Minia J. of Agric. Res. & Develop. Vol. (30) No. 2 pp 243 -25 !

20"0
FACULTY OF AGRICULTURE

ANTIOXIDANT ACTIVITY, PHENOLIC COMPONENTS AND NUTRITIONAL EFFECT OF SOME AROMATIC PLANTS

Magda A.A. Seleim Food Sci. and Tech. Dept., Fac. of Agric. Assiut University, Assiut. Received 6 Dec. 2010 Accep ed 12 Dec. 2010

A!STRACT
I" #e p$e%e" % &d'( e%%e" ial )il% *e$e )+ ai"ed ,$)m ,ive %elec ed a$)ma ic pla" %( "amel'- ,e""el( $)%ema$'( gi"ge$( #'me a"d ci""am)". T#ei$ c)" e" % ), ) al p)l'p#e")l% *e$e .&ali a ivel' a"d .&a" i a ivel' de e$mi"ed &%i"g /0LC a"al'%i%. A" i)1ida" ac ivi ie% *e$e de e$mi"ed *i # a Ra"cima appa$a &% c)mpa$i"g *i # %'" #e ic a" i)1ida" 2!/T3. T#e "& $i i)"al e,,ec )" $a % %e$&m lipid% *a% al%) % &died. A$)ma ic pla" e1 $ac % c)&ld +e a p) e" ial %)&$ce ), "a &$al a" i)1ida" % a"d ca" +e added ) ,))d% ) $eplace %'" #e ic a" i)1ida" %( mi"imi4i"g )il pe$)1ida i)". Ge"e$all'( #e )+ ai"ed $e%&l % %#)*ed #a #e % &died a$)ma ic pla" % *e$e $ic# i" p#e")lic c)mp)"e" % +& $)%ema$' #ad #e #ig#e% level a"d #'me c)" ai"ed #e l)*e% . T#e%e p#e")lic c)mp)&"d% dem)"% $a ed g))d a" i)1ida" ac ivi ' a"d #e pla" %( $ic# i" p#e")lic acid% a"d ,lav)")id% c)&ld +e c)"%ide$ed a% a g))d %)&$ce ), "a &$al a" i)1ida" %. Al%)( e1ami"ed a$)ma ic pla" % c)&ld +e $ega$ded a% a g))d $ea me" ,)$ dec$ea%i"g %e$&m ) al c#)le% e$)l( LDL5c#)le% e$)l( 6LDL5 c#)le% e$)l a"d $igl'ce$ide%( a% *ell a% i"c$ea%i"g /DL5c#)le% e$)l level i" $a %e$&m lipid%.

Magda A.A. Seleim I7TRODUCTIO7 Oxidative degradation of lipids is a major factor limiting the shelf life of foods. The free radical reaction of lipid peroxidation is generally responsi!le for the deterioration of lipid containing foods. "t decreases nutritional and sensory properties of foods since it involves the loss of essential fatty acids and vitamins, the generation of toxic compounds, causing additionally, flavor, texture and color deterioration #$orrissey et al., %&&'(. The use of antioxidants during the manufacturing process can minimi)e the extent of lipid peroxidation #Shahidi and *anasundara, %&&+(. ,ecently, there is an increasing interest !oth at industry and scientific levels for the use of spices and aromatic plants !ecause of their strong antioxidant and antimicro!ial properties, -hich exceed many currently used natural and synthetic antioxidants. These properties could !e due to the presence of many su!stances, including some vitamins, flavonoids, terpenoids, carotenoids, etc., -hich, render spices and some aromatic plants or their antioxidant components as preservative agents in food #.alucci et al., +//0(. 1olyphenolic compounds are commonly found in !oth edi!le and non edi!le plants, and they have !een reported to have multiple !iological effects, including antioxidant activity #2ah3onen et al., %&&&(. Aromatic plants are used in many domains, including nutrition, flavoring, or as !everages #Djeridane et al., +//4(. $any aromatic plants have !een recogni)ed to have medicinal properties and !eneficial impact on health, e.g. antioxidant activity, antimicro!ial, hypolipidemic and anticarcinogenic potential #5uo et al., +//6(. .rude extracts of aromatic plants and other plant materials rich in phenolic compounds are of increasing interest in food industry !ecause they can retard oxidative degradation of lipids and there!y improve the 7uality and nutritional value of foods. The aim of this study -as to evaluate and compare the antioxidant activities of some aromatic plant oils namely8 fennel, rosemary, ginger, thyme and cinnamon using the ,ancimat method. Total phenolic content, 7ualitative and 7uantitative analysis of the major phenolics -ere determined using 915. analysis. The nutritional effect of the studied aromatic plants on rat serum lipids #triglycerides, total cholesterol, 9D5, 5D5 and :5D5 cholesterols( -as also investigated. MATERIALS A7D MET/ODS

