LIPID ANALYSIS(Ch.13-14)karaali/lipidanalysis.pdf · before analysis. 3- Derived lipids: These are...
Transcript of LIPID ANALYSIS(Ch.13-14)karaali/lipidanalysis.pdf · before analysis. 3- Derived lipids: These are...
LIPI
D A
NALY
SIS(
Ch.1
3-14
)I.
Qua
ntificat
ion
of li
pid
compo
nent
s
a.Ex
trac
tion
s (we
t vs
dry
;so
lid-l
iqvs
liq-l
iq.)
b.In
dire
ct m
etho
ds(N
MR,
X r
ays,
Con
duct
ance
)
II.
Ass
ays
for
furt
her
inve
stigat
ions
on
extr
acte
d oil
a.Pu
rity
crite
ria:
give
ind
icat
ions
of
wholes
omen
ess
or
adulte
ration
(ph
ysical:n
D20,d
ensity
; ch
emical:
IV,
SV,
FAM
E, s
tero
ls,
toco
pher
ols
compo
sition
s )
b.Q
ualit
y cr
iter
ia:
indica
tes
good
qua
lity
or d
eter
iora
ted
quality
b.1.
MIU
(M
oist
ure,
Impu
rities
, Uns
apon
ifiabl
es)
b.2.
Ranc
idity
: H
ydrt
olyt
ic:A
V an
d ff
a
Oxi
dative
:PV,
TBA,T
OTO
X e
tc
b.3.
Sto
rage
sta
bilit
y(she
lf-l
ife)
Lipi
ds:F
ats
and
fat
like
com
poun
ds(n
otpo
lym
ers)
that
ar
e so
lubl
ein
no
n-po
lar
orga
nic
solv
ents
(pet
role
umet
her)
, but
inso
lubl
e in
H2O
. Lip
ids
have
:
1. Cu
linar
y im
port
ance
: Gre
at e
ffec
ts in
pre
para
tion
of
food
ssi
mnc
e th
ey p
rovi
dehe
at t
rans
fer
med
ia, c
arry
flav
ours
, and
con
trib
ute
to p
alat
abili
ty(e
atin
g qu
alit
y)of
foo
ds.
2. N
utri
tion
al i
mpo
rtan
ce:
Prov
ide
bulk
of
calo
ries
in
diet
(1
g: 9
kca
l).
For
heal
thy
diet
s, %
30 p
erce
nt o
f to
tal c
alor
ies
have
to
com
e fr
om li
pids
.
3.
Phys
iolo
gica
l im
port
ance
: Th
ey
are
vita
l fo
r
bi
olog
ical
fu
ncti
onin
g of
ce
lls,
sinc
e th
ey
cont
ain
esse
ntia
l fat
ty a
cids
and
oil-
solu
ble
vita
min
s.
“LIP
ID”
1.Fa
tty a
cids
2.N
eutra
l fat
s and
oils
3.W
axes
4.Ph
osph
olip
id5.
Ster
ols
6.Fa
t sol
uble
vita
min
s
Lipi
ds in
foo
ds o
ccur
in t
hree
diff
eren
t fo
rms:
1-Si
mple
lipids-
Fats
,oils
are
triglyc
erides
-The
y ar
e th
e es
ter
of
fatt
y ac
ids
with
glyce
rol(a
ll ed
ible
fats
an
d oils); wa
xes
are
este
rs of
fa
tty
acids
with
lon
ger
chain
alco
hols.
2-Co
mpo
site
lipids:
Con
tain
compo
nent
s ot
her
than
lip
ids.
Ex:P
hosp
holip
ids,
Lipo
prot
eins
,Glyco
lipids:
Th
ese
are
foun
d in
boun
d fo
rm
with
ot
her
cons
titu
ents
;th
eir
ana
lyse
s is h
arde
rth
an s
imple
lipids,
bec
ause
you
hav
e to
hyd
rolyse
the
molec
ule
befo
re a
nalysis.
3-Der
ived
lipids:
The
se a
re s
ubst
ance
s de
rive
d fr
om
lipids
and
are
solubl
e in l
ipid-e
xtra
cting
solven
ts(
i.e.
fat
solubl
e pigm
ents
, st
erols,
ant
ioxida
nts
like
toco
pher
ols
and
Vita
mins
A,D
, E,
K(f
at-s
olub
le
vita
mins).T
hey
are
not
true
lipids
but
are
foun
d in
lipid
phas
e.
They
be
have
lik
e lip
ids.
In
mos
t an
alys
es y
ou w
ill g
et t
hem in
the
lipid f
ract
ion.
Fatty
Aci
ds
OR
C
O
H
#1 C
arbo
nA
cid
Gro
up
OR
C
O
H
Non
-pol
ar E
nd -
Hyd
roph
obic
End
(
Fat-s
olub
le ta
il) Po
lar E
nd -
Hyd
roph
ilic
End
Uns
atur
ated
Fat
ty A
cids
CH
3
CH
2
CH
2
CH
2
CH
2
CH
2
CH
2
C
OH
O 12
45
67
83
CH
3
CH
2
CH
2
CH
2
CH
2
CH
2
CH
2
C
OH
O 12
45
67
83
3 -O
cten
oic
Aci
d
3, 6
-O
ctad
ieno
icA
cid
Shor
t han
d:8:
1 (∆
3)
8:2
(∆3,
6)
Cis
9 -O
ctad
ecen
oic
Aci
d (o
leic
)
Tran
s 9 -
Oct
adec
enoi
cA
cid
(ela
idic
acid
)
OC
H3(
CH
2)7
C
C
(C
H2)
7
C
OH
HH
910
OC
H3(
CH
2)7
C
C
(C
H2)
7
C
OH
H
H
Cis
And
Tra
ns F
atty
Aci
ds
Poly
unsa
tura
ted
Fatty
Aci
ds
Lino
leic
acid
:C
is, c
is, 9
, 12
-Oct
adec
adie
noic
acid
Lino
leni
cac
id:
Cis
, cis
, cis
9, 1
2, 1
5 -O
ctad
ecat
rieno
icac
id
Ara
chid
onic
acid
:C
is, c
is, c
is, c
is5,
8, 1
1, 1
4 -E
icos
atet
raen
oic
acid
Lino
leic
Aci
d
Lino
leni
cA
cid
Ara
chid
onic
Aci
d
Nat
ural
ly-o
ccur
ring
fatty
aci
ds
R
CH
2
CH
C
H
CH
2
CH
C
H
CH
2
C
OH
O
76
54
3
1.C
isfo
rm
2.N
ot c
onju
gate
d --
-iso
late
d do
uble
bon
d.
3.Ev
en n
umbe
red
fatty
aci
ds.
FAT
AN
D O
ILS
Mos
tly T
rigly
cerid
es:
O
H2C
O
H
HC
O
H
H2C
O
H
HO
C
R
HO
C
R
HO
C
R
OO O
H2C
O
C
R
O OH
2C
O
C
R
HC
O
C
R
++
3 H
2O
Gly
cero
l 3
Fatty
Aci
ds
GLY
CER
IDES
Mon
ogly
cerid
e(α
-mon
oste
arin
)D
igly
cerid
e(α
, α' -
dist
earin
)
H2C
O
H
HC
O
H
H2C
O
O C
(CH
2)16
CH
3
H2C
O
HC
O
H
H2C
O
O C
(CH
2)16
CH
3
C
(CH
2)16
CH
3
O
Trig
lyce
ride
(β-p
alm
ityld
iste
arin
)
H2C
O
HC
O
H2C
O
O C
(CH
2)16
CH
3
C
(CH
2)16
CH
3
O O C
(CH
2)14
CH
3
(C18
)
(C16
)
(C18
)
α-o
leod
ipal
miti
n
1 -o
leod
ipal
miti
n
Ole
ic
Palm
itic
Palm
itic
OPP
α-L
inol
eyld
iole
in
1 -L
inol
eyld
iole
in
Ole
ic
Lino
leic
Ole
icLO
O
FATS
AN
D O
ILS
AR
E PR
IMA
RIL
Y T
RIG
LYC
ERID
ES (9
7-99
%)
Veg
etab
le o
il -
68%
of w
orld
supp
lyC
ocoa
but
ter -
solid
fat
Oil
seed
s -liq
uid
oil
Ani
mal
fat -
28%
(fro
m H
ogs a
nd C
attle
)
Mar
ine
oil -
4%W
hale
oil
cod
liver
oil
Fatty
Aci
dC
ompo
sitio
ns (%
) of F
ats a
nd O
ils
43
63
82
610
36
123
4414
1018
116
2611
412
16:1
71
18:0
156
32
18:1
297
1824
18:2
22
5354
Fatty
Aci
dsB
utte
rC
ocon
utC
otto
nsee
dSo
ybea
n
18:3
28
WA
XE
SFa
tty a
cids
+ L
ong
chai
n al
coho
l
Impo
rtant
in fr
uits
:
1.N
atur
al p
rote
ctiv
e la
yer i
n fr
uits
, veg
etab
les,
etc.
