Trace Nutrients Research 26 : 2933(2009)/ 62-$()! Crassostrea gigas "519#,%'&*+$;03$.7!48:"Y » Ã¹1&" ­ Á Í1&" Ï ² Ä1&"¼ _ v 2&"m ­  z2&%1&´ÂLUPNL%j&¨awo("2&l¦hchÅyh»ºyhwo((&Bioavailability of Zinc in a Novel Oyster Extract (Part 2)
Mamiko ABE1&, Yoshikazu MATSUDA1&, Nami KOMURA1&, Kenji FUKUNAGA2&and Munehiro YOSHIDA2&1&
Central Research Institute, Japan Clinic Co., Ltd.,
2&
Laboratory of Food and Nutritional Sciences, Faculty of Chemistry, Materials and Bioengineering, Kansai University

Summary
We previously prepared a new oyster extract (NWZ) from a mixture (WZ) of a hot water extract and a zinc-rich fraction obtained from oysters. In the present study, the availability of NWZ was investigated in comparison with WZ andzinc sulfate. Serum zinc concentration after a single oral zinc administration in rats was examined. Prior to the start ofthe experiment, ten-week-old male Wistar rats were given a zinc-deficient diet for 6 days. The rats were orally administered zinc-deficient diet suspension or one of three experimental diet suspensions containing two types of oyster extracts(NWZ and WZ) or zinc sulfate. A blood sample was taken from the caudal vein before and 1, 2 and 4 hours after the oral
administration. Two hours after administration, rats that received the diet suspension containing zinc as NWZ or WZ
showed a higher serum zinc concentration than those administered the diet suspension containing zinc as zinc sulfate.
The area under the curve at 4 hours (AUC4) calculated from the serum zinc concentration after oral administration ofNWZ was larger than that of those administered WZ or zinc sulfate. It is possible that zinc in NWZ is more absorbable in
the gastrointestinal tract than the zinc in WZ or zinc sulfate. The effects of differences in zinc source on the absorption
and tissue distribution of zinc were examined in rats fed a diet containing various concentrations of sodium phytate. Five-week-old male Wistar rats were given 0, 0.2 or 0.5 % sodium phytate-supplemented diets containing 20 ppm of zinc as
NWZ or zinc sulfate for 29 days. The rats fed a diet higher in sodium phytate showed lower tissue zinc concentrations in
the liver, kidney and tibia. The zinc concentrations in those tissues responded to a decrease in dietary sodium phytate.
Differences in dietary zinc source (NWZ or zinc sulfate) did not significantly influence zinc concentrations in serum orvarious other tissues in rats receiving equal sodium phytate concentrations. The regression line between sodium phytate
in the diet and tibial zinc concentration, the slope was obviously more gradual for rats receiving a diet containing zinc as
NWZ than in those receiving zinc sulfate. It is possible that the zinc in NWZ is more resistant to the influence of sodium
phytate. Zinc solubility was evaluated by pepsin-trypsin digestion in vitro . Zinc in NWZ diet showed higher solubility
than WZ or zinc sulfate diet in pepsin-trypsin digestion. Therefore, zinc in NWZ showed higher absorbability from thegastrointestinal tract than WZ or zinc sulfate. These findings suggest that NWZ has higher zinc bioavailability than WZor zinc sulfate.

SQTV<^ÉÆx="º<Ì"?{G4An¡f½9*ERIMWH±;°Ç26:/"

¨<q¢¾;B78]rH0E!>6"cnÊ%Î|"u"d«&X`·¯.Ë¨RIMW·¯;¤3Eº¨<q¢¾<]r=¦/+!X`<nÊ;Z¢28¬b28,D"X`<^ÉÆx.¬b3E1="[¸;= 30' :+GF8+E."X`<cht:Hgµ263&!

¥"ck³<q¢½"X`<^É¥;B78¿c3
E:+GF8+E!GFGF="TNO;X`:5<1, 2&[À"GFGF=1F>9JK% Crassostrea gigas &<Æx¾HiÈ3E6@;"JK-C¶©½:X`·$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$(}§#p®\ps£e´ª 101%)6168555&((}§#­~ª 3335%)5648680&! 29 !

