This document outlines a study to screen and analyze selected plant species for their antioxidant properties. The objectives are to:
1. Screen 3-4 plant species from forest regions for antioxidant properties.
2. Identify primary and secondary metabolites in plant extracts.
3. Isolate and quantify bioactive antioxidant compounds and determine medicinal value.
4. Compare antioxidant profiles between plant species.
Plants will be extracted using solvent extraction. Phytochemical analysis will test for compounds like alkaloids, flavonoids, tannins. Total phenolic content and flavonoid content will be determined colorimetrically. Antioxidant capacity will be evaluated using DPPH, ABTS, hydroxy
Oxygen is highly reactive atom that is capable of becoming part
of potentially damaging molecule commonly called “free radical.”
Free radicals are capable of attacking cells of the body, causing
them to lose their structure and function.
Free radicals have been implicated in the pathogenesis of at
least 50 diseases.
Free radial formation is controlled naturally by various compounds
known as antioxidants.
It is when the ability of antioxidant is limited that this damage can
become cumulative and debilitating.
Following criteria should be considered while selecting an antioxidant.
It should be able to produce desire redox reaction.
It should be physiologically and chemically compatible.
It should be physiologically inert.
It should be non-toxic both in the reduced and oxidized forms.
It should be effective in low concentration.
It should provide prolonged stability to the formulation.
Flavonoids classification, isolation and identificationMona Ismail
Flavonoids are groups of polyphenolic compounds which are found in fruits, flowers, seeds & vegetable.
(named from the Latin word flavus meaning yellow, their colour in nature)
Oxygen is highly reactive atom that is capable of becoming part
of potentially damaging molecule commonly called “free radical.”
Free radicals are capable of attacking cells of the body, causing
them to lose their structure and function.
Free radicals have been implicated in the pathogenesis of at
least 50 diseases.
Free radial formation is controlled naturally by various compounds
known as antioxidants.
It is when the ability of antioxidant is limited that this damage can
become cumulative and debilitating.
Following criteria should be considered while selecting an antioxidant.
It should be able to produce desire redox reaction.
It should be physiologically and chemically compatible.
It should be physiologically inert.
It should be non-toxic both in the reduced and oxidized forms.
It should be effective in low concentration.
It should provide prolonged stability to the formulation.
Flavonoids classification, isolation and identificationMona Ismail
Flavonoids are groups of polyphenolic compounds which are found in fruits, flowers, seeds & vegetable.
(named from the Latin word flavus meaning yellow, their colour in nature)
Soxhlet extraction is a continuous process of extraction with a hot organic solvent. Typically, Soxhlet extraction is used when the desired compound has a limited solubility in a solvent, and the impurity is insoluble in that solvent.
Soxhlet extraction is a continuous process of extraction with a hot organic solvent.
Typically, Soxhlet extraction is used when the desired compound has a limited solubility in a solvent, and the impurity is insoluble in that solvent.
Plants produce a vast and diverse organic compounds, which do not appear to participate directly in growth and development.These substances traditionally referred to as secondary metabolites which terpenes are one of them.
Phytopharmaceuticals: Occurrence, isolation and characteristic features (chemical nature, uses in pharmacy, medicinal and health benefits) of Quercetin
The importance of medicinal plants in the treatment of a variety of human ailments man has been dependent on the higher plants as a source of food and medicine.
Biosynthesis lectures by Dr. Refaat HamedRefaat Hamed
This is a series of five lectures for 4th year Pharmacy Students (Assiut University) as part of the "Applied Pharmacognosy" course. The lectures cover the biosynthesis of many classes of natural products (e.g. Alkaloids, Polyketides, Flavonoids,..etc. Special emphasis is on the recent trends in biosynthesis research.
การวิเคราะห์ฤทธิ์ต้านอนุมูลอิสระที่ระยะต่างกันในกล้วยเล็บมือนาง
Analysis of Antioxidant Activity at Different Stage in Musa (AA group) ‘Kluai Leb Mu Nang’
อดิศร จำรูญ
Soxhlet extraction is a continuous process of extraction with a hot organic solvent. Typically, Soxhlet extraction is used when the desired compound has a limited solubility in a solvent, and the impurity is insoluble in that solvent.
Soxhlet extraction is a continuous process of extraction with a hot organic solvent.
Typically, Soxhlet extraction is used when the desired compound has a limited solubility in a solvent, and the impurity is insoluble in that solvent.
Plants produce a vast and diverse organic compounds, which do not appear to participate directly in growth and development.These substances traditionally referred to as secondary metabolites which terpenes are one of them.