2.A
dded
in so
me
case
s for
app
eara
nce
and
prot
ectio
n.
Bee
swax
(myr
icyl
palm
itate
), Sp
erm
acet
i (ce
tylp
alm
itate
)
OC 3
0H61
O
C
C
15H
31
OC
16H
33
O
C
C15
H31
PHO
SPH
OLI
PID
Leci
thin
(pho
spha
tidyl
chol
ine)
O O
H2C
O
C
R
O
R
C
O
CH
H2C
O
P
O
C
H2
C
H2
N
+ CH
3
CH
3
CH
3
O_
Phos
phat
idic
Aci
dC
holin
e
STER
OLS
Mal
e &
fem
ale
sex
horm
ones
Bile
aci
dsV
itam
in D
Adr
enal
cor
ticos
tero
ids
Cho
lest
erol
Ph
ytos
tero
ls(i.
e. Β
-sito
ster
ol)
HO
H3C
H3C
H3C
CH
3
CH
3
12 3
45
678
910
1112
1314
151617
182122
19
20
Vita
min
D2:
Vita
min
E:
HO
CH
2
HH
H3
CH
3C
CH
3
CH
3 CH
3
OR
1
R2
HO
R3
CH
3 (CH
2CH
2CH
2CH
2)2C
H2C
H2C
H2C
H(C
H3)
2
CH
3
Fat
cont
ents
of s
elec
ted
food
s:
Liqu
id v
eget
able
oils
≅10
0%
Mar
gari
ne a
nd b
utte
r ≅
80%
→th
ey a
re e
mul
sion
s of
lipi
d in
wat
er
Alm
onds
:%55
, W
alnu
ts:%
65, C
erea
ls:3
-5%
Sunf
lowe
r se
eds,
hul
led
: ~6
0%
Choc
olat
e:~3
5% ,
Milk
:37
%, M
ilk p
owde
r : 2
7.5%
Tuna
fis
h: 4
% (
but
ther
e ar
e so
me
fatt
y fi
sh li
ke
anch
ovie
s co
ntai
ning
15%
fat
)
Ia. G
ener
al p
roce
dure
for
sol
vent
ext
ract
ions
:
1.Sam
ple
Prep
arat
ion:
Plant
or
anim
al t
issu
es s
hould
be h
omog
enized
by
grind
ing,
blen
ding
etc
, sinc
e pa
rticle s
ize
is
impo
rtan
t; it
has
to
be
idea
lpa
rticle
size
for
max
imum
ex
trac
tion
yields
(i.e.
0.4m
mφ)
. th
en it ha
s to
be
dr
ied
befo
re ex
trac
tion
, be
caus
e if a
ny m
oist
ure
is let
sta
y in t
he s
amples
, it int
erfe
res
with
ex
trac
tion
, an
d th
e so
lven
t's
extr
action
ca
pabi
lity
decr
ease
s.Also
hydr
olys
is is ne
eded
to
br
eak
down
an
y bo
und
lipids,
eithe
r by
aci
ds(H
Clor
H2S
O4)
or b
y en
zym
es l
ike
Taka
dias
tase
.
2.
Solve
nt
select
ion:
Yo
u ne
ed
diff
eren
t ty
pes
of
solven
ts
for
extr
acting
diffe
rent
typ
es o
f lip
ids:
(Chlor
ofor
m+m
etha
nol)
is id
eal
for
lipop
rote
ins,
wh
erea
s pe
troleu
met
her
frac
tion
s(C6
an
d C5
) ar
e idea
l fo
r tr
ue
lipid
s (triyg
lyce
ride
s),
and
ethy
l et
her
for
extr
acting
ox
idized
lip
ids.
So
met
imes
a t
erna
ry m
ixtu
re w
orks
bet
ter
(CHCl
3: m
etha
nol:H2O
) fo
r ex
trac
tion
fro
m a
nimal
tiss
ues;
wat
er-s
atur
ated
but
anol f
or
cere
al lipid
s .
LIPI
D C
ON
TEN
T A
NA
LYSE
S
1.G
ravi
met
ric M
etho
ds
(1) W
et e
xtra
ctio
n -R
oese
Got
tlieg
b&
Moj
onni
er.
(2) D
ry e
xtra
ctio
n -S
oxhl
etM
etho
d.
2.V
olum
etric
Met
hods
(Bab
cock
, Ger
ber M
etho
ds)
1. G
ravi
met
ric M
etho
ds
(1)
Wet
Ext
ract
ion
-Roe
seG
ottli
eb &
Moj
onni
er.
For M
ilk:
1) 1
0 g
milk
+ 1
.25
ml N
H4O
H m
ix. s
olub
ilize
spr
otei
n an
d ne
utra
lizes
.
2) +
10
ml E
tOH
-sha
ke.
Beg
ins e
xtra
ctio
n, p
reve
nts
gela
tion
of p
rote
ins.
3) +
25
ml E
t2O
-sh
ake
and
mix
.
4) +
25
ml p
etro
leum
eth
er, m
ix a
nd sh
ake.
(2)
Dry
Ext
ract
ion
-Sox
hlet
Met
hod.
Sam
ple
in th
imbl
e is
con
tinuo
usly
ext
ract
ed w
ith e
ther
us
ing
Soxh
letc
onde
nser
. A
fter e
xtra
ctio
n, d
irect
m
easu
rem
ent o
f fat
-eva
pora
te e
ther
and
wei
gh th
e fla
sk.
Indi
rect
mea
sure
men
t -dr
y th
imbl
e an
d w
eigh
thim
ble
and
sam
ple.
2. V
olum
etric
Met
hods
(Bab
cock
, Ger
ber M
etho
ds)
Theo
ry:
1.Tr
eat s
ampl
e w
ith H
2SO
4 or
det
erge
nt.
2.C
entri
fuge
to se
para
te fa
t lay
er.
3.M
easu
re th
e fa
t con
tent
usi
ng sp
ecia
lly c
alib
rate
d bo
ttles
.
Met
hods
:1.
Kno
wn
wei
ght s
ampl
e.2.
H2S
O4
-dig
est p
rote
in, l
ique
fy fa
t.3.
Add
H2O
so th
at fa
t will
be
in g
radu
ated
par
t of b
ottle
.4.
cent
rifug
e to
sepa
rate
fat f
rom
oth
er m
ater
ials
com
plet
ely.
Typ
es o
f Ex
trac
tion
:a1
.. "D
ry E
xtra
ctions
":Th
ese
are
basi
cally
sol
id-
liqui
dty
pe o
f ex
trac
tion
s,an
d th
e fi
nal
resu
lts
are
grav
imet
ric.
Th
ey
may
be
ei
ther
ba
tch,
m
ulti
ple-
batc
h,
cont
inuo
us
or
coun
terc
urre
nt
extr
acti
ons.
