By: Dr. H.

Satrijo Saloko

Courtesy of Aisha
 Toksikan : Subtansi toksik yang diproduksi oleh aktifitas
manusia (contoh??)

 Toksin : Subtansi toksik yang diproduksi secara

alami (contoh???)
 Kemampuan suatu zat kimia/xenobiotik dalam
menimbulkan kerusakan pada organisme baik saat
digunakan atau saat berada di lingkungan
 Subtansi kontak dgn permukaan tubuh misal melalui kulit,
mata, mukosa saluran cerna atau traktus respirasi
 Acrylamide
Polycyclic aromatic
Maillard reaction product amine
Toxicant from food N-nitrosamines
processing
Polycyclic aromatic
hydrocarbons Benzo[a]pyrene
Source :
 Charred or smoked meat and fish
 Cereals
 Flour
 Vegetables
 Fruits
 Water, soil, dust
 Cigarette smoke
 Vegetables oil

-
 the largest class of chemical compounds,
containing two or more fused aromatic rings
made up of carbon and hydrogen atoms. PAHs
are formed in incomplete combustion processes
which occur whenever wood, coal or oil are
burnt.
 Result of the environmental pollution but also as
a consequence of some thermal treatments
 Food processing (smoking, drying, roasting,
baking or frying) produces PAHs in certain
levels
 Formed mainly from carbohydrates cooked at
hight temp in the absence of oxygen.
 Broiling meat in hot ceramic or charcoal
briquettes  the melted fat will come into
contact with very hot surface.
 PAHs then produced in the ensuing reactions,
PAHs rise with resulting cooking fumes and are
 Polycyclic aromatic
hydrocarbon (PAH) that is a
byproduct of incomplete
combustion or burning of
organic (carbon-containing)
items
 The most commonly known
carcinogenic PAH.
 Heating starch at :
370-390 0C =0.7 ppb
650 0C =17 ppb
 Many cooking processes: 370—
390 0C
 The surface temperature of
10 mg of BaP 3x a week: skin
tumors
100 mg of BaP in 10 weeks :
stomach cancers
 Benzo[a]pyrene is not mutagenic and carcinogenic
by itself, but must be first converted to active
metabolites.
 Metabolic conversion:
- A cytochrome’ P450-mediated oxidation produce a
7,8-epoxide.
- The 7,8- epoxide undergoes an epoxide hydrolase-
mediated hydration, produce the 7,8-diol
- 7,8-diol further oxidized by cytochrome P450,
produces diolepoxide.
- Diolepoxide: highly mutagenic without metabolic
activation and is also highly carcinogenic at the site
Case History: Benzopyrene from meat on a barbeque

Natural molecules can

become dangerous during
food preparation!
 Conventional  Modern method:
method: - Indirect smoking
- Direct smoking
(using external smoke
(smoke developed generator).
in the smoking - Used in modern
chamber, industrialized kilns
traditionally in - Operated
smokehouses) automatically under
carefully controlled
conditions
- Smoke can be washed
 Mean content of 15 PAHs in meat products
industrially smoked, μg/kg

PAHs Ham Cooked cured loin Medium ground

sausages
Externa Internal External Internal Extern Interna
l part part part part al part l part
Cyclopenta[c,d] 1.62 0.53 0.07 n.d 11.39 7.07
pyrene
Benzo[a]anthrac 8.88 0.41 6.48 0.38 8.78 1.29
ene
Chrysene 6.29 0.56 4.46 0.48 6.63 0.55
5-metylchrysene 1.72 0.28 2.73 0.30 2.06 0.36
Benzo[j]fluoroan 0.31 n.d 0.31 n.d 1.2 n.d
thene
Benzo[a]pyrene 0.37 0.28 0.30 0.28 0.89 0.32
Ciecierska, Marta. 2007. INFLUENCE OF SMOKING PROCESS ON POLYCYCLIC AROMATIC HYDROCARBONS’ CONTENT
IN MEAT PRODUCTS* . Acta Sci. Pol., Technol. Aliment. 6(4) 2007, 17-28
 Mean content of 15 PAHs in meat products
traditionally smoked, μg/kg

