I tried to quantify the total flavonoid content in I using the NaNO2, ALCl3 and NaOH method with Catechin as a standard. I got a pink/red color with standard but i don't get the same color with my sample although i did a qualitative analysis for my sample with different method and they gave a positive results. I developed TLC for these samples with MeOH/H2O and I got yellow band on TLC which indicates the presence of flavonoids compounds. So, can any one help me to determine TFC for my samples.
Is anyone have an idea to solve this problem and thanks.
i would like to ask you question to clarify
the problem, mention the method you followed for extraction.
Thanks for all, actually i got a yellow colour in the first time and when i tried to increase the sample conteration i got orange colour. The absorbance meauserments were low such as 0.17, 0.2 and 0.27. I used AlCl3 assay with NaNO2 and NaOH assay while Catechin was used as standard. So, should i change the method???
I did the extraction by maceration with MeOH.
But the extraction done by MeOH and i got various fractions by use different solvent on silica gel and now i trying to determine total flavonoid content of each fraction but the problem i don't get the pink colour to measure the absorbance at 510 nm.
Conduct preliminary analysis using laboratory chemical test methods. Then run TLC to know the type of fractions. After that Column Chromatography with HPLC, after isolated pure fraction run FT-IR, LC-MS, HNMR, CNMR for structure elucidation. You can read my papers too.
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