Download - Carbohydrate
CARB
OHYD
RATE
ANALY
SIS(
Ch. 11
+12)
I. A
naly
sis
of S
ugar
sA
-Qua
litat
ive
Det
ecti
on M
etho
dsB-
Qua
ntit
ativ
e A
naly
ses
II-
Ana
lysi
s of
Non
suga
rs(p
olys
acch
arid
es)
A-
Nut
rien
t Po
lisac
char
ides
(sta
rch,
de
xtri
ns,
glyc
ogen
etc
.) -Q
ualit
ativ
e an
alys
es-Q
uant
itat
ive
anal
yses
B-D
ieta
ryFi
ber
orSt
ruct
ural
Po
lysa
ccha
ride
s (c
ellu
lose
, hem
icel
lulo
seet
c.)
a. G
ravi
met
ric
met
hods
: 1.C
rude
fib
er2.
Det
erge
nt m
etho
ds3.
AO
AC
met
hod
b. C
hem
ical
Met
hods
Com
posi
tion
and
Occ
urre
nce
Carb
ohyd
rate
s ar
e th
e m
ost
abun
dant
and
mos
t wi
dely
con
sum
ed(>
%70
)m
acro
fo
od
com
pone
nt.
They
prov
ide
sati
ety,
an
dm
odif
yhu
man
phys
iolo
gica
lpro
cess
es.T
hey
are
a ve
ry h
eter
ogen
eous
gro
up in
str
uctu
re-
all
cont
ain
carb
on,
hydr
ogen
and
oxyg
en,
and
are
alm
ost
excl
usiv
ely
of
plan
tor
igin
(exc
epti
ons:
lact
ose
of m
ilkan
dgl
ycog
enin
mea
t).
CLA
SSIF
ICA
TIO
N:
I. M
ono
and
Olig
osa
chha
ride
s(S
ugar
s):
Suga
rs h
ave
nutr
itio
nal
and
met
abol
ic f
unct
ions
;the
ygo
vern
rhe
olog
ical
prop
erti
es,
caus
e br
owni
ng
reac
tion
s,
have
ro
le
in
food
st
abili
ty,
ar
e ra
w m
ater
ials
fo
r fe
rmen
tati
ons.
a. M
onos
acha
ride
s: P
olyh
ydro
xyal
dose
san
d ke
tose
s
Pent
oses
: Xyl
ose,
arab
inos
e(C 5
)
Hex
oses
: G
luco
se, f
ruct
ose(
C 6 )
OH
OH
CHH
CO
H
CH
H
HOH
C CH 2O
H
O
C
OH
O OH
OH
HO
CH 2O
H
1
23
456
b. O
ligos
acch
arid
es:
Som
esu
gar u
nits
(>1,
<10)
are
cond
ense
d to
geth
er
form
ing
di-,
tri-,
tetra
-sa
ccha
rides
: S
acha
rose
, lac
tose
, ra
ffino
se, s
tach
yose
O OH
OH
HO
CH
2O
HC
H2O
H
OC
H2O
H
O
HO
OH
H1
23
4
5 6
II. Non
suga
rs: P
olys
achh
arid
es
a.
Nut
rien
t po
lysa
char
ides
: Th
ey
are
met
abol
ic
rese
rves
of
plan
ts a
nd a
nim
als
,and
are
bas
ical
ly a
ll po
lym
ers
of g
luco
se (d
extr
ins,
sta
rch,
gly
coge
n)
b. S
truc
tura
l po
lysa
char
ides
: Th
ey c
onst
itut
e th
e ri
gid
mec
hani
cal
cell
wall
stru
ctur
e in
pl
ants
. Ce
llulo
se( p
olym
er o
f b-
D-G
luco
se b
(1, 4
) lin
kage
.
Repe
atin
g ce
llobi
ose
moi
ety)
O OH
OHCH
2O
H
OH
OO
O
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
n
O OH
OHCH
2O
H
OH
OO
O
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
O OH
OHCH
2O
H
Pect
in(g
alac
turo
nic
acid
and
met
hylg
alac
turo
nate
poly
mer
)
Hom
oglic
ans
are
poly
mer
s of
sam
e su
gar
(i.e.
xyla
n:xy
lose
poly
mer
) H
eter
oglic
ans
are
poly
mer
s of
mor
e th
an o
ne s
ugar
gro
up(i.
