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Analysis of Antioxidant Activity
at Different Stage in Banana
Musa (AA group) ‘Kluai Leb Mu
Nang’
By
Adisorn Jumroon
๔
KMITLโนโลยีพระจอมเกล้าเจ้าคุณทหารลาดกระบัง
วิทยาเขตชุมพรเขตรอุดมศักดิ์
Introduction
- ‘Kluai Leb Mu Nang’
- Free radical
- Antioxidant
- Phenolic compound
- Flavonoid
Family
Genus
Section
Genome
Cultivar
Musaceae
Enset Musa
Australimusa Callimusa Rhodochlamys Eumusa Ingentimusa
AA AAA AAAA AB AAB ABB ABBB
‘Kluai Leb Mu Nang’‘Kluai Leb Mu Nang’
Taxonomic classification
Shyamala et al. (2011)
Botanical characteristicsBotanical characteristics
1 Stem
8 Leaves
2, 3, 4 Flower
5, 6, 7 Fruit
Wendy (2001)
8
Analysis of Antioxidant in banana
Analysis of Antioxidant in banana
Analysis of Antioxidant in banana
Analysis of Antioxidant in banana
Analysis of Antioxidant in banana
Phenolic compound
Analysis of Antioxidant in banana
FlavonoidsFlavonoids
Analysis of Antioxidant in banana
1. To study the antioxidant activity at the
different stages in ‘Kluai Leb Mu Nang’
2. To Study total phenolic content and
total flavonoids at the different stages in
‘Kluai Leb Mu Nang’
Objectiv
es
Objectiv
es
SamplesSamples
Kluai Leb Mu Nang 3 different stages
- Mature banana (Green)
- Ripe banana (Full yellow)
- Overripe banana (Dark yellow)
Materials and methodsMaterials and methods
Evaluating parameters
- Color analysis
- Extraction process
- Antioxidant activity assay
- DPPH scavenging assay
- Ferric reducing antioxidant
power
assay
- Total flavonoid content analysis
- Total phenolic compound analysis
Use a chomameter Reported values
L* a* b*
Color analysisColor analysis
Extraction
process
50°C, 12 h
4500 rpm, 15 min
Add 80% methanol
filtration
• DPPH (2,2- diphenyl-1-picrylhydrazyl)
• Methanol
• BHT (butylated hydroxytoluene)
• UV-visible spectrophometer
• Micropipette
DPPH scavengingDPPH scavenging
assayassay
Materials and ChemicalsMaterials and Chemicals
OD 515 nm
DPPH
DPPH scavengingDPPH scavenging
assayassay
•C2
H3
NaO2
3H2
O (Sodium acetate
trihydrate)
•C2
H4
O (Acetaldehyde )
• Acetic acid
• FeCl3
6H2
O (Ferric chloride
hexahydrate)
•TPTZ (2,4,6-Tripyridyl-s-Triazine)
• HCl
•Trolox (6-hydroxy-2,5,7,8-
tetramethylchroman-
2-carboxylic acid)
Materials and Chemicals
Ferric Reducing AntioxidantFerric Reducing Antioxidant
PowerPower
OD 593 nm
Ferric ReducingFerric Reducing
AntioxidantAntioxidant
PowerPower
•Sodium nitrite (NaNO2
)
•Aluminum chloride (AlCl3
)
• Sodium hydroxide (NaOH)
•Catechin ((2R,3S)-2-(3,4-dihydroxyphenyl)-3,4-
dihydro-2H-chromene-3,5,7-triol
•UV-visible spectrophometer
• Micropipette
Materials and Chemicals
Total flavonoid content
Aluminium chloride colorimetric
method
0.1 g banana crude extract
+ 5 ml distilled water + 5% NaNO2
5 min
Add 10% AlCl3 0.6ml
Add 1M NaOH 2 ml
Adjust volume with DW to 10 ml
5 min
Fatemeh et al. (2012) and Baskar et al. (2011)
Total flavonoid content
Aluminium chloride colorimetric
method
Measuring of TFC at OD510 nm
Note: catechin was used as standard solution to
plot the calibration curve
Total flavonoid content
Aluminium chloride colorimetric
method
•Sodium carbonate (Na2
CO3
)
•Folin–Ciocalteur reagent
•Gallic acid (3,4,5-trihydroxybenzoic acid)
•UV-visible spectrophometer
• Micropipette
Materials and Chemicals
Total phenolic compound content
Folin- cioculture procedure
500 µl banana crude extract + Folin-ciocaltue reagent 1 ml
5 min
Add Na2CO3 7.5% 3 ml
60 min
Measuring of TPC at OD 760 nm
Note: Gallic acid was used as
standard solution to plot the
calibration curve(Sulaiman et al., 2010; Shian et al., 2012 and Montelongo et al., 2010)
Total phenolic compound content
Folin- cioculture procedure
Statistical analysisStatistical analysis
Data were subject to ANOVA and the
means were compare by Duncan’s New Multiple
