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d
r
o
g
en
ato
m
s
to
s
tab
ili
z
e
f
r
ee
r
ad
icals.
T
h
ey
ar
e
b
en
ef
icial
at
lo
w
c
o
n
ce
n
t
r
atio
n
s
b
u
t
lo
s
e
e
f
f
icac
y
as
lev
els r
is
e
d
u
e
to
a
d
v
er
s
e
ef
f
ec
ts
with
n
o
n
-
p
er
o
x
y
l
s
p
ec
ies.
I
d
en
tify
in
g
th
e
id
ea
l
an
ti
o
x
id
an
t
co
n
te
n
t
o
f
to
co
p
h
er
o
l
s
in
v
eg
eta
b
le
o
ils
r
em
ain
s
d
if
f
icu
lt
[
4
]
.
T
o
co
p
h
e
r
o
ls
an
d
to
c
o
tr
ien
o
ls
,
k
n
o
wn
as
to
co
ls
o
r
v
itam
in
E
,
a
r
e
a
g
r
o
u
p
o
f
n
at
u
r
ally
-
o
cc
u
r
r
in
g
ch
em
icals
th
at
ar
e
s
o
lu
b
le
in
f
at.
T
h
is
g
r
o
u
p
c
o
n
s
is
ts
o
f
eig
h
t
co
m
p
o
n
en
ts
,
n
am
ely
α
-
,
β
-
,
γ
-
,
an
d
δ
-
to
c
o
p
h
e
r
o
l
(
T
)
;
α
-
,
β
-
,
γ
-
,
an
d
δ
-
to
co
tr
ie
n
o
l
[
5
]
.
Vitam
in
E
is
th
e
m
o
s
t
p
o
wer
f
u
l
lip
id
-
s
o
lu
b
le
an
tio
x
id
an
t
f
o
u
n
d
in
th
e
p
lasma
o
f
h
u
m
an
s
.
T
h
e
d
ietar
y
co
m
p
o
n
e
n
ts
th
at
p
o
s
s
ess
an
tio
x
id
an
t
ac
tiv
ity
s
im
ilar
to
v
i
tam
in
E
ar
e
α
-
,
β
-
,
γ
-
,
an
d
δ
-
to
co
p
h
er
o
ls
,
as
well
as
α
-
,
β
-
,
γ
-
,
an
d
δ
-
to
co
tr
ien
o
ls
[
6
]
.
T
h
ese
m
o
lecu
les
co
n
s
is
t
o
f
a
ch
r
o
m
an
o
l
r
in
g
c
o
n
n
ec
te
d
to
a
s
atu
r
ate
d
p
h
y
ty
l
(
to
co
p
h
er
o
ls
)
o
r
u
n
s
atu
r
ated
(
to
co
tr
ien
o
ls
)
g
r
o
u
p
,
a
n
d
th
e
y
d
if
f
er
in
th
e
n
u
m
b
er
o
f
m
eth
y
l
g
r
o
u
p
s
o
n
th
e
ch
r
o
m
an
o
l
r
in
g
[
7
]
.
Var
i
o
u
s
tech
n
iq
u
es
h
av
e
b
ee
n
u
s
ed
s
o
f
ar
to
e
x
tr
ac
t,
s
ep
ar
ate,
an
d
p
u
r
i
f
y
v
itam
in
E
,
esp
ec
ially
f
r
o
m
v
e
g
etab
le
f
atty
ac
id
d
is
till
ate
(
FAD)
,
wh
ich
is
a
l
u
cr
ativ
e
b
y
p
r
o
d
u
ct
o
f
th
e
o
il r
ef
in
in
g
p
r
o
ce
s
s
[
8
]
.
T
h
is
is
d
u
e
t
o
th
e
r
ec
o
g
n
ized
h
ea
lth
b
e
n
ef
its
o
f
v
itam
in
E
.
T
h
e
v
itam
in
E
co
n
ten
t
in
s
o
y
b
ea
n
o
il
FAD
is
th
e
g
r
ea
test
,
r
an
g
in
g
f
r
o
m
1
0
-
1
4
%.
Ma
ize
o
il
f
o
llo
ws
with
a
v
alu
e
o
f
7
-
1
0
%,
wh
ile
co
tto
n
s
ee
d
o
il
a
n
d
s
u
n
f
lo
wer
o
il
h
a
v
e
co
n
te
n
ts
o
f
6
-
1
0
an
d
5
-
8
%
r
esp
ec
tiv
el
y
[
9
]
.
