DETECTION OF PORCINE AND

BOVINE DERIVATIVES IN FOOD

AND COSMETICS PRODUCTS FOR
HALAL AUTHENTICATION BY
POLYMERASE CHAIN REACTION
Dr. RASHA , MR IBRAHIM , Dr . SACHIE
NAKATANI , JAH HANINI , NOPRATILOVA ,
ZIANOR AZREEN

INTRDUCTION

AIM
ANDOBJECTIVES
OBECTIVES
AIM
AND

AIM

To detect any haram or prohibited

ingredient ,that are not allowed to be
consume by Muslims, in any formulation of
food cosmetic product for the market of
Muslims consumers.

To detect the porcine derivatives in

the food product and cosmetic
formulation.

OBJECTIV
ES

To provide knowledge for the

Muslims consumer regarding the
halal and haram food and cosmetic
product.
To enable authenticity of the food
and cosmetic product that are free
from
any
source
of
haram
ingredient for Muslim market

Muslim
Judaism
Hinduism
consumers
Half sales in
Middle East
(Saudi Arabia
USD 2.1
billion)

HALAL
means
permissible
and lawful

Halal Food
& Cosmetics

Halal
Cosmetic
Market (USD
5-14 bilion
sales per
anum)

HARAM

TOYYIBAN
means clean,
safe,
nutritionous,
healty ,
balanced

Means
prohibited.
(Hunter, 2012)

HARAM OR FORBIDDEN
CATEGORIES

C
Land
Swine
animals
(Pig) (Frog,
Animal slaughtered
without
pronouncing
theSnake)
name of God or
Prey
birds and
a
pronounce
the
name
other
than
God
r
C r
a i
r o
nI n
i n
vt o
Br
oo
l
r x
i d
oo
oe
uc
da
sa
nd
at
ns a
n
i
(MIAN & MUHAMMAD,
m i
a m

2004)

POLYMERASE- CHAIN
REACTION (PCR)
Useful in
detection
of animal
species

Specific &
Sensitive

Suitable to
control food
authenthicit
y
Identification
of procine
DNA content
in
pharmaceutic
al product
(gelatin)

Detection of
porcine DNA
in food
product

No study on
Halal market
surveillance in
pharmaceutic
al product and
cosmetic

(AZMI et al, 2011)

(SAHILAH et al, 2012)

LITERATURE REVIEW

Shah & Yusof.,2014, mentioned the concept of Halal(permissible or

lawful) and Toyyib (clean, good quality, and healthy) are grouped together.
The food or pharmaceutical product made of pig derivatives, unslaughter
animal or other sources that are not permissible according to Islamic law
are haram or forbidden to use.

Azmi et al.,2011, stated the usage of polymerase chain reaction (PCR)

technique for detection of the meat product and pig derivatives for halal
authentication is useful and practically proven in detection of animal
derivatives in food product. The advantage of been highly specified and
sensitive accompanied with the rapid processing are major useful tools in
the detection of various animal species which ideal for the purpose of
quality control in order to avoid species adulteration
for Halal
authentication and verification.

Tasara et al., 2005, mentioned the disability of species-specific DNA in

recognized for the origin of the gelatin DNA. The extensive target DNA
degradation from production and other DNA cross-reaction from various
species are as expected whereby the highly process material are not
feasible for DNA quantitation.

MATERIALS : CONTROLED
SAMPLES
Porcine reverse
ATGAAAGAGGAAATAGATTTT
(U15J265232)

CG
GCCTAAATCTCCCCTCAATGG
TA

Porcine forward
(U15J265232)

LEMON SKIN
PORK MEAT

POSITIVE
CONTROLS

NEGATIVE
CONTROLS

Bovine reverse

GTAGGCTTGGGAATAGTACG
(U15J265231)
A
GCCATATACTCTCCGGTGACA

Bovine forward
(U15J265230)

BEEF
FISH SKIN

MATERIALS : TEST SAMPLES

BOVINE
PURIFIED
GELATIN

BOVINE
OSEIN

PROCESSE
D FOODS
BOVINE
GELATIN

FISH
OSEIN
(SCALES
)