e.ar
abin
ogal
acta
ns)
O OH
OH
OH
OO
O
O OH
OH
O OH
OH
O OH
OH
O OH
OH
O OH
OH
CO
OH
CO
OC
H3
CO
OC
H3
CO
OH
FUN
CTI
ON
AL
PRO
PER
TIES
OF
CH
O’s
-Sto
rage
mat
eria
lfor
carb
onan
den
ergy
in
live
syst
ems
-Com
pone
ntof
Stru
ctur
e:
1.V
isco
sity
con
trol
2.Te
xtur
e co
ntro
l
3.Em
ulsi
fyin
g ag
ent
4.St
abili
zer d
ueto
wat
erbi
ndin
gca
paci
ty
Carb
ohyd
rate
con
tent
s of
som
e fo
ods
Hon
ey: 7
5-82
% f
ruct
ose
+ gl
ucos
e +
sach
aros
e
App
les:
15%
(fru
ctos
e+gl
ucos
e+sa
ccha
rose
+st
arch
)
Dri
ed f
igs:
30%
fruc
tose
+ 4
2%gl
ucos
e
Dat
es: 5
0% s
acch
aros
e
Seed
less
rai
sins
: 70%
fruc
tose
+glu
cose
; Gra
pes:
16-
17%
Mel
ons,
pea
ches
, tan
geri
nes
: <10
% s
acch
aros
e
Carr
ots:
10%
CHO
Ches
tnut
s: 3
3% s
tarc
h
Whi
te f
lour
: 70
% s
tarc
h , W
hite
Brea
d:50
% C
HO
Corn
-pat
atoe
s: %
15 s
tarc
h; C
orn
Flak
es: 8
6% C
HO
Chic
ken
brea
st: 0
%
Shor
t-cu
t to
CHO Q
uant
ificat
ion:
In g
ener
al(in
mos
t fo
od c
ompo
siti
on t
able
san
dfo
rnu
trit
iona
lla
belli
ng),
it is
ass
umed
tha
t:
Tot
alCH
O(w
ithf
iber
)=10
0-[M
oist
ure%
+Pro
tein%+F
at%+A
sh%]
or
CHO w
itho
ut c
rude
fiber
=N-f
ree
extr
act
In
calc
ulat
ing
the
calo
rifi
c va
lues
of
fo
ods,
su
gars
an
d nu
trie
nt p
olys
acch
arid
es o
f CH
O’s
are
gene
rally
ass
umed
as
havi
ng 4
cal/
g an
d st
ruct
ural
pol
ysac
char
ides
(i.e
.cru
de f
iber
) as
ha
ving
0
cal/
g.
H
owev
er,
nove
l “D
iete
tic
food
in
gred
ient
s”no
w in
clud
e co
mpo
nent
s wh
ich
are
part
ially
m
etab
oliz
ed
and
ther
efor
e do
prov
ide
som
eca
lori
es
(i.e.
olig
osac
char
ides
:1.2
kca
l/g;
sug
ar a
lcoh
ols:
2.4
kcal
/g)
thus
co
ntri
buti
ng t
o a
rath
er n
ew c
once
pt, “
diet
ary
fibr
e”.
The
alte
rnat
ive
mea
ns o
f qu
anti
fica
tion
of c
alor
ies
is b
y us
e of
a B
OM
B Ca
lorimet
er,
whic
hm
easu
res
hea
t pr
oduc
edwh
ensa
mpl
eis
bur
nt.
Tota
lCar
bohy
drat
es
CH
O’s
are
dest
roye
dby
heat
and
acid
. C
ontin
ued
heat
ing
in p
rese
nce
of a
cid
lead
sto
prod
ucts
that
cond
ense
with
phen
olic
com
poun
dsto
prod
uce
colo
red
com
poun
ds,th
enqu
antif
ied
byU
V-V
issp
ectro
phpo
tom
etric
read
ings
of a
bsor
banc
eat
490
nm
.
Ex: P
heno
l-Sul
furic
Aci
dM
etho
d(p.
172-
3)
.A
naly
ses
of S
ugar
s(W
ater
-sol
uble
CHO
’s)SA
MPL
E PR
EPA
RATI
ON
:Sa
mpl
e ha
s to
be
hom
ogen
ized
bef
ore
anal
yses
. -R
emov
al o
f “L
ipid
s an
d Pr
otei
ns"
is r
equi
red
befo
re s
tart
ing
anal
yses
, sin
ce t
hey
caus
e tu
rbid
ity
duri
ng a
naly
ses
whic
h in
terf
eres
wit
h po
lari
zati
on a
nd r
efra
ctio
n:Li
pids
are
extr
acte
d wi
th p
etro
leum
eth
er, b
ut c
are
shou
ld b
e ta
ken
not
to e
xcee
d 50
°C, s
ince
hig
her
tem
pera
ture
s m
ight
so
lubi
lize
star
ch.
Prot
eins
are
prec
ipit
ated
with
hea
vy m
etal
s us
ing
[Pb
acet
ate]
or
[ZnO
Hor
Zn
ferr
ocya
nide
]. Pr
otei
ns a
re c
opre
cipi
tate
din
co
lloid
al f
orm
. Ca
rrez
solu
tion
(1+1
) [%
15 Z
nSO
4 +
%30
K fe
rric
yani
de) i
s an
oth
er a
lter
nati
ve.
-Neu
tral
izin
gan
y ac
idit
y (b
y Ca
CO3)
, sin
ce a
cids
may
cau
se
hydr
olys
is o
f su
gars
. -R
emov
al o
f pi
gmen
ts b
y ad
sorp
tion
an
char
coal
-Ina
ctiv
atin
gen
zym
es [H
gCl 2
inac
tiva
tes
enzy
mes
]
Sam
ple
prep
arat
ion
and
extra
ctio
nsc
hem
e(fi
g.11
.1)
-Dry
sam
ple
-Fro
mde
hydr
ated
mat
eria
l, e
xtra
ctlip
idsw
ith95
:5
Chl
orof
orm
-met
hano
l
-Ext
ract
resi
due
with
80%
etha
nol
-Rem
ove
any
inte
rfer
ance
(i.e.
by
ion-
exch
ange
, Se
ppak
cartr
idge
etc)
Mon
o an
ddi
sacc
harid
esar
eob
tain
ed.
I.a.
Qua
litat
ive
Det
ecti
on M
etho
ds f
or s
ugar
s:1-
Tota
lsug
ars
::Met
hods
bas
ed o
n co
lour
reac
tion
s of
sug
ars.
