Structure and Chemical and Physicochemical Properties of Jobs

Tear (Coix lacryma-jobi L.) Kernels and Flours

unnin! head" Structure and Properties of Jobs Tear Kernels and Flours
Jiraporn Chaisiricharoen#ul$ Sunanta Ton!ta$
%
and Kano#&'rn
(ntarapichet
School of Food Technology, Institute of Agricultural Technology, Suranaree
University of Technology, a!hon "atchasi#a $%%%%, Thailand, Tel& '(( ))
**)*((+ Fa,& '(( )) **)$-.& /0#ail& s0tongta1g2sut2ac2th
34orresponding author
)bstract
(n Jobs tear$ the !erm is enfolded inside the #ernel and comprises ca. one third
of the *hole !rain. To assess the effect of !erm on flour properties$ de!ermed
flour *as manually prepared from !rains of *hite and blac# hus# culti+ars. The
protein of *hole !rain and de!ermed flours *as not si!nificantly different
(p,-.-.). The antio/idant properties$ *hich *ere total phenolic content$ 0PP1
radical sca+en!in! and reducin! po*er$ and coi/enolide of *hole !rain and
de!ermed flours of both culti+ars *ere in the ran!es of 2.33&4.54 m!6)78!$ ..9-&
2..3:$ ;..<&;.44 and -.-;&-..3 =!8!$ respecti+ely. The Jobs tear starch !ranules
sho*ed a round and poly!lonal shape and their a+era!e si>e *as 55.<4&5;.;? =m.
The !elatini>ation temperature ran!e of *hite and blac# Jobs tear starch *as
<2&45
o
C and the retro!radation beha+ior of Jobs tear starch occurred after
stora!e at 9
o
C for 3? days as monitored by differential scannin! calorimetry. The
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pea# +iscosity of the *hite Jobs tear flours *as !reater than those of the blac#
culti+ar flours but the setbac# and pastin! temperature *as lo*er. The s*ellin!
po*er of the *hite Jobs tear flours *as hi!her than that of the blac# culti+ar
flours$ but their solubility *as lo*er and the opposite result *as found in the
starch.
Key*ords" Jobs tear$ structure$ antio/idant properties$ physicochemical
properties
(ntroduction
Coix lacryma-jobi 52 is a distant relative of #ai6e in the 7aydae tribe of the grass
fa#ily, Poaceae or 8ra#inaeae2 It is co##only na#ed Jobs tear, adlay, #ayuen,
4hinese pearl barley and hato#ugi2 Jobs tear seeds are #ainly produced in /ast and
South0/ast Asia, including 4hina, Japan, the Philippines, 9ur#a, and Thailand2 The
seeds of Jobs tear are oval or egg shaped :ith ; ## dia#eter and have a #il!y :hite
to blac! outer surface after the dehulling process2 Jobs tear has long been used in
traditional 4hinese #edicine and as a nourishing cereal2 It is added in soups and
broths in the for# of flour or :hole grain2 In Japan and Thailand, a non0dairy drin!
fro# Jobs tears is available in the #ar!et as an alternative health food2 Ani#al and
hu#an clinical trials de#onstrated that the consu#ption of dehulled flour and seed of
Jobs tear can i#prove lipid #etabolis#, thereby decreasing the ris! of heart diseases2
In addition, it could reduce liver fat accu#ulation and protect fro# tu#or sti#ulating
co#pounds <4hang et al2, *%%$+ =u et al2, *%%;>2 So#e bioactive co#pounds in Jobs
tear, especially coi,enolide, inhibited tu#ors, prevented cancer and protected against
viral infection <?ung and 4hang, *%%$+ 4hun et al2, *%%)>2
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At present, Jobs tear is used as polished grains for food applications2 An
increase in Jobs tear utili6ation can ta!e place if #illing is e#ployed to transfor# it
into Jobs tear flour, :hich could be incorporated into several food products as a
#a@or food ingredient2 "ecently, the consu#ption of :hole grain has beco#e popular
due to its retention of functional co#ponents2 ?o:ever, this full0fat flour has
li#itations in #a!ing food products2 The use of defatted Jobs tear flour is necessary
to #a!e Jobs tear #ore a#enable to several food applications2 Therefore, the
investigation of the functional properties of this flour is essential2 In addition, a
structural study of Jobs tear !ernel, :hich is i#portant for #illing process, has not
yet been published2 There are * i#portant cultivars of Jobs tear that are co##ercially
available in Thailand, one :ith :hite hus! and one :ith blac! hus!2 Therefore, the
ob@ective of this study :as to investigate the structure of the grain, che#ical
co#positions, and che#ical and physicoche#ical properties of :hole grain flour,
deger#ed flour, and starch of :hite and blac! hus! Jobs tear2
@aterials and @ethods
@aterials
The blac! hus! Jobs tear :as a gift fro# 44P orthern 4o2, 5td, Phayao,
Thailand2 The :hite hus! Jobs tear :as purchased fro# =ongsa:adpudpol Aang
Saphung, 4o2, 5td, 5oei, Thailand2 All che#ical reagents :ere of analytical reagent
grade2 The en6y#es, porcine pancreatin <P0B.;%>, a#yloglucosidase <A0.%C;>,
cellulase <40BB-)>, al!aline protease <P0)-(>, and P8D en6y#e assay !it <P0.BBC>
:ere purchased fro# Sig#a0Aldrich 4he#icals, Inc2 <St2 5ouis, 7D, U2S2A2> and
isoa#ylase :as purchased fro# 7ega6y#e International Ireland 5td2 <Aic!lo:,
Ireland>
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Structure of 6rains
The Jobs tear grains :ere steeped in distilled :ater for ) hr2 The grain :as
sliced in longitudinal and cross0sectional planes, placed on a slide, and then dyed :ith
4ongo red2 The slide :as placed on a stereo #icroscope <i!on S7E0*T, i!on
4orp2, Japan>2 The i#ages :ere captured :ith a color 44F ca#era <#odel 7TG0
(*GBP, 7/IJI, Japan>2
The sliced grain sa#ples :ere placed on a stub and coated :ith gold using an
