Evaluation carotenoids content in South African potato, tomato and wheat cultivars Charlotte Mashaba...

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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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

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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