Why are UV (colorimetric) spectrophotometric methods used in determining Total phenolic content/Antioxidant activity of fruits? Is HPLC possible? what are the differences these methods?
Possible, if you have HPLC with UV is DAD detectors. See link below.
https://scialert.net/fulltext/?doi=rjphyto.2011.209.215
But spectrophotometric methods more easy and reliable if you interested in total content without distinguishing individual phenolic compounds. HPLC is better if want to know really which phenolic compounds in your sample.
Total phenolic content and antioxidant activity both are quantitative analysis of a specific analyte present in the herbal extract/sample.
UV spectrophotometric is a type of absorption spectroscopy in which light of the ultra-violet region is absorbed by the molecule.
It helps in determining the quantity of specific secondary metabolites present in the sample and essential to estimate the scope of the sample as an antioxidant.
The main purpose of using HPLC is for identifying, quantifying, and purifying the individual components of the isolated mixture.
Kindly check https://www.hindawi.com/journals/ecam/2014/253875/
Also check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6524357/
Dear Nurul Ain Natasha,
Please read these articles.
Best regards,
Siroos Iravani
HPLC is used to identify and quantify the amounts of certain compounds. Commonly, you can't identify all compounds in a mixture. HPLC does not measure the total phenol content and antioxidant activity. What is the cost of HPLC instrument?
Tube assays are designed to quantify certain properties of a mixture of unknown compounds. UV-vis instrument is cheap and available in the most of laboratories.
The values measured by UV-vis correlates with values you want to measure. You have to understand what is the chemistry behind of an assay and correctly interpret the data
HPLC is better if want to know really which phenolic compounds in your sample because you used standard compounds that gave rate times which can compared with sample time.
Essam Fadel Alwan Al-Jumaily Are you sure that you can quantify all phenolics in a complex mixture of plant extracts? What is a cost of characterization of a single sample?
phenolic compounds may undergo many colorimeric reactions like ( coupling agent in oxidative - coupling reactions and in diazotization reaction) ,like (oxidation- reduction reactions with suitable oxidizing agents , like ion pair may occurs also -----
but this depends on the substituted functional groups, so not all phenolic compounds in this ease
HPLC more detectable and sensitive but you already need to optimize the conditions of the analysis
Hana Shuker Mahmood , HPLC does not measure "Total phenolic content/Antioxidant activity of fruits." The colormetric assays are commonly designed to minimize a contribution of light absorbance of extracts. Also, see my previous post.
UV method depend upon aromatic structure and conjugated double bond of phenolic compounds, so the measure give the total antioxidant efficiency. HPLC is used to identify the members of phenolic compounds and their contents and more accurate.
Ahmed Aboul-Enein The colorimetric FC and antioxidant assays are not based on measurements of UV-vis absorbance phenolics in a sample. Basically, they are a kind of redox titration with an colored indicator.
Ramakrishnan Baskar et al. :Antioxidant potential of peel extracts of banana varieties (Musa sapientum)
- Badges
- Science topic