Methanol and dichloromethane extracts (1.0 mg) of wild and micropropagated plants were redissolved in 1 ml of methanol. HPTLC Silica gel plates 60 F-254 pre-coated with aluminum (Merck, Germany 10 × 10 cm2, and thickness 250 μm) were used. The samples were applied with 100 mL syringe (Hamilton, Bona-duz Schweiz, Camag, and Switzerland) using a Linomat V (Camag, Switzerland) system. 20 μL of sample were applied in bands of 10 mm on the slopes of the plates. The plates were developed vertically in a glass chamber (20 × 10 cm2, Camag, Muttenz, and Switzerland), using the mobile phase hexane-acetone 7:3 at 25 ± 2 °C. After development, the plates were dried and the components were visualized by irradiation of energy at different UV wavelengths, and sprayed with anisaldehyde to visualize the complete profile. The chromatograms were integrated using win CATS evaluation software (Version 1.4.4.6337; Camag, Muttenz, Switzerland). The analysis was carried out in duplicate.
Glass chamber
Manufactured by CAMAG
Sourced in Switzerland, Germany
The Glass Chamber is a laboratory equipment piece designed to provide a controlled environment for various analytical and experimental procedures. Its primary function is to create a contained, isolated space with regulated atmospheric conditions, such as temperature and humidity, to facilitate precise and reproducible results.
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Lab products found in correlation
3 protocols using glass chamber
1
HPTLC Analysis of Plant Extracts
2
Lipid Quantification from Membrane Samples
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Samples were resolved in pH 7 buffer (50 mM tris and 150 mM NaCl) or pH 4 buffer (50 mM sodium acetate and 150 mM NaCl). Lipid extraction was done according to Bligh and Dyer (45 (link)). Briefly, 100 μl of a buffer-resolved MLV sample was mixed with 200 μl of CHCl3/MeOH [1:1 (v/v)] for 30 s. Subsequently, phase separation was obtained by centrifugation at 20°C and 10,000g for 5 min. The CHCl3 phase containing the lipids was placed into a vacuum centrifuge to evaporate the organic solvent. For lipid quantification, the lipid film was resolved in CHCl3 and spotted on an high-performance TLC (HPTLC) silica gel 60 plate (Merck KGaA, Darmstadt, Germany) using a CAMAG Linomat 5 sample application system (CAMAG, Berlin, Germany) in addition to a lipid calibration standard on the same HPTLC plate. TLC was developed in a glass chamber (CAMAG, Berlin, Germany) with a mobile phase for lipid separation: chloroform/ethanol/water/triethylamine [30:35:7:35 (v/v/v/v)] for either 2 to 3 hours (PC lipids) or 45 min (PE/PG lipids). Lipid spot visualization was achieved by primuline staining in 200 ml of acetone/H2O [4:1 (v/v)] and subsequent excitation under ultraviolet light. The lipid concentration in the MLV sample was lastly calculated using the calibration curve (nonlinear model) recorded with an appropriate lipid standard.
3
Quantitative TLC Analysis of Pharmaceuticals
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Samples were spotted as separate bands on 20 × 10 cm aluminum sheets pre-coated with silica gel 60 F254 (Merck, Darmstadt, Germany), using sample applicator, Linomat 5 (CAMAG, Muttenz, Switzerland) supplied with a 100.0 µL microsyringe. Bands are separated from one other and from the top and bottom-end of the plate by10 mm with band width of 6.0 mm. Using a CAMAG glass chamber previously saturated with 60 mL mobile phase composed of ethyl acetate–ethanol–water–ammonia (8.5:1:0.5:0.3, by volume), a Linear ascending development was performed over a distance of 8 cm then removed from the chamber and left to dry at room temperature. The separated bands were scanned using a CAMAG TLC scanner at 220.0 nm for AML, HCT, DSA and CT and 295.0 nm for TIM. Scanning was performed using reflectance-absorbance mode of measurement with deuterium lamp as a radiation source and slit dimension kept at 3 × 0.45 mm at scanning speed of 20 mm/s. WinCATS® software was used for densitometric evaluation and the output consist of a densito-grams and integrated peak area.
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