T
h
e
cu
r
r
en
t
m
eth
o
d
s
cr
ea
ted
to
p
r
etr
ea
t a
n
d
ex
tr
ac
t
v
itam
in
E
in
o
r
d
e
r
to
en
h
a
n
ce
its
ex
tr
ac
tio
n
y
ield
in
clu
d
e
m
o
lecu
lar
d
is
till
atio
n
,
ester
if
icatio
n
,
s
ap
o
n
if
icatio
n
,
ch
r
o
m
ato
g
r
ap
h
y
,
cr
y
s
tallizatio
n
,
s
o
lv
e
n
t
ex
tr
ac
tio
n
,
m
ac
er
atio
n
,
m
atr
ix
s
o
lid
-
p
h
ase
d
is
p
er
s
io
n
,
p
r
ess
u
r
ized
liq
u
id
ex
tr
ac
tio
n
,
s
u
p
er
cr
itic
al
f
lu
id
ex
tr
ac
tio
n
(
SFE
)
an
d
u
ltra
s
o
n
ic
-
ass
is
te
d
ex
tr
ac
tio
n
,
a
n
d
co
m
b
i
n
atio
n
s
th
er
eo
f
[
1
0
]
.
E
ac
h
a
p
p
r
o
ac
h
h
as
ad
v
an
ta
g
es
an
d
d
is
ad
v
a
n
tag
es,
th
er
ef
o
r
e
th
e
d
ec
is
io
n
is
d
eter
m
in
ed
b
y
th
e
p
h
y
s
ical
an
d
ch
em
ical
f
ea
tu
r
e
s
o
f
th
e
s
am
p
le,
as
we
ll
as
th
e
av
ailab
le
r
eso
u
r
ce
s
an
d
eq
u
ip
m
en
t
[
1
1
]
.
Ne
v
er
th
eless
,
th
e
ex
tr
ac
tio
n
tec
h
n
iq
u
es
n
o
w
u
s
ed
in
in
d
u
s
tr
y
,
s
u
c
h
as
ester
if
icatio
n
f
o
llo
wed
b
y
d
is
till
atio
n
,
n
ec
e
s
s
itate
th
e
ap
p
licatio
n
o
f
elev
ated
tem
p
er
atu
r
es
[
1
2
]
.
Un
f
o
r
tu
n
ately
,
th
ese
h
ig
h
tem
p
er
atu
r
es
ca
n
lead
to
th
e
d
eter
io
r
atio
n
o
f
th
er
m
o
s
en
s
itiv
e
p
h
y
to
n
u
tr
ien
ts
,
in
clu
d
in
g
v
itam
in
E
[
1
2
]
.
Vitam
in
E
ac
ts
as
an
an
tio
x
id
an
t
s
p
ec
if
ically
tar
g
etin
g
ce
ll
m
em
b
r
an
e
lip
id
s
,
cir
c
u
latin
g
lo
w
-
d
en
s
ity
lip
o
p
r
o
tein
(
L
DL
)
,
lu
n
g
s
,
liv
e
r
,
an
d
ad
r
en
al
tis
s
u
e.
Vitam
in
E
s
e
r
v
es
a
s
an
an
tio
x
id
an
t,
p
r
im
ar
ily
p
r
o
tectin
g
ag
ain
s
t
h
ar
m
f
u
l
o
x
y
g
en
,
lip
id
p
er
o
x
id
es,
an
d
f
r
ee
r
ad
icals
[
1
3
]
.
I
t
h
alts
th
e
p
r
o
g
r
ess
io
n
o
f
f
r
ee
r
ad
icals
an
d
s
af
eg
u
ar
d
s
ag
ain
s
t
d
eo
x
y
r
ib
o
n
u
cleic
ac
id
(
DNA
)
d
am
ag
e
th
at
lead
s
to
m
u
tatio
n
s
.
Ad
d
itio
n
ally
,
it
h
elp
s
m
an
ag
e
L
DL
lev
els.
T
h
e
co
n
s
eq
u
en
ce
s
o
f
f
r
ee
r
ad
ic
al
ass
au
lt
in
clu
d
e
ac
ce
ler
at
ed
ag
in
g
,
ca
n
ce
r
,
ath
er
o
s
cler
o
s
is
,
an
d
o
th
er
r
ela
ted
co
n
d
itio
n
s
[
1
4
]
.
I
n
co
r
p
o
r
a
tin
g
a
h
i
g
h
er
in
tak
e
o
f
t
o
c
o
p
h
er
o
l
-
r
ich
m
ea
ls
ca
n
y
ield
ad
v
an
tag
es
s
u
ch
as
th
e
tr
ea
tm
en
t
a
n
d
p
r
e
v
en
tio
n
o
f
ca
r
d
io
v
ascu
lar
ailm
e
n
ts
,
in
clu
d
in
g
a
n
g
in
a
,
h
y
p
er
ten
s
io
n
,
an
d
ath
er
o
s
cler
o
s
is
[
1
5
]
.
