investigating fruit juice authenticity using ms - waters corporation food research

©2015 Waters Corporation 1

Investigating Fruit Juice Authenticity using

MS and ‘Omics approaches

http://rds.yahoo.com/_ylt=A0PDoS1TMnBNwlMAnq.JzbkF;_ylu=X3oDMTBxZWd1N2R0BHBvcwMxMQRzZWMDc3IEdnRpZANJMTM2Xzg2/SIG=1j9ndvfs3/EXP=1299227347/**http:/images.search.yahoo.com/images/view?back=http://images.search.yahoo.com/search/images?_adv_prop=image&va=pineapple&fr=yfp-t-311&w=322&h=595&imgurl=omahanightlife.com/images/1/media/2009/05/07/pineapple.jpg&rurl=http://omahanightlife.com/articles/pineapplexpress_drinkoftheweek&size=28KB&name=Arsenal&


Presentation overview

Project Background

Workflow

Results o UV

o MS

Sample Comparison o Visual inspection

o Software interpretation

Why are they different? o Are there any marker compounds?

o Can we ID marker compounds?

Ion mobility for discovery of natural compounds


Project background

Activities in food fraud and authenticity has increased in

importance over the last decade

– Estimated to cost $10-$15 billion per year

Fruit juices (orange and apple juice) were in the top 7 foods

reported from 1980 to 2010 as the most common targets for

adulteration

Food & beverage products are adulterated in many different ways:

– Be harmful to health

– Misleading to the consumer

o …includes unacceptable enhancement, dilution and/or substitution with

less expensive ingredients, failure to declare contamination and

inaccurate or misleading labelling of a product or ingredient

Reported cases of adulterated fruit juices have been economically

driven, e.g. inclusion of high fructose corn syrup, other fruit juices…


Workflow Overview How did we perform the analysis?


Results: PDA & MS (exact mass) Data

Time2.00 3.00 4.00 5.00 6.00 7.00

%

1

2.00 3.00 4.00 5.00 6.00 7.00

%

3

2.00 3.00 4.00 5.00 6.00 7.00

AU

0.0

2.5e-2

5.0e-2

PA_24_2_010 4: Diode Array 269

Range: 7.395e-2

4.81

1.30

4.29

1.75 2.533.99

5.66

PA_24_2_010 1: TOF MS ES- BPI

2.09e4

2.85

1.33

1.83

5.244.44

3.91

5.68

PA_24_2_010 2: TOF MS ES- BPI

9.19e3

5.04

1.33

5.68MS –

product ions

MS – precursor ions

PDA


Sample Comparison Visual inspection – Study Samples:

Time1.00 2.00 3.00 4.00 5.00 6.00 7.00

AU

0.0

5.0e-2

1.00 2.00 3.00 4.00 5.00 6.00 7.00

AU

0.0

5.0e-2

1.00 2.00 3.00 4.00 5.00 6.00 7.00

AU

0.0

5.0e-2

1.29 4.80

4.56

2.731.73 3.27

5.61

1.304.80

4.55

2.70 3.265.61

4.811.30

4.44

2.71 3.27

5.65


-600

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-100

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-1200 -1000 -800 -600 -400 -200 0 200 400 600 800 1000 1200

t[2]

t[1]

Scores Comp[1] vs. Comp[2] colored by Sample Group

S10S11S12

S12

S12

S12

S12

S12

S12S12

S11

S11S11S11S11S11S11S11

S10

S10S10S10S10

S10

S10

S10S10

EZinf o 2 - Negativ e_Pineapple_no_QC_3_samples (M8: PCA-X) - 2011-02-28 20:50:30 (UTC-5)

Sample Comparison Software inspection – Study Samples


Sample Comparison Software inspection – Study & Supermarket samples

Supermarket samples

Study samples


Identify Markers for Observed Differences OPLS-DA Model: 207 / 316 & 102


Identify Markers for Observed Differences S-Plot: 207 / 316 & 102

-1.0

-0.8

-0.6

-0.4

-0.2

-0.0

0.2

0.4

0.6

0.8

1.0

-0.2 -0.1 -0.0

p(c

orr

)[1]P

(C

orr

ela

tio

n)

w*[1]P (Covariance)

S-Plot (S10 = -1, S11_S12 = 1)

EZinf o 2 - Negativ e_Pineapple_S10_VS11S12 (M6: OPLS-DA) - 2011-03-11 09:49:12 (UTC-5)

Rt 5.03- m/z 649.2495

Rt 5.55-m/z 579.1712

Rt 5.67-m/z 609.1616

Rt 5.10-m/z 711.2862


Identify Markers for Observed Differences Trend Plot for the markers in all samples

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Sample Group

Variables

DS1: 5.55_579.1712DS1: 5.67_609.1816DS1: 5.03_649.2495DS1: 5.10_711.2862

EZinf o 2 - Negativ e_Pineapple_w_QC (M6: PCA-X) - 2011-03-11 10:35:34 (UTC-5)


Why are there differences?


