微流液相色谱-质谱法定量测定容积有限的人体血浆中的氧脂类物质

IF 3 3区 生物学 Q2 BIOCHEMICAL RESEARCH METHODS
Bingshu He, Rawi Ramautar, Marian Beekman, P Eline Slagboom, Amy Harms, Thomas Hankemeier
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引用次数: 0

摘要

氧脂素是众所周知的各种炎症中的脂质介质。它们的内源性浓度从低皮摩尔到纳摩尔不等,因此越来越多的病理研究需要测定它们在低容量基质中的浓度,包括血液、动物疾病模型的脑脊液、婴儿和微型采样设备。大多数已发表的全面分析草脂素的定量方法仍然需要 50 µL 以上的血浆作为起始容量,才能检测到这些低水平的草脂素。我们的研究旨在开发一种灵敏可靠的方法,用于定量检测受体积限制的人体血浆样本中的氧脂素。我们建立并验证了一种微液相色谱-质谱/MS 方法,该方法只需 5 µL 人体血浆即可测定 66 种草脂素。优化后的微液相色谱-质谱/质谱方法采用内径为 0.3 毫米的色谱柱,流速为 4 微升/分钟。该方法的进样量为 3 µL,检出限为 0.1 至 91.9 pM,定量限为 0.3 至 306.2 pM。根据 51 种化合物的物理化学性质,与传统流动相比较,灵敏度提高了 1.4 至 180.7 倍。经过验证后,该方法被用于分析一项健康老龄化研究中的 40 份血浆样本,以证明其稳健性和灵敏度。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A Micro-Flow Liquid Chromatography-Mass Spectrometry Method for the Quantification of Oxylipins in Volume-Limited Human Plasma.

Oxylipins are well-known lipid mediators in various inflammatory conditions. Their endogenous concentrations range from low picomolar to nanomolar, and there are growing demands to determine their concentrations in low-volume matrices for pathological studies, including blood, cerebrospinal fluids from animal disease models, infants, and microsampling devices. Most of the published quantification methods for comprehensive profiling of oxylipins still require more than 50 µL plasma as a starting volume to detect these low levels. The aim of our study is to develop a sensitive and reliable method for the quantification of oxylipins in volume-limited human plasma samples. We established and validated a micro-liquid chromatography (LC)-mass spectrometry (MS)/MS method that requires only 5 µL of human plasma for the determination of 66 oxylipins. The optimized micro-LC-MS/MS method utilized a flow rate of 4 µL/min with a 0.3-mm inner diameter column. With an injection volume of 3 µL, our method provides limits of detection in the range from 0.1 to 91.9 pM, and limits of quantification range from 0.3 to 306.2 pM. The sensitivity enhancement compared to conventional flow ranged from 1.4 to 180.7 times for 51 compounds depending on their physical-chemical properties. After validation, the method was applied to analyze 40 plasma samples from a healthy aging study to demonstrate robustness and sensitivity.

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来源期刊
ELECTROPHORESIS
ELECTROPHORESIS 生物-分析化学
CiteScore
6.30
自引率
13.80%
发文量
244
审稿时长
1.9 months
期刊介绍: ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.
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