Natu
r
al
f
o
r
m
s
o
f
to
c
o
p
h
er
o
l a
r
e
b
etter
ab
s
o
r
b
ed
b
y
t
h
e
b
o
d
y
co
m
p
ar
ed
to
s
y
n
th
etic
v
er
s
io
n
s
,
h
ig
h
li
g
h
tin
g
th
e
n
ee
d
to
in
v
esti
g
ate
d
ietar
y
s
o
u
r
ce
s
,
am
o
u
n
t
s
,
an
d
s
tab
il
ity
o
f
to
co
p
h
er
o
ls
to
b
etter
u
n
d
er
s
ta
n
d
th
eir
p
o
ten
tial h
ea
lth
b
en
ef
i
ts
an
d
u
s
es [
2
]
.
T
h
e
p
r
im
a
r
y
b
io
ch
e
m
ical
r
o
l
e
o
f
t
o
co
p
h
er
o
ls
is
to
s
af
e
g
u
ar
d
p
o
ly
u
n
s
atu
r
ated
f
atty
a
cid
s
f
r
o
m
p
er
o
x
id
atio
n
[
1
6
]
.
T
h
e
e
f
f
ec
ti
v
en
ess
o
f
to
c
o
p
h
er
o
ls
m
ay
b
e
in
f
lu
en
ce
d
b
y
th
e
r
atio
o
f
t
o
tal
to
co
p
h
er
o
ls
to
p
o
ly
u
n
s
atu
r
ated
f
atty
ac
id
s
.
T
o
co
p
h
e
r
o
ls
ar
e
cr
u
cial
in
in
h
i
b
itin
g
th
e
o
x
id
atio
n
o
f
lip
id
s
in
alm
o
n
d
s
,
wh
ich
ca
n
ex
ten
d
th
e
s
to
r
a
g
e
tim
e
o
f
th
e
k
er
n
els.
I
n
r
ec
en
t
tim
es,
t
h
e
b
ea
u
t
y
in
d
u
s
tr
y
an
d
clin
ica
l
d
er
m
ato
lo
g
y
h
av
e
s
h
o
wn
in
cr
ea
s
ed
in
te
r
est
in
to
co
ls
b
ec
au
s
e
o
f
t
h
eir
p
h
o
to
p
r
o
tectio
n
an
d
an
tio
x
i
d
an
t
ch
a
r
ac
ter
is
tics
[
1
7
]
.
An
an
tio
x
id
an
t
is
class
if
ied
as
h
av
in
g
v
er
y
s
tr
o
n
g
ac
tiv
ity
wh
en
its
I
C
5
0
v
alu
e
is
less
th
an
5
0
µg
/m
L
(
I
C
5
0
<5
0
µg
/m
L
)
,
s
tr
o
n
g
ac
tiv
ity
wh
en
th
e
I
C
5
0
v
alu
e
is
b
etwe
en
5
0
an
d
1
0
0
µg
/m
L
(
5
0
µg
/m
L
<I
C
5
0
<1
0
0
µg
/m
L
)
,
m
o
d
er
ate
ac
tiv
ity
wh
en
th
e
I
C
5
0
v
alu
e
is
b
etwe
en
1
0
0
an
d
1
5
0
µg
/m
L
(
1
0
0
µg
/m
L
<
I
C
5
0
<1
5
0
µg
/m
L
)
,
wea
k
ac
tiv
ity
wh
en
th
e
I
C
5
0
v
alu
e
is
b
et
wee
n
1
5
0
a
n
d
2
0
0
µ
g
/m
L
(
1
5
0
µg
/m
L
<I
C
5
0
<2
0
0
µg
/m
L
)
,
an
d
v
er
y
wea
k
ac
tiv
ity
wh
e
n
t
h
e
I
C
5
0
v
alu
e
is
g
r
ea
ter
th
a
n
2
0
0
µg
/m
L
(
I
C
5
0
>2
0
0
µ
g
/m
L
)
[
1
8
]
.
Hea
lth
y
s
k
in
is
ch
ar
ac
ter
ized
b
y
its
ab
ilit
y
to
s
h
ield
its
elf
f
r
o
m
t
h
e
h
ar
m
f
u
l
ef
f
ec
ts
o
f
s
u
n
ex
p
o
s
u
r
e
an
d
p
r
ev
en
t
p
r
e
m
atu
r
e
ag
in
g
[
1
9
]
.
Me
lan
in
h
as
a
cr
u
cial
f
u
n
ctio
n
in
s
af
eg
u
ar
d
i
n
g
ag
ain
s
t
u
ltra
v
io
let
(
UV)
r
ad
iatio
n
[
2
0
]
.
Nev
er
t
h
eless
,
an
o
v
er
a
b
u
n
d
an
ce
o
f
m
ela
n
in
s
y
n
th
esis
ca
n
r
esu
lt in
eith
er
h
y
p
er
p
ig
m
e
n
tatio
n
o
r
h
y
p
o
p
ig
m
en
tatio
n
o
f
th
e
s
k
i
n
.
