Fougère Louise, Moreau Hortense, Mirande-Bret Cécile, Fouillen Laetitia, Boutté Yohann
Univ. Bordeaux, CNRS, Laboratoire de Biogenèse Membranaire, UMR 5200, F-3314015 Villenave d'Ornon, France.
Bio Protoc. 2025 Sep 5;15(17):e5434. doi: 10.21769/BioProtoc.5434.
Phospholipids are major structural and regulatory elements of biological membranes and are involved in many different cellular and physiological processes. In this protocol, we provide an easy, cost-effective, and efficient method to obtain an overview of the phospholipid composition using high-performance thin layer chromatography (HPTLC). While the currently known phospholipid separation methods based on HPTLC display co-migration of certain lipid classes, the method we describe here allows the separation of all phospholipid classes, including anionic phospholipids in plant samples. This protocol combines elements of the classical Vitiello and Touchstone solvent systems to optimize phospholipid separation in a scaled pattern. Here, we provide a full characterization of this method, including statistical analyses of the retention factor of each phospholipid to show the robustness of the method and its efficiency in separating all phospholipid classes of a biological sample. Key features • Analysis of phospholipid composition through an easy, fast, robust, and cost-effective HPTLC method. • Separation of anionic phospholipids from the other phospholipid species. • Full overview of all phospholipids categories, including anionic phospholipids. • Qualitative and quantitative approach.
磷脂是生物膜的主要结构和调节成分，参与许多不同的细胞和生理过程。在本方案中，我们提供了一种简单、经济高效的方法，使用高效薄层色谱（HPTLC）来全面了解磷脂组成。虽然目前已知的基于HPTLC的磷脂分离方法存在某些脂质类别共迁移的情况，但我们在此描述的方法能够分离所有磷脂类别，包括植物样品中的阴离子磷脂。本方案结合了经典的维泰洛（Vitiello）和托奇斯通（Touchstone）溶剂系统的元素，以规模化模式优化磷脂分离。在此，我们对该方法进行了全面表征，包括对每种磷脂的保留因子进行统计分析，以展示该方法的稳健性及其在分离生物样品中所有磷脂类别的效率。关键特性• 通过简单、快速、稳健且经济高效的HPTLC方法分析磷脂组成。• 将阴离子磷脂与其他磷脂种类分离。• 全面了解所有磷脂类别，包括阴离子磷脂。• 定性和定量方法。
