evaluation carotenoids content in south african potato, tomato and wheat cultivars charlotte mashaba...
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Evaluation carotenoids content in South African potato, tomato and wheat cultivars
Charlotte Mashaba
Eugenia Barros
© CSIR 2007 www.csir.co.za
Introduction
• Malnutrition - 50% of deaths in children < 5
• Food insecurities - Community-based intervention projects
• Screening for high micronutrient crops
Carotenoids
• Lipid soluble molecules – produced by plants and photosynthetic organism
• Disease prevention and reduction
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Carotenoids Disease/condition References
β-carotene, β-cryptoxanthin, lycopene and lutein
Reduce risk of Breast cancer developing by up to 50%
Dorgan et al., 1998; Toniolo et al., 2001
Lycopene Reduced risk of coronary heart disease
Sesso et al., 2004
Lutein and zeaxanthin
Protective role against AMD
Gale et al., 2003
α-carotene, β-carotene, β-cryptoxanthin
Pro-vitamin A - surface linings, bone growth, immune system
Semba, 1998During and Harrison, 2007Unlu et al., 2005
Choice of Crops
• Why them?
→ Wheat
→ potato
→ Tomato
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© CSIR 2007 www.csir.co.za
Objectives
• Optimize method
• Quick protocol
Platforms used for analysis
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HPLC
Crude extraction
• Quantify known carotenoid
Results - Standard curves
Fig 1 Standard curves to determine carotenoids content
© CSIR 2007 www.csir.co.za
© CSIR 2007 www.csir.co.za
PotatoCrop Cultivar
(n=3)Lutein Zeaxanthin Canthaxanthin β-Carotene Lycopene
Potato Mondial 0.265 ± 0.008 0.441 ± 0.01 ND 0.0201 ± 0.004 ND
BP1 0.333 ± 0.003 0.120 ± 0.003 ND 0.0142 ± 0.005 ND
Tomato Salad 10.801 ± 0.028 0.303 ± 0.008 0.065 ± 0.006 28.940 ± 0.55 387.000 ± 5
Roma 10.867 ± 0.056 0.212 ± 0.001 0.080± 0.01 25.197 ± 1.37 443.090 ± 8.908
Wheat Mathabas 0.544 ± 0.017 0.138 ±0.004 ND 0.034± 0.003 ND
Elands 0.268 ± 0.017 0.154±0.011 ND ND ND
Moni 0.302±0.0339 0.082±0.006 ND ND ND
Orania 0.307±0.008 0.036±0.001 ND ND ND
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Tomato Crop Cultivar(n=3)
Lutein Zeaxanthin Canthaxanthin β-Carotene Lycopene
Potato Mondial 0.265 ± 0.008 0.441 ± 0.01 ND 0.0201 ± 0.004 ND
BP1 0.333 ± 0.003 0.120 ± 0.003 ND 0.0142 ± 0.005 ND
Tomato Salad 10.801 ± 0.028 0.303 ± 0.008 0.065 ± 0.006 28.940 ± 0.55 387.000 ± 5
Roma 10.867 ± 0.056 0.212 ± 0.001 0.080± 0.01 25.197 ± 1.37 443.090 ± 8.908
Wheat Mathabas 0.544 ± 0.017 0.138 ±0.004 ND 0.034± 0.003 ND
Elands 0.268 ± 0.017 0.154±0.011 ND ND ND
Moni 0.302±0.0339 0.082±0.006 ND ND ND
Orania 0.307±0.008 0.036±0.001 ND ND ND
Wheat
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Crop Cultivar(n=3)
Lutein Zeaxanthin Canthaxanthin β-Carotene Lycopene
Potato Mondial 0.265 ± 0.008 0.441 ± 0.01 ND 0.0201 ± 0.004 ND
BP1 0.333 ± 0.003 0.120 ± 0.003 ND 0.0142 ± 0.005 ND
Tomato Salad 10.801 ± 0.028 0.303 ± 0.008 0.065 ± 0.006 28.940 ± 0.55 387.000 ± 5
Roma 10.867 ± 0.056 0.212 ± 0.001 0.080± 0.01 25.197 ± 1.37 443.090 ± 8.908
Wheat Mathabas 0.544 ± 0.017 0.138 ±0.004 ND 0.034± 0.003 ND
Elands 0.268 ± 0.017 0.154±0.011 ND ND ND
Moni 0.302±0.0339 0.082±0.006 ND ND ND
Orania 0.307±0.008 0.036±0.001 ND ND ND
Tomato Chromatograms
Roma
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min5 10 15 20 25 30 35
Norm.