-Wit
h st
rong
m
iner
al
acid
s (H
2SO
4, H
Cl,
H3P
O4
etc.
) EX
:CH
O+P
heno
l-H
2SO
4ye
llow-
oran
geco
lour
due
to c
onde
nsat
ion
of d
egra
ding
sug
ar p
rodu
cts
: 490
nmac
ids+
hex
oses
HM
F in
dica
tor
acid
s+pe
ntos
esfu
rfur
ol-W
ith
ant
hron
ein
con
c. H
2SO
4,ch
arac
teri
stic
blu
e-gr
een
colo
ur: 6
20nm
2-Spe
cific
Ana
lyse
sof
Ind
ividua
lSug
ars:
M
etho
dsba
sed
on r
educ
ing
prop
erti
es o
f m
onos
acch
arid
esas
wel
l as
red
ucin
g di
sacc
hari
des
like
lact
ose
and
mal
tose
(Cu+
+ is
red
uced
toCu
+):
A-F
EHLI
NG
Reac
tion
( Ben
edic
t’s T
est,
Mun
son
Wal
ker-
Grav
imet
ric;
La
neEy
non-
Volu
met
ric,
Som
ogyi
-Nel
son:
spe
ctro
phot
omet
ric)
:Fe
hlin
gA
:35g
CuS
O4
/500
ml;
Fehl
ing
B:[1
73g
NaK
C 4H
406(
Roch
elle
sal
t)+5
0g N
aOH
]/50
0 m
l
OC
uC
uSO
2su
gar
redu
cing
4
→
+
++→Cu
Cu
B. –
Rea
ctio
n wi
th T
olle
n's
reag
ent:
[]
oA
g]
Ag[
NH
suga
rsre
duci
ngio
n2
3
→
+
Silv
er i
ons
is
redu
ced
to t
o m
etal
lic s
ilver
, wh
ich
is
depo
site
d on
a g
lass
sur
face
, for
min
g a
“mir
ror”
.
C. F
orm
atio
n of
“osa
zone
s”:
Oxi
dati
ve c
leav
age
of h
ydro
xyl
grou
ps o
f su
gars
and
phen
ylhi
draz
ine
fo
rm
beau
tifu
l cr
ysta
lline
ye
llow
com
poun
ds, c
alle
d os
azon
es(“
diph
enyl
hydr
azon
e”)
3. P
aper
Chr
amot
ogra
phy:
I
s by
far
t
he b
est
and
sim
ples
t qu
alit
ativ
em
etho
d to
dis
ting
uish
qua
litat
ivel
y be
twee
n va
riou
s su
gars
in f
oods
.Ex
: Su
gar
solu
tion
is
spot
ted
on F
ilter
pap
er;f
or d
evel
opin
g so
lven
t sy
stem
:
[Phe
nol-H
2O,
80:2
0]
or
[H2O
:but
anol
:ace
tic
acid
, 5:
1:4]
or
man
y ot
her
alte
rnat
ive
solv
ent
syst
ems
can
be
used
. Pe
culia
riti
es o
f di
ffer
ent
suga
rs l
ead
to d
iffe
renc
es i
n M
igra
tion
rat
es, r
esul
ting
in
diff
eren
t bu
t ty
pica
l R f
valu
e fo
r di
ffer
ent
suga
rs in
a g
iven
sol
vent
sys
tem
. Gen
eral
ly:
Hig
her
Rfva
lues
indi
cate
hig
her
mob
ility
, sm
alle
r M
W :
Pent
ose>
hexo
se>d
isac
char
ides
>tri
sach
arid
es>
And
ket
ohex
ose>
aldo
hexo
se
Then
the
chr
omat
ogra
m i
s sp
raye
d w
ith
a vi
sual
isat
ion
spra
y-re
agen
t (E
x. p
-ani
nisi
nein
n-b
utan
ol) o
r ph
loro
gluc
inol
in H
Cl, a
nd
afte
rdr
ying
for
5-10
min
, is
put
in
an o
ven
at
105°
C fo
r vi
sual
isat
ion.
ABC
AB
S
Solv
ent
Rf =
A S
For S
pot A
Thin
Lay
er c
hrom
atog
raph
ySi
lica
gel a
s sta
tiona
ry p
hase
(250
µm
)Q
uant
itativ
e by
den
sito
mer
(tim
e va
ries w
ith so
lven
t, ge
nera
lly
over
nigh
t):
ABC
AB
S
Solv
ent
Rf =
A S
For S
pot A
Gas
and
Liqu
id C
hrom
atog
raph
yGC
and
HPL
C: F
or q
uant
ific
atio
n of
indi
vidu
al s
ugar
com
pone
nts,
HPL
C yi
elds
mos
t sa
tisf
acto
ry r
esul
ts.A
lso,
GC
follo
wing
for
mat
ion
ofvo
lati
le
sily
lder
ivat
ives
of
suga
rs w
ith
TMS
(tri
met
hyls
ilyle
ther
) can
be
used
fo
r qu
anti
tati
ve a
naly
ses
of in
divi
dual
sug
ars.
This
is a
req
uire
men
t, s
ince
com
poun
ds m
ust
be v
olat
ile.