PAHs Ham Cooked cured loin Medium ground

sausages
Externa Internal External Internal Extern Interna
l part part part part al part l part
Cyclopenta[c,d] 3.26 0.49 1.27 n.d 12.17 7.78
pyrene
Benzo[a]anthrac 6.74 0.40 9.25 0.22 5.86 1.51
ene
Chrysene 6.20 0.68 9.12 0.42 5.07 0.57
5-metylchrysene 1.28 0.28 2.98 0.31 1.39 0.38
Benzo[j]fluoroan 0.59 n.d 0.43 n.d 0.83 n.d
thene
Benzo[a]pyrene 0.43 0.27 0.37 0.29 0.40 0.31
Ciecierska, Marta. 2007. INFLUENCE OF SMOKING PROCESS ON POLYCYCLIC AROMATIC HYDROCARBONS’ CONTENT
IN MEAT PRODUCTS* . Acta Sci. Pol., Technol. Aliment. 6(4) 2007, 17-28
 1. Traditional method of smoking affected the higher
contamination PAHs than industrial proceess.
2. Industrial smoking process influenced higher PAHs
content only in case of exteriors of medium-ground
sausages.
3. For all products smoked using both methods, interiors
had a significantly lower PAHs contamination than
exteriors of the same products.
4. Benzo[a]pyrene’s content was much lower than
maximum tolerable limit of 5 μg·kg-1, which was set for
smoked meat products in Commission Regulation (EC)
No. 208/2005. Therefore industrial and even traditional
smoking
Ciecierska, Marta. of meat
2007. INFLUENCE is aPROCESS
OF SMOKING safe ON process.
POLYCYCLIC AROMATIC HYDROCARBONS’ CONTENT
IN MEAT PRODUCTS* . Acta Sci. Pol., Technol. Aliment. 6(4) 2007, 17-28
 1912, L. C. Maillard hypothesized
reaction of brown pigments and
polymers produced from the reaction
of amino group of amino acid and
carbonyl group of sugar.
 Nonenzymatic browning reaction:
 Reaction of amines and carbonyl
under heat treatment  in vivo and
biological systems damage
 Reaction products: brown color,
characteristic roasted or smoky
odors, pro- and antioxidants,
mutagens, carcinogens.
 Occurs between reducing
sugars and amines at high
temperatures
 Produces flavor-aroma
 Produces color
 Produces antioxidant
products
 Produces toxic products
 Destroys nutrients (lysine)
 Melanoidins
Ammonia Brown colour
Amine
Phospholipids
Amino acids
Proteins HEAT
Amino Carbonyl
Interaction Volatile compounds
Aldehydes
Ketones
(Amadori product)
Carbonyl
Reducing sugars
Polysaccharides
Carbonyls
Oxidised lipids
Esters
Amide (acrylamide)
Heterocyclic compounds
  Source: produced during the cooking of protein-rich
food; beef, chicken, broiled seafood, hamburger,
 Mutagens , esp on the surface layers where most
pyrolysates are found.
Imidazoquin Heating a mixture of
oline (IQ)- creatine/creatinine, amino acids,
type and sugar
PAA Cyclization of creatinine to form
the imidazole moiety due to heat
treatment

Non-IQ Pyrolysis products formed

type from tryptophan
 Revertants of TA98
Food 2500C 3000C 4000C
Beef 178 11400
Chicken 661 15120
Egg 121 4750
Hairtail 849 12320
Eel 309 6540
Squid, dried 269 8000 4490
Skipjack tuna, 1,220 24300 6200
dried
Seaweed, nori 260 3040
The IQ-Type

