I have problem with this compound. It is colorless so I've stained it with a anisaldehyde stain. After staining, I can see the spot, but it is not clear at the bottom. After I adjusted the mobile phase, there was no spot at all. I only can see an orange layer at top of the TLC plate (no spot). Why does this happen? It's because my mobile phase?
I have performed it in a precoated silica gel HPTLC 60 F254 (20× 10 cm) plate of 0.20 mm layer thickness. The samples and standards were applied on the plate as 6 mm wide bands with an automatic thin layer chromatography (TLC) sampler (ATS4) under a flow of nitrogen gas, 10 mm from the bottom, 10 mm from the side, space between two spots was 6 mm of the plate, and application speed was 150 nm/s. HPTLC plates were developed in a CAMAG twin-trough chamber (20 × 10 cm), which was pre-saturated with 50 ml mobile phase ethyl acetateethanol-water (80:20:12, v/v/v) for 30 min at room temperature (25± 2 °C) and 50± 5% RH. The chromatogram was run upto 9 cm from the base. TLC plates were dried in air drier in adequate ventilation conditions. The flow of air in the laboratory was maintained unidirectional (laminar flow, towards exhaust). Spots were visualized by spraying with acetic anhydride: sulphuric acid: ethanol (1:1:10, v/v/v) followed by heating on CAMAG HPTLC plate heater at 110 °C for 2 min.
Ok thank you hitesh kumar. Do you think i can replace plate heater to normal microwave?
get and read the following papers
http://nopr.niscair.res.in/bitstream/123456789/10287/1/IJNPR%201%283%29%20267-286.pdf
TLC and HPLC methods for steviol diterpene glycoside
You may heat your plat using hair dryer
- Badges
- Science topic
- Similar topics
- Biological Science
- Anatomy
- Tissue