Thin layer chromatographic analyses were performed using TLC silica gel, pore size 60 Å 60, F254 on aluminum sheets (Merck Millipore, Darmstadt, Germany). For column chromatographic analyses (CC.), various stationary phases were used, including silica gel, pore size 60 Å 60, mesh 70–230 (Merck Millipore, Darmstadt, Germany), and reversed-phase octadecyl (RP-C18) silica gel (Merck Millipore, Darmstadt, Germany). Various mobile phases were composed of reagent-grade solvents (Loba Chemie Pvt. Ltd., Mumbai, India). Then, 1- and 2-D NMR spectra were recorded on a Bruker UltraShield Plus 500 MHz spectrometer (Rheinstetten, Germany), with CD3OD as the solvent. Chemical shifts (δ) were obtained in part per million (ppm) and the coupling constants (J) were measured in hertz (Hz). Electrospray ionization mass spectrometry (ESI-MS) was obtained using a Thermo Scientific UPLC RS Ultimate 3000-Q Exactive hybrid quadrupole-Orbitrap mass spectrometer (Waltham, MA, USA).
Tlc silica gel
Manufactured by Merck Group
Sourced in Germany
TLC silica gel is a type of thin-layer chromatography (TLC) stationary phase material. It is composed of silica gel particles coated onto a support, typically a glass or plastic plate. The silica gel acts as a sorbent, allowing for the separation and identification of chemical compounds based on their relative affinities for the stationary phase and the mobile phase used in the TLC process.
Automatically generated - may contain errors
Lab products found in correlation
Aluminum sheets, by Merck Group (1 mentions)
Uplc rs ultimate 3000 q exactive hybrid quadrupole orbitrap mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Silica gel, by Merck Group (1 mentions)
Boom stand stemi 2000 stereo microscope, by Zeiss (1 mentions)
Gentamicin, by Bioanalyse (1 mentions)
Tetracycline, by Bioanalyse (1 mentions)
Eca 500 mhz spectrometer, by JEOL (1 mentions)
Ft ir 6100 spectrometer, by Jasco (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
Hplc grade, by Tedia (1 mentions)
7 protocols using tlc silica gel
1
Chromatographic and Spectroscopic Analyses
2
Purification and Characterization of Organic Compounds
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All reagents were obtained from Sigma and Acros Organics. Anhydrous 1,2-dichloroethane and EtOAc were obtained by distillation over P2O5. TLC was performed on Merck TLC Silica gel 60 F254 plates eluted with the specified solvents and samples were made visual with a UV lamp, VL-6. LC (Vilber Lourmat, Eberald Zell, Germany). Acros Organics (Acros Organics BV, Geel, Belgium) silica gel (Kieselgur 60–200 µm, 60 A) was used for column chromatography. Yields refer to spectroscopically (1H and 13C NMR) homogeneous material s. Melting points were determined in glass capillaries on a Mel-Temp 3.0 (Laboratory Devices Inc., Auburn, CA, USA).
3
Monosaccharide Composition Analysis of EPS
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To identify the monosaccharide composition of EPS through TLC, 10 mg of EPS was hydrolyzed with 1 mL sulfuric acid (2 N) at 100 °C for 4 h. The residual sulfuric acid was neutralized with the enough BaCO3 for 12 h. After EPS hydrolysate was adjusted to PH 7, it was lyophilized for the analysis.
EPS hydrolysate was treated on TLC Silica gel (Merck, Darmstadt, Germany) and migrated with the buffer composed with n-butanol:methanol:25% ammonia solution:DW (5:4:2:1). To visualize the composition of EPS, the gel was soaked with aniline-diphenylamine reagent and baked in the oven at 110 °C for 5 min.
To conduct Benedict’s test, EPS and EPS hydrolysate were mixed with the same quantify of Benedict’s reagent (BIOZOA Biological Supply, Seoul, Korea) and then heated in boiling water.
EPS hydrolysate was treated on TLC Silica gel (Merck, Darmstadt, Germany) and migrated with the buffer composed with n-butanol:methanol:25% ammonia solution:DW (5:4:2:1). To visualize the composition of EPS, the gel was soaked with aniline-diphenylamine reagent and baked in the oven at 110 °C for 5 min.