ion sputtering device <JF4 0BB%/, J/D5 5td2, Japan>2 Then, they :ere e,a#ined :ith
a scanning electron #icroscope <S/7> <JS70()%%, J/D5 5td2, Japan> operated at B%
!G2
Sample Preparation
Jobs tear :hole grain flour :as prepared by dry #illing2 The deger#ed Jobs
tear flour :as obtained fro# #anually deger#ed grains and then dry #illed2 The
Jobs tear starch :as isolated by the #ethod of Puchong!avarin et al2 <*%%;>2 The
cellulase :as added into a slurry of deger#ed flour at p? ; and ;%
o
42 After it :as
centrifuged for ; #in, the pellet :as resuspended in distilled :ater2 Then, the al!aline
protease :as added2 After centrifugation, the supernatant and dar! tailing layer :ere
discarded and the residual pellet :as dried and #illed to yield the starch2 All sa#ples
:ere passed through a (% #esh sieve2
Chemical Composition
The #oisture, protein and ash contents :ere deter#ined according to ADA4
<*%%%>2 The conversion factor of protein :as ;2.2 5ipid content :as analy6ed :ith an
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auto fat e,traction syste# <*%;% So,tec, Foss Tecator, S!Hne lIn, S:eden>2 The
a#ylose content :as deter#ined by Julianos a#ylose0iodine co#ple, #ethod
<BC.C>2 All deter#inations :ere conducted in triplicate2
The starch fractions, :hich are rapidly digested starch <"FS>, slo:ly digested
starch <SFS>, and resistant starch <"S>, of the sa#ples :ere #easured by the #ethods
of /nglyst et al2 <BCC*> :ith slight #odification2 The sa#ple <)%% #g> and guar gu#
<;% #g> :ere suspended in *% #5 of %2B M acetate buffer <p? ;2*>, #i,ed by
vorte,ing, and incubated at $.J4 for ); #in in a sha!ing :ater bath <SA**, Julabo
5abortechni! 879?, Seelbach, 8er#any>2 Then, B2( #5 of a #i,ture of en6y#es
<porcine pancreatin and a#yloglucosidase, B2;&B> :as added2 After *% and B*% #in of
incubation, a %2) #5 aliKuot :as re#oved into - #5 of absolute ethanol, #i,ed :ell
and centrifuged at $,%%%,g for ; #in2 The glucose content in the supernatant :as
deter#ined :ith the P8D en6y#e !it2 The glucose contents at *% and B*% #in :ere
designated 8*% and 8B*% respectively2 The "FS is defined as the glucose released
after *% #in2 The glucose released in the second period <after a further B%% #in
incubation> is defined as SFS2 The "S :as #easured as the starch that re#ained
unhydrolysed after B*% #in of incubation2
Chain&Len!th 0istribution of )mylopectin
Starch <(0. #g> :as dissolved in B% #5 FI :ater and debranched :ith
isoa#ylase <.%% units> in %2%B M sodiu# acetate buffer <p? $2;>2 The starch solution
:as incubated at *.
o
4 overnight and the en6y#e :as inactivated by boiling for ; #in2
The chain0length distribution of a#ylopectin :as analy6ed by a high0perfor#ance
anion e,change chro#atography on a Fione, I4S0$%%% eKuipped :ith a pulsed
a#pero#etric detector <?PA/40PAF> <Fione,, Sunnyvale, 4A, USA>2 The sa#ple
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:as filtered and in@ected onto a *;% , $ ## Fione, 4arboPac PA0*%% colu#n :ith a
;% , $ ## guard colu#n <;% , $ ##>2 The sa#ple :as eluted :ith a gradient
bet:een eluent A <B;% #M sodiu# hydro,ide> and eluent 9 <B;% #M sodiu#
hydro,ide containing ;%% #M sodiu# acetate> at a flo: rate of %2; #5L#in2 The
proportion of eluent 9 :as changed as follo:s& *%M at % #in, )%M at B% #in, ;%M at
*% #in and .%M at ;% #in2
@orpholo!y and Particle Si>e 0istribution of Starch 6ranules
The starch :as coated :ith gold as described for the grains2 The #icrostructure
of the starch granules :as e,a#ined :ith a JS70()%% S/7 <J/D5 5td2, Japan> operated
at *% !G2 The particle si6e distribution :as deter#ined :ith a 7astersi6er S diffraction
particle si6e analy6er <7alvern Instru#ents 5td2, 7alvern, UK> in a :et0cell #ode
using :ater2
A&ray 0iffraction (A0)
The N"F analysis :as perfor#ed on a 9ru!er F;%%;N0ray diffracto#eter
<9ru!er 8#b?, 8er#any> :ith B2;) O 4u K
P
radiation2 The sa#ple :as e,posed to
the N0ray bea# :ith the N0ray generator running at )% !G and )% #A2 The 9raggs
angle <*Q> :as scanned fro# ) R $%
o
2 The relative degree of crystallinity of the sa#ple
:as Kuantitatively esti#ated according to ?er#ans and Aeidinger <BC(B>2
)ntio/idant Properties
Sample extraction. The sa#ple e,tracts for the deter#ination of antio,idant
activities :ere prepared by the #odified #ethod of Tseng et al2 <*%%(>2 Appro,i#ately
B% g of finely ground sa#ple :as e,tracted in B%% #5 of #ethanol on a sha!er at
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*;J4 for *) h2 The e,tract :as filtered through Ahat#an no2) filter paper2 The
residue :as then re0e,tracted :ith t:o additional B%% #5 portions of #ethanol by the
sa#e procedure2 The co#bined filtrate :as rotary evaporated to dryness at $;
o
42 The
dried e,tract :as re0dissolved in #ethanol to a concentration of *% #gL#5 and stored
at )
o
4 for further uses2
Determination of total phenolic content. The total phenolic content <TP4> of
sa#ple :as deter#ined using the Folin04iocalteu assay <7arinova et al *%%;>2 A %2*
#5 of the e,tract or standard solution of gallic acid <*%0*%% #gLl> :as added to test
tubes containing B2- #5 of distilled deioni6ed <FI> :ater2 A reagent blan! of FI :ater
:as prepared2 Folin0 4iocalteus phenol reagent <%2* #5> :as added to the #i,ture and
sha!en2 After ; #in, * #5 of .M a
*
4D
$
solution :as added to the #i,ture2 The
solution :as diluted to ;#5 :ith FI :ater and #i,ed2 After incubation for C% #in at
*;
o
4, the absorbance at .;% n# :as deter#ined2 The TP4 of sa#ple :as e,pressed as