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A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" % Ma e$ial%Five selected aromatic plants, namely8 fennel, rosemary, ginger, thyme and cinnamon -ere o!tained from local mar3et. Sunflo-er oil -as donated from ;l <ile .ompany for oils and soaps, Assuit, ;gypt. E1pe$ime" al a"imal%Fourty t-o male of al!ino rats -ere o!tained from Animal 9ouse, Faculty of $edicine, Assiut University and -ere randomly divided into seven groups #each group consisted of six rats( of similar total -eight. The rats in each group -ere assigned to the corresponding experimental diet and -ere housed individually in cages in a controlled environment. Diets and -ater -ere supplied ad li!tum throughout the study. Die %!a%al Die 2!D3- <ormal diet provided from animal house. Ric# c#)le% e$)l die 2RCD3- =asal diet>% g !rain #gavage( according to Al Sharja!i #+//?(. ,at groups -ere fed during experimental period as follo-s8
All rats groups 6+ rats

.ontrol " =asal Diet group #4 rats(

,ich cholesterol diet group for %/ days #04 rats( Treatment groups for 0/ days #0/ rats( Fennel group #4 rats( Ainger group #4 rats( .innamon group #4 rats( ,osemary group #4 rats( Thyme group #4 rats(

.ontrol "" =asal Diet group for 0/ days #4 rats(

Fig. 1- Fl)* %#ee diag$am ), di,,e$e" $a g$)&p% d&$i"g "& $i i)"al pe$i)d. Me #)d%A$)ma ic pla" )il e1 $ac i)"The aromatic plant oils -ere extracted using the -ater distillation method descri!ed !y ,avindran et al. #+//@(.

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Magda A.A. Seleim A" i)1ida" ac ivi 'Antioxidant activities of the studied aromatic plant oils compared -ith synthetic antioxidant #=9T( -ere determined -ith a ,ancimat apparatus #$etrohm, 9erisau, S-it)erland( !y measuring the induction period of oils containing the antioxidant, according to the method descri!ed !y 9asenhuettl and -an #%&&+(. The antioxidant index -as calculated as8

Antioxidant index =

"nduction period of oil -ith extract "nduction period of oil alone

T) al p)l'p#e")l% c)" e" Total polyphenol content -as demonstrated using Folin .iocalteu colorimetric method as descri!ed !y 9uang et al. #+//@(. The a!sor!ance of the resulting !lue color -as measured at @4? nm -ith a shimad)u U: spectrophotometer. Buantification -as done -ith respect to the standard curve of gallic acid. The results -ere expressed as mg gallic acidC%// g dry -eight. 0#e")lic c)" e" Bualitative and 7uantitative analysis of major phenolics -ere determined !y using 915. analysis as descri!ed !y Aa!y et al. #+//6(. !l))d a"al'%i%=lood sample -as ta3en from each group of rat for the determination of8 Serum total cholesterol level #mgCd5(. Serum high density lipoprotein 9D5 level #mgCd5(. Serum triglyceride level #mgCd5(. T) al c#)le% e$)l c)" e" Total cholesterol content -ere determined colorimetrically -ith commercially availa!le 3its #.holesterol . test, ;5"T;.9 diagnostics, French( according to Allain et al. #%&@6(. /DL c#)le% e$)l c)" e" -