Phytopharmaceuticals: Occurrence, isolation and characteristic features (chemical nature, uses in pharmacy, medicinal and health benefits) of Quercetin
The importance of medicinal plants in the treatment of a variety of human ailments man has been dependent on the higher plants as a source of food and medicine.
Biosynthesis lectures by Dr. Refaat HamedRefaat Hamed
This is a series of five lectures for 4th year Pharmacy Students (Assiut University) as part of the "Applied Pharmacognosy" course. The lectures cover the biosynthesis of many classes of natural products (e.g. Alkaloids, Polyketides, Flavonoids,..etc. Special emphasis is on the recent trends in biosynthesis research.
การวิเคราะห์ฤทธิ์ต้านอนุมูลอิสระที่ระยะต่างกันในกล้วยเล็บมือนาง
Analysis of Antioxidant Activity at Different Stage in Musa (AA group) ‘Kluai Leb Mu Nang’
อดิศร จำรูญ
This is an Engg Biotechnology project based on medicinal plant i.e singapore cherry or jamaican cherry tree (scientific name Muntingia calabure ), we did in 2013 in GMIT college Davangere, karanataka, India. i have complete project detail what we did..,
An antioxidant is a molecule capable of inhibiting the oxidation of other molecules.
Oxidation reactions can form free radicals and these start chain reactions that damage cells .
Antioxidants terminate these chain reactions by removing free radical intermediates and inhibit other oxidation reactions
Extraction of Secondary Metabolites from Roots of Acanthus Ilicifolius L and ...inventionjournals
The root extracts of Acanthus ilicifolius L finds a prominent place in folk medicine. In this study, we
extracted alkaloid, flavonoid, tannin and total phenols in benzene, ethyl acetate, acetone, methanol and
ethanol, their antibacterial activity and antioxidant activity was evaluated. The antioxidant activity is executed
by FRAP assay and agar well diffusion method is done to study the antibacterial activity against Enterobacter
aerogenes, Enterobacter cloacae, Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Streptococcus
pyogenes. The antibacterial activity of all the extracts was compared with standard antibiotic gentamicin.
The Minimum Inhibitory Concentration [MIC] was determined by serial dilution method. Alkaloids are rich in
acetone and Flavonoids are high in methanol extracts. The acetone extract showed higher antioxidant activity,
while benzene extract was identified to contain lower antioxidant activity. The extent of inhibition by the root
extracts diverge between the solvents used, among them ethanol extracts exhibited higher level of inhibition
against the gram positive test cultures compared to gram negative test cultures employed. Whereas, the acetone
extracts efficacy is more on gram negative test cultures than the gram positive cultures. The MIC was found to
be between 1mg/100µl to 5mg/100µl. This study gives the source for purification and characterization of
bioactive principles that possess antioxidant and antibacterial action from the root of Acanthus ilicifolius.
Evaluation of the Antioxidant Activities of Organic Extracts from Ammodaucus ...CrimsonAlternativemedicine
Evaluation of the Antioxidant Activities of Organic Extracts from Ammodaucus leucotrichus Coss & Dur Fruit Part Harvested from the Algerian Sahara by Imad Abdelhamid El Haci in Advances in Complementary & Alternative medicine
Aromatic and medicinal plants are a good source of natural preparations containing effective bioactive compounds which can be used for different applications. This work aims to evaluate the antioxidant activity of some organic extracts of Ammodaucus leucotrichus Coss & Dur fruit part. The whole plant was collected from the region of Beni Abbas (Bechar-Algeria).
For more open access journals in Crimson Publishers please click on link: https://crimsonpublishers.com/
For more articles in open access Complementary Medicine journals please click on link: https://crimsonpublishers.com/acam/
Evaluation of the Antioxidant Activities of Organic Extracts from Ammodaucus ...CrimsonAlternativemedicine
Aromatic and medicinal plants are a good source of natural preparations containing effective bioactive compounds which can be used for different applications. This work aims to evaluate the antioxidant activity of some organic extracts of Ammodaucus leucotrichus Coss & Dur fruit part. The whole plant was collected from the region of Beni Abbas (Bechar-Algeria). Five organic extracts were obtained and the evaluation of the antioxidant activity was performed by six conventional methods. Polar organic extracts exhibited more antioxidant power then non polar extracts. The level of phenolic compounds was moderate in all extracts. The investigation of the antioxidant activity of organic extracts from fruit part of Ammodaucus leucotrichusrevealed a moderate activity tested by six conventional methods.