T
h
e
s
u
p
p
r
ess
io
n
o
f
ty
r
o
s
in
ase
is
th
e
m
o
s
t
co
m
m
o
n
ly
em
p
l
o
y
ed
tech
n
iq
u
e
f
o
r
s
k
in
d
ep
ig
m
en
tatio
n
[
2
1
]
.
T
y
r
o
s
in
ase
is
th
e
p
iv
o
tal
en
zy
m
e
in
th
e
m
elan
in
p
r
o
d
u
c
tio
n
p
r
o
ce
s
s
[
2
2
]
.
T
o
co
p
h
e
r
o
l
f
r
o
m
co
r
n
o
il
a
n
d
o
th
er
p
lan
t
o
ils
h
as
b
ee
n
wid
ely
s
tu
d
ied
[
2
3
]
–
[
2
5
]
,
b
u
t
n
o
o
n
e
h
as
y
et
co
n
d
u
cte
d
r
esear
ch
o
n
to
c
o
p
h
er
o
l
co
r
n
o
il
f
r
o
m
Do
m
p
u
.
T
h
e
p
u
r
p
o
s
e
o
f
th
is
s
tu
d
y
was
th
e
id
en
tific
atio
n
o
f
to
co
p
h
er
o
ls
f
r
o
m
Do
m
p
u
co
r
n
o
il
b
y
UV
s
p
ec
tr
o
p
h
o
t
o
m
et
r
y
,
n
u
clea
r
m
a
g
n
etic
r
eso
n
an
ce
(H
-
NM
R
)
,
h
ig
h
-
p
er
f
o
r
m
an
ce
liq
u
i
d
ch
r
o
m
ato
g
r
ap
h
y
(
HPLC)
,
an
d
li
q
u
id
c
h
r
o
m
ato
g
r
ap
h
y
-
tan
d
em
m
ass
s
p
ec
tr
o
m
etr
y
(LC
-
MS)
an
aly
s
is
.
An
ti
-
ag
in
g
test
in
g
o
f
th
e
to
co
p
h
er
o
l
co
n
te
n
t
o
f
co
r
n
o
il
u
s
in
g
th
e
en
zy
m
e
ty
r
o
s
in
ase
was
a
s
ec
o
n
d
ar
y
o
b
jectiv
e
o
f
th
is
s
tu
d
y
.
Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Ad
v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
I
d
en
tifi
ca
tio
n
a
n
d
a
n
tio
xid
a
n
t
a
ctivity
test
o
f β
-
to
co
p
h
ero
l fro
m
Do
mp
u
co
r
n
o
il a
s
a
n
ti
-
a
g
in
g
(
A
d
i P
erma
d
i
)
177
2.
RE
S
E
ARCH
M
E
T
H
O
D
2
.
1
.
T
im
e
a
nd
pla
ce
o
f
re
s
ea
rc
h
T
h
is
r
esear
ch
was
co
n
d
u
cted
d
u
r
in
g
th
e
p
er
io
d
o
f
A
u
g
u
s
t
to
Dec
em
b
er
2
0
2
3
.
R
esear
ch
ac
tiv
ities
wer
e
ca
r
r
ied
o
u
t
in
two
m
a
in
lo
ca
tio
n
s
,
n
am
ely
th
e
De
p
ar
tm
en
t
o
f
C
h
em
ical
E
n
g
in
ee
r
in
g
,
Facu
lty
o
f
I
n
d
u
s
tr
ial
T
ec
h
n
o
lo
g
y
,
Un
iv
er
s
i
tas
Ah
m
ad
Dah
lan
,
Yo
g
y
ak
ar
ta,
an
d
E
B
M
SciTe
ch
,
I
n
s
titu
t
T
ek
n
o
lo
g
i
B
an
d
u
n
g
(
I
T
B
)
.
T
h
e
s
elec
tio
n
o
f
th
ese
two
lo
ca
tio
n
s
was
b
ased
o
n
th
e
av
ailab
ilit
y
o
f
ad
e
q
u
ate
f
ac
ilit
ies
an
d
th
e
s
u
p
p
o
r
t
o
f
a
p
p
r
o
p
r
iate
ex
p
er
ts
to
s
u
p
p
o
r
t th
e
s
u
cc
ess
o
f
th
e
r
esear
ch
.
2
.
2
.
Resea
rc
h t
o
o
ls
a
nd
m
a
t
er
ia
ls
T
h
is
s
tu
d
y
u
tili
ze
d
co
r
n
g
r
ain
s
f
r
o
m
Do
m
p
u
,
Su
m
b
awa
,
W
est
Nu
s
a
T
en
g
g
ar
a
,
I
n
d
o
n
esia.
