A convenient method for the relative and absolute quantification of lipid components in liposomes by 1H- and 31P NMR-spectroscopy

IF 3.4 3区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemistry and Physics of Lipids Pub Date : 2024-04-12 DOI:10.1016/j.chemphyslip.2024.105395

Andreas H. Franz, Nataliya M. Samoshina, Vyacheslav V. Samoshin

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引用次数: 0

Abstract

Objective

Liposomes are promising delivery systems for pharmaceutical applications and have been used in medicine in the recent past. Preparation of liposomes requires reliable characterization and quantification of the phospholipid components for which the traditional cumbersome molybdate method is used frequently. The objective was to improve relative and absolute quantification of lipid components from liposomes.

Methods

A reliable method for quantification of lipid composition in liposome formulations in the 1–10 μmol range with ¹H- and ³¹P NMR spectroscopy at 600 MHz has been developed. The method is based on three crystalline small-molecule standards (Ph₃PO₄, (Tol)₃PO₄, and Ph₃PO) in CDCl₃.

Results

Excellent calibration linearity and chemical stability of the standards was observed. The method was tested in blind fashion on liposomes containing POPC, PEG-ceramide and a pH-sensitive trans-aminocyclohexanol-based amphiphile (TACH).¹ Relative quantification (percentage of components) as well as determination of absolute lipid amount was possible with excellent reproducibility with an average error of 5 %. Quantification (triplicate) was accomplished in 15 min based on ¹H NMR and in 1 h based on ³¹P NMR. Very little change in mixture composition was observed over multiple preparative steps.

Conclusion

Liposome preparations containing POPC, POPE, DOPC, DPPC, TACH, and PEG-ceramide can be reliably characterized and quantified by ¹H NMR and ³¹P NMR spectroscopy at 600 MHz in the μmol range.

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利用 1H 和 31P NMR 光谱对脂质体中的脂质成分进行相对和绝对定量的简便方法

目的脂质体是一种前景广阔的药物输送系统，近年来已被用于医药领域。脂质体的制备需要对磷脂成分进行可靠的表征和定量，而传统的钼酸盐法操作繁琐，因此被频繁使用。我们的目标是改进脂质体中脂质成分的相对和绝对定量方法。我们开发了一种可靠的方法，利用 600 MHz 的 1H- 和 31P NMR 光谱定量脂质体制剂中 1-10 μmol 范围内的脂质成分。该方法基于 CDCl3 中的三种结晶小分子标准物质（Ph3PO4、(Tol)3PO4 和 Ph3PO）。该方法在含有 POPC、PEG-肉桂酰胺和对 pH 值敏感的反式氨基环己醇双亲化合物（TACH）1 的脂质体上进行了盲法测试。利用 1H NMR 和 31P NMR 分别在 15 分钟和 1 小时内完成了定量（一式三份）。结论含有 POPC、POPE、DOPC、DPPC、TACH 和 PEG-纤维酰胺的脂质体制剂可通过 600 MHz 的 1H NMR 和 31P NMR 光谱在 μmol 范围内进行可靠的表征和定量。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Chemistry and Physics of Lipids 生物-生化与分子生物学

CiteScore

7.60

自引率

2.90%

发文量

审稿时长

40 days

期刊介绍： Chemistry and Physics of Lipids publishes research papers and review articles on chemical and physical aspects of lipids with primary emphasis on the relationship of these properties to biological functions and to biomedical applications. Accordingly, the journal covers: advances in synthetic and analytical lipid methodology; mass-spectrometry of lipids; chemical and physical characterisation of isolated structures; thermodynamics, phase behaviour, topology and dynamics of lipid assemblies; physicochemical studies into lipid-lipid and lipid-protein interactions in lipoproteins and in natural and model membranes; movement of lipids within, across and between membranes; intracellular lipid transfer; structure-function relationships and the nature of lipid-derived second messengers; chemical, physical and functional alterations of lipids induced by free radicals; enzymatic and non-enzymatic mechanisms of lipid peroxidation in cells, tissues, biofluids; oxidative lipidomics; and the role of lipids in the regulation of membrane-dependent biological processes.