ED73.

9002: Processing Effects on the functional components of food

Acrylamide: Thermally induced Toxicant in foods

Yakindra Prasad Timilsena (111332) Jyotsna Shrestha Khanal (111328)
5 Nov, 2010
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By-

Presentation Outline
Introduction

Mechanisms of formation Toxicology Amounts in different food sources Regulations & allowable limits Methods of Determination Preventive measures Conclusion
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Less tasty side to these culinary delights:

Thermally induced harmful chemicals (Carcinogens)
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What is Acrylamide ?
Synthesized for the first time in 1949 Unsaturated & highly reactive amide White odorless crystalline solid, soluble in water, ethanol, ether, and chloroform Formation: baking, roasting, toasting, grilling, frying Common foods: French fries, potato chips, bread,biscuits, coffee, roasted cereals

Properties of Acryl amide

S.N. Properties Values 1. IUPAC name Prop-2 enamide 2. Molecular formula C3H5NO 3. MW 71.08 g mol1

4. 5. 6.
7. 8.

Density Melting point

Color Boiling point

1.13 g/cm3 84.5 C

white crystalline solid 136 C

Solubility in water 2.04 kg/L (at 25 C)

Acrylamide in industrial use

1950s: Synthetic Polyacrylamide-based flocculants 1960s: Synthetic coagulants in mineral processing industries 2002: Swedish researchers announced the discovery of acrylamide in food

PAM

Acrylamide production in foods

Concentrations & availability of Precursor molecules AA Formed by Maillard reaction between asparagine and reducing sugars (precursors) Factors (temp., heat intensity, water activity) in processing technologies

Cereals, potato & coffee : variety and growing factors

Cereals : reducing sugars are abundant conc. Of Asparagine AA Potato : asparagine is abundant conc. of reducing sugar AA 7

Impact of Potato variety on AA Levels

(Source: D. Mottram, U. Reading)

Sample

Baking potatoes raw Boiled Chipped & fried King Edward potatoes raw boiled Chipped & fried Frozen frying chips as sold Cooked Over cooked

Acrylamide concentration (g/kg) LC-MS-MS GC-MS <10 <10 310 <10 <10 2800 200 3500 12800 Nd Nd 350 Nd Nd 3500 100 3500 12000

SNFA result (g/kg)

<30

Acrylamide formation influenced <30 by starting raw material

Heat

Acryl amide formation

Carbonyl source

L-Asparagine source

Removing or changing one of the parameter prevents formation.

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Mechanism for formation

From Asparagine and Reducing sugar H2N C CH2 CH O Asparagine H2N C CH2 CH O COOH
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NH2

+
COOH

Glucose N

HO OH OH

OH OH

Mechanism for formation

From Asparagine and Reducing sugar

H2N
C CH2 CH

N Maillard reaction Products

O
- CO2 H2N C CH2 CH2 O

COOH

H2N N O C CH = CH2

Acrylamide
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Mechanism for formation

From Triglycerides Triglycerides Heat, O2

Acrolein NH3

Acrylic acid

Acrylamide
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Variation in concentration of Acrylamide with temperature and time

Acrylamide (ppm)

2.5 min 3.5 min

Temperature (0C)

Source: Gertz and Klosterman (2002) Eur. J. Lipid Sci. Technol. 104:762-771.

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AA Formation as a function of Temperature

Potato chips Frying time: 15 minutes
20000

AA = 442.3 * e Acrylamide (ppb) 10000 15000 5000

(0.07930*(Temp-383))

380

400

420

440
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Temperature (Kelvin)

Effect of pH on Acrylamide Formation

AA (ppb)

120 C, 40 min 150 C, 15 min

pH
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Effect of browning in AA formation

(Source: D. Mottram, U. Reading)

Sample

Acrylamide concentration (g/kg) GC-MS LC-MS-MS <10 <10 310 <10 <10 2800 200 3500 12800 nd nd 350 Nd Nd 3500 100 3500 12000

SNFA result (g/kg)

Baking potatoes raw Boiled Chipped & fried King Edward potatoes raw boiled Chipped & fried Frozen frying chips as sold Cooked Over cooked

<30

Yield of acrylamide increases <30 substantially with browning

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Amount of Acrylamide in Foods (ppm)

Category Breads Crispbread Crackers and Biscuits Cereal Other Grains Potato Chips Other Salty Snacks French Fries Other Potato Products Other Veg and Fruit Products European Data 12-3200 <30-1670 <30-2000 <30-2300 <30 150-1280 122-416 85-1104 <20-12400 10-<50 FDA Data <10-364

26-504 52-266
117-2762 12-1168 20-1325

<10-70

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Amount of Acrylamide in Foods (ppm)

Category
Meats Candy and Dessert items Cookies Coffee and Tea Other Nonalcoholic Beverages Alcoholic Beverages Dairy Products Baby Food and Formula Dry Soup Mixes Gravy and Seasonings

European Data FDA Data < 30-64 < 10-116 < 20-110 < 10-909 36-199 170-700 175-351
< 30 30 10-100 40-120