T
y
r
o
s
in
ase
en
zy
m
e,
k
o
jic
ac
id
,
ch
e
m
icals
,
an
d
an
aly
tical
m
ater
ials
f
o
r
test
in
g
wer
e
p
r
o
v
id
ed
b
y
E
B
M
SciTe
ch
,
R
esear
ch
C
en
ter
f
o
r
B
io
s
cien
ce
an
d
B
i
o
tech
n
o
lo
g
y
I
T
B
,
B
an
d
u
n
g
.
T
h
e
in
s
tr
u
m
en
ts
u
tili
ze
d
in
t
h
is
s
tu
d
y
co
n
s
is
ted
o
f
m
ac
er
atio
n
e
x
tr
ac
tio
n
eq
u
i
p
m
en
t,
UV
-
s
p
ec
tr
o
p
h
o
to
m
eter
,
r
ad
ial
c
h
r
o
m
at
o
g
r
a
p
h
y
(
E
R
SC
H
-
I
T
B
)
,
H
-
NM
R
(
NM
R
B
r
u
k
er
Neo
5
0
0
MH
z
m
ad
e
in
Ger
m
an
y
)
,
AC
QUI
T
Y
UPLC
H
-
clas
s
s
y
s
tem
with
PDA
d
etec
to
r
an
d
X
ev
o
-
T
QD
MS
(
W
ater
s
,
US
A)
,
an
d
L
C
-
MS.
All
o
f
th
e
an
aly
s
is
in
s
tr
u
m
en
t
s
wer
e
f
ac
ilit
ated
b
y
E
B
M
SciTe
ch
I
T
B
.
2
.
3
.
Resea
rc
h pro
ce
du
re
2
.
3
.
1
.
O
il e
x
t
r
a
ct
io
n f
ro
m
c
o
rn
g
ra
ins
1
k
ilo
g
r
am
o
f
d
e
h
y
d
r
ate
d
m
aize
k
er
n
els
s
o
u
r
ce
d
f
r
o
m
D
o
m
p
u
,
Su
m
b
awa
,
W
est
Nu
s
a
T
en
g
g
ar
a,
I
n
d
o
n
esia
was
m
ix
ed
to
g
eth
e
r
an
d
th
e
r
ea
f
ter
s
u
b
jecte
d
to
m
ac
er
atio
n
u
s
in
g
7
0
%
eth
an
o
l
f
o
r
a
d
u
r
atio
n
o
f
2
4
h
o
u
r
s
.
Su
b
s
eq
u
e
n
tly
,
th
e
p
r
o
ce
s
s
o
f
f
iltra
tio
n
was
ca
r
r
ie
d
o
u
t
u
s
in
g
f
ilter
p
ap
er
t
o
ac
q
u
ir
e
a
f
iltra
te.
T
h
e
s
o
lv
en
t
in
th
e
f
ilter
was
ev
ap
o
r
ated
u
s
in
g
an
e
v
ap
o
r
at
o
r
in
o
r
d
er
to
g
et
a
co
n
ce
n
tr
ated
e
x
tr
ac
t.
Su
b
s
eq
u
en
tly
,
th
e
p
r
o
ce
s
s
o
f
ev
ap
o
r
atio
n
w
as
p
r
o
lo
n
g
e
d
b
y
em
p
l
o
y
in
g
an
o
v
en
,
r
esu
ltin
g
in
th
e
ac
q
u
i
s
itio
n
o
f
a
30
-
g
r
am
q
u
an
tity
o
f
ex
tr
ac
t w
ith
a
y
ello
w
o
il
-
lik
e
ap
p
ea
r
an
ce
.
2
.
3
.
2
.
Co
nd
uct
ing
a
s
pect
ro
s
co
pic a
na
ly
s
is
T
h
e
an
aly
s
is
tech
n
iq
u
es
u
tili
ze
d
in
th
e
s
tu
d
y
en
c
o
m
p
ass
UV
s
p
ec
tr
o
p
h
o
to
m
et
r
y
,
r
ad
ial
ch
r
o
m
ato
g
r
ap
h
y
,
H
-
NM
R
,
an
d
ad
v
an
ce
d
s
y
s
tem
s
s
u
ch
as
UPLC
H
-
C
las
s
wi
th
d
etec
to
r
,
X
ev
o
-
T
QD
MS,
an
d
LC
-
MS.
T
h
ese
m
eth
o
d
s
p
r
o
v
id
e
co
m
p
r
e
h
en
s
iv
e
d
ata
o
n
m
o
lecu
lar
co
m
p
o
s
itio
n
an
d
s
tr
u
ctu
r
al
elu
cid
atio
n
,
en
h
an
cin
g
th
e
ac
c
u
r
ac
y
o
f
a
n
aly
tical
o
u
tco
m
es.
T
h
e
ap
p
r
o
a
ch
alig
n
s
with
th
e
tech
n
iq
u
es
d
escr
ib
ed
b
y
Sain
i
an
d
Keu
m
[
1
1
]
,
d
em
o
n
s
tr
atin
g
th
eir
r
eliab
ilit
y
an
d
r
elev
a
n
c
e
in
m
o
d
er
n
r
esear
ch
.
