Chlamydomonas strain CC-125 cells were cultured to late mid-log phase (∼6 × 106 cells mL-1) in TAP medium. Cells were labeled with 0.2 μCi mL-1 of [14C]acetate for 2 h, washed twice with acetate free TAP medium, and then treated with fenpropimorph at a concentration of 10 μg mL-1 for 1 h. Total lipids were extracted using chloroform/methanol/formic acid (10:10:1, by volume; Härtel et al., 2000 (link)). After centrifugation (2000 g; 10 min), the bottom phase was directly loaded onto a thin-layer chromatography (TLC) plate (TLC Silica gel 60, MERCK) and separated using a two-phase solvent mixture; with a first phase of [chloroform/methanol/acetic acid/water, 90/15/10/3 by volume] and a second phase of [hexane/diethyl ether/acetic acid, 80/30/1 by volume]. Neutral lipid separation was performed using the first phase solvent mixture. After completely drying the TLC plate, polar lipids were separated using the second phase solvent mixture. The plate was then exposed to an imaging plate (BAS-TR 2040S, Fujifilm) and the relative strength of the radioactivity of [14C]acetate was visualized with an image analyzer (FLA-2000, Fujifilm). The neutral lipid spots containing TAG and DAG, and polar lipid spots were then scraped off the plate and recovered separately, and the radioactivity of each spot was determined by liquid scintillation counting (Tri-Carb 2910 TR, Perkin Elmer).
Tlc silica gel 60
Manufactured by Merck Group
Sourced in Germany, United States
TLC Silica gel 60 is a type of thin-layer chromatography (TLC) adsorbent material. It is a porous, high-purity silica gel that is commonly used in TLC analysis and separation techniques. The silica gel 60 adsorbent has a particle size range of 0.040-0.063 mm and is designed for general-purpose TLC applications.
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Lab products found in correlation
Arx 400, by Bruker (2 mentions)
600 lcms, by Waters Corporation (2 mentions)
Av 3 hd 500, by Bruker (2 mentions)
Ods a column, by YMC (2 mentions)
Drx 500, by Bruker (2 mentions)
Inova300, by Bruker (2 mentions)
Siliaflash f60 230 400 mesh, by Silicycle (2 mentions)
Cyclone plus storage phosphor system, by PerkinElmer (2 mentions)
20 cm aluminum sheets, by Merck Group (2 mentions)
2695 separation module system, by Waters Corporation (2 mentions)
Empower 3, by Waters Corporation (2 mentions)
Dd2 600, by Agilent Technologies (2 mentions)
Ge typhoon fla 7000 phosphorimager system, by GE Healthcare (2 mentions)
Topcount nxt scintillation counter, by PerkinElmer (2 mentions)
Fla 2000, by Fujifilm (1 mentions)
Tri carb 2910 tr, by PerkinElmer (1 mentions)
Vertex 70 spectrometer, by Bruker (1 mentions)
Microtof q 2 mass spectrometer, by Bruker (1 mentions)
Avance 3 spectrometer, by Bruker (1 mentions)
Savant spd131dda speedvac concentrator, by Thermo Fisher Scientific (1 mentions)
48 protocols using tlc silica gel 60
1
Lipid Profiling in Chlamydomonas
2
Saponin Identification via TLC Analysis
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Thin-layer chromatography (TLC) was performed to analyze the saponins fractions. 10 mg/mL of the all the samples was dissolved in 80% ethanol. 10 mL of each sample was applied to a normal-phase TLC plate (TLC Silica gel 60, glass plates 10 × 20 cm, Merck, Germany), and n-butanol, water, and acetic (84:14:17) were used as development solution. The developed plates were visualized by dipping in 10% H2SO4 and heated in an oven at 115°C for 13 min to visualize the saponin bands.
3
Anhydrous Reaction Conditions and Analytics
4
Bacterial AHL Detection by TLC-Biosensor
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Bacterial strains were grown in KB medium for 24 h at 27°C. AHLs were extracted with an equal volume of ethyl acetate and detected using C18 reversed-phase thin layer chromatography (TLC Silica gel 60, Merck, Darmstadt, Germany) and the biosensor C. violaceum CV026 (Taguchi et al., 2006 (link)) as described in Ichinose et al. (2020).
5
Purification of Nostoc sp. Bioactive Compounds
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Dried crude extract of Nostoc sp. MGL001 was dissolved in methanol and purified by TLC (TLC silica gel 60, Merck, Darmstadt, Germany). In this process, carbon tetrachloride: methanol (9:1) was used as the mobile phase and silica was used as the stationary phase. The UV-illuminated orange bands on the TLC plate were designated as A, B, C, D, E, and F, which were dissolved separately in 100% methanol (1 mL). Further each elute was then subjected to TLC purification using ethyl acetate: n-hexane (1:1). Only the potent designated bands of the first step were subjected to the second step.