Th
ere
are
also
au
tom
ated
pr
oces
ses,
like
FOSS
LET
extr
acto
rs, w
here
the
pr
edet
erm
ined
num
ber
of s
tage
s is
(4
-5),
and
cart
ridg
esar
e us
ed t
o co
ntai
n th
e sa
mpl
e.In
co
mpl
ex m
atri
ces,
dig
esti
on m
ay b
e re
quir
ed t
o lib
erat
e bo
und
lipid
s by
hy
drol
ysis
be
fore
star
ting
the
ext
ract
ion.
Solven
t Ex
trac
tion
Met
hods
:a.
1.1.
Cont
inuo
us(G
oldf
ish)
or
semicon
tinu
ous(Sox
hlet
) or
disc
ontinu
ous(M
ajon
nier
) so
lven
t ex
trac
tion
met
hods
: Solve
nt
from
a
boiling
flas
k co
ntinuo
usly f
lows
ove
r th
e sa
mple
held in
a th
imbl
e.Fa
t co
nten
t is m
easu
red
by w
eigh
t of
fa
t re
mov
ed.
a.1.
2.El
evat
ed
pres
sure
/Tem
pera
ture
met
hods
: SFE
(Sup
ercr
itical
fluid
extr
action
s):inc
e it
avoids
th
e us
e of
ha
zard
ous
orga
nic
solven
ts,
and
uses
ine
rt,
inex
pens
ive,
and
non
toxi
c ga
ses
like
CO2, i
t is g
aining
mor
e po
pularity
.
a.2.
“W
etno
nsolve
nt e
xtra
ctions
":
are
pref
erre
d f
or
liqui
d sa
mpl
es, s
ince
the
y in
volv
e liq
-liq
extr
acti
on
(par
titi
onin
g of
mat
eria
l bet
ween
two
imm
isci
ble
liqui
ds)
and
yiel
d vo
lum
etri
c re
sult
s.
Ex: G
erbe
ran
d Ba
bcoc
k ty
pe o
f ex
trac
tion
of
milk
are
ex
ampl
esof
wet
ext
ract
ion.
In
milk
, the
fat
glo
bule
s ar
e pr
esen
t in
the
for
m o
f an
em
ulsi
onof
“oil
in w
ater
”, su
rrou
nded
by
a th
in f
ilm o
f pr
otei
n wh
ich
has
to b
e re
mov
ed o
r br
oken
by
acid
tre
atm
ent
befo
re s
epar
atin
g fa
t fr
om w
ater
pha
se.
Gerb
erPr
oced
ure:
11
ml m
ilk a
nd 1
ml i
soam
ylal
coho
l are
m
ixed
to
impr
ove
phas
e se
para
tion
, by
incr
easi
ng t
he w
ater
ph
ase.
Then
H2S
O4
(Sp.
gr.
:1.8
2 fo
r br
eaki
ng d
own
the
lipid
-pro
tein
bar
rier
) is
adde
d an
d m
ixtu
re is
cen
trif
uged
fo
r 5
min
,sub
mer
ged
in H
2O b
ath
(65°
C, 5
min
). V
olum
es
of t
wo r
espe
ctiv
e ph
ases
giv
es f
at-w
ater
rat
io, o
r th
e up
per
laye
r gi
ves
fat
cont
ent.
I.b.
Ind
irec
t M
etho
dsfo
r Q
uant
ific
atio
n of
Lip
ids
Espe
cial
ly g
enet
icis
ts i
n pl
ant
bre
edin
g re
sear
ch s
tudi
es
requ
ire
“non
-des
truc
tive
”ty
pes
of li
pid
anal
yses
. Fo
r th
is
purp
ose,
man
y “in
stru
men
tal s
yste
ms”
are
deve
lope
d:
*NM
R:Lo
w re
sool
utio
nnu
clea
rm
agne
tic
reso
nanc
e:Th
e m
easu
red
NM
R va
lue
is r
elat
ed t
o th
e to
tal
num
ber
of
prot
ons
in o
il fr
acti
on o
f se
ed ,
inde
pend
ent
of p
roto
ns in
no
n-oi
l fra
ctio
n, s
ince
pro
tons
ha
ve a
dif
fere
nt m
agne
tic
reso
nanc
e in
oil
and
in
wate
r ph
ase.
It
give
s ve
ry g
ood
appr
oxim
atio
n of
lipi
d le
vels
. Cal
ibra
tion
tab
les
or c
urve
s ar
e us
ed f
or d
iffe
rent
H 2
0 le
vels
(fig
. 13-
8).
*X-r
ay
abso
rbti
on:
Is
an
appr
oved
m
etho
d fo
r m
eat
carc
ass
sam
ples
. Le
an (
yağs
ız)
mea
t ab
sorb
s m
ore
x-ra
y th
an f
atty
tis
sues
(car
cass
-lipi
ds)
Cond
ucta
nce:
Lean
mea
t is
20-
fold
bet
ter
cond
ucto
r fo
r el
ectr
ical
cur
rent
than
fat
.
Den
sito
met
ric
Met
hods
:D
ensi
ty
of
seed
s (i.
e.
Flax
seed
) is
pro
port
iona
l to
its
oil c
onte
nt. L
ikew
ise,
de
nsit
y of
ext
ract
ing
solv
ent
chan
ges
prop
orti
onal
to
the
fat
cont
ent
it
has
extr
acte
d(i.e
.per
chlo
roet
hyle
ne
in
FOSS
LET)
. Fo
r m
eat
, the
re is
a m
etho
d us
ing
a cy
lindr
ical
con
tain
er
with
a p
isto
n pl
ate
that
hol
ds m
eat
unde
r co
nsta
nt
hydr
aulic
pre
ssur
e. M
easu
red
impr
essi
on d
epth
is
rela
ted
to f
at c
onte
nt.
Refr
acto
met
ry:
η D o
f th
e m
ixtu
re (
brom
onap
htal
ene
and
fat
extr
acte
d by
it f
rom
cho
cola
te)
is u
sed
for
appr
oxim
atio
n of
fat
in c
hoco
late
.
II. A
ssay
s on
Ext
ract
ed L
ipid
s-F
AT
CHA
RACT
ERIS
ATI
ON
(Ch.
14):
II.a
. Pu
rity
Crite
ria:
For
auth
enti
city
tes
ting
an
d de
term
inat
ion
of
adul
tera
tion
. M
ost
anal
ytic
al v
alue
s of
cer
tain
oils
and
fat
s ar
e qu
ite
char
acte
rist
ic:
a.1.
Phys
ical
purity
crite
ria
are
refr
acti
ve
inde
x,
mel
ting
po
int,
co
lour
,de
nsit
y,.
By
mea
suri
ng t
hese
cri
teri
a, w
e wi
ll fi
nd v
alue
s wi
thin
cer
tain
acc
epte
d lim
its
and
rang
es.
But
thes
e al
one
are
not
eno
ugh
for
auth
enti
city
ch
arac
teri
sati
on.
Col
or:E
ither
Lov
ibon
d co
lor s
cale
usi
ng
stan
dard
red-
yello
w-b
lue
filte
rs, o
r de
term
inat
ion
of “
phot
omet
ric c
olor
inde
x-PC
I)*
is re
quire
d fo
r obj
ectiv
e de
scrip
tion
of
colo
r.
Ref
ract
ive
Inde
x:R
I may
als
o be
use
d fo
r co
ntro
lling
hyd
roge
natio
n, si
nce
RI w
ill
decr
ease
line
arly
as u
nsat
urat
ion
decr
ease
s.
Mel
ting
poin
t, sm
oke,
flas
h an
d fir
e po
ints
)(4.
56)
(2.41
)(7.
69)
(29.1
*67
062
055
046
0A
AA
APCI
−+
+=
Effe
cts o
f Dou
ble
Bon
ds o
n th
e M
eltin
g Po
ints
16:0
6016
:11
18:0
6318
:116
18:2
-518
:3-1
120
:075
F. A
.M
. P. (
0 C)
20:4
-50
M.P
.