#g gallic acid eKuivalents <8A/>Lg e,tracted sa#ple2
DPPH radical scavenging ability. The B,BS0diphenyl0*0picrylhydra6yl <FPP?>
radical scavenging ability of sa#ple :as esti#ated follo:ing the #ethod of 4hoi et
al2 <*%%.>2 AliKuots of %2* #M FPP? in #ethanol <%2- #5> :ere #i,ed :ith %2* #5
of the e,tracts2 The #i,tures :ere vigorously sha!en and left to stand for B% #in in
the dar!2 The absorbance at ;B. n# :as #easured against :ater and #ethanol as the
blan! and control respectively2 The scavenging ability :as calculated as follo:s&
scavenging ability <M> T U< of control0 of sa#ple>L of controlV N B%%+ :here
T absorbance of sa#ple or control R absorbance of blan!2
Redcing po!er. The reducing po:er of sa#ple e,tracts :as deter#ined
according to Dyai6u <BC-(>2 The e,tract in #ethanol <*2; #5> :as #i,ed :ith *2; #5
of *%% #7 phosphate buffer <p? (2(> and *2; #5 of BM potassiu# ferricyanide2 The
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#i,ture :as incubated at ;%
o
4 for *% #in2 Then, the *2; #5 of B%M trichloroacetic
acid <:Lv> :as added to the #i,ture and it :as centrifuged at *%%,g for B% #in2 The
supernatant <; #5> :as then #i,ed :ith an eKual volu#e of FI :ater and ferric
chloride solution <%2BM :Lv>2 The absorbance at .%% n# :as #easured against :ater
and #ethanol as the blan! and control respectively2
Coi/enolide )ssay
The coi,enolide content :as deter#ined by #easuring *,$0butanediol
liberated fro# coi,enolide by acid0cataly6ed transesterification according to the
#ethod of =ang et al2 <*%%)> :ith slight #odification2 The crude oil of sa#ple :as
e,tracted in a So,hlet apparatus using diethyl ether according to the ADA4 <*%%%>
#ethod2 The crude oil of sa#ple <B g> :as dissolved in *% #5 of .M <:L:>
#ethanolic ?4l solution, and reflu,ed in a :ater bath at B%%
o
4 for ) h2 The #i,ture
:as cooled and neutrali6ed :ith $%M #ethanolic sodiu# #etho,ide2 The salt :as
then precipitated and filtrated and the filtrate :as rotary evaporated to a final volu#e
of * #5, and it :as #i,ed :ith B,; pentanediol <Sig#a 4he#ical 4o, St2 5ouis, 7D,
USA> as an internal standard2 The concentrated #ethanolic solution :as analy6ed
using a gas chro#atograph eKuipped :ith fla#e ioni6ation detector <Garian 4P$-%%,
Garian, Inc2, 7iddelburg, etherland>2 A 4P.)*%, A4DT fused silica, 4P0select 49
for FA7/ B%% # , %2*; ##, %2*; W# fil# thic!ness colu#n :as used2 The colu#n
te#perature :as progra##ed to increase fro# B%% to *;%
o
4 at ;
o
4L#in2 The in@ector
and detector te#peratures :ere set at *;%
o
42 The coi,enolide content :as calculated
fro# the *,$0butanediol content by co#paring the area under the pea! :ith that of the
internal standard2
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S*ellin! Po*er and Solubility
S:elling po:er and solubility :ere deter#ined according to 5i and 4or!e
<BCCC> :ith #odification2 The sa#ple <%2$ g dry basis> and B; #5 of distilled :ater
:ere added into a centrifuge tube :ith scre: cap2 The centrifuge tube :as heated at
(;, .;, -; and C;
o
4 for $% #in in a sha!ing :ater bath <SA9*%, "ate! Instru#ents
Pty2 5td2, Australia> and then centrifuged at *%%% , g for B; #in2 The clear supernatant
:as re#oved into a pre:eighed dish and dried at B%;
o
4 until a constant :eight :as
obtained2 The s:ollen sedi#ent and dried supernatant :ere :eighed to deter#ine the
s:elling po:er and solubility as follo:s& S:elling po:er <gLg> T Usedi#ent :eight L
<initial sa#ple :eight , <B%%M 0 Msolubility>>V , B%%+ Solubility <M> T Udry
supernatant :eight L initial sa#ple :eightV , B%%
Pastin! Properties
The pasting properties :ere #easured :ith a "apid Gisco Analy6er <"GA,
e:port Scientific, Aarrie:ood, Australia>, according to 5i and 4or!e <BCCC> :ith
slight #odification2 A $ g of sa#ple <db> :as :eighed into a "GA canister and FI
:ater :as added to obtain the total sa#ple :eight of *- g2 The ti#e0te#perature
profile :as& holding for B #in at ;%
o
4, heating to C;
o
4 in -2$ #in, holding for ; #in
C;
o
4, cooling to ;%
o
4 in .2. #in, and holding for * #in at ;%
o
4, :hich :as a standard
profile *2
0ifferential Scannin! Calorimetry (0SC)
The gelatini6ation and retrogradation properties of the sa#ples :ere
investigated by FS4 <FS4 ., Per!in/l#er Inc2, Shelton, 4T, USA>2 The sa#ple <B%
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#g> :as :eighed into a (% Wl stainless steel pan and distilled :ater :as added to
obtain starch0:ater suspension containing .;M :ater2 Indiu# and an e#pty stainless
steel pan :ere used for the standard and reference respectively2 The pans :ere heated
fro# *; to B*;
o
4 at a heating rate of B%
o
4 L #in2 After heating, the sa#ple :as aged at
)
o
4 for ., B), *B, *-, and $C days to #onitor retrogradation2 The retrograded sa#ples
:ere reheated again :ith the sa#e procedure2 The onset te#perature <T
o
>, pea!
te#perature <T
p
>, co#pletion te#perature <T
c
>, gelatini6ation te#perature range <T
c
0
T
o
> and enthalpy <X?> :ere deter#ined :ith Pyris soft:are2
Statistical )nalysis
A co#pletely rando#i6ed design <4"F> :as perfor#ed2 Analysis of variance
<ADGA> :as perfor#ed :ith SPSS version B$ <SPSS Inc2, I5, USA>2 /ach
e,peri#ent :as conducted in triplicate2 The differences bet:een #ean values :ere
established by Funcans #ultiple0range test at the C;M significance level2
esults and 0iscussion
Structural @orpholo!y of Jobs Tear 6rain
The si6e of :hite hus! Jobs tear is slightly bigger than that of the blac! hus!
