3/13/2013

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POSTHARVEST AND PROCESSING EFFECTS ON HUMAN HEALTH

PROPERTIES

Bhimu Patil

B-patil@tamu.edu

Overview

• Necessity of processing……

• Current trends: Consumer

• Evidences of health benefits– What are phytochemicals?

– Why do we need them?

• Post-harvest & Processing effects on phytochemicals– Degreening, Packaging

– Innovative technologies: HPP, Irradiation

• Future Outlook and Challenges

Population growth

< 9.0 B

~ 6.8 BDemand for food

will increase

Strategies for sustainability….

Food Processing

29,000 companies in USA (over-all)

Employ 1.4 million people

Top 5 food processing states:1) California

2) New York

3) Texas

4) Illinois

5) Florida

Trends in the food processing industry

Top 3 growth areas

1) Organics and naturals

2) Specialized beverages: (High phytochemicals)

3) Ready to eat: (Minimally processed)

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Light stress Climate stressClimate stress

Water, nutrients, saltstress

Disease,Insectsstress

Fruits and Vegetables: Phytochemicals

REACTIVE OXYGEN SPECIES

http://www.drblayney.com/

“Extranutritional constituents that typically occur in small quantities in foods”

- Kris-Etherton

Phytochemicals

Carotenoids

Limonoids

Tanins

Fruits and Vegetables: Phytochemicals

Flavonoids Capsiacinoids

Anthocyanins

•Smoking

•Pollution

•UV rays

•Ionizing Radiation

•Stress

•Pesticide application

Cancer Cardiovascular diseases          Inflammation

Free radical

DNA

Phytochemicals

National Cancer Institute

Mechanism of action

Social wellbeing

Protect the environment

Provide significantly

to the economyAesthetics

Reduce stress

Culinary

Heart

AgeingSkin HealthOsteoporosis

CancersColonPancreasBreastProstate…….

Evidences of F&V role in human health

Food pyramid/My Plate

2011: MyPlateUSDA’s food guide pyramid 2005

WHY FRUITS AND

VEGETABLES ?

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• Global mortality attributed to inadequate of F&V is 2,635 million death per year.

• Increase 600 g/day could reduce worldwide burden of coronary heart disease by 31% and stroke by 19%(1)

• Higher F&V consumption- savings of Medicare charges $2,000 (2)

1. Lock et al., Bull World Health Org 2005. 100-1082. Daviglus et al., J. Am Diet Assoc 2005, 1735-1744.

Cost of not consuming F&V

Current consumer trends: 2012

“Consumers prefer to obtain their nutrients naturally and their health benefits naturally, versus fortified foods or vitamin and mineral supplements”

Elizabeth Sloan, Editor of IFT's Food Technology magazine

Top 10 functional food trends of 2012

Elizabeth Sloan, IFT's Food Technology magazine

1.Real food nutrition2.Mini managers3.Bioavailability4.Protein power5.Plant based6. Gourmet nutrition7. The new risks (stroke, heart attack prevention)8. First aid (relief from sore throat, constipation, sleeplessness)9. Kids, dads and grannies (appealing to these groups)10. Liquidification

People are making an effort to serve meals that are naturally

higher in vitamins and nutrition

Overview

• Necessity of processing……

• Current trends: Consumer

• Evidences of health benefits– What are phytochemicals

– Why do we need them

• Processing effects on phytochemicals– Degreening, Packaging

– Innovative technologies: HPP, Irradiation

• Future Outlook and Challenges

Waxing, washing

UV and Gamma

Irradiation

Heat treatmentMAP

Postharvest treatments

Processing

Storage period &

temperature

Controlledatmosphere

storage

DegreenedNon degreened

Destroys chlorophyll

Carotenoids accumulate

Red / Orange color develops

Degreening - Ethylene

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Degreening room

•Ethylene Concentration•Duration •Temperature•Relative Humidity•CO2 %•Air circulation & ventilation

Factors affecting degreening

Effect of degreening on external appearance

Days after degreening

0 D

Control Degreening

7 D

14 D

21 D

28 D

35 D

Control Degreening

0

20

40

60

0 7 14 21 28 35

mg/

100g

dry

wt

Storage (days)

DNAG ND DG

a ab a

a

ba a a

b

a

b

0

20

40

60

80

100

0 7 14 21 28 35

mg/

100g

dry

wt

Storage (days)

