Does anyone one have protocol for total polyphenol and total flavonoid estimation by 96 well plate method.
gallic acid and quercetin is used as standard.
FOr TPC, you can follow this protocol (Gull, 2014)
Briefly 50 mg of crude extract was mixed with 0.5 mL of Folin-Ciocalteu reagent and 7.5 mL deionized water. The mixture was kept at room temperature for 10 minutes, and then 1.5 mL of 20% sodium carbonate (w/v) was added. The mixture was heated in a water bath at 40 ºC for 20 minutes and cooled in an ice bath. The solution (250 µL) of each sample was taken in 96 well-plate and absorbance was measured at 755 nm using microplate reader (Bio Tek Instrument, Jnc., VT, USA). The amount of TPC was calculated using gallic acid calibration curve and reported as mg g-1 of dry matter, measures as gallic acid equivalent (GAE).
For TFC (Gull, 2014)
Briefly, extracted solution (250 µL) containing 25 µL sample extract was placed in 96 well-plate and mixed with 125 µL of distilled water, 7.5 µL of 5% NaNO2 was added. After 5 minutes, 15 µL of 10% AlCl3 was added. Further, 50 µL of 1M NaOH was added after 50 minutes and volume was made up to 250 µL with distilled water. Absorbance was read at 510 nm by spectrophotometer (Bio Tek Instrument, Jnc., VT, USA). TFC were calculated using a calibration curve for catechin. The amount of TF were calculated and reported as mg/g of dry matter, measured as catechin equivalent(CE).
Thank you, But does anyone have the protocol to estimate same TPC and TFC for honeys samples.
There are several modifications on the protocols for TPC and TFC determinations. See also IOSR J Environ Sc. Toxico. and Fd Tech.Vol. 8, Issue 7 Ver. I (July. 2014), PP 104-111 on the article:
Phenolic content and antioxidant activity of some under-utilized Nigerian yam (Dioscorea spp.) and cocoyam (Xanthosomamaffa (scoth)) tubers by Ukom et al.
On honey, dilutions may be required after which you multiple result with dilution factor.
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