Pre and post harvesting strategies and processing modulation to enhance anthocyanin content in food...

Pre and post harvesting strategies and processing modulation to enhance anthocyanin content in food

A Predmore, G Sigurdson, and N AhmadianiaThe Ohio State University

Introduction

Chemical properties

◦ Flavonoid, C6C3C6 carbon skeleton

◦ Water soluble pigments

◦ Capable of producing red, purple, and blue colors

◦ Glycosylation at C3 and C5

◦ Acylation of sugar with aliphatic or aromatic acids

O

O

OH

HO

O

R1

O

OHO

HO HO

OH

HOO

OHOH

O

R2

A

B

3

5

OH

Introduction Natural antioxidant with possible health

benefits

◦ Reduce the risk of

Cardiovascular disease

Cancer

Obesity

◦ Protective properties

Anti-inflammatory

Diabetes

(Heins and others 2001; He and Giusti 2011; Pereira and others 2009; Andersen and Markham 2006)

Introduction

Anthocyanins biosenthesisActivate the genes necessary to

anthocyanin synthesis Initiating step involves

Phenylalanine Ammonia LyasePAL the rate limiting enzyme Other in increased activity of the

pentose-phosphate pathway and decreased protein synthesis

Not always intermediate compounds carried through to completion

Pre Harvest Techniques

Intrinsic effects ◦ Plant varieties

◦ Maturation stage

Extrinsic effects◦ Light

◦ Nutrients (N, P, …)

◦ pH of soil

◦ Irrigation system

◦ Wounding

◦ Pathogen infection

◦ Fungal elicitors

Pre Harvest Techniques Variety

ANC content varies between different varieties

Solomon A, Golubowicz S, Yablowicz Z, Grossman S, Bergman M, Gottlieb He, Altman A, Kerem Z, Flaishman Ma. 2006. Antioxidant Activities and Anthocyanin Content of Fresh Fruits of Common Fig (Ficus carica L.). J Agric Food Chem 54: 7717-7723.

Pre Harvest Techniques Harvest Time

Two peaks in the anthocyanins production during plant maturation

Two varieties of apple ‘Elstar’ and ‘jonagold’

Comparison of the anthocyanin contents in the skin

Anthocyanin content high early and late in the season

Awad MA, Jager AD, Plas LH, Krol AR. 2001. Flavonoid and chlorogenic acid changes in skin of `Elstar' and `Jonagold' apples during development and ripening. J Hortic Sci 90: 69-83.

Pre Harvest Techniques Light

Affect photomorphogenic reactions in plant Photomorphogenic phenomenon depends on photoreceptors Photoreceptors helps in the induction of anthocyanins

◦ Phytochrome◦ Blue/UV light photoreceptor◦ Cryptochrom◦ Protochlorophyllide

Photoreceptors alter the expression of the anthocyanin regulatory and structural genes

Resulted in enhancement of the rate of anthocyanin synthesis

http://www.youtube.com/watch?v=mJBv-vPLJn0&feature=related




Production of anthocyanins studied in Lollo Rosso lettuce ‘Revolution’

Plastics that block progressively at 20-nm intervals across the UV region

Total anthocyanins were greatest when both UVA and UVB were present

Tsormpatsidis E,∗, Henbest R.G.C, Davis F.J, Battey N.H, Hadley P, Wagstaffe A. 2008. UV irradiance as a major influence on growth, development and secondary products of commercial importance in Lollo Rosso lettuce ‘Revolution’ grown under polyethylene films. Environ exp bot (63) 232–239


Carrot cells were culturedIrradiated by white fluorescent

lamp at various intensities Cells were collected by

centrifugeExtracted with acidified

methanolIncreased sharply (0- 2000 Ix)Slow increase (2000-8000 Ix)

Takeda J. 1988. Light-Induced Synthesis of Anthocyanin in Carrot Cell in Suspension. Exp bot (39) 1065-1077.

Pre Harvest Techniques Nutrients

Different level of (N, P, K, Mg and Ca)One nutrient given at different levels;

remaining four nutrient at normal levelFruits randomly collected at commercial

harvest timeNutrient analyzed in the leafsNegative correlation between N and

ANC concentration“Carbon-Nutrient Balance” hypothesis

hold trueLow availability of N in organic fertilizers

could increase the ANC concentration Awad MA, Jager AD. 2002. Relationship between fruit nutrients and concentration of flavonoids and chlorogenic acid in “Elstar” apple skin. J Hortic Sci 92: 265-276.

Pre Harvest Techniques pH

Color of flowers affected by the pH of soil

Grape cell culturesLow pH caused more

anthocyanin production

Suzuki M. 1995. Enhancement of Anthocyanin Accumulation by High Osmotic Stress and Low pH in Grape Cells (Vitis hybrids). Plant Physiol 147: 152-155.

Post Harvest TechniquesStorage Techniques

◦Temperature

◦Light exposure

◦Environmental Gases

Preservation & Maturation Techniques◦CA, MAP, SO2, Hexanal/Hexenal, Ozone

◦Ethylene

◦Methyl Jasmonate

Post Harvest TechniquesStorage

Temperature ◦Influences enzymatic activity

◦High heat denatures enzymes

◦Freezing slows enzyme activity

Abiotic stress increases production of secondary metabolites

Minimize PPO activity◦Lessen exposure to oxygen


Freezing◦Small increase in ANC content with

time

Cooler Conditions (0-10 °C)◦High increases, in may products

◦71% increase in pomegranate at 10 °C after 6 wks

◦2-6 time increase in strawberry and raspberry between 10-20 °C after 10 days


Light◦Required for metabolite production

◦Effects vary with type: UV or Visible

◦Production of chromophores to protect against oxidative effects


Exposure of light equivalent of a normal day◦>double amount of ANC content in most