E
ac
h
te
ch
n
iq
u
e
co
n
tr
i
b
u
tes
u
n
iq
u
ely
to
i
d
en
tify
in
g
a
n
d
q
u
an
tify
in
g
co
m
p
o
u
n
d
s
,
e
n
s
u
r
i
n
g
r
o
b
u
s
t
a
n
aly
tical
v
alid
atio
n
.
T
h
is
c
o
m
b
in
atio
n
o
f
tech
n
iq
u
es r
ep
r
esen
ts
a
g
o
l
d
s
tan
d
ar
d
f
o
r
s
tu
d
ies r
e
q
u
ir
in
g
p
r
ec
is
e
ch
em
ical
an
d
m
o
lec
u
lar
an
aly
s
is
.
2
.
3
.
3
.
T
y
ro
s
ina
s
e
enzy
m
e
inh
ibi
t
io
n t
est
pro
ce
du
re
R
ea
g
en
ts
u
s
ed
in
clu
d
e
i
)
Ph
o
s
p
h
ate
B
u
f
f
e
r
0
.
1
M
p
H
6
.
8
c
o
n
s
is
tin
g
o
f
0
.
0
5
1
0
4
M
NaH
2
PO4
.
H2
O
(
3
5
2
.
2
mg
)
an
d
0
.
0
4
8
9
6
Na
2
HPO4
(
3
4
7
.
5
m
g
)
in
5
0
m
l
o
f
d
is
till
ed
wate
r
;
ii
)
1
2
.
5
m
M
L
-
DOPA
s
o
lu
tio
n
(
s
u
b
s
tr
ate)
,
a
to
tal
o
f
2
.
4
6
4
m
g
L
-
DOPA
p
o
wd
er
was
d
is
s
o
lv
ed
in
1
m
l
p
h
o
s
p
h
ate
b
u
f
f
e
r
;
an
d
iii
)
5
0
0
U/m
l
ty
r
o
s
in
ase
en
zy
m
e,
2
0
µ
L
o
f
2
5
,
0
0
0
U/m
l
ty
r
o
s
in
ase
e
n
zy
m
e
s
to
ck
was
ta
k
en
a
n
d
9
8
0
µL
p
h
o
s
p
h
ate
b
u
f
f
e
r
was a
d
d
ed
.
T
h
e
wo
r
k
in
g
m
et
h
o
d
u
s
ed
wa
s
a
s
to
ck
s
o
lu
tio
n
o
f
th
e
s
am
p
le
o
r
co
m
p
ar
ato
r
(
k
o
jic
ac
id
)
p
r
ep
ar
e
d
at
a
co
n
ce
n
tr
atio
n
o
f
1
0
,
0
0
0
p
p
m
in
5
%
DM
SO
.
T
h
e
test
s
o
lu
tio
n
f
o
r
t
h
e
co
m
p
ar
ato
r
was
m
ad
e
f
r
o
m
d
ilu
tio
n
o
f
th
e
s
to
ck
s
o
lu
tio
n
with
a
co
n
ce
n
tr
atio
n
o
f
5
-
5
0
p
p
m
(
µg
/
m
L
)
wh
ile
th
e
test
s
o
lu
tio
n
f
o
r
b
eta
to
co
p
h
er
o
l
was
m
ad
e
in
th
e
r
a
n
g
e
o
f
1
0
-
1
6
0
p
p
m
a
n
d
2
0
0
-
7
0
0
p
p
m
f
o
r
r
o
y
al
jelly
ac
id
.
A
to
tal
o
f
4
0
µ
L
p
h
o
s
p
h
ate
b
u
f
f
er
,
4
0
µL
s
am
p
le
s
o
lu
tio
n
o
r
co
m
p
ar
ato
r
,
an
d
4
0
µL
ty
r
o
s
in
ase
en
zy
m
e
wer
e
p
u
t
in
to
9
6
wells
an
d
th
en
in
cu
b
ated
f
o
r
1
5
m
in
u
tes
at
3
7
°
C
.
A
to
tal
o
f
4
0
µL
o
f
L
-
DOPA
s
u
b
s
tr
ate
was
ad
d
ed
to
th
e
well
an
d
th
e
m
ix
tu
r
e
was
in
cu
b
ated
f
o
r
1
5
m
in
u
tes
at
r
o
o
m
tem
p
e
r
atu
r
e
a
b
s
o
r
b
an
ce
was
r
ea
d
u
s
in
g
a
s
p
ec
tr
o
p
h
o
t
o
m
eter
a
t
a
wav
elen
g
th
o
f
4
7
5
n
m
.
T
y
r
o
s
in
ase
in
h
ib
iti
o
n
ca
lcu
lated
by
(
1
)
.