To conduct Benedict’s test, EPS and EPS hydrolysate were mixed with the same quantify of Benedict’s reagent (BIOZOA Biological Supply, Seoul, Korea) and then heated in boiling water.
4
Microscopic Examination and TLC Analysis
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Morphological characteristics were examined under a microscope (Zeiss Boom Stand Stemi 2000 Stereo Microscope) and the components were analyzed by Thin Layer Chromatography (TLC). TLC analyses were performed according to standardized methods using solvent systems A and C, and Merck TLC silica gel with Lethariella cladonioides (Nyl.) Krog. as the control (Culberson, 1972 (link); Orange et al., 2001 ).
5
Optimizing Rhamnose Biosynthesis Pathway
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The RBS sequence upstream of rhaD was mutated by PCR with saturation primers (RBS-pET28-F/R) using plasmid pET28a-rhaD as the template. Then, the plasmids with the RBS mutation upstream of rhaD were digested by Dpn I at 37 °C for 1 h. After purification, the plasmids harboring mutated RBS sequences of rhaD were co-transformed with the plasmid pCDFDuet-yqaB-aldO into ClearColi to obtain the RBS mutation library of the rhaD gene. The RBS mutation libraries of aldO and yqaB genes were constructed with similar operations. The mutant colonies were randomly picked and grown in LB medium. According to the above high-throughput screening process, the culture supernatant containing rare sugars was spotted on TLC silica gel (20 × 20, Merck, Darmstadt, Germany) and stained with anisaldehyde staining. Soon afterwards, the TLC plates were heated until green spots appeared. The strains with a darker green color compared with the wild-type strain were selected during the primary screening and then further verified by HPLC in the second screening. The RBS mutant plasmids of rhaD were co-transformed with the RBS mutant plasmids of aldO into ClearColi to obtain the RBS mutation libraries of rhaD and aldO genes. Finally, the RBS mutant sequence of the highest strain was confirmed by Tianlin Biotechnology (Wuxi, China).
6
Spectroscopic Characterization of Antimicrobial Compounds
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All melting points were determined using an Electrothermal Capillary melting point apparatus and are uncorrected. Infrared (IR) spectra were recorded as thin film (for oils) in NaCl discs or as KBr pellets (for solids) with a JASCO FT/IR-6100 Spectrometer (Japan) and values are represented in cm−1. 1H NMR (500 MHz) and 13C NMR (125 MHz) spectra were recorded on a Jeol ECA 500 MHz spectrometer (Japan) using TMS as internal standard and chemical shift values were recorded in ppm on the δ scale. Silica gel TLC (thin layer chromatography) cards from Merck (silica gel precoated aluminium cards with fluorescent indicator at 254 nm) were used for thin layer chromatography. Visualization was performed by illumination with UV light source (254 nm). Column chromatography was carried out on silica gel 60 (0.063–0.200 mm) obtained from Merck. The mobile phase consisted of chloroform or chloroform/ethyl acetate 1/1 v/v. Tetracycline, gentamicin and ofloxacin standard antibiotic discs were purchased from Bioanalyse®, Turkey.
7
Comprehensive Analysis of Ginsenoside Profiles
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Pesticide standards were purchased from Chem Service (West Chester, PA, USA), and six different vegetable oils were purchased from a local grocery store. Ethanol (70%) ginseng extract devoid of pesticide residues was prepared by our laboratory. A LC-Florisil solid-phase extraction tube for pesticide purification was purchased from Supelco (St Louis, MO, USA). Silica gel TLC employed for the qualitative analysis of ginseng saponin was procured from Merck (Darmstadt, Germany). The organic solvents employed for the quantitative analysis of ginsenosides were HPLC grade (Tedia, Fairfield, OH, USA). Fourteen reference ginsenosides [Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, Rg2(S), Rg2(R), Rh1, Rh2(S), Rh2(R), Rg3(S), Rg3(R)] were kindly supplied by the Korea Ginseng Corporation (Seoul, Korea).
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