variety but their shapes are si#ilar <data not sho:n>2 The cross and longitudinal
sections of :hite and blac! hus! Jobs tear grains are sho:n in Figure B2 The !ernel
structural #orphology is si#ilar to that of barley and :heat in that a crease e,ists in
the #iddle of the !ernel2 The ger# is #ostly entrapped in the endosper# and #a!es
up ca. one third of the :hole grain, :hich is Kuite large, co#pared :ith other cereals2
Figure Ba and b illustrates the acrospire and root location in the ger# part2 The S/7
i#age sho:ed that the endosper# and ger# could clearly be distinguished by their
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different structural #orphologies <Figure *a, b>2 A net:or! appeared in the ger# part
that :as li!ely to be protein #atri, <Figure *c, d>, :hile the endosper# e,hibited
starch granules and protein bodies <Figure *e, f>2 It revealed that Jobs tear !ernels
contain a large portion of ger# e#bedded beneath the crease of the seed :here it is
difficult to separate2
Chemical Composition of Jobs Tear
The che#ical co#position of :hole grain flour fro# :hite and blac! hus! of
Jobs tear presented in Table B :as si#ilar to that of Jobs tear fro# 5aos, Gietna#,
and Tai:an <Au et al2, *%%.>2 The protein and lipid contents of blac! hus! Jobs tear
:ere higher than those of the :hite hus! variety but the ash content :as lo:er
<pY%2%;>2 After re#oving the ger#, the lipid and ash contents of the deger#ed Jobs
tear :ere lo:er than those of the :hole grain, de#onstrating that the deger#ing
process :as conducted properly2 ?o:ever, the protein content of the :hole grain :as
si#ilar to that of the deger#ed flour2 This indicated that the protein of Jobs tear
endosper# has a relatively high protein content, :hich is eKually distributed along the
:hole !ernel2
After Jobs tear starch :as isolated, the protein, lipid, and ash of the starch
fro# both cultivars :ere less than BM2 The a#ylose contents of starch fro# :hite
and blac! hus! Jobs tear :ere B%2$) and B.2%BM respectively2 "egarding starch
digested fractions, the slo:ly digested starch <SFS> content of the :hite starch <AS>
<$C2C$M> :as higher than that of the blac! starch <9S> <$(2*%M>, :hereas the
resistant starch <"S> content of the AS <)C2*CM> :as lo:er that of the 9S <;*2%%M>
<pY%2%;>2 The rapidly digested starch <"FS> content of both Jobs tear cultivars :as
not significantly different <B%2.-0BB2-%M> <pZ%2%;>2 As co#pared :ith potato starch,
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the "FS and SFS content of Jobs tear starch :as higher than that of potato starch
:hereas the "S content of Jobs tear starch :as lo:er <data not sho:n>2 This suggests
the structural integrity of Jobs tear starch granule :as :ea!er than that of potato
starch, a 90crystalline type2 This hypothesis :as supported by the result that Jobs tear
starch e,hibited an A0crystalline pattern <data not sho:n> :hich :as less pac!ed than
the 90type starch <Ehang et al2, *%%(>2 In addition, the granular surface of Jobs tear
starch contained pores as sho:n in Figure $, :hich could enhance the en6y#e
accessibility2
@orpholo!y$ Si>e 0istribution$ and A0 of Starch 6ranules
The starch granules of Jobs tears fro# both cultivars :ere round, polyglonal
in shape :ith porous surfaces <Figure $>2 The particle si6e distribution of Jobs tear
starch :as bi#odal <Figure )>2 The s#all granules of :hite and blac! Jobs tear starch
had dia#eters in the range of %2*;0$ and %2*0$ W# respectively and those of the large
granules :ere $0*; and $0$% W# respectively2 The average si6es of granules of :hite
and blac! Jobs tear starch :ere BB2(- and B*2*C W# respectively, so they #ay be
considered to have relatively s#all granules2
The N"F pattern of Jobs tear is si#ilar to that of rice starch <data not sho:n>,
:hich is an A0type starch, a typical pattern of cereal starch2 The relative crystallinities
of :hite and blac! Jobs tear starch :ere *)2;( and **2-(M respectively2
Chain&Len!th 0istribution of )mylopectin
The chain0length distribution of debranched a#ylopectin fro# both cultivars is
sho:n in Figure ;2 The #a,i#u# proportion of chain0length distribution of Jobs tear
:as found at degree of poly#eri6ation <FP> B$0*), :hich :as si#ilar to :heat,
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barley, and other A0type starches <Jane et al2, BCCC>2 The average chain0length
distribution of the AS and 9S :as FP *B2% and *%2- respectively :hich are shorter
than that reported in other A0type starches, such as :heat <*$2$>, barley <*;2.>, and
triticle <*$2-> <Ao and Jane, *%%.>2
)ntio/idant Properties
The TP4 of #ethanolic e,tracts of :hite and blac! hus! Jobs tear are sho:n
in Table *2 The TP4 of the :hite hus! Jobs tear :as less than that of the blac! hus!
one for both :hole grain and deger#ed flours <pY%2%;>2 As co#pared :ith other
cereals, the TP4 of Jobs tear :as lo:er than those of #illet <B$2-. #gLg>, rye <B%2*(
#gLg> and sorghu# <)B2*- #gLg> <"agaee et al2, *%%;> but higher than those of blac!