Abbreviation Z R1 R2 R3
IQ C H H H
MeIQ C Me H H
MeIQx N H H Me
4,8-DiMeIQx N Me H Me
The non-IQ-Type
 Trp-P-1 and Trp-P-2 are toxic to animals
 LD50 of Trp-P-1: 200 mg/kg in mice, 380 mg/kg in
hamsters, 100 mg/kg in rats. Over LD50 died in 1h.
 IQ and MeIQX in Beef extracts  metabolically
converted to active mutagens by liver tissue through N-
oxidation and O-acetylation  produce highly reactive
metabolites that for, DNA adducts
 Food with Trp-P-1 or Trp-P-2  tumor and hepatoma in
mice
 Highly potent mutagens and carcinogens
PAA Found in Revertants /µg
MeIQ Broiled sardine 661,000
IQ Fried beef 433,000
MeIQx Fried beef 1435,000
Trp-P-1 Broiled sardine 104,000
Glu-P-1 Glutamic acid
49,000
pyrolysates
Trp-P-1 Broiled sardine,
39,000
broiled beef
Glu-P-2 Broiled, dried
1,900
cuttlefish
 Formed from the interaction of nitrite and secondary,
tertiary amines.
 Found in wide variety of foods
 A significant source of nitrite: cured meat
 Nitrite ions in curing meat: antimicrobial (inhibit the
growth of C. botulinum, produces appealing red color to
meats from nitrosomyoglobin and nitrosylhemoglobin
pigments, gives a desirable cured flavor to meat
products
 Permissible levels of nitrite in cured foods :vary, 10 to
200 ppm
 nitrite in humans : from reduction of dietary nitrate to
nitrite by bacteria in the mouth and in the intestinal tract.
 pH dependent (3.4)
 Weakly basic amines are more rapid
 Several anions, such as halogens and thiocyanate,
promote the nitrosation process
 High temperature food heating , i.e bacon
 Diethylnitrosamine (DEN) and dimethylnitrosamine
(DMN) occur in the gastric juice of experimental animals
and humans fed diets containing amines and nitrite
 Heating of nitrite-treated foods  produce nitrosamines.
 Cured meats have all been shown to contain
nitrosamines, the higher levels appearing in cured meats
that have been subjected to relatively high heating.
 In Norway in 1962, epidemic of food poisoning in sheep,
extremely high levels of nitrosamines were detected in
herring meal treated with nitrite as a preservative. The
sheep suffered severe liver disease and many of them
died.
 formation of nitrosamine in nitrite-treated fish: dependent
on the temperature of preparation following the addition of
nitrite.
 Refrigerated fish treated with nitrite had no more
nitrosamine than fresh fish treated with nitrite,
 heat treatment of fish increased the rate of nitrosamine
formation following addition of nitrite  due to increased
concentrations of secondary amines resulting from protein
meat Nitrosamines Level (ppb)
Smoked Dimethylnitrosamin <6
sausages es <6
Diethylnitrosamine
s
frankfuters Dimethylnitrosamin 11-84
es
salami Dimethylnitrosamin 1-4
es
Fried bacon Dimethylnitrosamin 1-40
es 1-40
 Over 100 food substances assayed, approximately 80%
were shown to be carcinogenic in at least one animal
species.
 Dimethyl- and diethylnitrosamine are two of the most
potent carcinogens
 Administration of dimethylnitrosamine at 50 ppm in the
diet produces malignant liver tumors in rats in 26—40
weeks. Higher doses produce kidney tumors.
 As the dose of diethylnitrosamine is reduced below 0.5
mg/kg,. No clear threshold dose for carcinogenicity of
nitrosamines in the diet has yet been established.
 Addition of reducing agent (erythsorbate or ascorbate) to the
curing mix to reduce or eliminate nitrosamine formation
 Reduce food processing temperature
 Minimize cured meat consumption
 First report, Sweden, April 2002 Food Amount (ppb)
 Found in a wide range of foods,
including dietary staples Baby food 17-130
• Potato products French fries 70-1036
• Breakfast cereal Potato chips 117-2764
• Coffee Cereals 47-266
• Bakery products
• Snack foods
Bread and bakery 10-354
Snack foods 111-1168
coffe 275-351
Chocolate products 45-909
Dried foods 11-1184
 Formed from asparagine and
glucose via Maillard reaction.
 Acrylamide Formation
(heating at 1800C, 30 min)
▪ Asparagine 0.99 µg/g
▪ Asparagine + glucose 1200 µg/g
▪ Asparagine + Potato starch + glucose
9270 µg/g
 Asparagine and carbonyl compound
(glucose, glyceraldehide) play an
important role in acrylamide
formation in cooked foods.
 Effectiveness of Amino Acids and
Dextrose to Form Acrylamide
Model System
Amino acid fry
+ Reducing sugar
Variety of ingredients
Potato Starch + Water Measure Acrylamide