LimoninND DGaa

b ab

aa

aa a

a a

0

20

40

60

80

100

0 7 14 21 28 35

mg/

100g

dry

wt

Storage (days)

NomilinND DG

a a

ba a a

ba a

a ab

Effect of degreening on limonoids

Limonoids pathway

Deacetyl nomilinic acid (DNA)

( )

Deacetyl nomilinic acid glucoside

(DNAG)

Limonin

Nomilin

Effect of degreening on flavonoids

0

100

200

300

0 7 14 21 28 35

mg/

100

g dr

y w

t

Storage (days)

Narirutin ND DG

a aa a

a

b

a a a aa

b

0

300

600

900

1200

1500

0 7 14 21 28 35

mg/

100

g dr

y w

t

Storage (days)

Naringin ND DG

aa

a aa a

a aa a

a

b

0

25

50

75

100

0 7 14 21 28 35

mg/

100

g dr

y w

t

Storage (days)

Poncirin ND DG

aa

a a

a a aa

a

a

bb

0

100

200

300

400

0 7 14 21 28 35

µg/

g dr

y w

t

Storage (days)

DHB ND DGa a

b

a

b

a

aa

aa

a a

0

10

20

30

40

0 7 14 21 28 35

µg/

g dr

y w

t

Storage (days)

Bergamottin ND DGa

ab

a

b

a

a ab

a a

b

0

25

50

75

100

125

150

175

0 7 14 21 28 35

µg/

g dr

y w

t

Storage (days)

5G7MC ND DG

a a b

aa a

b

a a a

b

a

Effect of degreening on furocoumarins

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Temperature Conditioning

• Grapefruits are susceptible to chilling injury when stored at low temperature (< 10°C) for prolonged period

• Chilling injury symptoms – browning of flavedo, pitting, surface lesions, water soaked tissues

• Temperature conditioning – 7 days at 16C or 3 days at 21°C then stored at low temperature

• Other treatments used – Intermittent warming, waxes and vegetable oils, modified atmosphere packaging

External appearance

Storage (weeks)4

11°C

2°C

8 12 16

CD + 2°C

Decay and Chilling injury

0

10

20

30

40

4 8 12 16

11° C 2° C CD

Dec

ay (%

)

Storage (weeks)

0

10

20

30

40

4 8 12 16

11° C 2° C CD

Ch

illin

g in

jury

(%)

Storage (weeks)

Flavonoids

0

100

200

300

400

500

600

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Narirutin 11ºC 2ºC CD

a

bb b a b

a

b ba aa

0

500

1000

1500

2000

2500

3000

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Naringin 11ºC 2ºC CD

abc

b a b

a

b ba aa

0

10

20

30

40

50

60

70

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Didymin 11ºC 2ºC CD

a a aa

c a b bbb b

a

0

30

60

90

120

150

180

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Poncirin 11ºC 2ºC CD

a a a

a

b

bc a

b b

c

a

Furocoumarins

0

30

60

90

120

150

0 4 8 16

(µg/

g d

ry w

t)

Storage (weeks)

DHB 11ºC 2ºC CD

b

ab a a b

bc

a

a a a

0

30

60

90

120

150

0 4 8 16

(µg/

g d

ry w

t)

Storage (weeks)

Bergamottin 11ºC 2ºC CD

a a a c b ba b a ab ab

0

30

60

90

120

150

0 4 8 16

(µg/

g dr

y w

t)

Storage (weeks)

5G7MC 11ºC 2ºC CD

a a a b

a a a b bab b

a

Limonoids

0

20

40

60

80

100

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

DNAG 11ºC 2ºC CD

a aa a b aa a a

a

b b

0

40

80

120

160

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Limonin 11ºC 2ºC CD

c

a b

a ab ac

b

a

b

a a a

0

30

60

90

120

150

0 4 8 16

mg/

100g

dry

wt

Storage (weeks)

Nomilin 11ºC 2ºC CD

a aa

aa a a ab b a a

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Pancreatic cancer

Colon cancer

Breast cancer

Limonoids: Cancer

Inhibitory Role of Limonoids in Cancer

Patil et al., J Agric. Food Chem. 57: 10933‐10942

Neuroblastoma

Prostate cancer

Jayaprakasha et al., Food Chemistry, 131:184‐191. 

