Tlc silica gel plates

Initial tests were carried out using TLC plates (SiO₂, DC Alufolien Kieselgel 60 F₂₅₄ (0.2 mm thick), Merck, Darmstadt, Germany). The mobile phase contained a mixture of chloroform and methanol in a 9:1 (v/v) ratio. The plates were observed using a UV lamp (254 and 365 nm).
The scale-up biotransformation products were separated using 500 µm preparative TLC silica gel plates (Anatech, Gehrden, Germany). The mobile phase contained a mixture of chloroform and methanol in a 9:1 (v/v) ratio. Separation products were scraped out and extracted twice using ethyl acetate.

Initial assessments were conducted utilizing thin-layer chromatography (TLC) plates (SiO₂, DC Alufolien Kieselgel 60 F254 (0.2 mm thick), Merck, Darmstadt, Germany). The mobile phase consisted of a hexane and acetone mixture in a 2:1 (v/v) ratio. Visualization of the plates was performed under a UV lamp (254 and 365 nm). For the scale-up biotransformation products, 1000 µm preparative TLC silica gel plates (Anatech, Gehrden, Germany) were employed. The mobile phase comprised a hexane and acetone mixture in a 2:1 (v/v) ratio. Following separation, the products were scraped off and subjected to double extraction using ethyl acetate. The solvent was subsequently evaporated, and the remaining residue was analyzed using a combination of TLC, GC, HPLC, and various NMR spectroscopy techniques (¹H NMR, ¹³C NMR, COSY, HMBC, and HSQC) (Bruker, Billerica, MA, USA) (Table 1 and Table 7).