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A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" % 9D5 cholesterol content -ere determined colorimetrically -ith commercially availa!le 3its #9D5 cholesterol test, ;5"T;.9 diagnostics, French(. The 7uantitative estimation of 9D5 cholesterol -as made using 9D5 cholesterol precipitating reagent in com!ination -ith en)ymatic colorimetric assay 3it for total cholesterol, -here chylomicrons, very lo- density lipoprotein #:5D5( cholesterol, and lo- density lipoprotein #5D5( cholesterol fractions -ere precipitated from serum or plasma !y means of phosphotungstic acid and magnesium ions, according to 5opes :irella et al. #%&@@(. After centrifugation, high density lipoprotein #9D5( cholesterol -as then determined in the supernatant using a cholesterol reagent and the derived dilution factor in the calculation. T$igl'ce$ide% c)" e" Triglyceride concentrations -ere determined colorimetrically at ?64 nm -ith commercially availa!le 3its #Triglycerides test, ;5"T;.9 diagnostics, French(, according to =ucolo and David #%&@0(. E% ima i)" ), LDL a"d 6LDL c#)le% e$)l i" %e$&mThe concentration of 5D5 cholesterol -as calculated according to the e7uation of Friede-ald et al. #%&@+( as follo-s8
:LDL5c#)l; < :T) al c#)l; = :/DL5c#)l; = :TG;>9

All concentrations -ere in mgCd5. The 7uotient DTAEC? -ere used as a measure of :5D5 cholesterol concentration. "t -as assumed first, that virtually all of the plasma TA -as carried on :5D5, and second, that the TA8 cholesterol ratio of :5D5 -as constant at a!out ?8% #Friede-ald et al., %&@+(.

528?5

Magda A.A. Seleim RESULTS A7D DISCUSSIO7 A" i)1ida" ac ivi ' ), a$)ma ic )il% pla" %Antioxidant activities of studied aromatic plant extracts are presented in Ta!le %. F =9T -as presented for comparison. *ith the exception of ginger and thyme, the extracts sho-ed higher antioxidant activity in sunflo-er oil. Aromatic plant extracts contained phenolic structure -hich -ere capa!le of minimi)ing oil , protecting sunflo-er oil against autoxidation #Arouma et al., %&&+(. Ta+le 1- A" i)1ida" ac ivi ie% ), a$)ma ic pla" e1 $ac %. A" i)1ida" i"de1 i" 0la" e1 $ac %&",l)*e$ )il Fe""el 2.8 R)%ema$' 2.6 Gi"ge$ 1.6 T#'me 1.9 Ci""am)" 2.@ !/T 2.9 T) al p#e")lic% c)" e" The amount of total phenolics content, estimated !y Folin .iocalteu method of the studied aromatic plant samples are presented in Ta!le +. The amount of total phenolics varied -idely !et-een selected aromatic plants, ranged from +.6' to %@.4/ mg gallic acidC%//g of dry -eight. The highest level of total phenolics -as found in rosemary, -hile thyme contained the lo-est level. 1henolics contents can !e arranged in the decreasing order as follo-8 rosemary G fennel G cinnamon G ginger G thyme. The o!tained results sho-ed that aromatic plants had relatively high level of polyphenols, and the differences !et-een the results could !e due to genotypic or and environmental differences -ithin species #Shan et al., +//?(.

528A5

A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" % Ta+le 2- T) al p#e")lic c)" e" 2mg gallic acid>100 g d$' *eig# 3 ), a$)ma ic pla" %.
A$)ma ic pla" Fe""el R)%ema$' Gi"ge$ T#'me Ci""am)" T) al p#e")lic c)" e" 1@.6@ 1?.60 @.B1 2.8A 11.82

Ide" i,ica i)" ), p#e")lic c)mp)"e" %The major types and the representative components of phenolic compounds in the samples -ere analy)ed using 915. method compared -ith authentic phenolic standard #Ta!le 0(. .affeic acid, p coumaric acid, ferulic acid and neochlorogenic acid -ere identified as the major phenolic acids present in the studied samples, -hile, luteolin, apigenin, 3aempferol and isorhamnetin -ere identified as the major flavonoids. Ta+le @- C&a" i a ive a"al'%i% ), #e maD)$ p#e")lic c)mp)&"d% ), a$)ma ic pla" % 2mg>100g d*3.
0#e")lic acid% p5c)&ma$ic acid 7e)c#l)$)ge"ic acid Ca,,eic acid acidFe$&lic A$)ma ic pla" % L& e)li" Flav)")id% I%)$#am"e i" Eaemp,e$)l Apige"i"

