1. 10/20/2012 Minaleshewa Atlabachew 1

2. •
•
•
•
•
•
10/20/2012 Minaleshewa Atlabachew 2

3. • !
"
• #
• $ %are biosynthetically derived from
primary metabolites. They are more limited in
distribution being found usually in specific families.
They are not necessary for growth and development,
but may serve as pollination attractants,
environmental adaptations, or protection.
10/20/2012 Minaleshewa Atlabachew 3

4. • &
' ( ! #
(
• )
(
• '
* ,(
#+
' ! (
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5. -' '
( * ,
# # (.
* . +",
• # ( (
/ !
$
! )
" ' !
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6. 10/20/2012 Minaleshewa Atlabachew 6

7. •
( ) !
((
(( / (
(
!
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8. •
• (
,
• * (( ! ,
& (
!
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9. (
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10. Polyphenols Simple phenols Miscellaneous
Liganas
Tannins 0 1
Cumarins
$
Lignins
Resveratrol
Flavones
Flavonols
Flavan-3-ol
Isoflavones
Flavanols
Anthocyanins
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11. #
• (
• (
• (( (
# ( ! /
!
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12. # (
10/20/2012 Minaleshewa Atlabachew 12

13. • (
•
•
# ' ( ( ( )
(( (
( !
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14. Freeze-drying, Air drying, Oven drying,
Grinding, Homogenization
Sample pre-
treatment
! Solid-liquid extraction
Conventional Modern
Extraction
Percolation UAE, MAE, SFE, PLE,
Maceration PHWE ,
Soxhlet, Refluxing, Infusion
Clean- up -
UV, Isolation
EC, CC-fractionation, Filtration, Evaporation,
FLU, LLE, SPE, SPME, SDME
Instrumental
TLC
analysis
MS
GC
LC
LC
FID, ECD MS, MS/MS
UV , 10/20/2012 Minaleshewa Atlabachew
Electo.Chem., FLU, MS, MS/MS, NMR 14

15. 2)
• ) * 3 ) ,
• 4 ! (
5&
• 4 ( #
• 6 4
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16. -
• ' !
• !
( !
1-50 g
• 3
• / . Percolator
• # 7
• - 8
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17. 9 &
" #
Solvent ½ h -24 h
Filtration
Pooled
Up to 72 hr Solvent renewed
two-three
Solvent
Sample
Stirred
Shacken
Solvent
Sample
Universal
Prolonged ext. time improves efficiency but
10/20/2012 Minaleshewa Atlabachew 17

18. )
• -
• 0
• 4
• # 8 $:
• ; 8 . 3
• -
• (( ( )
• *# ,
/
(
•
10/20/2012 Minaleshewa Atlabachew
Heat 18

19. 5 + ! (
• - (!
• & ((
• 4
Heat
• # < (( ( )
• 3
&
((
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20. • = ( (
(
3 / (
)
0 ( (
3 (
> / (
! )
! (
-
>
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21. - ? ) * -2,
?
US irradiation
• ?
• (
> 2
> !
Recently , UAE is widely used
for all types of phenolic cpds
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22. @
• 2(( ( )
• # 8* . 1 ,
• # 8*7 A ,
• ? ! *9 ! ! ,
• ) 8 4 (
9 $
>
) ( )
10/20/2012 Minaleshewa Atlabachew 22

23. $%& '
• & -2
(& ! > *$:. & +/
( 4 ,
Heating
5
expansion and rupture of cell walls
Weakening van der Waals forces between solute & matrix
liberation of phenolics
Only polar solvents are used for MAE
10/20/2012 Minaleshewa Atlabachew 23

24. - (& -2
> (
) * B$ ,
) *( )
,
• + !
3 (( ( )
*" . ,
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25. @ C !
24 20 6 0.5 0.1 h
D
Plant flavonoids
10/20/2012
-4
Minaleshewa Atlabachew 25

26. !%" ! ( ' "(') *+%
• ( )
• $ ! 02
! *7 AE , *F7 ,
! )
>
• 3 ! 5((
SF
• + !
• #
• & ( G /
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27. 2)
• 3 !
• # : A
• 8$. :. &
• # *$ : ,
• # "
• $.
• .A 3 (
Liquid solvent
10/20/2012 Minaleshewa Atlabachew 27
& (

28. -
•
• 9 )
• 0 )
• > )
• > ( $
• ( (
• 3
• - .
• & / * ) < / ,
• + )
• 2) Disadvantage
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29. # %, - ./ ' ,.'01"'
• ! ' !
• #*. $ , * .& ,
# (( 6 '
6 (
( < (
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30. 2)
100 atm to 200 atm
5 G
10/20/2012
Multiple extraction ( for static)
Minaleshewa Atlabachew 30

31. @
+ (( )
A
9 ) +$ *> + +7 ,
# ) * $ H > + H 5&
+ G +7 ,
5& 4 5& )
# 8 7 I
+ #
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32. 2 + 6 2) * +6 2,G * +6 2,
• 4
• / / !
• # 8 7F: J < 8
# - 2 $
(! 4
5 *" $:,
(! (
)
0 * $ J , A
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33. • ( (
4
• ( 0
•
• 3
)
•
• 2 >
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34. @
• 332 ' +$ $
#
• 2* " " ) ,
• &
K G
) * & 2,
) * 5 & 2,
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35. &
10/20/2012 Minaleshewa Atlabachew 35

36. &2 SDME-GC-MS
SDME-in syringe derivatization
• &2D &
- Hexyl acetate
• 5 /
•
SDME-in syringe derivatization
10/20/2012 Minaleshewa Atlabachew
GC/MS 36

37. • ( &2 ( / !
/ '
( / 4 $. :
( 7 A
• (
)
• ' * )
) G ) ) G
/ ,
• ( 5& 2 ( ' * )
, (
( !
• + ( !
4
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38. K (
Derivatization
D
Individual
Determination
2
2)
+ 3
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39. -
• 9 ( )
• & ( 2
• ) *
& 2,
• )
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40. D
L1 % 234 / 5 1')
# L6 ( # 0! 784
# L6 1!
( L 1944 / !')
L
L1 , 1 (
# 1 %1 9*4
L 1' *7%
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41. • #
• # ! ! (
• ( ) $"
) ( 4 (
? 4
• $ # ( ! * >,
0 ( :
( # ) (
•
•
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42. #
• ? ' ; !M
• ? ' 9 -
!M
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43. - )
• ; <
• ( ) ' 2
/ '
• # ( )
(
• + ( )
( ! (
• # ( ) /
5 -
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44. • - ) '
( ( )
) ) )
• # (
) ( !
( ( (( ( (
(
10/20/2012 DPPH Minaleshewa Atlabachew
(2, 2 -diphenyl-1-pycryl-hydrazyl) 44

45. & 2#+ 5 5 2>-#
• ; )
) ((
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46. # 3 ) 4
4 (
(( !
)
)
# ) 4
(
/ 4
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47. 10/20/2012 Minaleshewa Atlabachew 47