Analysis of Antioxidant Activity at Different Stage in Banana

Musa (AA group ) ‘Kluai Leb Mu Nang’

ByAdisorn Jumroon

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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

Phenolic compound

FlavonoidsFlavonoids

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’

ObjectivesObjectives

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 valuesL* a* b*

Color analysisColor analysis

Extraction process

50C, 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 scavenging DPPH scavenging assayassay

Materials and ChemicalsMaterials and Chemicals

OD 515 nm

DPPH

DPPH scavenging DPPH scavenging assayassay

•C2H3NaO2 3H2O (Sodium acetate

trihydrate)•C2H4O (Acetaldehyde )• Acetic acid • FeCl3 6H2O (Ferric chloride hexahydrate)•TPTZ 2( ,4 , -6 Tripyridyl-s-Triazine )• HCl •Trolox (6-hydroxy-2,5,7,8-tetramethylchroman- 2-carboxylic acid ) •UV-visible spectrophometer • Micropipette

Materials and Chemicals

Ferric Reducing Ferric Reducing Antioxidant PowerAntioxidant Power

OD 593 nm

Ferric Ferric Reducing Reducing

Antioxidant Antioxidant PowerPower

•Sodium nitrite (NaNO2

)

•Aluminum chloride (AlCl3

)• Sodium hydroxide (NaOH )•Catechin ((2R,3 S)- -2 (3 , -4dihydroxyphenyl)-3 , -4 dihydro-2 H-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 (Na2CO3) •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 ml5 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 analysis Statistical 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 b Ripe 74.96 a 1.34 b 55.90 a 82.33 a -1.36 b 22.68 b 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

Conclusion Conclusion

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