Suga
rs f
orm
vol
atile
Trim
ethy
lsily
lEth
ers:
Ethe
r lin
kage
Trim
ethy
lchl
oros
ilane
SiSi
Cl
CH
3
CH
3
CH
3
O
CH
3
CH
3
CH
3
Liqu
id C
hrom
atog
raph
y
Car
bohy
drat
e A
naly
sis (
Wat
ers A
ssoc
s. H
PLC
840
380
Stat
iona
ry
Phas
e)
Solv
ents
yste
m:
H2O
/CH
3CN
Fruc
tose
Glu
cose
Mal
totri
ose
Mal
tose
•Th
e m
ost c
omm
only
use
d de
tect
or fo
r ca
rboh
ydra
te a
naly
sis
is th
e di
ffere
ntia
l re
fract
ive
inde
x (R
I) de
tect
or. B
ecau
se it
re
spon
ds to
all
anal
ytes
who
se re
fract
ive
inde
x di
ffers
from
that
of t
he m
obile
pha
se,
it fu
nctio
ns a
s a
univ
ersa
l det
ecto
r. It
is
pref
erre
d ov
er t
radi
tiona
l UV
det
ecto
r for
no
n-ch
rom
opho
ricco
mpo
unds
like
ca
rboh
ydra
tes.
En
han
ced
Se
nsit
ivit
y a
nd
Red
uce
d R
un
Tim
es U
sin
g G
rad
ien
ts
42
DE
Co
rn S
yru
p M
alt
ose
Olig
om
ers
•Col
umn:
Pre
vail™
Car
bohy
drat
e E
S,
•25
0 x
4.6m
m•M
obile
Pha
se:
Wat
er:A
ceto
nitri
le(3
8:62
)
•F
low
rate
:1.
0mL/
min
•Col
umn
Tem
p:A
mbi
ent
•Inje
ctio
n Vo
lum
e:5µ
L
Co
lum
n:
Prev
ail™
Car
boh
ydra
te E
S,
250 x
4.6
mm
Mo
bile P
hase
:A
:Ace
tonitrile
B:
Wat
er
Gra
die
nt:
Tim
e:
0 1
5
25
%B
:35 50 50
Flo
wra
te:
1.0
mL/
min
Co
lum
n T
em
p:
Am
bie
nt
Inje
ctio
n V
olu
me:
5µL
1.
Glu
cose
2.
Mal
tose
3.
Mal
totr
iose
4.
Mal
tote
trao
se5
.M
alto
pen
taos
e6
.M
alto
hex
aose
7.
Mal
tohep
taos
e8
.M
alto
octa
ose
9.
Mal
tonon
aose
10
.M
alto
dec
aose
11
.M
alto
undec
aose
12
.M
alto
dod
ecao
se1
3.
Mal
totr
idec
aose
9353
9354
Iso
crati
cG
rad
ien
t
•Col
umn:
Pre
vail™
Car
bohy
drat
e E
S, 2
50 x
4.
6mm
•Mob
ile P
hase
:W
ater
:Ace
toni
trile
(30:
70)
•Flo
wra
te:
1.0m
L/m
in•C
olum
n Te
mp:
Am
bien
t•In
ject
ion
Volu
me:
5µL
Su
gar
Alc
oh
ols
1.
Iso-
eryt
hrito
l2
.Xyl
itol
3.
Man
nitol
4.
Mal
tito
l
9352
•Col
umn:
Pre
vail™
Car
bohy
drat
e E
S,
250
x 4.
6mm
•Mob
ile P
hase
:W
ater
:Ace
toni
trile
(25:
75)
•Flo
wra
te:
1.0m
L/m
in•C
olum
n Te
mp:
Am
bien
t•In
ject
ion
Volu
me:
20µL
1.
Fruct
ose
2.
Glu
cose
3
.Sucr
ose •C
olum
n:P
reva
il™C
arbo
hydr
ate
ES
, 25
0 x
4.6m
m•M
obile
Pha
se:
A: A
ceto
nitri
leB
: Wat
er•G
radi
ent:
Tim
e:0
30
•%
B:
25 5
0•F
low
rate
:1.
0mL/
min
•Col
umn
Tem
p:A
mbi
ent
•Inje
ctio
n Vo
lum
e:20
µL
Ora
ng
e J
uic
eV
eg
eta
ble
Ju
ice
9331
9317
1.
Fruct
ose
2.
Glu
cose
3.
Sucr
ose
Co
lum
n:
Prev
ail™
Car
boh
ydra
te E
S, 250 x
4.6
mm
Mo
bile P
hase
:A
:Ace
tonitrile
B:
Wat
er
Gra
die
nt:
Tim
e:
0 1
5%
B:
25 40
Flo
wra
te:
1.0
mL/
min
Co
lum
n T
em
p:
Am
bie
nt
Inje
ctio
n V
olu
me:
5µL
Dete
cto
r:ELS
D 2
000
Clo
ver
Ho
ney
9365
I.b.
Qua
ntit
ativ
e Su
gar
Ana
lyse
s M
etho
dsTh
e fo
llowi
ng
met
hods
giv
e to
tal
suga
r co
nten
t an
d al
ways
req
uire
cal
ibra
tion
wit
h st
anda
rd s
ugar
sol
utio
ns.
b.1-
Chem
ical M
etho
ds:
Chem
ical
met
hods
are
bas
ed
on
redu
cing
pr
oper
ties
of
m
onos
acch
arid
es.
Free
al
dehy
deor
ket
ogr
oup
redu
ces
alka
line
solu
tion
s of
m
etal
lic s
alts
to
met
al o
xide
s(Cu
O) o
r fr
ee m
etal
(Ag)
.
