Lately i've been researching Nisin in a microsphere system. I have a problem in methods to quantify nisin from the system. I've tried using UV spectrophotometry but it seems the polymer matrix interferes the quantification.
Now i'm trying to use TLC methods to quantify nisin. I've read a research that they use eluent n-butanol : acetic acid : water = 4:1:1 and the Rf of Nisin was quite high (around 0.8) with spot detection using UV light at 254 nm (no stain coloring such as Ninhydrin).
I've tried the eluent above and my spot of nisin didn't go up at all (Rf = 0). The spot stayed at the same spot i applied on the plate. I am using Silica Gel GF254 plate from Merck if i recall correctly.
Any advice about another eluent or maybe i did something wrong that my spot didn't go up at all, or maybe my spot is too concentrated (my concentration is 1000 ppm vs 500 ppm at that research i said before, both applied 2 microliter)
Thanks in advance.
Hi, please try to find other TLC system in the Camag data base, maybe you can find better methods, I have found 9 hits for your compound, or you can meet me at my office, I can help you to solve your problems. best regards
http://www.camag.com/en/tlc_hptlc/customer_magazine_cbs/ccbs_database.cfm
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