I have prepared crude extract from the plant material and then fraction it with different solvent. How can I calculate the total flavonoid and total phenol in each fractions.
First I extract the material with methanol.
then I re-suspend the dried extract in water (Distilled).
Then fraction the above solution with n-hexane, CHCl3, Ethyl Acetate, n-Butanol and remaining water extract.
I have already prepared the standard plots; for phenol I have used gallic acid and quarcetine for flavonoids.
Dear Devendra
Calculate the content of your test sample on the basis total equivalent concentration of gallic acid for phenolics and total equivalent concentration of querecetin for flavanoids.
Means you make your unknown sample in concentration of any ranges between as utilized for the preparation of standard curve.
Then calculate the absorbance of your sample and calculate the yield as stated above.
I think you already have a method for estimation of phenolics and flavonoids since respective calibration curves plotted. For extract and fractions, you must use the same method on a single concentration for each sample. But, the concentration must in the range of calibration curves. Then, interpolate the absorbance of sample to the linear equation of the plotted calibration curve, to get the concentration of phenolics and flavonoids in the sample.
Dear Niraj and Sabandar,
I have already plot the standard curve for both total phenol and flavonoids and I have calculated both for crude, my confusion is regarding the fractions which I have collected from the crude extract.
Thanks for your answers.
Hope you may through some light on fractions and their calculations.
Dear Devendra
Calculation of content from fraction is as same as for extract. Try to take three different wide ranges of concentration of unknown fraction to quantify the content. Try to quantify your extract in lower concentration then your extract.
Dear
First of all you isolate the pure fraction from plant extracts using following techniques. First run TLC to know the type of fractions. After that Column Chromatography You can identify and quantify with HPLC, after isolated pure fraction run FT-IR, LC-MS, HNMR, CNMR for structure elucidation. You can read my papers too.
Hi Devendra
Pl see the attachments. I am sure you will get an answer for your query in these papers. Many methods have been described in these papers regarding your query.
Regards
Dear Alokji,
Thank you very much for sharing this research articles. But I need the formula to calculate total phenol and total flavonoids, which is not described in the articles you shared.
Thank you for your reply
Hi
Attached paper has a formula for calculation of tannin content. Hit and trial for phenolics can be done the same way. Here I would like to know the method that you have used to prepare the standard plots for gallic acid and quercetin.
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