-Gra
vim
etri
c m
etho
ds(C
u-M
unso
n-W
alke
r):
Wei
ghin
g pr
ecip
itat
ed m
etal
oxi
de-T
itri
met
ric
met
hods
a) L
ane-
Eyno
n:Su
gar
solu
tion
is a
dded
fro
m a
bur
etto
a
vigo
rous
ly
boili
ng
mix
ture
of
Fe
hlin
gA
+B
solu
tion
m
ixtu
re.
Nea
r th
e en
d po
int,
met
hyle
nebl
ue i
ndic
ator
wi
ll ch
ange
fro
m b
lue
to c
olor
less
in e
xces
s of
red
ucin
g su
gars
.b)
Alk
alin
e fe
rric
yani
dem
etho
d:2K
3Fe(
CN) 6
+2KI
2K 4
Fe(C
N) 6
+I2
Libe
rate
d io
dine
is t
itra
ted
with
Na 2
S 20 3
b.2-
Phys
ical
met
hods
-Ins
trum
enta
l met
hods
(p.18
4)
Mic
rosc
opy:
esp
ecia
llyfo
rst
arch
orig
inde
term
inat
ion
anal
ysis
Spec
ific
grav
ity:
hyd
rom
eter
sca
libra
ted
in B
rix
degr
ees
(suc
rose
% b
ywe
ight
)
Spec
trop
hoto
met
ric
met
hods
: Sin
ce in
the
ran
ge (0
-80µ
g )
the
abso
rban
ce v
alue
at
490n
m f
or r
educ
ed c
oppe
r is
in li
near
ra
nge,
spe
ctro
phot
omet
ric
read
ings
can
be
used
for
qu
anti
fica
tion
of
tota
l sug
ar c
onte
nt.
-Ref
ract
omet
ry:
Incr
easi
ng s
ugar
con
cent
rati
on i
ncre
ases
the
an
gle
of r
efra
ctio
n of
a s
olut
ion,
whi
ch i
s al
so a
ffec
ted
by
conc
entr
atio
n te
mpe
ratu
re.
Po
cket
an
d A
bbé
type
s of
re
frac
tom
eter
syi
eld
rati
o of
sol
uble
sol
ids.
Res
ults
are
pre
cise
on
ly f
or p
ure
suga
r so
luti
ons.
-Pol
arim
etry
(see
11.6
.5):
This
is
a
nond
estr
ucti
ve
and
rapi
d m
etho
d de
pend
ing
on t
he f
act
that
sug
ar s
olut
ions
, lik
eal
lco
mpo
unds
cont
aini
ngch
iral
carb
on,
have
the
abi
lity
to r
otat
e a
plan
e of
pol
ariz
ed li
ght
thro
ugh
an a
xis
para
llel t
o it
s di
rect
ion
of
prop
ogat
ion,
an
d th
e sp
ecif
ican
gle
of
rota
tion
be
ing
pr
opor
tion
al w
ith
suga
r m
olec
ular
stru
ctur
ean
dco
ncen
trat
ion.
D
extr
orot
ator
y -p
lane
pol
ariz
ed li
ght r
otat
ed c
lock
wise
(or
to th
e ri
ght)
Levo
rata
tory
-pla
ne p
olar
ized
ligh
t rot
ated
cou
nter
cloc
kwise
.
Inve
rt S
ugar
---w
hen
sucr
ose
has b
een
hydr
olyz
ed in
to g
luco
se a
nd fr
ucto
se,
the
rota
tion
chan
ges f
rom
dex
tror
otat
ory
(+66
.5) t
o le
voro
tato
ry (-
19.8
). S
o,
sucr
ose
solu
tion
is ca
lled
“Inv
ert S
ugar
”.IC
UM
SA-I
nter
nati
onal
Co
mm
issi
on
for
Uni
fied
M
etho
ds
of
Suga
r A
naly
sis
cite
s po
lari
met
ryin
sta
ndar
d m
etho
ds f
or
suga
r ın
dust
ry.
For
sach
aros
e=+6
6.5°
For
inve
rt s
ugar
sol
utio
n(gl
ucos
e+fr
ucto
se)=
-20°
-
b.3.
Enzy
mat
icM
etho
ds:G
luco
se O
xida
seSy
stem
Glu
cose
Oxi
dase
D-G
luco
se
+ O
2G
luco
nic
Aci
d +
H
2O2
Pero
xida
seH
2O2+
0
-Dia
nisi
dine
2 H
2O +
Oxi
dize
d 0-
Dia
nisi
dine
(Col
orle
ss)
(Bro
wn)
OC
H3
H3C
O
H2N
NH
2
H3C
OO
CH
3
HN
NH
II. A
. Ana
lysi
s of
Nut
rien
t Po
lisac
char
ides
STARC
H(1
1.4.
1):M
ost
abun
dant
CHO c
ompo
nent
of f
oods
.-Q
ualit
ativ
e m
etho
dsfo
rst
arch
dete
rmin
atio
n:
-Pol
ariz
ing
light
Mic
rosc
opy:
inta
ct s
tarc
h gr
anul
es c
an b
e id
enti
fied
as t
oth
eir
sour
ce:l
ike
pota
toe,
cor
n, w
heat
sinc
e gr
anul
esi
ze ,
shap
e, io
dine
stai
ning
char
acte
rist
ics
of e
ach
diff
er(s
ee11
.6.1,
p.18
4)
-Iod
ine
test
: lin
ear
stra
igh-
chai
n am
ylos
ere
acts
wit
h io
dine
tfo
rm
a bl
ue-b
lack
st
arch
-iodi
de
com
plex
, wh
erea
s th
bran
ched
ch
ains
of
amyl
opec
tins
form
pa
le
re
d vi
ole
com
plex
es.