 Acrylamide Formation
– Potato starch <50 ppb
– Potato starch + dextrose <50 ppb
– Potato starch + asparagine 117 ppb
– Potato starch + dextrose + asparagine 9270 ppb
Other Amino Acids
–Alanine <50 ppb Arginine <50 ppb
–Aspartic A. <50 ppb Cysteine <50 ppb
–Lysine <50 ppb Methionine <50 ppb
–Threonine <50 ppb Valine <50 ppb
–Glutamine 156 ppb Asparagine 9270 ppb
 Amino Acid Composition in Potatoes

Approximately 50% of amino acids are in the free state

(not incorporated into protein).

Asparagine is roughly half of the free amino acid content.

 Amide moiety
NH2
C O
CH2
H2O CO2
NH2 CH COOH
Carbonyl
Asparagine Schiff’s base Acrylamide
(glucose)
 3H2O NH3
From amino acid
(O)

2H2O Acrylic acid

Lipid Glycerol Acrolein .NH 2 Acrylamide

From amino acid

Radical
 Ingestion of heat-treated starch-rich foods (potato chips and
french fries)
 Acute toxicity :LD50 in rats: 159 mg/kg -300 mg/kg body weight
 Classified by the US EPA ( as a B2 (probable human carcinogen)
IARC (international Agency for research on
cancer) as a 2B (possible human carcinogen)
ACGIH (American Conference of Industrial
Hygiene) as A3 (confirmed animal carcinogen with unknown
relevance to human
Acrylamide - toxicology

 Proven neurotoxic compound in animals and in humans

 Effects range from drowsiness to incoordination,
hallucinations, confusion, abnormal sensation, muscle
weakness, incoordination
 Genotoxic compound with the potential to affect the
germinal cells thus leading to hereditary changes
 Causing cancer in laboratory animals (rats)
 Studies in humans (e.g. 8000 workers in China) which
were positive on neurotoxicity failed to prove
relationship with cancer in humans (too small
numbers ?)
POTATO PRODUCTS
 Acrylamide levels
increased with degree of
browning 45 µg/kg 76 µg/kg 262 µg/kg

 Brown color as measured

by “L” and “a” values
correlated highly with
acrylamide levels 516 µg/kg 866 µg/kg 1512 µg/kg
 Same acrylamide levels
found in French fries with
micrograms acrylamide/kg

10000

micrograms acrylamide/kg
similar degree of
10000
R2 = 0.8551
French fries

R2 = 0.8558
1000

French fries
browning (fried French
1000

100
100

fries) 10
45 55 65 75
10
5 10 15 20
"L" value
"a" value
 Food should not be cooked excessively (for too long or
at too high temp)
 Maintain cooking at temp under 1500C, no preferable
flavor chemicals
 FDA: Eat a balanced diet, choose a variety of foods that
are low fat and rich in high-fiber grains, fruits, and
veggies