Tyr
osin
a
s
e
in
hib
i
tion
(
%
)
=
(
(
−
)
−
(
−
)
−
)
×
100
(
1
)
W
h
er
e
Ab
s
A
is
a
b
s
o
r
b
an
ce
b
lan
k
(
p
h
o
s
p
h
ate
b
u
f
f
er
,
en
zy
m
e,
s
u
b
s
tr
ate
,
an
d
n
o
s
am
p
le)
,
Ab
s
B
is
th
e
a
b
s
o
r
b
an
ce
o
f
b
lan
k
(
p
h
o
s
p
h
ate
b
u
f
f
er
,
s
u
b
s
tr
ate,
with
o
u
t
s
am
p
le
an
d
en
zy
m
e
)
,
Ab
s
C
is
th
e
a
b
s
o
r
b
an
ce
o
f
s
am
p
le,
en
zy
m
e
,
an
d
s
u
b
s
tr
at
e
,
Ab
s
D
is
th
e
a
b
s
o
r
b
an
ce
o
f
s
am
p
le
an
d
s
u
b
s
tr
ate
with
o
u
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Evaluation Warning : The document was created with Spire.PDF for Python.
I
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N
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2
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3.
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r
ain
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r
e
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r
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a)
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d
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r
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1
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x
tr
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o
m
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o
r
n
g
r
ain
s
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a)
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r
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s
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b
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d
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co
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x
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r
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t
Fig
u
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e
s
2
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s
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ate
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e
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tific
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r
e
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ar
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s
s
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ic
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h
n
iq
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h
ese
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d
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a
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s
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tr
o
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h
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to
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y
p
r
o
f
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th
at
s
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t
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k
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at
m
a
x
im
u
m
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g
th
s
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in
d
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o
f
co
n
ju
g
ated
ch
r
o
m
o
p
h
o
r
e
b
o
n
d
s
(
Fig
u
r
e
2
).
Ad
d
itio
n
all
y
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th
e
UPLC
p
r
o
f
ile
r
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ea
l
s
a
ch
r
o
m
at
o
g
r
a
m
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o
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atin
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m
p
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n
d
p
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r
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o
f
less
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an
9
7
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Fig
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r
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3
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e
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R
p
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r
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ir
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'
s
d
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ct
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h
if
ts
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r
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4
)
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th
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m
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s
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g
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s
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tes
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m
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h
t
o
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e
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h
er
o
l
co
m
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n
d
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r
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5
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h
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ity
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ter
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lates
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ig
u
r
e
2
.
S
p
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tr
o
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h
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to
m
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p
r
o
f
ile
Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Ad
v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
I
d
en
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Fig
u
r
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4
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p
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is
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late
Evaluation Warning : The document was created with Spire.PDF for Python.
I
SS
N
:
2
2
5
2
-
8
8
1
4
I
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t J Ad
v
Ap
p
l Sci
,
Vo
l.
1
4
,
No
.
1
,
Ma
r
c
h
2
0
2
5
:
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g
s
p
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tr
o
p
h
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UV,
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NM
R
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an
d
LC
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MS,
wer
e
u
s
ed
to
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eter
m
i
n
e
th
e
ch
em
ical
s
tr
u
ctu
r
e.
T
h
e
s
p
ec
if
ic
d
ata
r
esu
lts
c
an
b
e
s
ee
n
in
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u
r
e
s
2
-
5
.
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n
o
r
d
er
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eter
m
in
e
th
e
r
ete
n
tio
n
d
u
r
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n
o
f
to
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p
h
er
o
ls
in
ch
r
o
m
ato
g
r
a
p
h
y
,
p
u
r
e
o
ils
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e
em
p
lo
y
e
d
.
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h
e
k
in
d
an
d
p
r
o
p
o
r
tio
n
o
f
to
co
p
h
er
o
ls
wer
e
d
eter
m
in
ed
b
y
a
n
aly
zin
g
th
e
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tio
n
d
u
r
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d
q
u
a
n
titi
es
o
f
th
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co
m
p
o
u
n
d
s
i
n
th
e
p
u
r
e
o
il
s
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les.
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ased
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n
th
e
UV
s
p
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tr
u
m
s
h
o
wed
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ea
k
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m
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x
im
u
m
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elen
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th
λ
=2
2
6
.
8
8
a
n
d
2
9
6
.
8
8
n
m
wh
ic
h
m
ea
n
s
th
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m
p
o
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n
d
h
as
a
co
n
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g
ated
ch
r
o
m
o
p
h
o
r
e
b
o
n
d
with
p
u
r
ity
<9
7
%
wh
en
v
iewe
d
o
n
UPLC
ch
r
o
m
ato
g
r
am
.