Thou
gh c
olor
int
ensi
ty i
s no
t sp
ectr
opho
tom
etri
cre
acti
on
is
very
se
nsit
ive(
0.00
2g/m
l) an
d ap
plic
able
t
star
ch,d
extr
ins
and
glyc
ogen
).
-Qua
ntit
ativ
e M
etho
dsfo
rto
talS
tarc
h:
Star
chHyd
rolysis:
Follo
wing
Carr
ezTre
atmen
t*fo
rre
mov
ing
inte
rfer
ing
subs
tanc
es,
star
chis h
ydro
lized
with
acids
(0.4
NH
2SO
4)
and/
or en
zymes
[
firs
twi
th
αan
d β-
amylas
e en
zymes
to br
eak
amylos
ech
ains
to
malto
se a
nd g
luco
se;t
hen
with
amylog
luco
sida
se(o
rgluc
oamylos
e) e
nzym
eto
bre
ak a
mylop
ectins
]to
gluco
se,
which
can
then
be
dete
rmined
co
lorimet
rica
llywi
th
anth
rone
reag
ent
or
quan
tified
by
any
othe
r met
hod
(See
Fig.
11-
7,p.
180.
)
*CARR
EZ T
REATM
ENT:F
irst
addi
nga
solution
of p
otas
ium
hexa
cyan
ofer
rate
K 4[F
e(CN
) 6] th
enfo
llowe
dby
firs
tZn
SO
4 an
dth
enNaO
Hso
lution
s, t
hus
brea
king
any
emulsion
s, p
recipita
ting
prot
eins
, th
enge
tting
rid
of p
igmen
tsaf
ter
which
the
clea
rfiltra
teis u
sed
dire
ctly
in s
pecific
enzy
me
cata
lyse
dre
action
sfo
rea
chclas
sof
CH
O..
Hyd
roco
lloid
sot
hert
han
Sta
rch:
Food
Gum
s•
Stru
ctur
alhe
tero
geni
ty:li
near
orbr
anch
ed,
neut
ralo
rani
onic
,diff
erin
gso
lubi
lity
degr
ees;
ther
efor
eve
ryco
mpl
exan
alyt
ical
sche
mes
have
been
deve
lope
d(F
ig.1
1-8,
p.1
82-3
)
•M
etho
dsde
pend
on e
xtra
ctio
nfo
llow
edby
fract
iona
tion
and
hydr
olys
isfo
llow
edby
spec
ific
iden
tific
atio
nte
sts
Pec
tin•
~80%
is D
-gal
actu
roni
cac
id.
•S
apon
ifyw
ithN
aOH
, the
nac
idify
and
add
Ca+
2
topr
ecip
itate
pect
in.
•C
olle
ctC
ape
ctat
e, w
ash,
dry
and
wei
gh.
•D
egre
eof
est
erifi
catio
n(D
E):
The
isol
ated
pect
inis
was
hed
with
acid
ified
alco
holt
oco
nver
tto
acid
. Afte
rwas
hing
, titr
atio
nw
ithN
aOH
solu
tion
give
sno
nest
erifi
edgr
oups
. Afte
rsap
onifi
catio
nof
pec
tins,
bac
ktit
ratio
nw
ithst
anda
rdis
edac
idso
lutio
nto
dete
rmin
eex
cess
base
give
sD
E.
“Die
tary
Fibe
r”Fo
odco
mpo
nent
sno
t di
gest
ible
bym
amm
als
(Ch.
12)
•H
elp
prot
ect
agai
nst
colo
nca
ncer
•H
elp
norm
aliz
ebl
ood
lipid
s•
Redu
ceca
rdio
vasc
ular
dise
ases
•Sl
owgl
ucos
eab
sorb
tion
and
insu
linse
cret
ion
•H
elps
prev
ent
cons
tipa
tion
•D
RV(D
ieta
ryRe
fere
nce
Valu
e): 2
5g/2
000k
cal
Stru
ctur
al P
olis
acch
arid
es:
Stru
ctur
al P
olys
acch
arid
es a
re c
ompo
nent
s of
cel
l wal
ls o
fpl
ants
Plan
t ce
ll wa
ll co
ntai
nsLi
gnin
(non
CHO
)Ce
lulo
seCr
ude
fibe
r**
Hem
icel
lulo
seD
ieta
ry f
iber
*Pe
ctin
Gu
ms
Muc
ilage
Prot
eins
&Lip
ids
Die
tary
Fibe
r*:
Sum
of
the
nond
iges
tibl
eco
mpo
nent
sof
foo
ds
a. I
nsol
uble
fibe
r: C
rude
Fibe
r**:
b.So
lubl
efib
er:P
ecti
n+Gu
ms+
Muc
ilage
(Hyd
roco
lloid
s)
Solubl
efib
er
Tota
l, s
olub
le, i
nsol
uble
diet
ary
fiber
con
tent
sof
sel
ecte
dfo
ods
g/10
0 g
of F
ood
16.9
29.
737.