For more open access journals in Crimson Publishers please click on link: https://crimsonpublishers.com/
For more articles in open access Complementary Medicine journals please click on link: https://crimsonpublishers.com/acam/
Pharmacognosy & Phytochemistry 2 unit 3.pptxPranita Sunar
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Terpenoids: Menthol, Citral, Artemisin Glycosides: Glycyrrhetinic acid & Rutin
Terpenes or terpenoids are a secondary metabolite compounds, majority of which are found in plant species and few are obtained from other sources such as fungi, algae and sponges. These are volatile substances which is also responsible for fragrance of some flowers and plants. Terpenoids are the polymers of isoprene units (C5H8)n. Hence, they are also known as Isoprenoids.
Tinospora Cordifolia the magical Herb (Giloy)Vedant Patel
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How to find antioxidant properties from selected plants
1. HOW TO DETERMINE TOTAL PHENOLIC,
FLAVONOID CONTENTS AND IN VITRO
ANTIOXIDANT
ACTIVITY OF SELECTED PLANTS
Vinars Dawane
2. Do
1. Screening of 3-4 plant species from forest region
having antioxidant properties.
2. Identify primary and secondary metabolite profile of
the plant spices of interest.
3. Isolate and quantify the bioactive antioxidant
compound and find out its medicinal values.
4. Determine the Comparative antioxidant profile
between screened plants species.
4. Extract preparation
• washing with tap water,
• shade dried and powdering the sample.
• then soxhelt apparatus with methanol as solvent (different
combinations of solvents) will be used for extraction.
5. Identification of primary and secondary metabolite.
Objective - 2
• Phytochemical Analysis:
• Phytochemical analysis to screen the plants for the presence of
alkaloids, glycosides, saponins tannins, flavonoids and carbohydrates,
will be performed according to the method described by Sofowora (1993)
and Evans (1998).
• Test for Carbohydrates:
• Add few drops of Molisch’s reagent to 2ml of each of the water extract in
two tubes. A small quantity of concentrated sulphuric acid will then add
and allowe to form a lower layer. A purple ring at the interface of the
liquids indicates the presence of carbohydrates. Each mixture will be
then shaken and allowed to stand for 2 minutes and diluted with 5ml of
water. A purple precipitate also showed the presence of carbohydrates
(Evans, 1989).
6. • Test for Alkanoids:
• Five test tubes will be used for each of the different drug sample
(ethanolic extract). Few drops of the following reagents manager’s
reagent, Drangendorff’s reagent. Wanger’s reagent, Hanger’s reagent
and 10% tannic acid solution will be added respectively to each of the
five test tubes. The presence of precipitate in at least 3 or all of the
above reagents indicates the presence of alkaloids (Evans 1989).
• Test for Saponins:
• A small quantity of the ethanolic extract wil be boiled. The mixture will be
filtered and 2.5ml of the filtrate will be added to 10ml of the distilled
water in a test tube. The test tube will be corked and shaken vigorously
for about 30 seconds, then it will be allowed to stand for half an hour. A
honeycomb forth will an indicator of the presence of saponins
(Sofowora, 1993).
7. • Test for Tannins (Ferric chloride test):
• A portion of the water extract will be diluted with distilled water in a ratio
of 1:4 and few drops of 10% ferric chloride solution was added. A blue
or green colour indicates the presence of tannins (Evans,1989).
• Test for Glycosides:
• To a portion of the aqueous extract will be added fehlings reagent and
boiled for 2 minutes. A brick red colouration indicates the presence of
glycosides.
8. Total Phenolic Contents (TPC)
determination
• Folin – Ciocalteu reagent method –
• Method is used for the colorimetric in vitro assay of phenolic
and poly-phenolic antioxidants.
• Reagent - a mixture of phospho-molybdate and phospho -
tungstate.
• Method - Samples (100μL) + 2 ml of 2% Na2CO3 + 100 μL of Folin-
Ciocalteu reagent + absorbance was measured at 743 nm against a
blank.
9. Presence of flavonoid
shinoda test –
• Four pieces of magnesium fillings (ribbon) are added to the
ethanolic extract followed by few drops of
concentrated hydrochloric acid.
If –
• A pink or red colour indicates the presence of flavonoid.
• Colours varying from orange to red indicated flavones.
• crimson to magenta indicated flavonones.
10. Total Flavonoid Content (TFC) Determination
• Aluminium Chloride Method –
• Steps –
• 0.5ml of sample (1mg/ml) mixed with
• 0.1ml of 10% aluminium chloride and 0.1ml of
potassium acetate (1M).