Fe""el R)%ema$' Gi"ge$ T#'me Ci""am)"

680 A?2 1B@ 160 912

81.0 @6.2 81.6 B0.2 82.6

22.6 2B.6 16.A @8.9 20.A

A.0 12.A 10.2 20.@ B.6

@21.0 620.0 A2.0 66.@ 102.0

28.0 @B.0 @0.2 21.0 16.2

1B.6 1A.6 81.9 @2.? 89.6

@6.? 12.A 22.@ 20.2 11.B

.onsidera!le variation -as found in the phenolic compounds of the different aromatic plants. The main phenolic acids in these plants -ere caffeic acid and p coumaric acid. 9o-ever, ferulic acid and neochlorogenic acid occurred in minor 7uantities. These results are in 528B5

Magda A.A. Seleim agreement -ith those reported !y 5uo et al. #+//6( and Shan et al. #+//?(. Aenerally, results sho-ed that the aromatic plants -ere rich in phenolic components and demonstrated good antioxidant activity. These plants, rich in phenolic acids and flavonoids could !e a good source of natural antioxidants. Therefore, the 7ualitative and 7uantitative analysis of major individual phenolics in the aromatic plants could !e helpful for explaining the relationships !et-een the total antioxidant capacity and the total phenolic contents in the aromatic plants. 7& $i i)"al e,,ec ), a$)ma ic pla" % )" $a %e$&m lipid%,ats -ere divided into seven groups and fed during experimental period #6/ days( as sho-n in Fig. %. At the first ten days, except group % -hich left as control all groups fed ,ich cholesterol diet to raised serum lipids level. Ta!le 6 sho-s that the higher increase -as in 5D5 cholesterol #@4.0H(, triglycerides and :5D5 cholesterol #?+.+H( then total cholesterol #0'.4H(, -hile, 9D5 cholesterol decreased !y +0.0H. Ta+le 8- C#a"ge% i" $a %e$&m lipid% d&$i"g e1pe$ime" al *i #)& a"' addi i)".
Ra %e$&m lipid T$igl'ce$ide C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" C)" G$)&p $)l I a 4e$) ime $)l I a e"d ime $)l II a 4e$) ime $)l II a e"d ime $)l I a 4e$) ime $)l I a e"d ime $)l II a 4e$) ime $)l II a e"d ime $)l I a 4e$) ime $)l I a e"d ime $)l II a 4e$) ime $)l II a e"d ime $)l I a 4e$) ime $)l I a e"d ime $)l II a 4e$) ime $)l II a e"d ime $)l I a 4e$) ime $)l I a e"d ime $)l II a 4e$) ime $)l II a e"d ime Mea"% 1@9.?6 1@?.28 206.9 1B9.22 11?.29 11B.10 162.6 19?.B8 @?.66 @A.8? 2A.B @2.28 92.88 9@.1B B2.8 A6.61 2?.19 2?.88 81.@ @B.09 F c#a"ge% 92.2

T) al c#)le% e$)l

@A.6

/DL5c#)le% e$)l

2@.@

LDL5c#)le% e$)l

?6.@

6LDL5c#)le% e$)l

92.2

The data presented in Figures #+ 4( sho- the effect of adding aromatic plants to rat diets on serum lipids.