71O
atbr
an
2.89
6.66
2.01
5.25
1.02
1.41
Gre
enB
eans
Dry
bean
s
3.93
2.81
1.10
Car
rots
9.29
4.17
5.07
Pru
nes
1.12
0.59
0.53
Apr
icot
s
Tota
lDF
Inso
lubl
eD
FS
olub
leD
FFo
od
Bot
anis
tscl
assi
fyfib
er a
s:
1.C
ellw
allP
olys
acch
arid
es2.
Non
-cel
lwal
lpol
ysac
char
ides
1.C
ellw
allP
:S.:
Cel
lulo
se(lo
ngch
ain
mol
ecul
esm
ade
upof
glu
cose
arra
nged
in
mic
rofib
rils,
with
par
alle
l gro
ups
form
ing
stro
ngbu
ndle
s)H
emic
ellu
lose
(het
erog
eneo
usm
olec
ules
cont
aini
nga
num
bero
f su
gars
,and
tyin
gce
llulo
sem
icro
fibril
s; s
olub
lein
dilu
teal
kali
but n
ot in
w
ater
, Pe
ctin
(uro
nic
acid
poly
mer
solu
ble
in h
otw
ater
and
form
sge
ls)
2. N
on-c
ellw
allP
.S.:P
olys
acch
arid
esno
t inv
olve
d in
cel
lwal
lstru
ctur
e an
d ar
e no
t nut
ritio
nal.
They
incl
ude
hydr
ocol
loid
ssu
chas
muc
ilage
s(gu
aran
dlo
cust
bean
sgu
ms
) pla
ntex
udat
egu
ms(
arab
ic, t
raga
cant
hgu
ms)
and
alga
lpol
ysac
char
ides
(alg
inat
es, a
gar.
Car
rage
enan
). G
ener
ally
they
for
m
visc
ous
solu
tions
in w
ater
, but
ins
olub
le in
alc
ohol
s.
Lign
inis
a n
on-C
HO
thre
edi
men
sion
alpo
lym
erw
ith~4
0 ph
enol
units
cova
lent
lylin
ked
tohe
mic
ellu
lose
.
Die
tary
Fibe
r ME
THO
DS
•1.
Gra
vim
etric
:Dig
estib
leC
HO
’s, l
ipid
s, p
rote
ins
are
sele
ctiv
ely
solu
biliz
edby
chem
ical
san
den
zym
es; u
ndig
estib
les
are
colle
cted
byfil
tratio
nan
dqu
antit
ated
byw
eigh
ing.
•2.
Che
mic
al:D
iges
tible
CH
O’s
are
rem
oved
byen
zym
atic
dige
stio
n, fi
ber c
ompo
nent
sar
ehy
drol
ysed
byac
id, a
ndm
onos
acch
arid
esin
the
acid
hydr
olys
ate
repr
esen
tfib
er.
Extr
actio
n Pr
oced
ures
for S
elec
ted
fiber
com
pone
nts:
Cellu
lose
: In
solu
ble
resi
due
from
ext
ract
ion
with
17
.5%
NaO
H; c
an b
e so
lubi
lized
by a
ddin
g 72
% H
2SO
4
Pect
in: E
xtra
ct w
ith h
ot w
ater
or a
queo
us a
mm
oniu
m o
xala
te
(0.5
%).
whi
ch c
hela
tes
calc
ium
and
sol
ubiliz
esot
herw
ise
inso
lubl
e C
a pe
ctat
em
olec
ules
. The
n pr
ecip
itate
with
95%
et
hano
l.
Hem
icel
lulo
ses:
Ext
ract
with
25%
KO
H o
r 10%
NaO
H; m
ake
extra
ct a
cidi
c by
add
ing
acet
ic a
cid.
The
n pr
ecip
itate
by
addi
ng 4
vol
umes
of e
than
ol
Lign
in:.
Del
igni
ficat
ion:
Tre
at w
ith C
lor s
ulfit
e to
bre
ak c
hain
a.Gr
avim
etric
Met
hods
:a.
1.
Crud
e Fi
ber
::Si
nce
1850
s, i
t is
bein
gde
term
ined
as
mat
eria
l ins
olub
le in
dilu
te a
cid
and
dilu
te a
lkal
i und
er s
peci
fied
co
ndit
ions
(Wee
nde
met
hod
). Se
quen
tial
extr
acti
onof
sam
ple
with
1.25
N H
2SO
4, 1.2
5NN
aOH
, the
inso
lubl
ere
sidu
ebe
ing
drie
dwe
ighe
d,
ashe
dto
corr
ect
for
min
eral
s. T
ecat
orIn
c. h
as a
utom
ated
the
W
eend
em
etho
d(F
iber
Tec)
.Th
isan
alys
isdo
esno
t de
tect
hem
icel
lulo
se,
pect
ins
and
hydr
ocol
loid
s,
but
only
cellu
lose
and
ligni
n.
Sout
hgat
eha
s de
velo
ped
the
met
hod.
a.2.