rice <$2B$ #gLg>, bro:n rice <%2;) #gLg>, barley <%2;% #gLg>, #ungbean <%2); #gLg>,
fo,tail #illet <%2). #gLg>, and proso#illet <%2*C #gLg> <4hoi et al2, *%%.>2 It could be
possible that the TP4 of each sa#ple depended on the type of cereal, cultivar,
co#position and technical assay <7aisuthisa!ul et al2, *%%->2 7oreover, co#parable
TP4 of :hole grain and deger#ed flour :ere observed for the * cultivars <pZ%2%;>2 It
could be #entioned that the phenolic co#pounds of Jobs tear grains are evenly
distributed in both the ger# and endosper# portions2 This could be due to the fact that
the #a@or phenolic co#pounds in the Jobs tear grains #ay have both hydrophilic and
hydrophobic properties in nature that are able to solubili6e both in the ger# and
endosper#, i2e2, coniferyl alcohol, syringic acid, and ferulic acid <Kuo et al2, *%%*>
The FPP? radical scavenging ability of #ethanolic e,tracts of both :hite and
blac! hus! Jobs tear flour :as not significantly different <pZ%2%;> <Table *>2
?o:ever, slightly better activity :as observed for the :hole grain blac! Jobs tear
flour and for the deger#ed flour of both cultivars2 As co#pared :ith other cereals, the
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FPP? radical scavenging ability of Jobs tear :as higher than that of #ungbean,
fo,tail #illet, and proso#illet, but lo:er than that of bro:n rice, blac! rice, sorghu#
and barley <4hoi et al2, *%%.>2 The FPP? radical scavenging ability is a #easure of
the conversion of the DPPH radical <FPP?[> to FPP?0? by an antio,idant giving
either an electron or hydrogen ato#2 Stratil et al2 <*%%.> reported that the total
phenolic content has a positive correlation :ith the free radical scavenging ability in
fruits and cereals2 Dbviously, the TP4 of the blac! hus! cultivar :as slightly higher
<pY%2%;> than that of the :hite one in both :hole and deger#ed flours resulting in
slightly higher FPP? activity2 In addition, it could be considered that the deger#ed
flours had a better antio,idant activity in ter#s of the ability to scavenge free radicals
of FPP? although significant differences :ere not found2 Antio,idant activity of
carbohydrate products could be affected by other co#ponents in the sa#ple as
reported by Ehang et al2 <*%%B>, Ehang et al2 <*%%)> and 4hirinang and Intarapichet
<*%%C> in that antio,idant activity of so#e #ushroo#s not al:ays solely depended on
the TP4 content only but also depended on other co#ponents of the #aterials such as
dietary fiber and a#ino acid contents2
Si#ilar to the results of the FPP? radical scavenging ability, the reducing
po:er of both cultivars of Jobs tear :as not significantly different <pZ%2%;> <Table *>2
4o#pared :ith other cereals, the reducing po:er of Jobs tear flours of both cultivars
:as lo:er than those of sorghu# and blac! rice, but greater than those of bro:n rice,
:hite rice, #ungbean, fo,tail #illet, and proso#illet <4hoi et al2, *%%.>2 Therefore,
:ith regard to reducing po:er, :hole grain and deger#ed flours of :hite and blac!
Jobs tears :ere co#parable2
Coi/enolide Content
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4oi,enolide is one of lipids in Jobs tear2 Jobs tear oil and coi,enolide :ere
reported to provide #any health benefits such as blood circulation i#prove#ent, a
reduction of infla##ation, purulence and pain, anti0cancer, and anti0tu#or <9ao et al2,
*%%;+ Fhara#anada, *%%.>2 The coi,enolide content of :hole grain :hite and blac!
Jobs tear :as %2%* WgLg2 After the deger#ing process, the coi,enolide of both Jobs tear
flour :as not detected due to a #inute content of oil2 4hang and 9ut <BC-.> reported
that the coi,enolide content in Jobs tear :as less than %2*;M2 In addition, so#e
processing steps for brea!ing the structure and e,traction such as #icrobial
fer#entation and supercritical fluid e,traction :ith or :ithout ultrasound #ay be
needed to i#prove the content of coi,enolide <=ang et al2, *%%)+ ?u et al2, *%%.>2
Physicochemical Properties
S*ellin! Po*er and Solubility
The s:elling po:er of Jobs tear flours and starches :as higher :ith
increasing the te#perature as sho:n in Figure (2 9eginning fro# the te#perature of
.;
o
4, the s:elling po:ers of :hole grain and deger#ed :hite Jobs tear flours :ere
higher than those of the blac! ones due to their lo:er protein, lipid and a#ylose
contents2 Protein in grain could enhance the #olecular interactions bet:een protein
and protein or protein and starch by heat2 It #ay obstruct the hydration of :ater and
could reduce or restrict the s:elling volu#e of starch granules <?a#a!er and 8riffin,
BCC$> A#ylose could interact :ith lipid to for# an a#ylose0lipid co#ple, :hich #ay
affect the bonding force :ithin starch granules, conseKuently decreasing the s:elling
volu#e <Adebo:ale et al2, *%%*>2 The s:elling po:er of the deger#ed flour :as
higher than that of the :hole grain flour due to the lo:er lipid content of the
deger#ed flour2 5ipid that surrounded the starch granules could hinder :ater
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penetration into the starch granules2 In contrast, the s:elling po:er of the AS :as
lo:er than that of the 9S2 It #ay be due to the difference in the crystallinity of the
starch of both cultivars as #entioned previously2 The ratio of crystalline to a#orphous
regions influences the interactions of :ater #olecules :ith the hydro,yl groups of
a#ylose and a#ylopectin in these regions+ thus, a higher proportion of the a#orphous
region or lo:er relative crystallinity contributes to a higher s:elling po:er <Tester
and 7orrison, BCC%>2
The solubility of Jobs tear flour and starches is sho:n in Figure .2 The lo:er
solubility of Jobs tear flours #ay be due to the protein0a#ylose co#ple, for#ation in
Jobs tear flour2 According to Po#eran6 <BCCB>, the for#ation of protein0a#ylose
co#ple, in native starch and flour resulted in a decreased s:elling po:er2 The
solubilities of the blac! Jobs tear flours :ere higher than those of the :hite ones due
to the higher content of a#ylose, lipid, and protein in the blac! flours2 "egarding the
:hole grain and deger#ed flours, the solubility of the :hole grain flour :as higher,
probably related to the greater lipid and ash of the :hole grain flour2 Aith increasing
te#peratures, the solubility of Jobs tear starches :as substantially increased up to ;-0
;CM, :hich is relatively high2 This #ight due to the fact that the Jobs tear starch
contained a high a#ount of s#all si6e starch #olecules and so#e big and s#all holes
appeared on the surface of the starch granules2 Upon s:elling, the surface holes #ay
allo: a nu#ber of the s#all starch #olecules to leach out into the :ater2
Pastin! Properties
Table $ illustrates that the "GA pasting profiles of the flours fro# :hite hus!
Jobs tear had a higher pea!, trough, brea!do:n, and final viscosities but lo:er
setbac! and pasting te#peratures than those of the blac! flours2 Since :hite hus!