Rang Test (DMRT) using SPSS Software with
the same software (SPSS version 17.0 for
window, SPSSInc.,Chicago,IL,.USA)
Results
and
Discussion
Peel Pulp
Stag L* a* b* L* a* b
Mature 50.19 b -18.73 c 34.05 b 83.58 a -0.84 a 22.71
Ripe 74.96 a 1.34 b 55.90 a 82.33 a -1.36 b 22.68
Overripe 72.72 a 4.80 a 53.88 a 82.65 a -0.77 a 25.96 a
F-test ** ** ** ns * *
Means sharing different letters in column are significantly from each other for each
product type (P≤ 0.05)
Color analysis
Table 1 Color value L* a* b* in Kuai Leb Mu Nang 3 different stages
DPPH assay FRAP assay
Analysis of antioxidant activity
317.81b
700.35a
679.75a
21884.26b
33641.87a 33198.12a
Fig.1 : Antioxidant activity by DPPH assay in Kuai
Leb Mu Nang 3 different stages
Fig.2 : Antioxidant activity by FRAP assay in Kuai
Leb Mu Nang 3 different stages
Total phenolic compound content
713.42a
334.20b
670.96a
Fig.3 : Total phenolic compound content in Kuai Leb Mu Nang
3 different stages
Total flavonoid content
2656.87a
2541.25a
2475.62a
Fig.4 : Total flavonoids content in Kuai Leb Mu Nang 3 different stage
ConclusionConclusion
1. Crude extract from all different stages in ‘Kuli Leb Mu
Nang’ showed free radical scavenging activity.
2. Phenolic compounds and flavonoids were found in ‘Kuli
Leb Mu Nang’.
3. Ripe and overripe stage s in ‘Kuli Leb Mu Nang’ showed
antioxidant activity higher than mature stage.
1. Crude extract from all different stages in ‘Kuli Leb Mu
Nang’ showed free radical scavenging activity.
2. Phenolic compounds and flavonoids were found in ‘Kuli
Leb Mu Nang’.
3. Ripe and overripe stage s in ‘Kuli Leb Mu Nang’ showed
antioxidant activity higher than mature stage.
Special thanks to Horticulture Program Division
of Agriculture of Technology King Mongkut's Institute of
Technology Ladkrabang Prince of Chumphon Campus
Chumphon Province.
The authors are thankful to Dr. Pannipa Youryon
(Advisor) and Central laboratory staff.
Acknowledments
Analysis of Antioxidant in banana

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Analysis of Antioxidant in banana

  • 1. Analysis of Antioxidant Activity at Different Stage in Banana Musa (AA group) ‘Kluai Leb Mu Nang’ By Adisorn Jumroon ๔ KMITLโนโลยีพระจอมเกล้าเจ้าคุณทหารลาดกระบัง วิทยาเขตชุมพรเขตรอุดมศักดิ์
  • 2. Introduction - ‘Kluai Leb Mu Nang’ - Free radical - Antioxidant - Phenolic compound - Flavonoid
  • 3. Family Genus Section Genome Cultivar Musaceae Enset Musa Australimusa Callimusa Rhodochlamys Eumusa Ingentimusa AA AAA AAAA AB AAB ABB ABBB ‘Kluai Leb Mu Nang’‘Kluai Leb Mu Nang’ Taxonomic classification Shyamala et al. (2011)
  • 4. Botanical characteristicsBotanical characteristics 1 Stem 8 Leaves 2, 3, 4 Flower 5, 6, 7 Fruit Wendy (2001) 8
  • 14. 1. To study the antioxidant activity at the different stages in ‘Kluai Leb Mu Nang’ 2. To Study total phenolic content and total flavonoids at the different stages in ‘Kluai Leb Mu Nang’ Objectiv es Objectiv es
  • 15. SamplesSamples Kluai Leb Mu Nang 3 different stages - Mature banana (Green) - Ripe banana (Full yellow) - Overripe banana (Dark yellow) Materials and methodsMaterials and methods
  • 16. Evaluating parameters - Color analysis - Extraction process - Antioxidant activity assay - DPPH scavenging assay - Ferric reducing antioxidant power assay - Total flavonoid content analysis - Total phenolic compound analysis
  • 17. Use a chomameter Reported values L* a* b* Color analysisColor analysis
  • 18. Extraction process 50°C, 12 h 4500 rpm, 15 min Add 80% methanol filtration
  • 19. • DPPH (2,2- diphenyl-1-picrylhydrazyl) • Methanol • BHT (butylated hydroxytoluene) • UV-visible spectrophometer • Micropipette DPPH scavengingDPPH scavenging assayassay Materials and ChemicalsMaterials and Chemicals
  • 20. OD 515 nm DPPH DPPH scavengingDPPH scavenging assayassay