< 10-43 < 10-130 < 10-1184 38-54 18

Asparagine !!!
Amino Acids Alanine Asparagine Level of Acrylamide Formation <50 ppb 9270 ppb

Aspartic Acid
Cysteine Glutamine Lysine Methionine Threonine

<50 ppb
<50 ppb 156 ppb <50 ppb <50 ppb <50 ppb
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Source : Barbara Petersen, Exponent, Inc. October 2002 JIFSAN workshop

Stability of acrylamide in various foodstuffs

Food product Interval (month)
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Acrylamide level (g/kg) Initial

238

Second
238

Breakfast cereal

Soluble coffee powder

Roasted barley Roasted coffee Dried chicory Roasted chicory Cocoa Chocolate with almond Soluble chocolate powder

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9 7 5 5 3 2 1

771
265 203 214 4015 180 94 54

256
225 147 174 3395 177 73 41
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Adapted from Delatour et al.

Asparagine in Various Crops

Cheese Asparagus Cocoa (raw) - roasted @ 1250C - roasted @ 1350C Potato Rye Wheat Corn 40 300 mg/100gm 5.4 108 mg/100gm 30.9 mg/100gm 14.5 mg/100gm 9.4 mg/100gm 500 1000 mg/100gm 0.2 2.8 mg/100gm 0.2 20 mg/100gm 0.6 1 mg/100gm

Source: Ellin Doyle, Ph.D., Food Research Inst., U. Wisc.

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Toxicology
Known neurotoxicant (IARC 2002; Manson et al. 2005) Peripheral neuropathy Tingling/numbness of extremities Loss of reflexes Chronic CNS dysfunction and neuropathy Reproductive toxicity Animal carcinogen (CNS, endocrine organs) EPA has classified acrylamide as a B2 carcinogen (probable human carcinogen) (IRIS2009). Biomarker adducts on amino acid valine of Hb.
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Maximum allowable dose level (MADL

2

micrograms per kg body weight per day

(Office of Environmental Health Hazard Assessment) Exposure at a level 1,000 times greater than the MADL is expected to have no observable effect No Observed Adverse Effect Level (NOAEL) of 15 mg/kg bw/day were identified in mice Legislation under formulation
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 Water extraction Analysis by GC-MS with or w/o bromination by LC-MS/MS by LC-MS or -UV after deriv. with mercaptobenzoic acid GC-Ion Trap MS
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GC-MS method

Acryl amide extracted with water, test portion homogenized, acidified to pH 4-5

Addition of Carrez I and Carrez II solution

Extraction with Ethylacetate hexane (80:20), Filtration over Na2SO4

Clean up with Flourisil elution of acrylamide with acetone

Evaporation, residue taken up in ethylacetate, triethylamine added

Filtration, injection into GC-MS

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Acrylamide Precursors Where to Intervene

Asparagine

Reducing Sugars - Glucose

- Fructose - Sucrose hydrolysis?

Factors affecting asparagine and reducing sugars - Variety of potato - Storage conditions

Acrylamide Possible Mitigation Strategies

1. Reduction or removal of sugars.
Selection of low sugar cultivar. Blanching and soaking to leach out sugars.

2. Reduction or removal of asparagines.

Use of enzymes to degrade asparagine during food processing. Blanching to leach out asparagine

3. Blocking formation of Acrylamide

Altering time-temperature of processing. Altering heat processing methods Changing pH of food by using acids Use of antioxidants
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Reduction of Acrylamide formation in Potatoes products

Improve agronomic practices Select potatoes variety with less asparagine Store potatoes tubers >10C Blanching (82C) Add antioxidants Dip slices in citric acid (0.75%) solution Dip in NaCl, CaCl2 solution Use of enzyme- asparaginase Addition of amino acids glycine.
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Reduction of Acrylamide in cereal products

Adjustment of time and temperature during baking. Extend fermentation times where feasible. Substitution of ammonium bicarbonate with alternatives Avoid or minimize use of reducing sugars Avoidance of very high baking temperature Use of asparaginase in dough to degrade asparagine.
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Enzyme mechanism

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Impact of treating with Asparaginase

By treating with asparaginase there was a 99% reduction in the levels of acrylamide in the potato mixture
Product characteristics fully preserved No change in process parameters Commercially viable and available ???

Relative Exposure to Acrylamide in U.S. Food

French Fries & Potatoes Breads Potato Chips Cereal

Biscuit / Cookies

Coffee Cakes Dried Foods Pop Corn Salty Snacks Chocolate Products Nuts/Seeds/Butters

All Other Foods

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Conclusion
Acrylamide formation in thermally processed foods is a major challenges for bakery, French fries and chip producing industries. challenges still remain in terms of the needs to develop simple and rapid test methods. different pre-frying treatments on the reduction of acrylamide (AA) formation are under investigation 33

What about these cereal based foods ?

Selroti

Puri

Chapati

Doughnut

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Foods should not be cooked excessively...for too long or at too high a temperature... However, all foodshould be cooked thoroughly to destroy food borne pathogens. Is it possible to cook food without forming at least some AA ?????

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