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o
co
n
f
ir
m
th
e
s
tr
u
ctu
r
al
f
o
r
m
o
f
th
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m
p
o
u
n
d
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1
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_
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r
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k
er
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z
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is
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r
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t
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o
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d
1
2
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ig
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p
s
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7
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r
b
o
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n
u
m
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er
s
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h
er
e
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n
e
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ig
n
al
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7
)
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th
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m
atic
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p
2
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s
h
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t
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6
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3
3
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s
,
1
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ig
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als
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o
r
m
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r
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r
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at
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H
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8
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m
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2
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h
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m
p
o
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as
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eth
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(
C
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at
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1
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9
3
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d
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6
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8
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n
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9
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1
2
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5
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3
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2
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H
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0
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6
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H
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2
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H
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2
7
)
.
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h
e
la
tter
is
th
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s
p
3
s
ig
n
al
f
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m
1
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4
5
-
0
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9
6
p
p
m
wh
ich
s
tr
etch
es
to
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o
r
m
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H2
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C
H
alip
h
atic.
B
ased
o
n
NM
R
d
ata
s
h
o
wed
th
at
th
is
co
m
p
o
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n
d
is
b
eta
-
to
co
p
h
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l
wh
ich
was
co
n
f
ir
m
ed
b
y
m
o
l
ec
u
lar
weig
h
t
an
aly
s
is
wh
ich
is
[
M+
H]
+
m
/z
4
1
7
.
T
h
e
ch
e
m
ical
s
tr
u
ctu
r
e
o
f
β
-
to
co
p
h
er
o
l c
an
b
e
s
ee
n
in
F
i
g
u
r
e
6
.
Fig
u
r
e
6
. β
-
to
c
o
p
h
e
r
o
l c
h
em
ic
al
s
tr
u
ctu
r
e
Evaluation Warning : The document was created with Spire.PDF for Python.
I
n
t J Ad
v
Ap
p
l Sci
I
SS
N:
2252
-
8
8
1
4
I
d
en
tifi
ca
tio
n
a
n
d
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n
tio
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ctivity
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s
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n
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a
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(
A
d
i P
erma
d
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)
181
β
-
to
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er
o
l
h
as
r
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ei
v
ed
les
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atten
tio
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ase
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T
h
e
ty
r
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ase
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it
o
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ter
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am
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le
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2
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Fig
u
r
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7
s
h
o
w
s
an
in
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v
it
r
o
test
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s
in
g
a
well
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icr
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etwe
en
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to
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s
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cted
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test
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th
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ty
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e
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p
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m
ai
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il.
Ko
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u
s
ed
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a
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m
p
ar
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is
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tu
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T
h
e
I
C
5
0
test
f
in
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f
o
r
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l
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h
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8
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at
th
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tio
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n
t
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s
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to
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n
t
ca
teg
o
r
y
[
1
8
]
,
[
2
8
]
.
Fig
u
r
e
7
d
ep
icts
an
in
-
v
itro
test
f
o
r
ty
r
o
s
in
ase
en
zy
m
e
i
n
h
ib
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n
ac
tiv
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u
s
in
g
a
well
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icr
o
p
late.
T
h
is
test
co
m
p
ar
es
th
e
in
h
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ito
r
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tiv
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f
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to
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r
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Do
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aize
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ase
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ly
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h
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o
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th
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in
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ited
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aid
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i
to
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m
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Fig
u
r
e
7
(
a
)
d
is
p
lay
s
m
icr
o
p
lates
lab
eled
with
s
am
p
le
co
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ati
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icatin
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lled
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p
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h
e
d
a
r
k
er
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p
ly
h
ig
h
er
am
o
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n
ts
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ela
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n
th
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t
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e
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ter
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icate
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o
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er
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ito
r
y
ef
f
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ts
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ay
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e
f
r
o
m
k
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n
t
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s
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ag
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h
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p
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io
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at
t
h
e
test
ed
ch
em
icals
p
r
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t
m
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in
s
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n
th
esis
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T
h
e
d
if
f
e
r
en
ce
i
n
co
lo
r
in
ten
s
i
ty
aid
s
in
d
eter
m
in
in
g
th
e
ef
f
icac
y
o
f
th
e
test
ed
in
h
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ito
r
s
.
Fig
u
r
e
7
(
b
)
s
h
o
ws
th
e
wells
ap
p
ea
r
clea
n
er
in
s
o
m
e
r
eg
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n
s
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in
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icatin
g
t
h
at
th
e
test
ed
ch
em
icals
p
r
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en
t
m
elan
in
s
y
n
t
h
esis
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T
h
e
d
if
f
e
r
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ce
i
n
co
lo
r
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ten
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ty
aid
s
in
d
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m
in
in
g
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e
ef
f
icac
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o
f
th
e
test
ed
in
h
ib
ito
r
s
.
(
a)
(
b
)
Fig
u
r
e
7
. I
n
-
v
itr
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test
u
s
in
g
a
well
m
icr
o
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late
of
(
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to
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ase
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s
e
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t
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[
2
9
]
–
[
3
1
]
.
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to
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