6
Radiochemical Purity Analysis of [18F]PSMA-1007
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TLC was performed using TLC Silica Gel 60 (Merck KGaA, Darmstadt, Germany), Approximately 1–2 microliters of IMP [18F]PSMA-1007 injection solution was spotted on the plate. The solvent for the development of the TLC plates was Acetonitrile/Water 3:2 v/v. The developed plate was analyzed by autoradiography on MS (MultiSensitive) storage phosphor screens and by a Cyclone Plus Storage Phosphor System (PerkinElmer).
7
Luminescent TLC Binder for Pressure Sensing
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In the present study, commercially available TLC plates (Merck, TLC Silica gel 60) were used as the binder for the TLC-PSP [13 (link)]. The pore diameter of this silica gel is 60 Å, and the particle diameter is approximately 10 µm. The dye was adsorbed on the TLC binder after spraying with a spray gun. The dye solvents were prepared by dissolving PtTFPP in toluene at a ratio of PtTFPP:toluene = 4 mg:20 mL and Ru(dpp) dissolved in dichloromethane at the same mixing ratio as the PC-PSP at Ru(dpp) :dichloromethane = 14 mg:20 mL.
8
Anticancer Compounds from Cyanobacteria
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The methanolic extracts (100 mg·mL-1) of the target cyanobacteria were subjected to TLC purification (TLC silica gel 60, Merck, Darmstadt, Germany) in a two-step process. The first step used carbon tetrachloride: methanol (9:1) as the mobile phase. The UV-illuminated orange bands on the plates were designated as A, B, C, D, and E and were dissolved in methanol (1 mL). In the second step, each eluate was subjected to TLC purification using hexane: ethyl acetate (1:1). Only the potent designated bands of the first step were subjected to the second step. Ultimately, the bands separated in the second TLC step were monitored for anticancer potential against HT29 and A498 cancer call lines. All experiments were performed in triplicate.
9
Comprehensive Analytical Characterization of Compounds
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All chemical reagents were of analytical grade and purchased from Sigma-Aldrich® and Oakwood Chemical® (USA) without any further purification. Column chromatography was carried out using silica-flash 230–400 mesh from Aldrich. Analytical preparative thin-layer chromatography was performed on aluminum sheets (1.0 mm thick) Merck TLC silica-gel 60.
Compounds were analyzed using a high-resolution mass spectrometer with electrospray ionization (HRMS-ESI) in the positive mode using a micrOTOF-QII mass spectrometer (Bruker Daltonics, Billerica, MA). Mass spectrum was recorded for each sample in methanolic solution (concentration of 500 ppb) with flow of 4.0 μL min−1 and capillarity of 3500 V. 1H, 19F, COSY 2D and HMBC NMR spectra were recorded with a Bruker Avance III spectrometer at 600 (1H) and 565 MHz (19F) respectively, using deuterated chloroform (CDCl3) as the solvent and tetramethylsilane (TMS) as the internal reference. The chemical shifts are expressed in δ (ppm) and coupling constants (J) are given in Hertz (Hz). The multiplicities are expressed as: s, singlet; d, duplet, and m, multiplet. Fourier-transform infrared spectroscopy (FT-IR) spectra was recorded on a Bruker Vertex 70 spectrometer equipped with diamond attenuated total reflectance (ATR) accessory in the 4000–400 cm−1 region.
Compounds were analyzed using a high-resolution mass spectrometer with electrospray ionization (HRMS-ESI) in the positive mode using a micrOTOF-QII mass spectrometer (Bruker Daltonics, Billerica, MA). Mass spectrum was recorded for each sample in methanolic solution (concentration of 500 ppb) with flow of 4.0 μL min−1 and capillarity of 3500 V. 1H, 19F, COSY 2D and HMBC NMR spectra were recorded with a Bruker Avance III spectrometer at 600 (1H) and 565 MHz (19F) respectively, using deuterated chloroform (CDCl3) as the solvent and tetramethylsilane (TMS) as the internal reference. The chemical shifts are expressed in δ (ppm) and coupling constants (J) are given in Hertz (Hz). The multiplicities are expressed as: s, singlet; d, duplet, and m, multiplet. Fourier-transform infrared spectroscopy (FT-IR) spectra was recorded on a Bruker Vertex 70 spectrometer equipped with diamond attenuated total reflectance (ATR) accessory in the 4000–400 cm−1 region.
10
NMR and HPLC Characterization of Compounds
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The methodology used in this study was based on that used by Ble-González et al. [83 (link)] but with some modifications. NMR spectra were recorded on an Agilent DD2-600 at 600 MHz for 1H and 150 MHz for 13C NMR, using CD3OD as the solvent. Chemical shifts are reported in ppm relative to TMS. Thin-layer chromatography (TLC) was performed using TLC Silica gel 60, F254, and 20 × 20 cm aluminum sheets (Merck KGaA, Darmstadt, Germany). High-performance liquid chromatography (HPLC, Waters, Milford, MA, USA) analyses were performed on a Waters 2695 Separation module system, equipped with a photodiode array detector (Waters Co. 2996) and Empower 3 software (Waters Corporation, Milford, MA, USA).
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