# D
oubl
e bo
nds
a.2.
Ch
emical
purity
cr
iter
ia:
Thes
e ar
e ve
ry
char
acte
rist
icfo
r ea
ch o
il an
d fa
t ty
pes:
•Sa
poni
fica
tion
valu
em
easu
ring
ave
rage
f.a
. m
olec
ular
we
ight
and
cha
in le
ngth
•I
odin
e va
lue
mea
suri
ngle
vel o
f un
satu
rati
on
•Fa
tty
acid
com
posi
tion
(FA
ME)
,
•In
divi
dual
ste
rola
nd t
ocop
hero
l com
posi
tion
s.
Wit
h an
inte
grat
ed e
valu
atio
n of
res
ults
of
puri
ty
crit
eria
ana
lyse
s,we
can
be
sure
of
iden
tity
and
sour
ce o
f fa
ts a
nd o
ils(R
efer
ence
met
hods
in
AO
CS,I
UPA
C an
d D
GF).
1. S
apon
ifica
tion
Val
ue(1
4.3.
9)
Sapo
nific
atio
n-h
ydro
lysi
s of e
ster
link
ages
und
er
alka
line
(KO
H)c
ondi
tion.
O
C
ROO C
R
C
RO
H2C
O
HC
O
H2C
O
KO
H
HHH
H2C
O
HC
O
H2C
O
R
C
OK
+3
+3
Milk
Fat
210-
233
Coc
onut
Oil
250-
264
Cot
ton
Seed
Oil
189-
198
Soyb
ean
Oil
189-
195
Fat
Sapo
nific
atio
nV
alue
Lard
190-
202
Sapo
nific
atio
nV
alue
of F
ats a
nd O
ils
Sapo
nific
atio
nV
alue
=-m
gs o
f KO
H r
equi
red
to sa
poni
fy1
g of
fat.
1.5
g in
250
ml E
rlen
mey
er.
2.50
ml K
OH
in E
rlen
mey
er.
3.B
oil f
or sa
poni
ficat
ion.
4.T
itrat
e w
ith H
Clu
sing
phe
nolp
htha
lein
.5.
Con
duct
bla
nk d
eter
min
atio
n.
B -
ml o
f HC
lreq
uire
d by
Bla
nk.
S -m
l of H
Clr
equi
red
by S
ampl
e.
SP#
= 56
.1(B
-S) x
N o
f HC
l
Gra
m o
f Sam
ple
Sapo
nific
atio
nV
alue
Det
erm
inat
ion
SV is
an
inde
x of
the
mea
n m
olec
ular
wei
ght
of t
he
tria
cylg
lyce
rols
in s
ampl
e oi
l or
fat.
The
mea
n m
olec
ular
wei
ght
of t
riac
ylgl
ycer
ols
when
di
vide
d by
thr
ee w
ill g
ive
an a
ppro
xim
ate
mea
n m
olec
ular
wei
ght
of f
atty
aci
ds p
rese
nt.
2. I
odin
e N
umbe
r (14
.3.8
)
Iodi
ne v
alue
(IV)
: Hal
ogen
s ad
d to
dou
ble
bond
s.
IV is
the
num
ber
of g
ram
s of
iodi
ne t
hat
adds
to
100
gram
of
oil.
If 1
g o
f the
fat a
dsor
bs1.
5 g
of io
dine
, the
n IV
= 15
0.
It is
a m
easu
re o
f sa
tura
tion
-uns
atur
atio
n.
Iodi
ne V
alue
= (m
l of N
a 2S 2
O3
volu
me
for
blan
k -
ml o
f N
a 2S 2
O3
volu
me
for
sam
ple)
×N
of N
a 2S 2
O3
×0.
127g
/meq
×10
0
Wei
ght o
f Sam
ple
(g)
CH
C
HC
H
CH
Cl
IIC
lIo
dine
chl
orid
e+
ICl
KI
KC
l
I 2
I 2
Na
2S
2O
3N
a2
S4
O6
NaI
+
+2
2+
+
Exc
ess u
nrea
cted
ICl
Iodi
ne V
alue
Det
erm
inat
ion
Iodi
ne N
umbe
rs o
f Sel
ecte
d F.
A.
Palm
itole
icA
cid
195
Ole
ic A
cid
186
Lino
leic
Aci
d2
173
Lino
leni
cA
cid
326
1
Fatt
y A
cids
# of
Dou
ble-
bond
sIo
dine
#
Ara
chid
onic
Aci
d4
320
If m
olec
ular
wei
ght a
nd io
dine
num
ber o
f f.a
. isk
now
n,
we
can
cal
cula
te th
e nu
mbe
r of d
oubl
e bo
nds:
IV/M
W
Dur
ing
oxid
atio
n,
ther
e wi
ll be
a
decl
ine
in
unsa
tuar
tion
and
the
refo
re in
IV:
The
rang
es o
f Io
dine
Val
uefo
r ol
ive
oil,
whic
h is
an
inde
x of
its
degr
ee o
f un
satu
rati
on,
is 7
5-94
in t
he C
odex
. F
or I
taly
thi
s va
lue
fluc
tuat
es
betw
een
79 a
nd 8
8, b
ut f
or T
unis
iath
e ra
nge
is
80-9
7.Th
ere
may
be
such
reg
iona
l or
clim
atic
va
riat
ions
, th
eref
ore,
ev
en
thou
gh r
esul
ts a
re
quit
e m
eani
ngfu
l, so
met
imes
add
itio
nal
test
s ar
e re
quir
ed f
or f
inal
con
firm
atio
n.
GC
Ana
lysi
s for
Fat
ty A
cids
(14.
6.1)
1.Ex
tract
fat.
2.Sa
poni
fy(h
ydro
lysi
s und
er b
asic
con
ditio
n).
3.Pr
epar
e “f
atty
aci
d m
ethy
l est
ers”
or F
AM
E (b
y So
dium
met
hoxi
de-
CH
3ON
a or
BF3
-met
hano
l rea
gent
).
4.C
hrom
atog
raph
y of
met
hyl e
ster
s.
5.D
eter
min
e pe
ak a
reas
of f
atty
aci
ds.
(Fat
ty a
cids
are
iden
tifie
d by
thei
r spe
cific
rete
ntio
n tim
es).
6.C
ompa
re w
ith re
spon
se c
urve
of s
tand
ard .
14
18:1
18:2
2018
:3
22
21:1
2416
18
Tim
e
Res
pons
e
For
anal
ysin
gFA
ME:
Th
e FA
ME
are
carr
ied
in a
nd o
ut o
f th
e GC
Col
umn
by
the
carr
ier
gas
and
each
will
leav
e th
e co
lum
n on
ly a
fter
a
spec
ific
ret
enti
on t
ime,
Rt, w
hich
is d
epen
dant
on
the
chai
n le
ngth
and
deg
ree
of s
atur
atio
n of
the
fat
ty a
cid.
Ge
nera
lly, a
s ca
rbon
num
ber
as w
ell a
s do
uble
bon
ds in
f.
atty
aci
ds i
ncre
ase,
the
rete
ntio
n ti
me
Rt w
ill
also
in
crea
se.
FAM
E:Th
e fa
tty
acid
com
posi
tion
det
erm
ined
by
GC
(gas
chr
omot
ogra
phy)
is a
real
"Fi
nger
prin
t" .
It
also
se
rves
fo
r “n
utri
tion
al
labe
lling
”: sa
tura
teds
,mon
ouns
atur
ates
, pol
yuns
atur
ates
, tra
ns f
.a.
etc.
For
auth
enti
city
tes
ting
of
butt
ers,
thi
s an
alys
isby
its
elf
may
not
som
etim
es b
esu
ffic
ient
, si
nce
som
e m
arga
rine
s m
ight
hav
e si
mila
r co
mpo
siti
on o
f FA
ME
with
but
ters
.