52905

A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" %
210

200

190 T$igl'ce$ide% 2mg>dL3

180

170

Fennal Rosemry Ginger Thyme Cinnamon

160

150

140 0 10 Time 2da'%3 20 30

Fig. 2- E,,ec ), addi"g a$)ma ic pla" % )" $a T$igl'ce$ide%

165

%e$&m

155

T) al c#)le% e$)l 2mg>dL3

145

135

Fennal Rosemry Ginger Thyme Cinnamon

125

115 0 10 Time 2da'%3 20 30

Fig.@- E,,ec ), addi"g a$)ma ic pla" % )" $a %e$&m ) al c#)le% e$)l. 52915

Magda A.A. Seleim

65

60

55 /DL5c#)le% e$)l 2mg>dL3

50
Fennal Rosemry Ginger Thyme Cinnamon

45

40

35

30

25 0 10 Time 2da'%3 20 30

Fig.8- E,,ec ), addi"g a$)ma ic pla" % )" $a %e$&m /DL5 c#)le% e$)l.
100 90

80 LDL5c#)le% e$)l 2mg>dL3

70

60

Fennal Rosemry Ginger Thyme Cinnamon

50

40

30 0 10 Time 2da'%3 20 30

Fig.9- E,,ec ), addi"g a$)ma ic pla" % )" $a %e$&m LDL5 c#)le% e$)l

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A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" %
42

40

38 6LDL5c#)le% e$)l 2mg>dL3

36

34

Fennal Rosemry Ginger Thyme Cinnamon

32

30

28 0 10 Time 2da'%3 20 30

Fig.6- E,,ec ), addi"g a$)ma ic pla" % )" $a %e$&m 6LDL5 c#)le% e$)l. Fig. + sho-s that the highest decrease in rat serum triglycerides -as in groups fed on rosemary specially after 0/ days of treatment, follo-ed !y fennel, cinnamon, ginger and thyme in the descending order. This could !e due to inhi!ition of hepatic triglyceride synthesis and stimulation of hepatic peroxisomal oxidation #,ui) Autierre) et al., %&&&(. Fig. 0 sho-s that the highest decrease in rat serum total cholesterol -as in rats fed cinnamon, possi!ly due to inhi!iting a!sorption and synthesis of cholesterol. *hereas there -as a decrease in lymphatic a!sorption of cholesterol accompanying an increase in fecal excretion of neutral, !ut not acidic sterioids, particularly -hen cholesterol enriched diet -as given #9irose et al., %&&%(. 9o-ever, adverse trend of rat serum 9D5 cholesterol level #Fig. 6( -as sho-n in 5D5 cholesterol. Fig. ? sho-s that the highest

529@5

Magda A.A. Seleim decrease in rat serum 5D5 cholesterol -as in rats fed cinnamon follo-ed !y those fed thyme. The trend of serum :5D5 cholesterol level in rats during the experimental period is sho-n in Fig. 4. They sho-ed the same trend as of triglyceride. "t -as assumed that first all of the plasma triglyceride is carried on :5D5, and second, that the #triglyceride8 cholesterol( ratio of :5D5 is constant at a!out ?8% #Friede-ald et al., %&@+(. ;xtracted natural antioxidants #aromatic plant extracts( could !e considered as a good treatment for decreasing serum total cholesterol, 5D5 cholesterol, :5D5 cholesterol and triglycerides, !ut it increase 9D5 cholesterol level of rats serum. "t could !e concluded that the studied aromatic plant samples -ere rich in phenolic components and demonstrated good antioxidant activity. $oreover, aromatic plants may contain polar products -hich -ould !e a!le to lo-er lipid concentrations in hyperlipidemia rats, and could !e !eneficial in preventing hyperlipidemia and related cardiovascular diseases. REFERE7CES Aa+'( E.G /va &m( E. a"d SH$ede( G. 220083. Analysis of flavonoids and other phenolic compounds using 915. -ith color metric array detection. ,elationship to antioxidant activity. Iournal of the Agricultural and Food .hemistry, ?+, 6?&? 64/0. Al5S#a$Da+i ( F.A. ( 220093 - =iochemical and nutritional studies on some oil seeds . 19.D. Thesis, Food Sci.J Techn. Dept., Faculty of Agric., Assiut, University , ;gypt . Allai"( C.C.G 0))"( L.S.G C#a"( C.S.G.G Ric#m)"d( I. a"d F&( 0.C. 21B?83. ;n)ymatic determination of total serum cholesterol. .lin. .hem., +/, 6@/ 6@?.