Dete
rgen
tM
etho
ds:
Two
met
hods
have
been
deve
lope
d:
1. A
cid
dete
rgen
tfi
ber
met
hod
: det
erm
ines
ligni
n+
cellu
lose
2.N
eutr
al-d
eter
gent
fibe
r= d
eter
min
esac
idde
terg
entf
iber
+ he
mic
ellu
lose
ND
F =
Neu
tral
-det
erge
nt f
iber
(hem
icel
lulo
se+c
ellu
lose
+lig
nin)
ND
F-A
DF
= he
mic
ellu
lose
(Nei
ther
incl
ude
pect
ins
orhy
droc
ollo
ids)
Grou
nd d
ry s
ampl
e(1
mm
)
Refl
ux w
ith
H2S
O4
and
CTA
B so
l’n
Cellu
lose
, lig
nin
Sepa
rate
all
solu
ble
com
pone
nts
Dry
wei
gh
Filt
er, w
ash
with
hot
wate
r an
d ac
eton
e
ACI
D D
ETER
GENT F
IBER
Fig
1: T
he a
cid
dete
rgen
t fibe
r met
hod
Det
erge
nt M
etho
d fo
r Dieta
ry F
iber
Grou
nd d
ry s
ampl
e(1
mm
)
Refl
ux w
ith
SLS*
+ED
TA s
ol’n
Filt
er, w
ash
with
hot
wate
r an
d ac
eton
e
Dry
,wei
ghin
solu
bles
(W1)
Det
erm
ine
ash
(W2)
by
Burn
ing
Sepa
rate
all
solu
ble
com
pone
nts
Cellu
lose
, lig
nin,
min
eral
sH
emic
ellu
lose
(W1)
+ash
NDF
= W
1 –
W2
Fig
2: T
he n
eutr
al d
eter
gent
(*SL
S:So
dium
Laur
ylSu
lfat
e)fibe
r met
hod
Enzy
mat
ic M
etho
d fo
r Dieta
ry F
iber
Grou
nd, d
efat
ted
sam
ple
Aut
ocla
ve, E
xtra
ctio
n,
Cent
rifu
gati
on
Cent
rifu
gate
Extr
act
Am
ylog
luco
sida
sean
d pa
ncre
atin
for
hydr
olys
ing
star
ch a
nd p
rote
ins
Filt
rati
on
Was
h, d
ry, w
eigh
Inso
lubl
e di
etar
y fi
ber
Mem
bran
e Fi
ltra
tion
Was
h, d
ry, w
eigh
Solu
ble
diet
ary
fibe
r
Fig
3:Det
erminat
ion
of d
ieta
ry f
iber
using
enz
ymat
ic(M
euse
r)met
hod
a.3.
The
AO
AC
Met
hod
•Th
ism
etho
dco
mbi
nes
crud
efi
ber,
det
erge
ntfi
ber
and
Sout
hgat
em
etod
olog
ies
(see
fig.
12.1,
and
Tabl
e12
-1, p
.192
-5
for
tota
ldie
tary
fibe
r an
alys
is).
•Pr
inci
ple:
D
ryfa
t-ex
trac
ted
grou
ndfo
odsa
mpl
esar
een
zym
atic
ally
dige
sted
with
α-am
ylas
e, a
myl
oglu
cosi
dase
, and
prot
ease
tore
mov
epr
otei
nsan
dst
arch
. Ins
olub
lefi
ber
is c
olle
cted
byfi
ltra
tion
. Sol
uble
fibe
r is
pre
cipi
tate
dby
brin
ging
filt
rate
to78
%
eth
anol
and
colle
cted
byfi
ltra
tion
. The
filt
ered
fibe
r re
sidu
esar
ewa
shed
with
etha
nola
ndac
eton
e, o
ven
–dri
edan
dwe
ighe
d.
The
fibe
r re
sidu
eis
ana
lyse
dfo
rpr
otei
n an
das
h.
Tota
lfib
er=
resi
due
fibe
r we
ight
-(w
tof
pro
tein
+ w
tof
ash
)
b. C
hem
ical
Die
tary
Fib
er A
naly
ses
ENGL
YST-
CUM
MIN
GS P
roce
dure
Def
init
ion:
Fi
ber=
Sum
of a
llno
n-st
arch
mon
osac
char
ides
+ lig
nin
Prin
cipl
eof
EN
GLYS
T-CU
MM
INGS
Pro
cedu
re:
Star
chis
fir
stge
lati
nise
dan
dth
enen
zym
atic
ally
(pul
lula
nase
and
amyl
oglu
cosi
dase
)dig
este
d; r
emai
ning
non-
star
chpo
lysa
ccha
ride
sar
ehy
drol
ysed
byH
2SO
4 to
liber
ate
free
mon
osac
char
ides
, whi
char
ede
term
ined
eith
erby
GC o
rby
colo
rim
etri
cm
etho
ds. T
here
sult
is t
otal
diet
ary
fibe
r.
Tom
easu
rein
solu
ble
fibe
r, e
xtra
ctth
ewa
ter-
solu
ble
fibe
r by
heat
ing
the
dige
sted
mix
ture
for
30 m
inut
esin
a b
oilin
gwa
ter-
bath
.The
prec
ipit
ate
afte
rce
ntri
fugi
ngis
inso
lubl
efi
ber.
So
cont
inue
the
sam
ewa
ywi
thth
ispr
ecip
itat
ewi
thth
ehy
drol
ysis
step
. So
lubl
efi
ber=
Tot
alfi
ber-
Inso
lubl
efi
ber
Dra
wbac
ks:li
gnin
and
resi
stan
tst
arch
are
not
mea
sure
dby
this
met
hod.
Any
suga
rin
itia
llypr
esen
tsh
ould
be e
xtra
cted
prev
ious
ly, o
rwi
llyi
eld
high
resu
lts.
. Se
eFi
g. 1
2-2,
p. 1
96-7
.