16
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Jobs tear had the lo:er protein, lipid, and a#ylose contents, these co#ponents
apparently affected gelatini6ation behavior and viscosity2 In addition, the protein
co#ponent of Jobs tear #ight be a factor related to the lo:er s:elling volu#e of
blac! Jobs tear that, in turn, affected pea! viscosity and pasting te#perature <7artin
and Fit6gerald, *%%*>2 The deger#ed flours of both cultivars sho:ed higher pasting
viscosity profiles but lo:er pasting te#peratures than those of the :hole grain flours2
This :as a result of the lo:er lipid content for the deger#ed flour2
The Jobs tear starches had the higher pea! viscosities and lo:er trough
viscosities, final viscosities, and pasting te#peratures co#pared :ith the flours
<pY%2%;>2 This indicated that the interactions :ith the lipid and protein co#ponents
affected starch gelatini6ation and the pasting properties of starch in the flours2
Therefore, a high level of ther#al treat#ent should be e#ployed to coo! Jobs tear
flours, especially for those #ade fro# the blac! cultivar2
6elatini>ation and etro!radation of Starch
The gelatini6ation para#eters of Jobs tear starches are sho:n in Table )2 The
T
p
, T
c
, and T
c
0 T
o
of the AS :ere significantly higher than those of the 9S <pY%2%;>2
Further#ore, the T
p
values of the AS and 9S :ere the sa#e as their pasting
te#peratures2 The X? of the AS :as slightly greater than that of the 9S, correlating
:ith their relative crystallinities2 A starch :ith a higher degree of crystallinity
#aintains structural stability+ conseKuently, the granule is #ore resistant to:ard
gelatini6ation and thus e,hibits a higher transition te#perature2 Song and Jane <*%%%>
reported that a starch :ith a longer branch chain0length developed into large
crystallites, :hich reKuired a higher te#perature to gelatini6e2 Therefore, the slightly
higher relative crystallinity and proportion of longer branch chain0length of the AS
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:as probably responsible for its higher gelatini6ation te#perature co#pared :ith the
9S2
The retrogradation te#peratures of :hite and blac! Jobs tear starch :ere in
the ranges of $C2*0;C2; and )%2%0;*2C
o
4 respectively <Table )>, :hich are si#ilar to
starch fro# #ost sources2 ?o:ever, the ther#al transition of retrogradation of both
starches :as observed on day $C at )
o
4, :hich is considerably slo:er than other A0
type cereal starches, such as :heat, barley, triticle, rice, and :a,y rice, at the sa#e
storage te#perature <Ao and Jane, *%%.+ 5in et al2, *%%B>2 The Jobs tear starches also
had a considerably lo:er setbac! value2 This :as probably related to the lo: a#ylose
content together :ith a higher proportion of short0branch0chain of a#ylopectin2
Conclusions
The ger# of Jobs tear is entrapped in the endosper# beneath the crease and
its si6e is relatively large2 Ahen the ger# :as re#oved, the protein content re#ained
high, indicating deger#ed Jobs tear flour is a good source of protein, especially that
fro# blac! Jobs tear flour2 The antio,idant activities, :hich :ere FPP? radical
scavenging and reducing po:er, and coi,enolide contents for both cultivars and types
of flour :ere not different2 ?o:ever, the TP4 of the blac! hus! Jobs tear :as slightly
higher for both types of flour as co#pared :ith the :hite ones2 The pasting profile of
the :hite Jobs tear flours :as higher but the pasting te#perature :as lo:er2 In
contrast, the gelatini6ation te#perature of the :hite Jobs tear starch :as higher2 The
granular si6e of Jobs tear starch fro# both cultivars :as s#all :ith the average si6e
of B* W#2 The average chain length distribution of its a#ylopectin :as relatively
short co#pared :ith other cereals2 These structural features contributed to the lo:
paste viscosity of Jobs tear starch and its slo: retrogradation2
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448
)c#no*led!ments
The authors :ould li!e to than! Suranaree University of Technology and the Thailand
"esearch Fund for support via 7aster "esearch 8rants <7A8 Aindo: II>2
eferences
Adebo:ale, K2D2, Afolabi, T2A2, and 5a:al, D2S2 <*%%*>2 Isolation, che#ical
#odification and physicoche#ical characteri6ation of 9a#barra groundnut
<"oand#eia sbterranean> starch and flour2 Food 4he#2, .-&$%;0$BB2
Ao, E2 and Jane, J252 <*%%.>2 4haracteri6ation and #odeling of the A0 and 90granule
starches of :heat, triticle and barley2 4arbohydr2 Poly#2, (.&)(0;;2
ADA42 <*%%%>2 Dfficial 7ethods of Analysis. B.
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ed2 Association of Dfficial
Analytical 4he#ists, Inc2, Aashington F42
9ao, =2, =uan, =2, Nia, 52, Jiang, ?2, Au, A2, and Ehang, N2 <*%%;>2 eutral lipid isolated
fro# endosper# of Jobs tears inhibits the gro:th of pancreatic cancer cells via
apoptosis, 8*L7 arrest, and regulation of gene e,pression2 8astroenterol2
?epatol2, *%&B%)(0B%;$2
4hang, ?242, ?uang, =242, and ?ung, A242 <*%%$>2 Antiproliferative and
4he#opreventive effect of adlay seed on lung cancer in vitro and in vivo. J2
Agric2 Food 4he#2, ;B&$(;(0$((%2
4hang, ?272 and 9ut, P2P2?2 <BC-.>2 Phar#acology and Applications of 4hinese
7ateria 7edica2 Golu#e *2 Aorld Scientific& Singapore, .*;p2
4hirinang, P2 and Intarapichet, K2 <*%%C>2 A#ino acids and antio,idant properties of
the oyster #ushroo#s, Plerots ostreats and Plerots sajor-caj2
ScienceAsia, $;& $*(0$$B2
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473
4hoi, =2, Jeong, ?2S2, and 5ee, J2 <*%%.>2 Antio,idant activity of #ethanolic e,tracts
fro# so#e grains consu#ed in Korea2 Food 4he#2, B%$<B>&B$%0B$-2
4hun, K2S2, 4hiang, A2, and Kuo, 7252 <*%%)>2 /ffect of adlay on a6o,y#ethane0
induced colon carcinogenesis in rats2 Food 4he#, To,icol2, )*&B$$C0B$).2
Fhara#anada, S2 <*%%.>2 /ffective use of #ild acting herbs GI2 4oi,0=i =i "en2
4hinese 7ed2 Ti#es2 *<)>& B0$2
/nglyst, ?22, King#an, S272, and 4u##ings, J2?2 <BCC*>2 4lassification and