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A" i)1ida" ac ivi ' a"d "& $i i)"al e,,ec ), a$)ma ic pla" % A$&)ma( O.I.G /alli*ell( !.G Ae%c#+ac#( R. a"d L)lige$( J. 21BB23. Antioxidant and pro oxidant properties of active rosemary constituents8 carnosol and carnosic acid. Keno!iotica ++, +?@ +4'. !&c)l)( G. a"d David( /. 21B?@3. Buantitative determination of serum triglycerides !y the use of en)ymes. .lin. .hem., %&, 6@4 6'+. Cal&cci( L.G 0i"4)")( C.G Ka"d)me"eg#i( M.( a"d Cap)cc#i( A. 2200@3. ;ffect of gamma irradiation on the free radical and antioxidant contents in nine aromatic her!s and spices. Iournal of the Agricultural and Food .hemistry, ?%, &+@ &06. DDe$ida"e( A.G Y)&%,i( M.G 7adDemi( !.G !)& a%%)&"a( D.G S )cHe$( 0. a"d 6idal( 7. 220063. Antioxidant activity of some Algerian medicinal plants extracts containing phenolic compounds. Food .hemistry, &@, 4?6 44/. F$iede*ald( I.T.G Lev'( R.I. a"d F$ed$icH%)"( D.S. 21B?23. ;stimation of the concentration of lo- density lipoprotein cholesterol in plasma -ithout use of the preparative ultracentrifuge. .lin. .hem., %', 6&& ?/+. /a%e"#&e l( G.L. a"d Ia"( 0.J. 21BB23. Temperature effects on the determination of oxidative sta!ility -ith the $etrohm ,ancimat. I. Am. Oil .hem. Soc., 4&, ?+? ?+@. /i$)%e( 7.G I")&e( T.G 7i%#i#a$a( E.G S&ga")( M.G AHim) )( E.G S#imi4&( S. a"d Yamada( /. 21BB13. "nhi!ition of cholesterol a!sorption and synthesis in rats. I. 5ipid ,es., 0+, 4+& 40'. /&a"g( I.Y.G Cai( Y.K.G Li"g( J.G C)$He( /. a"d S&"( M. 2200?3. A potential antioxoidant resource8 endophytic fungi isolated from traditional .hinese medicinal plants. ;conomic =otany, 4%, %60/. Ea#H)"e"( M.0.G /)pia( A.I.G 6&)$ela( /.J.G Ra&#a( J.0.G 0i#laDa( E. a"d E&Dala( T.S. 21BBB3. Antioxidant activity of plant extracts containing phenolic compounds. Iournal

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Magda A.A. Seleim of the Agricultural and Food .hemistry, 6@, 0&?6 0&4+. L)pe%56i$ella( M.F.G S )"e( 0.G Elli%( S. a"d C)i*eil( J.A. 21B??3. .holesterol determination in high density lipoproteins separated !y three different methods. .lin. .hem., +0, ''+ ''6. L&)( Y.G Cai( C.G S&"( M. a"d C)$He( /. 220083. Antioxidant activity and phenolic compounds of %%+ traditional .hinese medicinal plants associated -ith anticancer. 5ife Science, @6, +%?@ +%'6. M)$$i%%e'( 0.A.G S#ee#'( 0.J.A.G Galvi"( E.G Ee$$'( J.0. a"d !&cHle'( D.J. 21BBA3. 5ipid sta!ility in meat and meat products. $eat Science, 6&, S@0 S'4. Ravi"d$a"(0.7.( !a+& 7.E. a"d Siva$ama" E.2200?3. Turmeric8 The genus curcuma. .,. 1ress, <e- Lor3 . R&i45G& ie$$e4( 6.G 0e$e45E%pi")%a( A.G 6a4.&e4( C.M. a"d Sa" a5Ma$ia( C. 21BBB3. ;ffect of dietary fats on lipid composition and antioxidant en)ymes in rat liver. =ritish I. <utr. '+, +00 +6%. S#a#idi( F. a"d Ia"a%&"da$a( J.0.E.0.D. 21BB23. 1henolic antioxidants. .ritical ,evie-s in Food Science and <utrition, 0+, 4@ %/0. S#a"( !.G Cai( Y.K.G S&"( M. a"d C)$He( /. 220093. Antioxidant capacity of +4 spice extracts and characteri)ation of their phenolic constituents. Iournal of the Agricultural and Food .hemistry, ?0, @@6& @@?&.

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