Determination of Antioxidants in Calisia Fragrans Leaf Extract

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DETERMINATION OF TOTAL PHENOLICS AND TOTAL

FLAVONOIDS CONTENTS IN LEAVES EXTRACTS OF CALLISIA FRAGRANS

Student Name: Trinh Thai HaSupervisor: Assoc. Prof. Dr. Nguyen Dac Vinh

Hanoi, November 2009

Hanoi University of ScienceFaculty of Chemistry

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Contents

Antioxidants in Callisia Fragrans

Experiments

Results

Discussion

Conclusion

Q&A

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Callisia Fragrans

- was first grown in Europe,

particularly in Russia;- tradionally known as a miracle

plant;- is widely grown in Thanh Hoa

province, Vietnam;- is well- known with the ability

of curing many kind of ills, such

as asthma, tuberculosis, diabetes.

Callisia Fragrans Overview

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Callisia Fragrans

- Scientific Informationo Class : Liliopsida --

monocotylédones, monocotyledons

o Subclass: Commelinidae o Order : Commelinales o Family: Commelinaceae --

spiderwort

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Numerous researches have proved that Callisia

Fragrans has medicinal properties due to the high

contents of bioactive substances, the group of

flavonoids and plants (harmless) steroids.

The juice of Callisia fragrans contain elements which

are important to human life such as iron, chromium and

copper.

Flavonoids Over 4,000 flavonoids have been identified, many of which occur

in fruits, vegetables and beverages (tea, coffee, beer, wine and fruit

drinks).

Has potential beneficial effects on human health (have antiviral,

anti-allergic, antiplatelet, anti-inflammatory, antitumor and

antioxidant activities).

Callisia fragrans juice contains two type of flavonoids: quercetin

and kaempferol

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Quercetin Kaempferol

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Steroids A steroid is a terpenoid lipid characterized by its sterane core and

additional functional groups. The core is a carbon structure of four

fused rings: three cyclohexane rings and one cyclopentane ring.

Hundreds of bitch distinct steroids are found in plants, animals, and

fungi.

Steroids in plants mostly presents as Phytosterols and

Brassinosteroids.

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Experiments

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ExperimentsGoal:

◦ to determine the composition of polyphenolic and flavonoid

compounds which are 2 nominated antioxidants in plants.

◦ examine the effects of polarity of solvent on the antioxidant

composition of the extracts.

Subject: 5kg of fresh leaves of Callisia fragrans collected from

Thanh Hoa province, Vietnam.

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ExperimentsSampling:

◦ Fresh leaves were allowed to dry under the sunlight at the

temperature of about 32°C for 30 hours.

◦ Dried leaves were made up into tiny species using household

blender.

◦ Each proportion of 20g dried leaves was extracted in 4 different

solvents. Ethanol 100%

Ethanol 75%

Ethanol : Acetone (50:50 v/v)

Acetone 75%

◦ After 1 month, filter and obtain 4 different samples.

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ExperimentsDetermination of Total Phenolic Contents

0.4 ml of sample extract (or standard solution) was mixed with 2ml

of Folin-Ciocalteu reagent which was diluted 10 fold. After 3

minutes, 1.6 ml of 7.5% Na2CO3 solution was added, the reaction

solution was incubated for 30 minutes at room temperature.

Samples were allowed to measure colorimetrically at 765nm by

UV-Vis spectrophotometer. Total phenolics content was expressed

as mg gallic acid equivalent (GAE) /100g dried weigh.

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ExperimentsDetermination of Total Flavonoid Contents

Total flavonoid contents was measured by the aluminum chloride

colorimetric assay. An aliquot (1ml) of extracts or standard solution

of catechin (20, 40, 60, 80, 100mg/l) was added to 10ml volumetric

flask containing 4ml of distilled deionized H20. To the flask was

added 0.3 ml 5% NaNO2. After 5 minutes, 0.3ml 10% AlCl3 was

added. At 6th minute, 2 ml 1M NaOH was added and the total

volume was made up to 10ml with distilled deionized H2O. The

solution was mixed well and the absorbance was measured against

prepared reagent blank at 510nm. Total flavonoid content of fruits

and vegetables was expressed as mg catechin equivalents

(CE)/100g mass. Samples were analysed in duplicates.

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ResultsTotal Flavonoid Contents in Callisia fragrans leaves

extracts.

Total Phenolic Contents in Callisia fragrans leaves

extracts.

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ResultsTotal phenolic contents in Callisia fragrans leaves

extracts.

Sample Solvent

Total phenolic content (mg gallic acid/100g dried

weigh)#1 Ethanol 100% 25.4#2 Ethanol 75% 27.4#3

Ethanol/Acetone (50:50 v/v) 40.9#4 Acetone 75% 45.7

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ResultsTotal phenolic contents in Callisia fragrans leaves

extracts.

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extracts.

Sample Solvent Total flavonoid content (mg catechin/100g dried weigh)

#1 Ethanol 100% 89.33#2 Ethanol 75% 65.33#3 Ethanol/Acetone (50:50 v/v) 118.67

#4 Acetone 75% 196.00

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extracts.

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DiscussionAccording to the outcome of this study, it can be stated

that Callisia fragrans contains high content of phenolic

and flavonoid compounds which are common

antioxidants in plants.

We also determined the most effective solvent which

were used yielded the highest total phenolic and

flavonoid contents in the extraction of Callisia fragrans

leaves.

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Conclusion Things have been done so far in this research:

◦ Studied of literature background of antioxidants in food.

◦ Studied of the methods and results of determining antioxidant

activities in plants ,which were officially published on journals of

science.

◦ Executived spectrophotometric methods to measure the total

phenolic and flavonoid contents in leaf extracts of Callisia

Fragrans.

◦ Calculated and accessed the results of those experiments.

The positive outcome of the experiments is the good initiative for

further researchs on potential antioxdiant activities of Callisia

Fragrans and its use in medicinal treatment.

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Thanks for your attention !

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Q&A

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Determination of Antioxidants in Calisia Fragrans Leaf Extract