Total Lipid Extraction and TLC Analysis

Total lipids extraction and TLC analysis were performed according to the protocol described by Mu et al.^{23 (link)}. In brief, 10 mg of ENPs were mixed with equal volume of chloroform and methanol and centrifuged at 2000 rpm for 5 min at room temperature. The organic phase obtained was dried under a stream of nitrogen and suspended in chloroform. The extracted lipids were resolved through silica gel 60 F254 TLC plates (Merck) using chloroform/methanol/acetic acid (95:4.5:0.5, by volume) as a solvent system. Plates were dried at room temperature and sprayed with a solution containing 10% copper sulfate and 8% phosphoric acid. Lipid bands were visualized by charring the plates at 120 °C for 10 min.