Kim et al., Eur. J.Pharma. 670: 44‐48

Murthy et al., Food Chem. Tox.49:1616‐1625 Murthy et al.,In vitro Toxicology, 25: 859‐867

Patil et al., Microchem. J., 94:108‐117.

Somasundaram et al., J. App. Glob. Res., 3: 67‐72.

Jayaprakasha et al., J. Agric. Food Chem., 58: 180‐186

Leonardi et al., Exp. Bio. Med., 235:710‐717

Poulose et al, The FASEB Journal 2006. 20:A11‐A12

Gunasekara et al., Nutrition and Cancer 58:117‐121

Neuroblastoma: Limonoids & Caspase activity

0

40

80

120

0 6 12 18 24Hours

Caspase 3/7 activity 

Control

LGOG

NAGDNAG

CAMP

Poulose, Harris and Patil. J. Nutr. 2005 135: 870-877.

Neuroblastoma: Limonoids & Caspase activity

Featured on the cover page

Poulose, Harris and Patil. J. Nutr. 2005 135: 870-877.

Geranyloxy‐7‐MOC induces apoptosis in SW480 cells 

….KNC Murthy et al., Bioorganic & Medicinal Chemistry 2012

Other secondary metabolites from Citrus: Cancer prevention

Citrus limonoids: GST activity

0.00

0.25

0.50

0.75

1.00

Stomach Intestine

Control Mixture Nomilin Deacetyl Isoobacunoic

Perez et al. BMC Complementary and Alternative Medicine 2010, 10:51

GS

T A

ctiv

ity

(un

its

/ m

g p

rote

in)

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Citrus limonoids: GST and QR activity

QR

Act

ivit

y(u

nit

s/ m

g p

rote

in)

0.00

0.50

1.00

1.50

2.00

2.50

Liver Lung

Control

Mixture

Nomilin

Deacetyl

Isoobacunoic

Perez et al. BMC Complementary and Alternative Medicine 2010, 10:51

Defuran limonin Limonin 7‐methoxime

Perez et al., J. Agric. Food Chem. 2009, 57, 5279-5286.

38

Limonoids: induction of phase II enzymes..?

Percentages whPerez et al. BMC Complementary and Alternative Medicine 2010, 10:51en inductions statistically significant (p<0.05)

GS

T A

ctiv

ity

(un

its

/ m

g p

rote

in)

130%

51%

*

Perez et al., J. Agric. Food Chem. 2009, 57, 5279-5286.39

Modified Limonoids and GST activity

INTESTINE STOMACHLIVER LUNG

Limonoids and GST activity

Perez et al., J. Agric. Food Chem. 2009, 57, 5279-5286.40

Modified Limonoids and GST activity

Quarantine treatments

Fruit flies (Caribbean, Mediterranean, Mexican etc)

Temperature treatments– Exposure temperature and time

– Dry heat, vapor heat, hot water, hot air,

quick freezing temperature

Irradiation Dose and time

ColdCold HeatHeat

Irradiation

Gamma rays

UV rays

Electron beam

Approved doses (APHIS, USDA)Pest _ Common name Dose (Gy) Bactrocera dorsalis Oriental FF 250 Ceratitis capitata Mediterranean FF 225 B. cucurbitae Melon fly 210 Anastrepha fraterculus South American FF 150 A. suspensa Caribbean FF 150 A. ludens Mexican FF 150 Sternochetus mangiferae Mango seed weevil 300

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Naringin and Narirutin were significantly higher in freeze dried pulp from Irradiated fruits

Vanamala, Cobb, Turner, Lupton, Yoo, Pike and Patil 2005. J.Ag Food Chem. 3989-3995.

Nomilin significantly decreased in freeze dried pulp from Irradiated fruits

Vanamala, Cobb, Turner, Lupton, Yoo, Pike and Patil 2005. J.Ag Food Chem. 3989‐3995.

