Tlc silica gel 60 f254 plate

Normal phase thin layer chromatography (NP-TLC) was done on TLC silica gel 60F₂₅₄ plates (E. Merck, Darmstadt, Germany, # 1.05554). Additionally, the TLC silica gel 60F₂₅₄ plates (E. Merck, Germany, # 1.05570) were used for the robustness test. The plates were prewashed with methanol and dried for 24 h at room temperature. Before use, the plates used in NP-TLC were activated at 120 °C for 10 min.
The solutions of pharmaceutical samples and standards of active substances (5 μL) were spotted manually on the chromatographic plates. The mixture of the chloroform–ethanol (96%)-glacial acetic acid in a volume composition of 5:4:0.03 (v/v/v) was used as the mobile phase. A total of 50 mL of the mobile was used in all cases. After saturation of the chamber (20 cm × 20 cm) with the mobile phase vapor for 20 min, the plates were developed vertically at room temperature (20 °C) to a distance of 7.5 cm and then dried for 20 h at room temperature (20 °C) in a fume cupboard. Additionally, a twin-trough chamber of 20 × 10 cm (#0.222.5254, Camag, Muttenz, Switzerland) was used for the robustness test.

Normal phase thin layer chromatography (NP-TLC) was done on TLC silica gel 60F₂₅₄ plates (E. Merck, Germany, # 1.05554). Additionally, TLC silica gel 60F₂₅₄ plates (E. Merck, Germany, # 1.05570) were used for the robustness test. The plates were prewashed with methanol and dried for 24 h at room temperature. Before use, the plates used in NP-TLC were activated at 120 °C for 10 min.
The solutions of pharmaceutical samples and standards of active substances (5 μL) were spotted on the chromatographic plates. The mixture of cyclohexane:chloroform:methanol:glacial acetic acid (6:3:0.5:0.5, v/v) was used as mobile phase. Of mobile 50 mL was used in all cases. After saturation of twin-trough chamber of 20 cm × 10 cm (#0.222.5254, Camag, Muttenz, Switzerland) with the mobile phase vapor for 30 min., the plates were developed vertically at room temperature (20 °C) to a distance of 7.5 cm and then dried for 20 h at room temperature (20 °C) in a fume cupboard.

All reactions were performed under an argon atmosphere unless otherwise stated. Commercially available starting materials were generously provided by Apollo Scientific and used without further purification. Tetrahydrofuran (THF) was freshly distilled from Na/benzophenone. Toluene for the Suzuki–Miyaura cross-coupling reactions was redistilled prior to use. Some of the compounds described in this article are known and have been previously reported; for compounds 1a, 2a, 11a, 12a, 1b, 2b, see reference [7 (link)], for compound 6a—[25 (link)], 8a—[9 (link)] and 9a—[17 (link)].
NMR spectra were recorded on a Bruker Avance III spectrometer (400 MHz for ¹H NMR and 100 MHz for ¹³C NMR). Chemical shifts δ are reported with respect to the residual solvent peak and are given in ppm. GC-MS analyses were performed on a Shimadzu GCMS-QP2010 ultra plus system using a Restek Rxi-5 ms column (30 m, 0.25 mmID). Reactions were monitored by TLC using Merk TLC silica gel 60 F₂₅₄ plates and visualized using UV light (254 nm) or KMnO₄ stain. Column chromatography was performed on ZEOprep 60 silica gel (35–70 µm, Apollo Scientific). Melting points were measured in open capillaries using a Mettler Toledo FP90 central processor equipped with Mettler Toledo FP81HT MBC cell.