Hi guys, I have some ceramide and dihydroceramide (d18:0/14:0) dissolved in methanol : chloroform=1 : 1. I tried to run TLC for both of them and stain the plate with Iodine and potassium permanganate. The band for ceramide is very clear but the band for dihydroceramide is always invisible. For the amount I used 10 micromoles for both of them.
I wonder is there something wrong with my sample or it's that the dihydroceramide is really hard to stain. If it's the staining problem, is there a better way to stain it? Most of paper I read used LC-MS or radioactive labelled dihydroceramide in TLC to detect it.
The invisibility of the dihydroceramide band on your TLC plate may be due to its low reactivity with iodine and potassium permanganate; consider using alternative stains like orcinol or ceric ammonium molybdate for better detection. Additionally, employing techniques such as LC-MS can provide more sensitive visualization of dihydroceramides.
Dear Weiqi Gu, the link below gives a whole protocol and different staining reagents. My Regards
https://www.ncbi.nlm.nih.gov/books/NBK594049/
Dear Weiqi Gu
There are TLC plates that fluoresce when illuminated with 254 nm UV light. In these plates, when there is a band or point of some compound that absorbs light at that wavelength (most organic compounds) the plate looks dark in that area. What you would need is a 254 nm ultraviolet lamp. Another option is to reveal the plaque with sulfuric acid and heat. Regards
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