#easure#ent of nutritionally i#portant starch fractions2 /uro2 J2 4lin2 utr2,
)(&$$0;%2
?a#a!er, 92"2 and 8riffin, G2K2 <BCC$>2 /ffect of disulfide bond0containing protein
on rice starch gelatini6ation and pasting2 4ereal 4he#2, .%&$..0$-%2
?er#ans, P2?2 and Aeidinger, A2 <BC(B>2 Dn the deter#ination of the crystalline
fraction of polyethylenes fro# N0ray diffraction2 7acro#ol2 4he#2, *)&*)0$(2
?u, A2J2, Ehao, S2, 5iang, ?2, \iu T2\2, and 4hen, 82 <*%%.>2 Ultrasound assisted
supercritical fluid e,traction of oil and coi,enolide fro# adlay seed2 Ultrason2
Sonoche#2, B)&*BC0**)2
?ung, A242 and 4hang, ?242 <*%%$>2 7ethanolic e,tract of adlay seed suppresses
4DN0* e,pression of hu#an lung cancer cells via inhibition of gene
transcription2 J2 Agric2 Food 4he#2, ;B&.$$$0 .$$.2
Jane, J2, 4hen, =2=2, 5ee, 52F2, 7cPherson, A2/2, Aong, K2S2, "adosavl@evic, 72, and
Kase#su:an, T2 <BCCC>2 /ffects of a#ylopectin branch chain length and a#ylose
content on the gelatini6ation and pasting properties of starch2 4ereal 4he#2,
.(<;>&(*C0($.2
20
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20
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476
477
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479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
Juliano, 92D2 <BC.C>2 The che#ical basis of grain Kuality2 In& Proceedings of the
:or!shop on 4he#ical Aspects of "ice 8rain \uality, The Inter "ice "es2 Ins2
5os 9anos, 5aguna, Phillippines, ).Cp2
Kuo, 4242, 4hang, A2, 5iu, 82P2, 4hen, =252, 4hang, J2=2, 5ee, 42K2, 5o, J272, ?uang,
S252, Shih, 7242, and Kuo, =2?2 <*%%*>2 *,*
S
0Fiphenyl0B0picrylhydra6yl radical0
scavenging active co#ponents fro# adlay <Coix lachryma-jobi 52 Gar2 #a0
yuen Stapf2> hulls2 J2 Agric2 Food 4he#2, ;%&;-;%0;-;;2
5i, J2 and 4or!e, ?2 <BCCC>2 Physicoche#ical properties of nor#al and :a,y Jobs
tears <Coix lachrymal-jobi 52> starch2 4ereal 4he#2, .(<$>&)B$0)B(2
5in, =2, =eh, A2I, and 5ii, 42 <*%%B>2 4orrelation bet:een starch retrogradation and
:ater #obility :as deter#ined by differential scanning calori#eter <FS4> and
nuclear #agnetic resonance <7">2 4ereal 4he#2, .-&().0(;$2
7aisuthisa!ul, P2, Pasu!b, S2, and "itthiruangde@, P2 <*%%->2 "elationship bet:een
antio,idant properties and che#ical co#position of so#e Thai plants2 J2 Food
4o#pos2 Anal2, *B&**C0*)%2
7arinova, F2, "ibarova, F2, and Atanassova, 72 <*%%;>2 Total phenolics and total
flavonoids in 9ulgarian fruits and vegetables2 J2 Univer2 4he#2 Tech2
7etallurgy2, )%<$>&*;;0*(%2
7artin, 72, and Fit6gerald, 72A2 <*%%*>2 Proteins in rice grains influence coo!ing
properties2 J2 4ereal Sci2, $(&*-;0*C)2
Dyai6u, 72 <BC-(>2 Studies on products of bro:ning reaction& antio,idant activities of
products of bro:ning reaction prepared fro# glucosa#ine2 Jpn2 J2 utr,
))&$%.0$B;2
Po#eran6, =2 <BCCB>2 Functional Properties of Food 4o#ponents2 *
nd
/d2 Acade#ic
Press, =, ;(Cp2
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21
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505
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519
520
521
Punchong!avarin, ?2, Garavinit, S2, and 9ergthaller, A2 <*%%;>2 4o#parative study of
pilot scale rice starch production by an al!aline and an en6y#atic process2
StarchLStIr!e, ;.&B$)0B))2
"agaee, S2, Abdel0Aal, /2S272, and oa#an, 72 <*%%;>2 Antio,idant activity and
nutrient co#position of selected cereals for food use2 Food 4he#2, C-& $*0$-2
Song, =2 and Jane, J2 <*%%%>2 4haracteri6ation of barley starches of :a,y, nor#al and
high a#ylose cultivars2 4arbohydr2 Poly#2, )B& $(;0$..2
Stratil, P2, Kle@dus, 92, and Kub, G2 <*%%.>2 Feter#ination of phenolic co#pounds and
their antio,idant activity in fruits and cereals2 Talanta, .B&B.)B0B.;B2
Tester, "2F2 and 7orrison, A2"2 <BCC%>2 S:elling and gelatini6ation of cereal starches
I2/ffects of a#ylopectin, a#ylose and lipids2 4ereal 4he#2, (.& ;;B0;;.2
Tseng, =2 ?2, =ang, J20?2, 4hang, ?2 52, 5ee, =2052, and 7au, J2052 <*%%(>2 Antio,idant
properties of #ethanolic e,tracts fro# #onascal adlay2 Food 4he#2, C.& $.;0
$-B2
Au, T2T2, 4harles, A2 52, and ?uang, T2E2 <*%%.>2 Feter#ination of the contents of the
#ain bioche#ical co#pounds of adlay <Coix lachrymal-jobi>2 Food 4he#2,
B%)&B;%C0B;B;2
=ang, J2?2, Tseng, =2?2, 4hang, ?252, 5ee, =252, and 7au, J252 <*%%)>2 Storage
stability of #onascal adlay2 Food 4he#2, C%&$%$0$%C2
=u, =2T2, 5u, T2J2, 4hiang, 72T2, and 4hiang, A2 <*%%;>2 Physicoche#ical properties
of :ater0soluble polysaccharide enriched fractions of adlay and their
hypolipide#ic effect in ha#sters2 J2 Food Frug Anal2, B$<)>&$(B0$(.2
Ehang, 82, Ao, E2, and ?a#a!er, 92"2 <*%%(>2 Slo: digestion properties of native
cereal starches2 9io#acro#olecules, .&$*;*0$*;-2
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526
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528
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530
531
532
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536
537
538
539
540
541
542
543
544
545
Ehang, 72, 4heung, P242K2, and Ehang, 52 <*%%B>2 /valuation of #ushroo# dietary
fiber <nonstarch polysaccharides> fro# sclerotia of Plerots tber-regim
<Fries> Singer as a potential antitu#or agent2 J2 Agric2 Food 4he#2,
)C<B%>&;%;C0;%(*2
Ehang, 72, 4heung, P242K2, Ehang, 52, 4hiu, 4072, and Doi, G242/2 <*%%)>2
4arbo,y#ethylated ]0glucans fro# #ushroo# sclerotiu# of Plerots tber-
regim as novel :ater0soluble anti0tu#or agent2 4arbohydr2 Poly#2, ;., $BC0
$*;2
23
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546
547
548
549
550
551
552
553
Fi!ure 5. Cross (a$ b) and lon!itudinal (c$ d$ e$ f) sections of *hite (a$ c$ e) and
blac# (b$ d$ f) hus# Jobs tear #ernels$ as +ie*ed under a stereo
microscope
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24
554
555
556
557
558
559
Fi!ure ;. Scannin! electron micro!raph of lon!itudinal sections of *hite (a$ c$
e) and blac# (b$ d$ f) hus# Jobs tear #ernels. (a$ b" endosperm +s.