In t
his
case
,the
res
pect
ive
ster
ol c
ompo
siti
onsh
ould
als
o be
dete
rmin
ed. S
tero
l ana
lysi
s is
don
e by
TLC
fol
lowe
d by
GC
of
the
unsa
poni
fiab
lefr
acti
on
of l
ipid
s. (
Ther
e ar
e “P
hyto
ster
ols”
-st
igm
aste
rol,
cam
pest
erol
, β-
sito
ster
ol-
inth
e ve
geta
ble
ster
olfr
acti
on, b
ut o
nly
“cho
lest
erol
”in
an
imal
fat
s).
CH
OLE
STER
OL
DET
ERM
INA
TIO
N(1
4.6.
4)C
hole
ster
ol b
y G
LC:s
apon
ify e
xtra
cted
lipi
d,de
rivat
ize
G
C
µg/
ml C
hole
ster
ol
Abs
orpt
ion
at
440
nm
Spec
trom
ertic
Abs
orpt
ion
Stan
dard
Cur
ve o
f Cho
lest
erol
Enzy
mat
ic D
eter
min
atio
n: C
hole
ster
ol O
xida
se H
OO
H2O
2
Cho
lest
erol
Oxi
dase
etc.
+
H2O
2
CH
3OO
CH 3
H2N
NH 2
HN
NH
OC
H 3C
H3O
H2O
Pero
xida
se+
+
0-D
iani
sidi
neO
xidi
zed
0-D
iani
sidi
ne(C
olor
less
)(B
row
n co
lor)
At 4
40 n
m
TRIG
LYC
ERID
E A
NA
LYSI
S
BY
LIQ
UID
CH
RO
MA
TOG
RA
PHY
Soyb
ean
Oil
Solv
ent C
H3C
N/H
FC
olum
n 84
346
(Wat
ers A
ssoc
iate
s)
RES
PON
SE
RET
ENTI
ON
TIM
E
Ole
ate-
cont
aini
ng tr
igly
ceri
des i
n ol
ive
oil
OL
254
:544
O2L
54:4
46
OPL
52:3
46
O3
54:3
48
OSL
54:3
48
O2P
52:2
48
O2S
54:2
50
OPS
52:1
50
Fatt
y A
cid
Com
posi
tion
Tot
al A
cylC
arbo
ns:
Uns
atur
atio
nE
quiv
alen
t Car
bon
Num
ber
OS2
54:1
52
Furt
her
Spec
ific I
dent
ificat
ion
Test
s fo
r So
meF
ats
and
Oils
:( F
or d
etec
tion
of
adul
tera
tion
)
For
butt
er: R
eich
ert-
Mei
ssel
inde
x fo
r bu
tter
is a
mea
sure
of
its
con
tent
of
vola
tile
and
wat
er-s
olub
le b
utyr
ic a
cid;
>2
4.Fo
rol
ive
oil:
Squa
lene
(700
0 m
g/kg
) is
a
very
spe
cifi
c co
nsti
tuen
t of
un
sapo
nifi
able
par
t of
oliv
e oi
l, wh
erea
s in
ot
hers
, it
vari
es b
etwe
en 3
0-40
0ppm
.Fo
r co
tton
seed
oil:
Hal
phen
test
for
dete
ctin
g pr
esen
ce o
f cy
clop
rope
noic
aci
ds w
hich
for
m a
red
col
or o
n he
atin
g wi
th a
myl
alc
ohol
and
CS2
.Fo
r se
sam
esee
d oi
l: Vi
llave
chia
tes
t: u
nsap
onif
iabl
es o
f se
sam
esee
d oi
l onl
y wi
ll fo
rm p
ink
colo
r wi
th f
urfu
ral.
For
coco
butt
er: T
rigl
ycer
ides
C50
and
C54
are
dom
inan
t in
co
cobu
tter
but
not
in C
BE a
nd C
BS.
II b
. Qua
lity
Crit
eria
IIb.
1. M
I U
(M
oist
ure+
Inso
lubl
e Im
puri
ties
)(d
irt,
oil
seed
pa
rtic
les
in
crud
e oi
ls
and
soap
in
re
fine
d oi
ls)+
(uns
apon
ifia
bles
) :
They
are
nonf
atty
cons
titu
ents
of
oils
whic
h in
crea
se
refi
ning
co
sts
for
crud
e oi
ls
.
Ther
efor
e,
ever
y pe
rcen
t of
in
crea
se
in
MIU
co
nten
t br
ings
par
alle
l pr
ice
decr
ease
sin
cru
de o
il pu
rcha
ses.
Als
o so
ap>5
ppm
lef
t in
ref
ined
oils
will
be
det
rim
enta
l to
oil
qual
ity
.II
b.2
Fat
ranc
idit
yan
d st
abili
ty
Ranc
idit
y:“D
evel
opm
ent
of
obje
ctio
nabl
e fl
avou
rsan
d od
ours
resu
ltin
g fr
om l
ipol
ysis
or
lipid
bre
ak-d
own
(eit
her
due
to h
ydro
lyti
c or
to
oxid
ativ
e re
acti
ons)
lead
ing
to lo
wer
cons
umer
acc
epta
nce”
.St
abili
ty:
“Cap
abili
ty t
o re
mai
n fr
esh
or m
aint
ain
fres
h ta
ste+
odou
r”or
“re
sist
ance
to
spoi
lage
dur
ing
stor
age
or
“res
ista
nce
of li
pids
to
ranc
idit
y”
RANCI
DIT
Y(Hyd
rolytic
and
Oxi
dative
)
A-H
ydro
lyti
c Ra
ncid
ity:
Invo
lves
che
mic
al o
r en
zym
atic
(lipa
se
from
mic
roor
gani
sms
or p
lant
cel
ls)
hydr
olys
is o
f fa
ts in
to f
ree
fatt
y ac
ids
and
mon
o or
di-g
lyce
ride
s. R
elev
ant
Test
s:A
V an
d ff
aa.
A.V
.=Th
e nu
mbe
r of
mili
gram
sof
KO
H r
equi
red
to n
eutr
aliz
e 1
gram
of
fat
or o
il.T
and
aw
b.FF
A:
Free
fa
tty
acid
s=
here
, ac
idit
y is
ex
pres
sed
as
perc
enta
geof
wei
ght
of a
spe
cifi
c fr
ee f
atty
aci
d in
a f
at o
r oi
l,thu
s ta
king
into
con
side
rati
on t
he M
.W. o
f th
e in
divi
dual
f.a
.:*
In o
live
oil,
expr
esse
d as
ole
icac
id,
MW
= 1
82*I
n pa
lm o
il, e
xpre
ssed
as
palm
itic
aci
d,M
W =
256
* %
FFA
ole
ic a
cid=
0.5
03 x
Aci
d va
lue
* A
cid
valu
e= 1
.99
x %
FFA
ole
ic a
cid
100
)10
00(
%x
Weight
acids
fatty
MW
xV
FFA
=
AV
=
ml o
f KO
H x
N x
56
Wei
ght o
f Sam
ple
= m
g of
KO
H
Sign
ifican
ce o
f AV
and
ffa:
This
val
ue w
ill e
stim
ate
the
amou
nt o
f cr
ude
oil t
hat
will
be lo
st d
urin
g re
fini
ng, s
o wi
ll m
ean
high
er o
pera
ting
co
sts
for
refi
ning
pla
nts,
and
lead
s to
pri
ce d
ecre
ases
in
crud
e oi
l pur
chas
es.
In r
efin
ed o
ils,
high
val
ues
will
mea
n ei
ther
poo
r re
fini
ng o
r fa
t br
eakd
own
in s
tora
ge o
r us
e. I
n ca
ses
wher
e th
e li
bera
ted
ffa
are
vola
tile
, ff
a wi
ll be
a
mea
sure
of
hydr
olyt
ic r
anci
dity
.