0
200
400
600
800
1000
DAD1 A, Sig=450,10 Ref=400,10 (CAR1812\CARO0018.D)
Lutein
Zeaxanthin
Canthaxanthin
Beta-carotene
Lycopene
min5 10 15 20 25 30 35
Norm.
0
50
100
150
200
DAD1 A, Sig=450,10 Ref=400,10 (CAR1812\CARO0028.D)
Lutein
Beta-carotene
Lycopene
A
B
Absorbance
(mAU)
tr -Retention time (minutes)
min5 10 15 20 25 30 35
Norm.
0
10
20
30
40
DAD1 A, Sig=450,10 Ref=400,10 (CAR0412\CARO0027.D)
Lutein
Zeaxanthin
Beta-carotene
min5 10 15 20 25 30 35
Norm.
0
2.5
5
7.5
10
12.5
15
17.5
DAD1 A, Sig=450,10 Ref=400,10 (CAR0412\CARO0028.D)
Lutein
Zeaxanthin
Beta-carotene
Absorbance
(mAU)
Rt -Retention time (minutes)
A
B
Mondial
BP 1
min5 10 15 20 25 30 35
Norm.
0
10
20
30
40
DAD1 A, Sig=450, 10 Ref =400, 10 (CAR0412\CARO0029.D)
Lut
ein
Zea
xant
hin
Can
thax
anth
in
min5 10 15 20 25 30 35
Norm.
0
20
40
60
80
100
DAD1 A, Sig=450,10 Ref=400,10 (CAR0412\CARO0031.D)
Lutein
Zeaxant
hin
Canthax
anth
in
Beta-ca
rote
ne
A
B
Absorbance
(mAU)
tr -Retention time (minutes)
Bread wheat
Durum
wheat
Platforms used for analysis
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MALDI-get pic
Crude extraction
• Identify unknown
Identification of ‘unknown’ peaks
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Theoretical mass Formula Matched carotenoid
552.4331 C40H56O Cryptoxanthin
582.4073 C40H54O3 Adonixanthin;
596.4593 C42H60O2 Sprilloxanthin
598.4022 C40H54O4 Ertythroxanthin
TomatoTheoretical
mass Fomula Matched carotenoid
574.4750 C40H62O2 Chloroxanthin
596.4590 C42H60O2 Sprilloxanthin
598.4750 C42H62O2 Dihydrosprilloxanthin
600.4906 C42H64O2 Tetrahydrospirilloxanthin
716.3359 C40H53NaO8S Ketonostoxanthin 3-Sulfate
780.5693 C52H76O5 Siphonaxanthin
Wheat
Potato
Theoretical mass Fomula Matched carotenoid
574.4750 C40H62O2 Chloroxanthin
582.4073 C40H54O3 4-Ketozeaxanthin; Oxosaproxanthin
598.4022 C40H54O4 Halocynthiaxanthin;
Conclusion
• Methods used robust
→ More carotenoids were separated
→Sensitive to quantify low amounts
→ Can be used for more crops
→Develop SOP
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Thank you!
Acknowledge
• Judy Reddy
• Dr Stoyan Stoychec
© CSIR 2007 www.csir.co.za
AIMTheoratical
mass Formulae Matched carotenoid534.4226 C40H54 Didehydrolycopene; 536.4382 C40H56 alhpa/beta -Carotene; Lycopene536.4382 C40H56 alhpa/beta/ -Carotene; Lycopene540.4695 C40H60 Tetrahydro-beta-Carotene; Tetrahydrolycopene542.4851 C40H62 Phytofulene550.4175 C40H54O tetradehydolycopene550.4175 C40H54O 4-Keto-gamma-carotene; tetradehydolycopene552.4331 C40H56O -cryptoxanthin568.428 C40H56O2 Zeaxanthin; Lutein;
582.4073 C40H54O3 Adonixanthin; 596.4593 C42H60O2 Sprilloxanthin598.4022 C40H54O4 Ertythroxanthin
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Current & future work
• Analyze more cultivars and increase nutrients
PotatoComparison of Carotenoids content in Mondial and BP1 potato cultivarss
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min5 10 15 20 25 30 35
Norm.
0
10
20
30
40
DAD1 A, Sig=450,10 Ref=400,10 (CAR0412\CARO0027.D)
Lutein
Zeaxanthin
Beta-carotene
min5 10 15 20 25 30 35
Norm.
0
2.5
5
7.5
10
12.5
15
17.5
DAD1 A, Sig=450,10 Ref=400,10 (CAR0412\CARO0028.D)
Lutein
Zeaxanthin
Beta-carotene
Absorbance
(mAU)
Rt -Retention time (minutes)
A
B
Mondial
BP 1