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