For
Vir
gin
oliv
e oi
ls <
4 %
ole
ic a
cid,
and
for
ref
ined
oliv
e oi
ls <
0.3;
but
for
sho
rter
cha
in f
.a. l
ike
buty
ric,
capr
ic a
nd e
ven
laur
ic (C
4-C1
2), h
avin
g m
uch
lowe
r fl
avou
r th
resh
olds
(as
in c
hoco
late
), th
e pr
oduc
t
will
be in
edib
le a
fter
its
ffa
cont
ent
reac
hes
> 0.0
2 %
.
B-O
xida
tive
Ran
cidi
ty
“LIP
ID O
XID
ATI
ON
“M
ajor
flav
or p
robl
ems i
n fo
od d
urin
g st
orag
e ar
e m
ainl
y du
e to
the
oxid
atio
n of
lipi
d.
Lipi
d O
xida
tion
-fre
e ra
dica
l rea
ctio
ns.
1.In
itiat
ion.
2.Pr
opag
atio
n.
3.Te
rmin
atio
n.
Met
hods
for
Mea
suring
Pre
sent
Sta
tus
of
Lipid
Oxi
dation
:-P
erox
ide
num
ber:
The
amou
nt o
f io
dine
lib
erat
ed(N
a 2S 2
O3
titr
atio
n)fr
om s
atur
ated
KI
solu
tion
by
fat
, exp
ress
sed
in
meq
of
pero
xide
O2/
kg o
f fa
t(L
ea v
alue
=exp
ress
ed in
mill
imol
es o
f O
2).
<10
for
edib
le o
ils; m
easu
res
only
tra
nsie
nt p
rim
ary
oxid
atio
n pr
oduc
ts li
ke p
erox
ides
and
hyd
rope
roxi
des.
-Thi
obar
bitu
ric
acid
val
ue (
TBA
):Th
e pi
nk c
olor
for
med
by
seco
ndar
y ox
idat
ion
prod
ucts
like
mal
onal
dehy
des
with
TBA
re
agen
t i
s m
easu
red
at 5
23 n
m.C
orre
late
s be
tter
wit
h se
nsor
y an
alys
es.
-Ani
sidi
neva
lue:
Ano
ther
co
lor
reac
tion
of
alde
hyde
s(al
kena
ls
and
dien
als-
seco
ndar
y pr
oduc
ts
of
oxid
atio
n) w
ith
P-an
isid
ine.
-Tot
oxva
lue
:2
tim
es p
erox
ide
valu
e +
anis
idin
eva
lue
= To
tal
oxid
atio
n pr
oduc
ts (
2 is
bec
ause
per
oxid
es h
ave
2 ti
mes
mor
e ox
ygen
tha
n al
dehy
des.
)
In m
ost
case
s, r
anci
dity
is
due
to p
rodu
cts
of a
uto-
oxid
atio
n of
un
satu
rate
d
fatt
y ac
ids
by
prog
ress
ive
addi
tion
of
ox
ygen
to
lip
id
com
pone
nts.
A
utox
idat
ion
reac
tion
s ar
e ca
taly
sed
by p
rese
nce
of
proo
xida
nts
like
Cu, F
e, s
unlig
ht a
nd l
ipox
igen
ase
enzy
me.
The
shel
flif
e of
fat
s an
d oi
ls is
ass
umed
as
the
end
of t
he
indu
ctio
n pe
riod
, af
ter
whic
h th
e ra
te o
f fo
rmat
ion
of
oxid
atio
n pr
oduc
ts (
Pero
xide
s,O
xide
s, H
ydro
pero
xide
s)
incr
ease
s dr
asti
cally
. IP
is 1
.3 h
ours
for
C18
:3, 1
9 ho
urs
for
C18:
2, a
nd 8
2 ho
urs
for
C18:
1 at
40°
C.D
iffe
rent
te
sts
have
be
en
deve
lope
d to
m
easu
re
the
exte
nt o
f au
toxi
dati
on, b
ut e
ach
resp
ecti
ve t
est
is li
ke a
sn
apsh
ot a
t th
e ti
me
test
ed a
nd d
o no
t ne
cess
arily
sho
w th
e co
mpl
ete
pict
ure.
Qua
ntity
of
R
eact
ants
and
Prod
ucts
Qua
ntity
of
R
eact
ants
and
Prod
ucts
TIM
E
Indu
ctio
n pe
riod*
Seco
ndar
y ox
idat
ion
prod
ucts
Pero
xide
s
Prim
ary
oxid
atio
n pr
oduc
ts
*Ind
ucti
on
peri
od
is
the
leng
th
of
tim
e be
fore
de
tect
able
ra
ncid
ity,
af
ter
whic
h th
ere
is
an
acce
lera
ted
lipid
oxi
dati
on
AN
ALY
SIS
OF
FLA
VO
R Q
UA
LITY
& S
TAB
ILIT
Y O
F O
IL
Pero
xide
Val
ue
KI
C
H3
C
O
HH
I
CH
3
C
OK
OO
RO
OH
H
II 2
H
2O
RO
H
I 2
Na 2
S2O
3N
aI
Na 2
S4O
6
A.
B.
C.
+
+
+
+
+
+
+
2
2
2
Pero
xide
Val
ue =
m
l of N
a2S2
O3
×N
×10
00
(mill
iequ
ival
entp
erox
ide/
kg o
f sam
ple)
G
ram
s of O
il
Act
ive
Oxy
gen
Met
hod
(AO
M)
Det
erm
ined
the
time
requ
ired
to o
btai
n ce
rtain
pe
roxi
de v
alue
und
er sp
ecifi
c ex
perim
enta
l con
ditio
ns.
The
larg
er th
e A
OM
val
ue, t
he b
ette
r the
flav
or
stab
ility
of t
he o
il.
TBA
Tes
t
To d
eter
min
e th
e ra
ncid
ity d
egre
e of
mea
t or f
ish
prod
uct.
N
N
HS
OH
OH
C
CH
2
CO
HO
H
OH
OH
HS
N
N
OH
SH
N
N
CH
C
H
CHH
O
H2O
Col
ored
Pig
men
t
+
+2
Stor
age
stab
ility
(sh
elf-
life)
:Li
pids
var
y in
the
ir s
usce
ptib
ility
to
ranc
idit
y be
caus
e of
in
here
nt
prop
erti
es
like
unsa
tura
tion
, pr
esen
ce
of
anti
oxid
ants
, an
d so
me
exte
rnal
fac
tors
lik
e pr
oces
sing
an
d st
orag
e co
ndit
ions
.Acc
eler
ated
tes
ts a
re u
sed
for
pred
icti
ng f
utur
e be
havi
our.
For
exa
mpl
e, b
y ra
isin
g th
e te
mpe
ratu
re,
you
natu
rally
acce
lera
te
the
rate
of
de
teri
orat
ion.
At
the
end
of t
he in
duct
ion
peri
od,y
ou s
top
and
perf
orm
per
oxid
e de
term
inat
ion
anal
ysis
.En
d of
she
lf li
fe c
an b
e de
fine
d as
day
s at
20°
C to
rea
ch
P.V.
=20.
This
poi
nt i
s ge
nera
lly t
he e
nd o
f th
e in
duct
ion
peri
od,
and
the
end
of t
he t
est.
The
long
er t
he I
P, t
he
bett
er t
he s
tora
ge s
tabi
lity
and
the
long
er t
he e
xpec
ted
shel
flif
e.Fo
r th
is p
urpo
se m
any
diff
eren
t “a
ccel
erat
edsh
elf-
life”
met
hods
hav
e be
en d
evel
oped
:
-Sch
aal
Ove
nte
st:
Oil
is
kept
at
63
°C
till
ranc
idit
y is
de
tect
ed
or
till
the
end
of
incu
bati
on
peri
od(A
ssum
ptio
n:1
day
of
incu
bati
on a
t 63
°C c
orre
spon
ds t
o 10
day
s at
20
°C.)
