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- Posts: 1
- Joined: Wed Mar 06, 2019 7:22 pm
I've been trying to apply NMKL (NORDIC COMMITTEE ON FOOD ANALYSIS) 196 - 2013 for the detection of histamine in standart solutions.
I am reproducing exactly as mentioned. However, by increasing and decreasing the concentration of the samples, there's not a peak that reproduces the expected behavior for histamine.
What could I ve been doing wrong?
Mobile phases are Ammonium acetate 0.1M (A) and Acetonitrile (B). Flow rate 0.9 mL/min; UV detector 254 nm; gradient 50% B to 90% B in 19 minutes. My column is a C18 - 250mm*4,6mm*10um.
Derivatization is prepared with sodium hydroxide solution, saturated sodium bicarbonate, dansyl chloride solution, ammonia and ammonium acetate:acetonitrile mixture ( 4.8 ). Time and temperature for reaction are respected. Final filtration too.
Any ideas or tips? Doesnt anyone know a better method?