FISH
PURIFIED
GELATIN

GELATIN
MARSHMALLOW, GUMMY AND FOOD GELATIN

COSMETICS SAMPLE

C-Roland
(Lotion
containing
placenta)
Pure Beau
Essence
(Placenta based
Serum )

Alaisyah (Toner)
Goat milk based

DHC ( Collagen)
Elastin Collagen
Ceramide Placenta

METHOD

Stage 1: [ 94C , 4minutes

]
10ml food lysis
Water 6.15 l
Stage 2 : (a) [94C , 30
buffer
Buffer 1 l
seconds]
25l proteinase K
dNTP 0.8 l
(b) [58C ,
500l chloroform
Primer mix 1l
30seconds]
350l Buffer PB
Ex taq 0.05 l
(c) [72C , 3
500l Buffer AW2
Sample (20ng/l) 1l
minutes]
30l Buffer EB
3 : [ 4C , ]
POLYMERASE Stage
CHAIN
DNA EXTRACTIONS

REACTION

SAMPLE
2g

Dneasy mericon Food Kit

AGAROSE
GEL
ELECTROPOEISIS

SOFTWARE
DATA ANALYSIS

Bio-Rad UV
gel imaging

METHOD

1/2

40ng/L

SENSITIVITY RANGE
DETERMINATION
1/10
1/2
1/2
1/5

20ng/L

10ng/L

5ng/L

1ng/L

SERIAL DILUTION OF PORK MEAT

0.1ng/L

RESULTS 1: Positive Contol

40
1

20

10

0.1

0.1 ng/l DNA was detected

from Pork meat by PCR method

(ng/l)

DNA CONCENTRATION
800
700
600
500
400
300
200
100
0

DNA
CONCENTRATION

RESULTS 2: Positive & Negative Control

Bovine Primer
n f
o
sh em ee ork
i
F L
B P

Porcine Primer
n f
o
sh em ee ork
i
F L
B P

Bovine
Porcine

Porcine and bovine as positive control were detected,

fish and lemon as negative control were not detected
by PCR.

RESULTS 3: Processed Food Sample

and Food Gelatin
Bovine Primer

Porcine Primer

1
2
1
2
w
r
w
r
o
e
r
r
o
l
e
l
d
al y
de
de wd
al y
w
m m
ow po
h m m po pow
h
s
p
s
ar gum elly lly
ar um lly elly
J
m
J
m g Je
Je

Bovine
Porcine

Processed Food Sample were detected

and Food Gelatin samples were not detected porcine
and bovine by PCR.

RESULTS 4: Various Purified Processed Gelatin

ne
i
v
o
B

ne
o
B

ne
i
v
o
B

el

in
at
ne
i
v
o
B

ed
fi
ri
u
P

tin
a
el

sh
i
F

es
l
a
Sc
sh
i
F

ed
fi
ri
u
P

in
t
a
el

Gelatin samples were not detected

bovine by PCR method.

RESULTS 5: Cosmetic Samples

e
e
id
c
n
tin ram se
s
h
a
la ce
es
y
e
s
C en a eau
ai
l
on
H ag t b
i
`
t
D ll
ne
A
e
Lo
o
c lacur
p P

e
id

e
c
en
s
es

tin ram
s
la ce
h
e
u
a
n
a
y
C
e
on
is
H ag ta be
i
t
a
D ll
l
n re
e
Lo
o
A`
c lac Pu
p

Cosmetic samples were not detected

porcine and bovine DNA by PCR method.

CONCLUSION
In this study , commercial QIAGEN DNA food
extraction kit and commercial conventional PCR is
used and it has revealed that porcine and bovine
DNA can be sensitively detected in processed food
such as marshmallow and gummy . However,
detection of such derivatives in highly purified,
gelatin and cosmetics products are not effective . It
is potentially reliable , fast and sensitive technique
for the detection of porcine derivatives in food
products but not in highly purified gelatin and
cosmetics .

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