Why are there differences? Marker ID – using R.T, MS and UV


Time1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

100

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

100

1.50 2.00 2.50 3.00 3.50 4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50

%

0

100

PA_24_2_063a 1: TOF MS ES- 609.182 0.0030Da

8.09e3

PA_24_2_062a 1: TOF MS ES- 609.182 0.0030Da

8.09e3

5.66

PA_24_2_061a 1: TOF MS ES- 609.182 0.0030Da

8.09e3




m/z50 100 150 200 250 300 350 400 450 500 550 600 650

%

0

100301.0706

609.1822


nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

2.0e-2

2.5e-2

nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

2.0e-2

2.5e-2

Time3.00 4.00 5.00 6.00 7.00

AU

0.0

1.0e-2

2.0e-2

3.0e-2

4.0e-2

5.0e-2

3.00 4.00 5.00 6.00 7.00

AU

0.0

1.0e-2

2.0e-2

3.0e-2

4.0e-2

5.0e-2

PA_24_2_096a 4: Diode Array 283

Range: 2.583e-2

5.66


Range: 5.559e-2

4.83

4.31

2.733.993.33

5.03

5.66

Confirm marker Identification Marker ID – using R.T, MS and UV


Confirm marker Identification Marker ID – using R.T, MS and UV

Time1.00 2.00 3.00 4.00 5.00 6.00 7.00

%

0

100

1.00 2.00 3.00 4.00 5.00 6.00 7.00

%

0

100

PA_24_2_098a 2: TOF MSMS ES- 609.182 0.0050Da

1.96e3

5.67

PA_24_2_093a 2: TOF MSMS ES- 609.182 0.0050Da

3.57e3

5.67

m/z100 200 300 400 500 600

%

0

100

m/z100 200 300 400 500 600

%

0

100301.0707

609.1819

301.0706

609.1821


Hesperidin Sources


Identifying other markers highlighted....



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S12

S12

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Sample Group

Variables

DS1: 5.55_579.1712DS1: 5.67_609.1816DS1: 5.03_649.2495DS1: 5.10_711.2862



Time2.00 3.00 4.00 5.00 6.00 7.00

%

0

100

2.00 3.00 4.00 5.00 6.00 7.00

%

0

100

2.00 3.00 4.00 5.00 6.00 7.00

%

0

100

PA_24_2_070a 1: TOF MS ES- 579.171 0.0030Da

4.12e3

PA_24_2_071a 1: TOF MS ES- 579.171 0.0030Da

4.12e3

5.54

PA_24_2_072a 1: TOF MS ES- 579.171 0.0030Da

4.12e3

Identify Markers for Observed Differences Potential Compound ID’ed as Naringen


Time2.00 4.00 6.00

%

0

100

2.00 4.00 6.00

AU

0.0

1.0e-2

2.0e-2


Range: 3.084e-2

5.57

PA_24_2_097a 1: TOF MS ES- 579.171 0.0030Da

1.73e4

5.59

Identify Markers for Observed Differences

Potential Compound ID’ed as Naringen

Same elemental composition C27H31O14

Time2.00 4.00 6.00

%

0

100

2.00 4.00 6.00

AU

0.0

2.0e-2

4.0e-2

6.0e-2


Range: 6.675e-2

1.314.80

4.28

2.703.95

3.31

5.64

PA_24_2_062a 1: TOF MS ES- 579.171 0.0030Da

8.08e3

5.54


Identify Markers for Observed Differences Potential Compound – Naringen ”like”

m/z100 200 300 400 500 600

%

0

100

m/z100 200 300 400 500 600

%

0

100579.1720

271.0604

151.0029

459.1152

271.0603

151.0029 579.1714

Naringin like component

nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

nm260 280 300 320 340

AU

0.0

5.0e-3

1.0e-2

1.5e-2

JA-11S2-102


C27H32O14

MW:580.54



0

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Sample Group

Variables

DS1: 5.55_579.1712DS1: 5.67_609.1816DS1: 5.03_649.2495DS1: 5.10_711.2862




0

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S12

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S12

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S11

S11

S11

S11

S11

S11

S10

S10

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S10

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S10

S10

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Sample Group

Variables

DS1: 5.55_579.1712DS1: 5.67_609.1816DS1: 5.03_649.2495DS1: 5.10_711.2862


Rt 5.10 min, m/z 711.2862 Nomilinic acid 17-beta-D-glucopyranoside (NAG)

Rt 5.03 min, m/z 649.2495 Limonin 17-beta-D-glucopyranoside (LG)


Conclusions


Conclusions

Using the data processing tools we were able to identify the

presence of citrus components in JA-11S2-102.

Flavone-O-glycosides

– Hesperidin

– Naringin “like” compound possibly narirutin

Tentative identification of two citrus limonoids (no standards

available)

– Limonin 17-beta-D-glucopyranoside (LG)

– Nomilinic acid 17-beta-D-glucopyranoside (NAG)