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Analysis of Lipid Adsorption on Nanoparticles by Nanoflow Liquid Chromatography-Tandem Mass Spectrometry

机译:纳流液相色谱-串联质谱法分析纳米颗粒上的脂质吸附

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摘要

Nanoparticles (NPs) tend to adsorb matrix molecules like proteins and lipids incubated with biological fluids, forming a biological corona. While the formation and functions of protein corona have been studied extensively, little attention has been paid to lipid adsorption on NPs. However, lipids are also abundantly present in biological fluids and play important roles in processes like cell signaling and angiogenesis. Therefore, in this study, we established the analytical procedure for study of lipid adsorption on three different types of NPs in two matrices: human serum and heavy cream, using nanoflow liquid chromatography-mass spectrometry (nanoflowLC-MS). Serum was chosen to represent the common environment the NPs would be present once entering human body; and heavy cream was the representative food matrix NPs may be added to improve the color or taste. Steps of liquid-liquid extraction were established and optimized to achieve maximum recovery of the adsorbed, standard lipids from the NPs. Then, the LC-MS/MS method was developed to attain base-line separation of the standard lipids that represent the major lipid classes. At last, the lipid adsorption profiles of the three NPs were compared. We found that the lipid adsorption profile on the same type of NP was significantly different between the two matrices. The established method will help us investigate lipid adsorption on additional NPs and reveal how it could be affected by the physiochemical properties of NPs and the presence of proteins and other components in the biological matrix.
机译:纳米粒子(NPs)倾向于吸附与生物流体一起孵育的蛋白质和脂质等基质分子,从而形成生物电晕。虽然已经广泛研究了蛋白质电晕的形成和功能,但对脂类在NP上的吸附却很少关注。然而,脂质也大量存在于生物流体中,并在诸如细胞信号传导和血管生成的过程中起重要作用。因此,在这项研究中,我们使用纳流液相色谱-质谱法(nanoflowLC-MS)建立了用于研究脂质在两种基质(人血清和浓乳膏)中三种不同类型NP上吸附的分析程序。选择血清来代表NP进入人体后将存在的常见环境。可以添加重奶油作为代表的食品基质NP,以改善颜色或味道。建立并优化了液-液萃取步骤,以实现从NP中最大程度地回收吸附的标准脂质。然后,开发了LC-MS / MS方法以实现代表主要脂质类别的标准脂质的基线分离。最后,比较了三个NP的脂质吸附曲线。我们发现,在两种基质中,相同类型的NP上的脂质吸附曲线显着不同。既定的方法将帮助我们研究脂质对其他NP的吸附,并揭示它如何受到NP的理化特性以及生物基质中蛋白质和其他成分的存在的影响。

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