Opportunities for Colon Cancer Prevention

10 - 20 years

Chronic Inflammation

iNOS and COX-2

Image source: http://personalweb.sunset.net/~mansell/polyp.htm

http://medicalimages.allrefer.com/large/structure-of-the-colon.jpg

Colon Cancer Development

Pitot, Ailment Pharmacol Ther, 2000

Initiation

Promotion

Progression

Tumor formation

DNA Damage

Proliferation

Apoptosis

Early Lesions

Facing the open, flat surface of colon

– Small round openings are normal crypts

– Larger openings are aberrant crypts

– Clusters of aberrant crypts are ACF

This is a high multiplicity ACF

Diets Containing GFPP, IGFPP, Naringin, and

Limonin significantly reduced AC and HMACF

GFPP – Grapefruit Pulp Powder

IG – Irradiated Grapefruit Pulp Powder

Vanamala et al., 2006. Carcinogensis

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Proliferating Cells

• Slide of a rat colon– Slice through

– Proliferating cells are usually located in the bottom part

– Indicated by brown stain generated in PCNA assay

Proliferative IndexProliferative Index

(% of Basal)

All diets reduce proliferation with the maximum reduction occurring with the GFPP and limonin diets.

Vanamala et al., 2006. Carcinogensis

P = 0.02a

b

bb

b

Cyclooxygenase - 2

COX-2 Protein Level

aa

bb

ab

P < 0.032

COX-2 levels were lowest in rats consuming the GFPP and limonin diets.

Vanamala et al., 2006. Carcinogensis

Nitric Oxide Synthase

iNOS Protein Levels

a

bb b

b

P < 0.003

Rats consuming all experimental diets had less iNOS protein than those consuming the basal diet.

Vanamala et al., 2006. Carcinogensis

Control

1

2.5

5

10

3

2.5

2

1.5

1

0.5

0

Vitam

in C (mg / ml) 

E‐beam doses (kGy)

Rio Red Marsh White

Girennavar et al., 2008. J.Agric Food Chem. 56:10941‐10946

Electron beam Irradiation on Naringin

Control

1

2.5

5

10

E‐beam doses (kGy)

Rio Red Marsh White

4.5

4

3.5

3

2.5

2

1.5

1

0.5

0

Na

rin

gin

con

c(m

g /

ml)

Girennavar et al., 2008. J.Agric Food Chem. 56:10941‐10946

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Electron beam Irradiation on Nomilin

Control

1

2.5

5

10

E-beam doses (kGy)Rio Red Marsh White

60

50

40

30

20

10

0

No

mil

inco

nc

(µg

/ m

l)

Girennavar et al., 2008. J.Agric Food Chem. 56:10941‐10946

Industrial processing techniques

High pressure processingThermal processing

Commercially processed HPP Juice

In US, HPP-processed fruit juices are commercially marketed by retail giants such as Starbucks.

Annual market for HPP processed foods is estimated at ~$3.4B. Picture source: Starbucks/ Evolution fresh

Color

L*

(A) Lightness (B) Redness

a*

7 0 14 21 7 0 14 21

7 0 14 21

b*

(C) YellownessDays after storage at 4° C Days after storage at 4° C

Thermal processing increased the lightness

and yellowness, indicating juice color

turning paler

Ascorbic acid

7 0 14 21 Days after storage at 4° C

mg/

100

mL

a ab bc bc

dc

e e

bc

ef f

Thermal processing degraded

ascorbic acid

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HPP maintains the antioxidant activity in GFJ

GFJ processed by HPP at 600 MPa higher antioxidant activity (DPPH assay)than TP processed juice

HPP maintains aroma compounds

-CadineneD-LimoneneGermacrene D

-Copaeneß-Caryophylleneoxide

ß-Myrecene

-Caryophyllene -Selinene

Ethyl octanoate

Control

HPP processed juice

Thermal processed juice

HPP inactivates Pectinmethylesterase(PME) activity in GFJ

HPP treatment at 600 MPacompletely inactivated PME activity

Objectives:

Different household practices

Blending Juicing Squeezing

Household processing effect

Household processing techniques: Flavonoids

mg/ 100 m

L

Household processing effect: Flavonoids

Uckoo et al., J. Food science 2012, Vol. 77, Nr. 9, 2012

Household processing techniques: Limonoids

mg/ 100 m

L

c

b

a

Household processing effect: Limonoids

Uckoo et al., J. Food science 2012, Vol. 77, Nr. 9, 2012

3/13/2013

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mg/ 100 m

La

a

a

b

cc

c

b

b

Household processing techniques: Furocoumarins

Household processing effect: Furocoumarins

Uckoo et al., J. Food science 2012, Vol. 77, Nr. 9, 2012

Household processing techniques: Citric acid and Ascorbic acid

mg/ 100 m

L

Citric acida

ab

b

Ascorbic acida

abb

Household processing effect: Ascorbic & Citric acid

Uckoo et al., J. Food science 2012, Vol. 77, Nr. 9, 2012

Summary:

BLENDING JUICING SQUEEZING

Flavonoids

Limonoids

Furocoumarins

Citric acid

Ascorbic acid

Household processing effect: Summary

Uckoo et al., J. Food science 2012, Vol. 77, Nr. 9, 2012

Quality Nutrition

Balance between quality and nutrition

Media Impact

Consumers need to be:PROACTIVE,ATTENTIVE

& INTELLIGENT

?$$

Future outlook…….. PERSPECTIVES & CHALLENGES

Increase consumption

of F&V

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Challenges

1.Systems wide approach:• Consumer to farm

2.Optimization3.Bioavailability4.Microbioata5.Toxicity6.Consumers trend

FARM ANALYTICALPROCESSING

BIOASSAY/ISOLATION

BIOLOGICAL STUDIESCONSUMER

CONSUMER to

FARM

FARMto

CONSUMER

74

Bioavailability

Increasing the bioavailability is the single most

important challenge in

obtaining optimum health

benefits derived from

F&V

Case study: Bioavailability of carotenoids

Food matrix

Food-preparation techniques

Dose

Amounts of fat

Type ofprocessing

Amounts offiber

(Haskell, Am J Clin Nutr , 2012)

Increasing bioavailability of carotenoids

+

Low bioavailability

More bioavailability

Bioavailability: Plasma Limonin conc. in subjects at different times after Limonin glucoside intake

0

1

2

3

4

5

6

1 3 6

0.250.512

LG dose conc. (g)Bioavailability of a) other bioactivesb) interaction effectc) probable synergistic activity

STILL UNKNOWN

Bioavailability of a) other bioactivesb) interaction effectc) probable synergistic activity

STILL UNKNOWN

Manners et al., J. Agric. Food Chem. 2003, 51, 4156‐4161

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Human Microbiome:

Source: http://articles.courant.com

100,000,000,000,000

How diet/microbiome affects health

• 178 participants

• 4 groups– GROUP 1: general community

– GROUP 2: day hospitals

– GROUP 3: short-term hospital care

– GROUP 4: long-term nursing homes

Nature 492, S14–S15 (06 December 2012)

Results: -GROUP 1 & 2 had microbial profiles similar to those of 13 younger adult controls.

- GROUP 3 & 4 had a higher proportion of bacteria from the phylum Bacteroidetes, and a lower proportion of Firmicutes

“The microbiota appear to be driven by whatpeople eat” : O’Toole

Ancient Knowledge/ Epidemiological studies Ancient Knowledge/ Epidemiological studies

In vitro / In vivo -Animal studiesIn vitro / In vivo -Animal studies

Molecular mechanisms-(Cell culture studies…)Molecular mechanisms-(Cell culture studies…)

Human Clinical trials (Randomized Controlled, Case Control and Cohort)

Human Clinical trials (Randomized Controlled, Case Control and Cohort)

Meta analysisMeta analysis

compound/moleculeSuccessful

compound/molecule

Millions Millions

Very Few

Hundreds

Food and Bioactives ladder: Road to success

Consumer Preferences

Markettrends

Sensory Evaluation

Color, Flavor, Quality

Nutrition (healthful nutrients)

Enhancing bioactive compounds, shelf life, Bioaccessability and

bioavailability

Quality (Color, Flavor), Nutrition

Impact on quality and nutrition[CA, TX, OSU, FL, AZ, NH]

Outreach

Crop Management and Food Safety

Breeding, Genetics, Genomics

Scientific Advisory Board

Nutrition Science Medical Science

Characterization of Bioactives

Biotechnology Breeding and Genetics

Plant Physiology

Post-Harvest & Processing

INTEGRATION OF SCIENCES

INTEGRATION OF SCIENCES

Education and Economics

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A-W Produce Co., Inc.

Bagley Produce Co.

C&V Supply Inc.

Corey Brothers Inc.

Costa Oro, LLC

Frio Valley Farms

Fruit World

Judy Jackson

Lark Seeds International

Patty Leander

Pan american produce

Super Foods Extracts LLC

VFIC Industry Partners

VFIC Family

86

VFIC FAMILY