products than dark storage

UV-C◦20% ANC increase in blueberries compared

to visible light treatment

◦Reduction of ripe-rot, shelf life extension

Gamma Radiation◦Conflicting evidence, generally lowered ANC


Environmental Gases: Carbon Dioxide◦Slows microbe growth

◦Reduces PPO and other enzyme activity

◦Inverse relationship with PAL Increase CO2 leads to decrease of PAL

activity

◦Reversible action ANC increases after moving to normal air

storage

Post Harvest TechniquesPreservation Treatment

CA, MAP, SO2, Hexanal/Hexenal, Ozone◦Reduce microbial activity, esp. fungi

◦Alters respiration and enzyme activity

◦Control senesce

Generally caused ANC losses◦8 μL hexanal MAP treatment for 38 days: no

loss

◦Intermittent ozone shock treatment: no loss

Post Harvest TechniquesMaturation Treatment

Ethylene Gas◦Produced by fresh produce during

ripening

◦Induces ripening: flavor & secondary metabolite production

◦Levels as low as 15 μL/L of air, increase PAL activity

◦No significant changes in already ripened fruit

Post Harvest TechniquesMaturation Treatment

Methyl Jasmonate◦Two-fold mechanism

Stress/Injury Reaction

Causes increased ethylene production

◦Linear trend of anthocyanin increases with MJ concentration Increases of 3x-100x, ANC type

dependant in apples

◦Application of light still required

Processing ModulationsTemperaturepHOzoneEffect of EnzymesHigh pressure processingInteraction with ascorbic acid

Processing ModulationsTemperature

Anthocyanins very sensitive to heating◦Anthocyanin content in elderberries was

decreased by 50% after 3h at 90⁰C

◦Anthocyanin content in blackberry and strawberry puree significantly decreased after 70oC treatment for 2 min

Logarithmic pattern of destruction as temperature increases Sadilova, E., Stintzing, F. C., & Carle, R. (2006). Thermal degradation of acylated and nonacylated

anthocyanins. Journal of Food Science, 71, C504eC512. Patras, A., Brunton, N. P., Gormely, T. R., & Butler, F. (2009). Impact of high pressure processing on antioxidant activity, ascorbic acid, anthocyanins and instrumental colour of blackberry and strawberry puree. Innovative Food Science and Emerging Technologies, 10(3), 308e313.

Processing ModulationspH

•As the pH increases, typically the stability of anthocyanins decreases

•High pH in grapes can have effects on the color and stability of the pigments

Morris, J.R., W.A. Sistrunk, J. Junek and C.A. Sims, (1986). Effects of maturity, juice storage, and juice extraction and temperature on quality on concord grape juice. J. Am. Sci, 111: 742-746. Laleh, G. H., Frydoonfar, H., Heidary, R., Jameei, R., & Zare, S. (2006). The effect of light temperature, pH and species on stability of anthocyanin pigments in four berneris species. Pakistan Journal of Nutrition, 5, 90–92.

Processing ModulationsOzone

Ozone is a very powerful oxidantAnthocyanins are very sensitive

to ozone treatment In a strawberry juice model,

treatment with 7.8% w/w ozone ◦92.8% reduction in anthocyanin

content

Tiwari, B. K., O’Donnell, C. P., Patras, A., Brunton, N. P., & Cullen, P. J. (2009). Effect of ozone processing on anthocyanins and ascorbic acid degradation of strawberry juice. FoodChemistry, 113(4),1119e1126.

Processing ModulationsEffect of Enzymes

Polyphenoloxidase (PPO)◦Causes browning in fruits

When PPO completely inactivated, stability of pelargonidins was intact during storage

β-glucosidase and peroxidase also play roles in anthocyanin degradation during storageGarcia-Palazon, A., Suthanthangjai, W., Kajda, P., & Zabetakis, I. (2004). The effects of high hydrostatic pressure on b-glucosidase, peroxidase and polyphenoloxidase in red raspberry (Rubus idaeus) and strawberry (Fragaria x ananassa). Food Chemistry, 88, 7e10.

Processing ModulationsHigh Pressure Processing

Method of preserving and sterilizing food using very high pressure

Anthocyanins are stable during HPPVarying stability after storage

◦Based on MPa of treatment

◦Amount of PPO inactivationCyanidin 3-glucoside and cyanidin 3-

sophoroside had higher rates of stability during storage after 800MPa treatment than 400 or 600MPa in raspberries

Suthanthangjai, W., Kajda, P., & Zabetakis, I. (2005). The effect of high hydrostatic pressure on the anthocyanins of raspberry (Rubus idaeus). Food Chemistry, 90, 193e197.

Processing ModulationsInteraction with Ascorbic Acid

Studied extensivelyMay occur by oxidation or

condensation of the ascorbic acid with the pigments from the anthocyanins

Causes numerous interactions including:◦Degradation of both compounds

◦Decrease in color

◦Decrease in the quality of the product Markakis, P., Livingstone, G. E., & Fillers, G. R. (1957). Quantitative aspects of strawberry pigment degradation. Food Research, 22, 117e130.

ConclusionMany factors influence anthocyanin content

and stability including: ◦ Pre-harvest factors

Varity affects the ANC production

ANC content varies during maturation

White light increase ANC concentration

Higher concentration of Nitrogen could decrease ANC

pH of soil could affect the ANC production

◦ Post-harvest factors

◦ Processing modulations Most processing conditions degrade anthocyanins

More research is needed for alternative methods

Pre and post harvesting strategies and processing modulation to enhance anthocyanin content in food...

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