  • 21. •C2 H3 NaO2 3H2 O (Sodium acetate trihydrate) •C2 H4 O (Acetaldehyde ) • Acetic acid • FeCl3 6H2 O (Ferric chloride hexahydrate) •TPTZ (2,4,6-Tripyridyl-s-Triazine) • HCl •Trolox (6-hydroxy-2,5,7,8- tetramethylchroman- 2-carboxylic acid) Materials and Chemicals Ferric Reducing AntioxidantFerric Reducing Antioxidant PowerPower
  • 22. OD 593 nm Ferric ReducingFerric Reducing AntioxidantAntioxidant PowerPower
  • 23. •Sodium nitrite (NaNO2 ) •Aluminum chloride (AlCl3 ) • Sodium hydroxide (NaOH) •Catechin ((2R,3S)-2-(3,4-dihydroxyphenyl)-3,4- dihydro-2H-chromene-3,5,7-triol •UV-visible spectrophometer • Micropipette Materials and Chemicals Total flavonoid content Aluminium chloride colorimetric method
  • 24. 0.1 g banana crude extract + 5 ml distilled water + 5% NaNO2 5 min Add 10% AlCl3 0.6ml Add 1M NaOH 2 ml Adjust volume with DW to 10 ml 5 min Fatemeh et al. (2012) and Baskar et al. (2011) Total flavonoid content Aluminium chloride colorimetric method
  • 25. Measuring of TFC at OD510 nm Note: catechin was used as standard solution to plot the calibration curve Total flavonoid content Aluminium chloride colorimetric method
  • 26. •Sodium carbonate (Na2 CO3 ) •Folin–Ciocalteur reagent •Gallic acid (3,4,5-trihydroxybenzoic acid) •UV-visible spectrophometer • Micropipette Materials and Chemicals Total phenolic compound content Folin- cioculture procedure
  • 27. 500 µl banana crude extract + Folin-ciocaltue reagent 1 ml 5 min Add Na2CO3 7.5% 3 ml 60 min Measuring of TPC at OD 760 nm Note: Gallic acid was used as standard solution to plot the calibration curve(Sulaiman et al., 2010; Shian et al., 2012 and Montelongo et al., 2010) Total phenolic compound content Folin- cioculture procedure
  • 28. Statistical analysisStatistical analysis Data were subject to ANOVA and the means were compare by Duncan’s New Multiple Rang Test (DMRT) using SPSS Software with the same software (SPSS version 17.0 for window, SPSSInc.,Chicago,IL,.USA)
  • 30. Peel Pulp Stag L* a* b* L* a* b Mature 50.19 b -18.73 c 34.05 b 83.58 a -0.84 a 22.71 Ripe 74.96 a 1.34 b 55.90 a 82.33 a -1.36 b 22.68 Overripe 72.72 a 4.80 a 53.88 a 82.65 a -0.77 a 25.96 a F-test ** ** ** ns * * Means sharing different letters in column are significantly from each other for each product type (P≤ 0.05) Color analysis Table 1 Color value L* a* b* in Kuai Leb Mu Nang 3 different stages
  • 31. DPPH assay FRAP assay Analysis of antioxidant activity 317.81b 700.35a 679.75a 21884.26b 33641.87a 33198.12a Fig.1 : Antioxidant activity by DPPH assay in Kuai Leb Mu Nang 3 different stages Fig.2 : Antioxidant activity by FRAP assay in Kuai Leb Mu Nang 3 different stages
  • 32. Total phenolic compound content 713.42a 334.20b 670.96a Fig.3 : Total phenolic compound content in Kuai Leb Mu Nang 3 different stages
  • 33. Total flavonoid content 2656.87a 2541.25a 2475.62a Fig.4 : Total flavonoids content in Kuai Leb Mu Nang 3 different stage
  • 34. ConclusionConclusion 1. Crude extract from all different stages in ‘Kuli Leb Mu Nang’ showed free radical scavenging activity. 2. Phenolic compounds and flavonoids were found in ‘Kuli Leb Mu Nang’. 3. Ripe and overripe stage s in ‘Kuli Leb Mu Nang’ showed antioxidant activity higher than mature stage. 1. Crude extract from all different stages in ‘Kuli Leb Mu Nang’ showed free radical scavenging activity. 2. Phenolic compounds and flavonoids were found in ‘Kuli Leb Mu Nang’. 3. Ripe and overripe stage s in ‘Kuli Leb Mu Nang’ showed antioxidant activity higher than mature stage.
  • 35. Special thanks to Horticulture Program Division of Agriculture of Technology King Mongkut's Institute of Technology Ladkrabang Prince of Chumphon Campus Chumphon Province. The authors are thankful to Dr. Pannipa Youryon (Advisor) and Central laboratory staff. Acknowledments