-Act
ive
oxgy
enm
etho
d (A
OM
):O
2is
bub
bled
th
roug
h dr
ied
and
filt
ered
oil
to
furt
her
acce
lera
te a
utox
idat
ion
at 1
10°C
, thi
s is
car
ried
ou
t ti
ll PV
incr
ease
s to
20
for
lard
and
100
for
ve
geta
ble
oils
. Th
e in
duct
ion
peri
od
is
dete
rmin
ed b
y di
scon
tinu
ous
mea
sure
men
ts o
f pe
roxi
de v
alue
or
sens
ory
eval
uati
on o
f ra
ncis
od
or.
OXID
ATIV
E ST
ABI
LITY
INDEX
(OSI
)O
xida
tive
Stab
ility
Inde
x te
st, u
nlik
ete
sts s
uch
as p
erox
ide
valu
e w
hich
det
erm
ine
oil q
ualit
y on
ly a
t the
tim
e of
the
test
, ha
spre
dict
ive
valu
e, th
ereb
y al
low
ing
qual
ity d
ecis
ions
to
be m
ade
befo
re sp
oila
ge b
ecom
es a
pro
blem
. Th
e 0S
I is a
n in
valu
able
qua
lity
cont
rol t
ool f
or e
limin
atin
g in
ferio
r ing
redi
ents
(for
exa
mpl
e ha
zeln
uts f
or c
hoco
late
m
anuf
actu
re),
for m
easu
ring
and
com
parin
g th
e st
abili
ty o
f fin
ishe
d oi
l-fat
pro
duct
sas w
ell a
s lo
w m
oist
ure
snac
ks li
ke
pata
to c
hips
, for
eva
luat
ing
antio
xida
nt a
ctiv
ity, f
or
mon
itorin
g th
e de
terio
ratio
n of
coo
king
oils
with
use
, and
fo
r man
y ot
her a
pplic
atio
ns w
here
oxi
dativ
e ra
ncid
ity is
the
caus
e of
pro
duct
failu
re. B
ecau
se th
e O
SI h
as p
redi
ctiv
e va
lue,
pro
blem
s are
det
ecte
d an
d el
imin
ated
bef
ore
they
ca
use
reje
ctio
n, u
nlik
e ot
her t
ests
whi
ch m
erel
y in
dica
te th
at
prob
lem
s hav
e oc
curr
ed a
fter i
t is t
oo la
te to
cor
rect
them
.
-RA
NCI
MA
TM
etho
d:Ra
ncim
at v
alue
at
Tem
pera
ture
T°
is t
he n
umbe
r of
hou
rs,t
, bef
ore
the
oil r
each
es
the
end
of it
s in
duct
ion
peri
od.T
his
devi
ce p
erfo
rms
shel
f lif
e an
alys
is f
or a
t le
ast
2 pr
esel
ecte
d hi
gh
tem
pera
ture
s (i.
e. 1
00 °
and
130
° o
r 14
0°C)
.
For
this
pur
pose
, yo
u pu
t th
e sa
mpl
e in
the
dev
ice,
in
crea
se t
he t
empe
ratu
reto
the
pre
sele
cted
T1,
and
the
prod
ucts
of
oil o
xida
tion
und
er t
hese
con
diti
ons
are
colle
cted
by
dist
illat
ion
into
a w
ater
tra
p(a
spec
ial c
olle
ctin
g ve
ssel
con
tain
ing
deio
nize
d wa
ter)
. Th
e co
nduc
tivi
ty o
f wa
ter
is c
onti
nuou
sly
mea
sure
d an
d r
ecor
ded.
The
abr
upt
cha
nge
in c
ondu
ctiv
ity
mar
ks t
he e
nd o
f th
e in
duct
ion
peri
od,t
.
Ass
umpt
ions
:Q
10=
rate
at
(T+1
0) °
C/ r
ate
at T
°C
= sh
elfl
ife
at T
°C
/ sh
elfl
ife
at (T
+10)
°C
Grap
hs w
here
Co
nduc
tanc
e va
lue(
1/R)
is p
lott
ed
agai
nst
tim
e(t)
are
use
d fo
r sh
elfl
ife
pred
icti
on.
A p
lot
of lo
g t
(tim
e pe
riod
cor
resp
ondi
ng t
o en
d of
she
lf-
life)
ver
sus
T°C
thro
ugho
ut a
ran
ge o
f te
mpe
ratu
res
will
be
a st
raig
ht li
ne. T
he t
wo p
oint
s co
rres
pond
ing
to lo
gt1
and
log
t2 v
alue
s at
tem
pera
ture
s T
1an
d T 2
resp
ecti
vely
wi
ll
be p
lott
ed a
nd t
he t
wo p
oint
s so
con
nect
ed a
s to
in
terc
ept
both
x-a
xis
and
y-ax
is. T
he s
helf
life
at
a th
ird
tem
pera
ture
T 3
ca
n th
en
be
read
fr
om
grap
h by
ex
trap
olat
ion.
1/R
T(tim
e)
For
refi
ned
edib
le v
eget
able
oils
, Ra
ncim
at v
alue
s ca
n be
use
d in
the
gen
eral
eq
uati
on o
f lin
e to
de
duce
exp
ecte
d sh
elf-
life:
D20
≅7.4
0 R
100-
7.9
(her
e, r
=0.9
33)
Whe
re D
20=d
ays
at 2
0°C
to re
ach
PV=2
0
And
R10
0=R
anci
mat
val
ue in
hou
rs a
t 100
°C
Phys
ical
Met
hods
-The
rmal
Met
hods
*Diff
eren
tial s
cann
ing
calo
rimet
er*T
herm
ogra
vim
etric
and
pres
sure
di
ffer
entia
l cal
orim
etric
met
hods
*Oxi
dativ
e st
abili
ty-le
ngth
of t
he
incu
batio
n pe
riod-
time
to m
ax w
eigh
t ga
in a
nd ra
te o
f wei
ght i
ncre
ase-
time
of m
ax e
ntha
lpy
chan
ge
Ther
mal
Met
hod-
Che
milu
min
esce
nce
*Det
erm
ine
the
cont
ent o
f ef
fect
iven
ess o
f ant
ioxi
dant
s in
oils
and
fats
*cor
rela
tes w
ith th
e pe
roxi
de
valu
e an
d an
isid
ine
valu
es a
nd
flavo
ur*e
mitt
ed d
urin
g pe
roxi
date
brea
kdow
n
Indi
cato
rs o
f Qua
lity
of H
eate
d O
ils *Det
erio
tive
chan
ges-
ther
mal
oxid
atio
n,hy
drol
ysis
,po
limer
izat
ion
*Dee
p fa
t fry
ing
pota
toes
with
fast
turn
over
in s
unflo
wer
oi
l-the
rmox
idat
ive
chan
ges
*A d
ram
atic
rise
in p
olar
com
poun
ds; M
ore
than
25-
27%
po
lar l
ipid
s sh
ould
be
reje
cted
*NIR
S-d
eter
min
e th
e su
m o
f dim
eran
d po
lym
er
trigl
ycer
ides
and
aci
d va
lue
to e
valu
ate
fryin
g oi
lC
hang
es in
die
lect
ric c
onst
ant
Tox
ic C
onta
min
ants
and
A
dulte
rant
s
-Con
tam
inat
ion
by w
eed
seed
s du
ring
harv
estin
g-S
tora
ge o
f saf
e oi
l in
unsa
fe
cont
aine
r
Con
tam
inat
ion
due
to F
aulty
Sto
rage
*Con
tam
inat
ion
of v
eget
able
oi
ls b
y m
iner
al o
ils-
dete
rmin
ing
the
sapo
nific
atio
nva
lues
of t
he lu
bric
atin
g oi
l sa
mpl
es*
Indi
ces o
f Adm
ixtu
res,
Ble
nds,
Con
tam
inan
ts a
nd
Adu
ltera
nts-
One
Fat
to
Ano
ther
*Sta
ble
carb
on is
otop
e ra
tio
anal
