การวิเคราะห์ฤทธิ์ต้านอนุมูลอิสระที่ระยะต่างกันในกล้วยเล็บมือนาง Analysis of Antioxidant Activity at Different Stage in Musa (AA group) ‘Kluai Leb Mu Nang’ อดิศร จำรูญ

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

10. Phenolic compound

12. FlavonoidsFlavonoids

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)

29. Results
and
Discussion

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