• Addition of 4.3ml of 80% methanol.
• Absorbance measure at 415nm.
12. Antioxidant capacity
• DPPH: Quenching Activity method.
• α, α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging method.
• This method is based on the reduction of DPPH in methanol solution in the
presence of a hydrogen donating antioxidant due to the formation of the
non radical form DPPH-H.
• 0.1mM solution of DPPH in methanol + added to 3 ml of extract + mix.
• Absorbance measure at 517 nm. And Ascorbic acid was used as the
reference.
• DPPH scavenging effect (% inhibition) = {(A0 – A1)/A0)*100}.
• Where, A0 is the absorbance of the control reaction.
and A1 is the absorbance in presence of all of the extract samples
and reference.
13. • ABTS – Cation Decolorizing Test.
• ATBS Cation production –
• 7mM ABTS solution + 2.45 mM potassium Per sulphate + mix in
dark room.
• The ABTS + Solution were diluted with ethanol to an absorbance of
0.70+0.02 at 734 nm.
• 100μL of sample + to 3.9 mL of diluted ABTS+ Solution + absorbance
at 734 nm.
• ABTS radical cation activity = {(A0 –A1)/A0)*100}.
where , A0 is the absorbance of the control reaction, and A1 is the
absorbance in presence of all of the extract samples and reference.
14. Hydroxyl Radical ScavengingActivity method.
• Deoxyribose assay was used to determine the hydroxyl radical
scavenging activity in an aqueous medium.
• Extract + FeCl 3 (100 µM) + EDTA (104 µM) + H2O2 (1 mM) and 2-
deoxy- D-ribose (2.8 mM) + mix them + phosphat buffer for makeup.
• Mixture + 95 0 C in water bath for 15 min + 1 ml each of TCA (2.8%)
and TBA (0.5% TBA in 0.025 M NaOH containing 0.02% BHA).
• Mixture will be cooled on ice and centrifuged at 5000 rpm for 15 min.
• Absorbance of supernatant was measured at 532 nm.
15. Potassium Ferricyanide Reduction Method for Reducing
Capacity testing.
• Various concentrations of the extracts and standards (50, 100, 250,
500, 750, 1000 mg/ml) .
• Then add to 2.5 ml of (0.2 M) sodium phosphate buffer (pH 6.6) and
2.5 ml of potassium ferricyanide [K 3Fe 3 (CN) 6 ] (1%) solution and
vortex.
• Incubation at 50°C for 20 min + 2.5 ml of TCA + centrifuged at Χ3000
g for 10 min.
• Mix with deionized water (5 ml) and then 1 ml of FeCl 3 (1%).
• Formation of Perl's Prussian color was measured at 700 nm.
Note - Increased absorbance of the reaction mixture indicated
increasing reducing power.
16. In vitro hemolysis of methanolic extract method for
AntihemolyticActivity testing.
• Inhibition of H 202 induced red blood cell (RBC) haemolysis of
methanolic extract are examined by this method.
• Haemolytic activity of the extract can investigate by measuring the lysis
of a 10% (v/v) human red blood cells suspension in a
spectrophotometric assay.
• In this experiment, method will be used same as -
Naim M, Gestetner B, Bondi A, Birk Y. Antioxidative and antihemolytic activities of soybean
isoflavones. J Agric Food Chem 1976;24:1174-7.
17. References
• (Silva MRO, Almeida AC, Arruda FVF, Gusmão N (2011) Endophytic fungi from brazilian
mangrove plant Laguncularia racemosa (L.) Gaertn. (Combretaceae): their antimicrobial
potential. Science against Microbial Pathogen: Communicating Current Research and
Technological Advances; 2011, 2:1260-1266).
• Patra JK, Thatoi HN. Metabolic diversity and bioactivity screening of mangrove plants: a
review. Acta Physiol Plant 2011; 33: 1051-1061.
• N. Bunyapraphatsara et al. Pharmecological studies of plants in the mangrove forest. Thai
Journal of Pharmacy; 2003, Vol.10 (2).
• Vagi E, Rapavi E, Hadolin M, Vasarhelyine Peredi K, Balazs A, Blazovics A, et al. Phenolic
and triterpenoid antioxidants from Origanum majorana L. herb and extracts obtained with
different solvents. J Agric Food Chem. 2005; 53:17–21. [PubMed: 15631502].
• Jonathan Yisa. Phytochemical Analysis and Antimicrobial Activity Of Scoparia
Dulcis and Nymphaea Lotus. Australian Journal of Basic and Applied Sciences,
3(4): 3975-3979, 2009.