Lipid determination in bone marrow and mineralized bone tissue: From sample preparation to improved high-performance thin-layer and liquid chromatographic approaches

• Published in: Journal of Chromatography A 2017-09, Vol.1515, p.232-244
• Main Author: During, Alexandrine
• Format: Article
• Language: English
• Subjects: Bone marrow; Fatty acids; HP-TLC; HPLC; Mineralized tissue; Phospholipids
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2017.08.004

•Procedure to isolate the two bone marrow and mineralized tissue compartments.•A multi-one-dimension HP-TLC method to study the major polar and neutral lipids.•Demineralization facilitates phosphatidylserine and cholesterol ester extractions.•The mineralized tissue seems to be more metabolically active than the bone marrow. In view of their key roles in the bone physiology (e.g., in the biomineralization process) and their potential implication in bone pathologies, an approach to study lipids in situ is needed. The aim of the present paper is to propose an original procedure to characterize lipids in both bone marrow (BM) and mineralized tissue (MT) compartments, taking into consideration sample preparation, lipid extraction and analytical issues, when using small sample size (≤ 0.5g of rat femurs). The potential contamination of the MT by marrow lipids and the poor accessibility of certain lipids from the MT − two major issues in bone handling − were taking care, respectively by performing two cleaning steps after BM removal and by adding a demineralization step to the overall lipid extraction protocol. For lipid analyses, a multi-one-dimensional HP-TLC method was developed to analyze the major neutral and polar lipids at once and showed an excellent resolution (for 15 standards) and a good precision (inter-day RSD