Tlc silica gel 60 f254 aluminum sheet

The chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), Honeywell Fluka (Morristown, NJ, USA), Kanto Chemical (Tokyo, Japan), Nacalai Tesque (Kyoto, Japan), Tokyo Chemical Industry (Tokyo, Japan), and FUJIFILM Wako (Osaka, Japan). The commercially available reagent-grade chemicals were used without further purification. The reaction progress was monitored by thin-layer chromatography (TLC) on TLC silica gel 60 F₂₅₄ aluminum sheets (Merck, Darmstadt, Germany). The flash column chromatography was performed on Silica Gel 60N (40–100 mesh, Kanto Chemical, Tokyo, Japan). Melting points (mp) were determined on a Yanaco melting point apparatus (Kyoto, Japan) and were uncorrected. ¹H and ¹³C NMR spectra were recorded on a Bruker Avance 600 spectrometer (600 MHz) (Billerica, MA, USA). Mass spectrometry (MS) and high-resolution mass spectrometry (HRMS) data were recorded on a JEOL (Tokyo, Japan) JMS-DX303HF using positive fast atom bombardment (FAB) technique with 3-nitrobenzyl alcohol as the matrix.

Melting points were determined using a Mel-Temp from Electrothermal. TLC was performed using TLC Silica gel 60 F254 aluminum sheets from Merck. ¹H-NMR and ¹³C-NMR spectra were measured using a Bruker Avance 300 (300 MHz, 75 MHz) (Rheinstetten, Germany), a Bruker NEO 500 (500 MHz, 125 MHz) (Rheinstetten, Germany), or a Bruker Avance 600 (600 MHz, 150 MHz) NMR spectrometer (Rheinstetten, Germany). The solvents (CDCl₃ or CD₂Cl₂) were used as standard for calibrating chemical shifts. Signals were assigned by two-dimensional methods (HSQC). IR spectra were recorded in KBr pellets using a Nicolet Avatar 370 FT-IR spectrometer (Madison, WI, USA) from Thermo Electron Corporation. Optical rotations were measured on a JASCO P-1020 digital polarimeter (Tokio, Japan) at 589 nm. Elemental analysis was performed on a Vario EL III elemental analyzer (Elementar, Langenselbold, Germany). HRMS spectra were measured at a GC-MS Trace DSX II spectrometer (Dreieich, Germany). UV spectra were recorded with an Analytik Jena Specord S600 spectrometer (Analytik Jena, Jena, Germany). HPLC was performed with a Chrom Tech, Chiral-AGP column (100 mm × 3.00 mm, 5 μm), eluent hexane/ethyl acetate 10:1. All starting materials were used as purchased without further purification. Trichloroacetimidate gluco-6 was synthesized according to the literature [54 (link)].

Thin layer chromatographic (TLC) analysis of CHF was performed on TLC Silica gel 60 F₂₅₄ Aluminum sheets (MERCK, Germany) with n-hexane:chloroform:methanol (4 : 4 : 2) as the mobile phase. UV spectra of the CHF were recorded in methanol solution using the BECKMAN double beam spectrophotometer in the Central Science Laboratory, University of Rajshahi, and the data are given in λ_max. IR spectra of the CHF were recorded using the PERKIN ELMER 1600 FTIR spectrophotometer in the Central Science Laboratory, University of Rajshahi, and the data are given in cm⁻¹.

All reagents used were commercial products. Ethyl 2,4-dioxovalerate, hydrazine hydrate, 2-hydrazinopyridine, 2-hydrazinobenzothiazole, lithium aluminum hydride, nicotine, sparteine, and anhydrous tetrahydrofuran (THF) were obtained from Sigma-Aldrich (St. Louis, MO, USA). Anhydrous ethyl alcohol and all other solvents were of p.a. grade, obtained from Avantor Performance Materials Poland S.A. (Gliwice, Poland) and were used without further purification. The commonly used analgesic, consisting of paracetamol, propyphenazone, and caffeine, was obtained from the local pharmacy.
Synthesis of pyrazole derivatives used in this work as analytes for the TLC-FAPA technique has been described in detail in our previous publication [25 (link)]. For all TLC-FAPA experiments, the Merck Millipore TLC silica gel 60 F₂₅₄ aluminum sheets was applied.

Thin-layer chromatography (TLC) was applied to separate bioactive compounds from the crude ethyl acetate. The crude compounds were spotted on TLC silica gel 60 F254 aluminum sheets (Merck, Darmstadt, Germany) and left for drying. The TLC plate was vertically placed in a developing tank containing chloroform and hexane (9.5:0.5 v/v). The solvent was allowed to run until it moved up to 80% of the TLC plate. The chromatogram was left to dry and visualized under UV light at 254 nm. The antibacterial compounds on the chromatogram were identified with the contact bioautography method.

The chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), Kanto Chemical (Tokyo, Japan), Nacalai Tesque (Kyoto, Japan), Tokyo Chemical Industry (Tokyo, Japan), and FUJIFILM Wako (Osaka, Japan). The commercially available reagent-grade chemicals were used without further purification. The reaction progress was monitored by thin-layer chromatography (TLC) on TLC silica gel 60 F254 aluminum sheets (Merck, Darmstadt, Germany). The flash column chromatography was performed on Silica Gel 60N (40–100 mesh, Kanto Chemical). ¹H-NMR and ¹³C-NMR spectra were recorded on a Bruker Avance 600 spectrometer (600 MHz) (Billerica, MA, USA). Mass spectrometry (MS) and high-resolution mass spectrometry (HRMS) data were recorded on a JEOL (Tokyo, Japan) JEOL JMS-700M Station using a positive fast atom bombardment (FAB) technique with 3-nitrobenzyl alcohol as the matrix.

All commercially available chemicals of the appropriate purity were purchased from Sigma (St. Louis, MO, USA) or Merck (Kenilworth, NJ, USA). The ¹H-NMR and ¹³C-NMR spectra were recorded using an AGILENT DD2-500 MHz (Santa Clara, CA, USA) spectrometer. Chemical shifts were reported in δ (ppm) and signals were given as follows: s, singlet; d, doublet; t, triplet; m, multiplet. Melting points (mp) were determined with a MEL-TEMPII apparatus, Laboratory Devices, Sigma-Aldrich (Milwaukee WI, USA) and were uncorrected. The microanalyses were performed on a Perkin-Elmer 2400 CHN elemental analyzer (Waltham, MA, USA). Thin-layer chromatography (TLC silica gel 60 F254 aluminum sheets, Merck (Kenilworth, NJ, USA) was used to follow the reactions and the spots were visualized under UV light.