G60 f254 plates

The NMR spectra (¹H-NMR, ¹³C-NMR, DEPT 135°, COSY, HMQC, HMBC) of 8-prenylnarinenin were recorded at 500 MHz on a Bruker Ultra Shield TM Plus instrument in acetone-d₆. UV spectra (Cintra 303 spectrofotometer; GBC, Braeside, Australia) were recorded in methanol. HR ESI-MS was taken on a Bruker micrOTOF-Q spectrometer. Elemental analysis was done with an EA 1108, Carlo Erba analyzer. IR spectra were measured on a Mattson IR 300 Thermo-Nicolet spectrophotometer (Mattson, Warszawa, Poland).
Analytical TLC was carried out on silica gel G 60 F₂₅₄ plates (Merck, Darmstadt, Germany) with a mixture of CH₃Cl:MeOH in various ratios as developing systems. Compounds were detected by spraying the plates with 1% Ce(SO₄)₂ and 2% H₃[P(Mo₃O₁₀)₄] in 10% H₂SO₄, followed by heating to 120–200 °C. The product was separated by column chromatography using silica gel (SiO₂, Kieselgel 60, 230–400 mesh, 40–63 µm, Merck).

The total lipid analysis for TAG was carried out by thin layer chromatography (TLC) and Fourier-transform infrared (FTIR) spectroscopy. The chromatograms for TLC were developed by using G-60 F254 plates, Merck, India (0.25 mm thick silica gel) with hexane/diethyl ether/acetic acid (85:15:1, v/v/v) solvent system. Methanolic MnCl₂ solution (0.63 g MnCl₂·4H₂O, 60 ml water, 60 ml methanol, and 4 ml concentrated sulfuric acid) was sprayed over the plate and kept for charring at 120 °C for 10 min. For the detection of FAMEs by TLC, the samples were spotted on TLC plate and developed by the dual-solvent system, firstly with hexane/tert-butyl methyl ether/acetic acid (50:50:0.5, v/v/v), and then air dried, redeveloped to 8 cm from the origin with hexane/tert-butyl methyl ether/acetic acid (97:3:0.5, v/v/v). The spots were visualized by spraying sulfuric acid 50% (w/w) followed by charring the plates at 135 °C. The extracted lipids from the water mold were further analyzed by FTIR spectrometer (Thermo Nicolet NEXUS, Maryland, USA).