!erm$ c$ d" !erm$ e$ f" endosperm)
25
25
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561
562
563
564
Fi!ure 3. Scannin! electron micro!raph of starch !ranules of *hite (a) and
blac# (b) Jobs tear
26
26
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566
567
568
Fi!ure 9. Particle si>e distribution of *hite (a) and blac# (b) Jobs tear starches
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27
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571
Fi!ure .. Chain len!th distribution of *hite and blac# Jobs tear amylopectin
28
28
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573
574
575
Fi!ure <. S*ellin! po*er of *hole !rain and de!ermed flours and starch of Jobs
tear. (B6BF$ *hole !rain *hite Jobs tear flourC B6DF$ *hole !rain
blac# Jobs tear flour$ 06BF$ de!ermed *hite Jobs tear flourC
06DF$ de!ermed blac# Jobs tear flourC BS$ *hite Jobs tear starch$
DS$ blac# Jobs tear starch)
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577
578
579
580
581
582
583
Fi!ure 2. Solubility of *hole !rain and de!ermed flours and starch of Jobs tear.
(B6BF$ *hole !rain *hite Jobs tear flourC B6DF$ *hole !rain
blac# Jobs tear flour$ 06BF$ de!ermed *hite Jobs tear flourC
06DF$ de!ermed blac# Jobs tear flourC BS$ *hite Jobs tear starch$
DS$ blac# Jobs tear starch)
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590
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593
Table 5. Chemical composition of *hole !rain and de!ermed Jobs tear flours
Composition
Bhole !rain flour 0e!ermed flour
Bhite Dlac# Bhite Dlac#
Protein <M,db> B$2;)^%2B*
b
B(2-;^%2*-
a
B$2B-^%2*%
b
B(2)(^%2$$
a
5ipid <M,db> )2-(^%2B)
b
;2$;^%2B(
a
%2CB^%2%*
c
B2%C^%2%B
c
Ash<M,db> B2.)^%2%*
a
B2B.^%2%B
b
%2;C^%2%*
c
%2*$^%2%B
d
a$ b$ c$ d
Different letters !ithin the same ro! indicate a significant difference %p&'.'().
31
31
594
595
596
Table ;. )ntio/idant properties of methanolic e/tracts of *hole !rain and
de!ermed Jobs tear flours
Sample
)ntio/idant Properties
Total Phenolic
content
(m!6)78!)
0PP1 radical
sca+en!in! acti+ity
(:)
educin! po*er
()
2--
)
A8AF
B
.2$$^%2B)
b
;2)%^B2%%
b
*2--^%2*C
A89F
*
.2C$^%2B-
a
(2)C^B2B.
ab
*2;(^%2*-
F8AF
$
.2%;^%2)B
b
(2;(^%2)B
ab
*2(C^%2*$
F89F
)
-2B-^%2$C
a
.2;$^%2-;
a
*2()^%2*$
a$ b
Different letters !ithin the same colmn indicate a significant difference %p&'.'().
*
!hole grain !hite +ob,s tear flor
-
!hole grain blac. +ob,s tear flor
/
degermed !hite +ob,s tear flor
0
degermed blac. +ob,s tear flor
32
32
597
598
599
600
601
602
603
604
Table 3. Pastin! properties of *hole !rain$ de!ermed flours and starches of
Jobs tear
Sample
Pastin! parameter (EF) Pastin!
temperature
(
o
C)
Pea#
+iscosity
Trou!h
+iscosity
Drea#do*n
Final
+iscosity
Setbac#
A8AF
B
;)2*;
c
;%2;%
b
$2.;
e
()2$B
c
B$2-%
d
..2%*
c
A89F
*
BC2.;
e
B-2)*
d
B2$$
f
)B2%-
d
**2(.
c
C$2B*
a
F8AF
$
-(2$B
b
.)2*-
a
B*2%$
c
B%$2B$
a
*C2%$
b
.;2C;
d
F89F
)
)-2;(
d
)$2*%
c
;2$(
d
.C2;%
b
$(2$%
a
--2-.
b
AS
;
B%B2(B
a
0)2*-
f
B%;2-C
a
0B2B.
f
$2BB
f
.$2$%
e
9S
(
.%2C*
b
0%2-(
e
.B2.-
b
.2%(
e
.2C*
e
.B2$$
f
a$ b$ c$ d$ e
Different letters !ithin the same colmn indicate a significant difference %p&'.'().
*
!hole grain !hite +ob,s tear flor
-
!hole grain blac. +ob,s tear flor
/
degermed !hite +ob,s tear flor
0
degermed blac. +ob,s tear flor
(
!hite +ob,s tear starch
1
blac. +ob,s tear starch
33
33
605
606
607
608
609
610
611
612
613
614
615
Table 9 6elatini>ation and retro!radation properties of Jobs tear starches
Starch
Thermal transition parameters
T
o
(
o
C) T
p
(
o
C) T
c
(
o
C) T
c
&T
o
(
o
C) G1(J8!)
6elatini>ation
:hite Jobs tear (-2*-^%2B$ .$2C-^%2..
a
-%2C)^B2.%
a
B*2((^B2(%
a
B;2%%^%2*)
blac! Jobs tear ((2-*^%2** .%2C-^%2%C
b
.(2.)^%2*C
b
C2C*^%2%C
b
B)2**^%2(;
etro!adation at 9
o
C for 2$ 59$ ;5$ and ;4 days
:hite Jobs tear nd nd nd nd nd
blac! Jobs tear nd nd nd nd nd
etro!adation at 9
o
C for 3? days
:hite Jobs tear $C2%*^%2(% ).2%*^B2**
a
;C2)-^B2.%
a
*%2.C^B2)*
a
-2*%^B2;.
blac! Jobs tear )%2*%^%2*% );2.;^%2BB
a
;*2C(^*2)B
b
B*2.(^)2%C
b
B%2B.^$2$$
a$ b
Different letters !ithin the same colmn indicate a significant difference %p&'.'().
nd2 not detectable
34
34
616
617
618
619
620
621