ysis
-est
ablis
hing
the
auth
entic
ity o
f oils
*Fou
rier t
rans
form
infra
red
spec
trosc
opy
for a
uthe
ntic
ity
of e
xtra
virg
in o
live
oil
Adm
ixtu
re o
f Veg
etab
le O
ils
with
Oth
er V
eget
able
Oils
Bio
synt
hesi
sof
Fat
-Spe
cies
spe
cific
-Det
ectio
n of
adu
ltera
tion
can
be b
ased
on
the
basi
s of
var
ious
con
stitu
ents
in
the
fat a
dmix
ture
Adm
ixtu
re o
f Veg
etab
le O
ils
with
Oth
er V
eget
able
Oils
-Fat
ty A
cid
Com
posi
tion
-Trig
lyce
ride
Ana
lysi
s-U
nsap
onifi
able
Frac
tion
of O
il*S
tero
l ana
lysi
s*T
ocop
hero
lana
lysi
s*P
heno
lics
and
Alco
hols
Fatty
Aci
d C
ompo
sitio
n*W
ithou
t bei
ng d
etec
ted
by th
e ro
utin
e ph
ysic
al a
nd c
hem
ical
cha
ract
eris
tics-
GLC
an
alys
is o
f fat
ty a
cids
pro
ve u
sefu
l-pr
esen
ce o
f so
ybea
n oi
ls in
oliv
e oi
l *I
OO
C fo
r oliv
e oi
l pur
ity-s
atur
ated
fatty
ac
id c
onte
nt a
t pos
ition
-2*P
ancr
eatic
lipa
se a
llow
s th
e di
ffere
ntat
ion
betw
een
virg
in a
nd e
ster
ified
oliv
e oi
ls-
Sat
urat
ed fa
tty a
cid
leve
l <1,
5% in
virg
in
oliv
e oi
l
Fatty
Aci
d C
ompo
sitio
n*C
ocon
ut o
il -d
eter
min
atio
n of
sap
onifi
catio
nan
d IV
-det
ect g
ross
and
sim
ple
adul
tera
tion
*Ana
lysi
s of
met
hyl e
ster
s of
fatty
aci
ds b
y G
C-
ratio
sof
cer
tain
fatty
aci
ds-d
etec
tion
of 5
%
rape
seed
oil
in p
eanu
t oil(
C22
-C16
)P
alm
ker
nel a
nd c
ocon
ut o
ils in
adu
ltera
ted
coco
a bu
ttera
s w
ell a
s ad
ditio
n of
coc
onut
or
palm
ker
nel o
il to
but
ter (
high
laur
ican
d m
yris
ticac
id c
onte
nt).
*Lin
olen
icac
id-d
etec
tion
of 5
% g
rape
seed
oil o
r <5
% s
unflo
wer
oil
in o
live
oil
Uns
apon
ifiab
leFr
actio
n of
Oils
*The
pre
senc
e or
abs
ence
of
certa
in in
divi
dual
uns
apon
ifiab
leco
mpo
nent
s ca
n he
lp in
es
tabl
ishi
ng th
e id
entit
y of
eac
h oi
l (a
uthe
ntic
ity o
r ge
nuin
enes
s)an
d al
so in
det
ectio
n of
adm
ixtu
re w
ith
anot
her o
il.
Uns
apon
ifiab
leFr
actio
n of
Oils
Ster
ol A
naly
sis
*Ana
lysi
s of
ste
rols
from
uns
apon
ifiab
lefra
ctio
n -a
dulte
ratio
n ol
ive
oil w
ith s
oybe
an o
il*S
tero
l com
posi
tion
obta
ined
by fr
actio
n of
the
unsa
poni
fiabl
em
atte
r is
char
acte
ristic
of v
irgin
ol
ive
oils
*The
ratio
of t
he c
onte
nt o
f β-s
itost
erol
to th
e su
m o
f (s
tigm
aste
rola
nd c
ampe
ster
ol)i
s 25
-30
for v
irgin
oliv
e oi
l.V
irgin
oliv
e oi
l has
β-s
itost
erol
cont
ent o
f 94%
or
hig
her
Uns
apon
ifiab
leFr
actio
n of
Oil
Toco
pher
olA
naly
sis
*Con
cent
ratio
n of
γ-to
coph
erol
sin
so
lven
t ext
ract
ed c
otto
nsee
d oi
ls
incr
ease
d on
con
tam
inat
ion
with
so
ybea
n oi
l*C
onta
min
atio
n of
oliv
e oi
l with
pe
anut
oil
-det
ecte
d fro
m
ingr
ease
dβ-
toco
pher
olle
vels
*Qua
ntita
tive
dete
rmin
atio
n of
δan
d γ-
toco
pher
ols
-a m
etho
d fo
r de
term
inin
g th
e ad
ulte
ratio
n of
ol
ive
oil w
ith le
ss e
x pen
sive
oils
Uns
apon
ifiab
leFr
actio
n of
Oil
Phen
olic
sand
Alc
ohol
s
*Rev
erse
pha
se H
PLC
with
UV
de
tect
ion
and
GC
-MS
-qua
ntifi
catio
n of
ph
enol
s*O
live
oil p
ress
cak
e in
oliv
e oi
l can
be
dete
cted
on
the
basi
s of
the
pres
ence
of
hig
her f
atty
alc
ohol
s in
the
form
er
Ble
nds o
f Veg
etab
le a
ndM
arin
e /A
nim
alFa
tsFa
tty A
cid
Com
posi
tion
*GLC
ana
lysi
s of f
atty
aci
ds c
ompo
sitio
n-de
tect
adm
ixtu
re
of m
arin
e oi
ls in
veg
etab
le o
ils(i
ncre
ased
myr
istic
, pa
lmito
leic
, C20
and
C22
aci
d co
nten
ts).
*Pur
ity a
nd c
onta
min
atio
n of
veg
etab
le o
ils w
ith la
rd-v
alue
s of
met
hyl e
ster
s of C
8-C
20 fa
tty a
cids
Fatty
Aci
d C
ompo
sitio
n
*Ani
mal
fats
con
tain
tetra
deca
noic
and
hexa
deca
noic
acid
s in
gre
ater
am
ount
s th
an d
oes
oliv
e oi
l*A
vera
ge m
olec
ular
wei
ght o
f the
di
still
ed fr
actio
n fro
m p
ure
oliv
e oi
l ha
s a
sapo
nific
atio
nnu
mbe
r les
s th
an 1
91-in
crea
ses
with
add
ing
anim
al fa
ts
Uns
apon
ifiab
leFr
actio
n
*Diff
eren
ces
in th
e st
erol
co
mpo
sitio
n of
ani
mal
and
pla
nt
fats
-an
effic
ient
met
hod
for
dete
ctin
g ad
mix
ture
*Adu
ltera
nt a
nim
al fa
t at %
10
in
vege
tabl
e oi
l-de
tect
ed o
n ba
sis
of
chol
este
rol
Det
ectio
n of
Mix
ture
s of A
nim
al
Fats *D
etec
tion
for a
dulte
ratio
n of
bee
f fa
t by
pig
lard
-fatty
aci
d co
mpo
sitio
n an
d ph
ysic
al
char
acte
ristic
s *D
etec
tion
foin
ters
peci
es m
eat
adul
tera
tioni
sve
ry o
ften
on th
e di
ffere
nces
in fa
t cha
ract
eris
tics
Sens
ory
Qua
lity
of O
ils“G
loba
l” qu
ality
eva
luat
ion
of v
irgin
ol
ive
oil t
he IO
OC
prop
osed
(198
9) th
follo
win
g eq
uatio
n:O
.Q.I
= 2,
55 +
0,9
1SE
-0,
78A
V7,
35K
270
-0,0
66P
V O
.QI:
Ove
r-allq
ualit
yin
dex
SE
:Sen
sory
eva
luat
ion
(from
3,9-
9,0)
AV
:Aci
d va
lue
(from
0,1
-3,3
)K
270
:Abs
orba
nce
at 2
70 n
m
PV
:Per
oxid
e va
lue(
from
1,0
-20,
0)