Abel, Cascant-Vilaplana Mari Merce, Pinilla-González Alejandro, Torrejón-Rodríguez Laura, Balas Laurence, Durand Thierry, Quintás Guillermo, Kuligowski Julia, Cernada María
Neonatal Research Group, Health Research Institute Hospital La Fe (IIS La Fe), Avenida Fernando Abril Martorell 106, 46026, Valencia, Spain; Servicio de Análisis de Vesículas Extracelulares (SAVE), Health Research Institute Hospital La Fe (IIS La Fe), Avda Fernando Abril Martorell 106, 46026, Valencia, Spain.
Neonatal Research Group, Health Research Institute Hospital La Fe (IIS La Fe), Avenida Fernando Abril Martorell 106, 46026, Valencia, Spain.
Anal Chim Acta. 2025 Oct 8;1370:344373. doi: 10.1016/j.aca.2025.344373. Epub 2025 Jun 30.
Conventional sampling methods for clinical analysis make use of plasma or serum and require large blood volumes, cold storage, and specialized handling. This renders them impractical and invasive, especially when dealing with vulnerable groups of patients and the analysis of low-abundant and unstable compounds such as oxylipins. Dried blood micro-sampling methods are designed for collecting minimally invasive small-volume blood samples (<100 μL), providing advantages in stability, handling, and storage. The aim of this study is to develop and compare micro-sampling approaches that can be readily implemented in clinical analysis circumventing the barriers of conventional sampling methods. (96) RESULTS: This study assessed the performance of Pre-Cut Dried Blood Spots (PCDBS) and Volumetric Absorptive Micro-Sampling (VAMS®) devices for determining oxylipins, using small-volume liquid whole blood sampling as reference. A Liquid Chromatography - tandem Mass Spectrometry method was successfully developed and validated. Both PCDBS and VAMS® were prepared using 30 μL of spiked blood and dried for 2 h before extraction and analysis. Optimal results were achieved by pre-treating devices with antioxidant before sample deposition and avoiding the internal standard to undergo evaporation steps during preparation of micro-sampling devices. Long-term storage effects were assessed over a three-month period, and the micro-sampling devices were tested on umbilical cord blood samples from 35 newborns. For PCDBS and VAMS®, eight and nine oxylipins, respectively, showed comparable results to those obtained from liquid whole blood samples with mean concentrations ranging between 0.7 and 6.3 nM (140) SIGNIFICANCE: This study is the first to analyse oxylipins in umbilical cord blood using VAMS® and PCDBS micro-sampling devices, demonstrating their effectiveness for quantification at nM concentrations. It highlights the impact of sample collection, storage, and internal standard handling on lipid profiles. Stability is critical and cold storage (-20 °C) is mandatory. Future research focusing on the standardization of protocols for comparability among laboratories and enhancing sensitivity with improved LC-MS/MS techniques to expand accessible oxylipins are needed. (75).
临床分析的传统采样方法使用血浆或血清，需要大量血液、冷藏和专门处理。这使得它们不切实际且具有侵入性，尤其是在处理弱势群体患者以及分析低丰度和不稳定化合物（如氧化脂质）时。干血微量采样方法旨在采集微创小体积血样（<100 μL），在稳定性、处理和储存方面具有优势。本研究的目的是开发和比较可在临床分析中轻松实施的微量采样方法，以规避传统采样方法的障碍。（96）结果：本研究评估了预切干血斑（PCDBS）和体积吸收微量采样（VAMS®）装置用于测定氧化脂质的性能，以小体积液体全血采样作为参考。成功开发并验证了一种液相色谱 - 串联质谱方法。PCDBS和VAMS®均使用30 μL加标血液制备，干燥2小时后进行提取和分析。在样品沉积前用抗氧化剂预处理装置，并避免内标在微量采样装置制备过程中经历蒸发步骤，可获得最佳结果。在三个月的时间内评估了长期储存效果，并在35名新生儿的脐带血样本上测试了微量采样装置。对于PCDBS和VAMS®，分别有8种和9种氧化脂质的结果与液体全血样本相当，平均浓度范围在0.7至6.3 nM之间（140）意义：本研究首次使用VAMS®和PCDBS微量采样装置分析脐带血中的氧化脂质，证明了它们在nM浓度下定量的有效性。它突出了样品采集、储存和内标处理对脂质谱的影响。稳定性至关重要，必须冷藏（-20°C）。未来需要开展研究，重点是实现实验室间可比性的方案标准化，并通过改进的液相色谱 - 串联质谱技术提高灵敏度，以扩大可检测的氧化脂质范围。（75）