È%'ZRF(TéÖ7"Êÿ7A2=46(!6OF"OE=C/PB+Q!5ROG·@T"w}î5A 1 mg/kg

C ZRF G¯¬%'x WZ(0OæÒ!B7Ew}

áÇ"9ES>·@î5A 2.5 g/kg áÇFEQN-F"

T«÷íF29QçÉ% NWZ TéÖ7"<G2¨ÒF

Í=ÐF¡â7"regFbvfT8,A©î57@,A¢ñ7A2=!<G=!^vgu%tHw},-·@TÍ=ÐF¡â7="kVdv²î5regF/,A"ÎGZ[çÉ%B+Q NWZ æGw}H"Æ:LGTò6F©î57=LGC7=!î5ÚCî5 1"2"

GZ[çÉ%æGw}M>²w}Fû7AkVdv²G

4 ¸§FregGü×.0O±|TÁ4F33"ÂÒ1«,øÒ1+Q5CTº°2#Ex. 2

7=!

Table 1 Fº7=w},·@F 0"0.2"0.5* GkVd

7=3(!
-£BH"À$GZ[çÉ%æGw}C>²w}G2v²hgsWpTatu%`Gz#Cå7AäëFì¨ÒT"regF/,AÀ$Gw}Tã©î57=C

7= 3 ÀG·@TéÖ7=!5RO 3 ÀGkVdv²

2GÔw}÷íG¸("/NI"4 Äà7A2w},·@Fw}÷í 20 ppm FEQN-Fw}¦TkVdv²Cw}Tî57=C2Gw}G¤04GÂ<ì7=!·@Fì9Qw}¦C7A"Z[çÉ%B+Mw}Gáó&"ED"kVdv²FNQw}Â9Ñ0Q NWZ /NI>²w}T8,=!

OG$¨áÇ 116&132 g G 5 ÄCG Wistar 4reg 36 ýTT¶þF7A¢ñT.=GB*­9Q!

6 ý:@ 6 F&4"Ì 6 ÀG·@Tî57"»3Ø¿)'/0+B 29 õ·y7=!{@ÐHÍ=ÐC7=!E/"NWZ
GoirtÛÓHÌ Table 2 Fº7=C/PB+Q!

1!$%,*.

·yµ 22&29 õ1G 8 õFS=?A'T±¿7A-½¥B´87=Z[çÉ%CH"Z[GöÐçÉ%Cw}÷íTßê7"w}G¤04GÂ<TK=!·y<G³DNPçÉ7=w}÷È%'ZRF(G¯¬%'WZ(Ã?§"Þ"Ù"FÞ"ÏÞ"E®"ÕÜ"/NI"WZ 0OçÉ7=æÒ!G pH B7Ew}Tá/"Ëè"0èT±¿7=!ËèH 8 ð&7"Åô¶è«÷íF29QçÉ%'NWZ(G 2 ÀBGçÉ%B+

FÝï9Qa]qvg 1'Ì#("èFÝï9Qa]qQ!

vg 3'æ#("èFÝï9Qa]qvg 8'#(T´Ô"87=!
2!-.)'
3! In vitro +&('"Ex. 3#

1#Ex. 1
Table 1 Fº9;ùUtlovTcvj\¼¦C7=w

Table 1 Fº7=w},·@F 0"0.25"0.50* GkV}A 0.6 ppm Gw},·@'YsXvctª)¾Ö(dv²hgsWpTatu%`Gz#Cå7AäëF

B"áÇ 280&330 g G 10 ÄCG Wistar 4regT+ì7= 3 ÀG·@TéÖ7=!5RO 3 ÀG·@Fw}

O08K 6 õ·y7=§"4 F&4=!¹F"w}

÷í 200 ppm FEQN-Fw}¦Tì7=!·@Fì,-·@Fw}÷í1 400 ppm FEQN-F"w}¦

9Qw}¦C7AH"Z[çÉ%B+Q NWZ"WZ /Tì7·@TéÖ7=!·@Fì9Qw}¦C7A"NI>²w}T8,=!

Z[çÉ%B+Q NWZ"WZ /NI>²w}T8,=!

Ì 6 ÀG·@ 1 g F"200 mg Gnm_vTJ 0.1 N

E/"NWZ /NI WZ GoirtÛÓH Table 2 Fº7

G~² 100 mL T."37 )B 3 ¸ú6;=!úÃTable 1 Composition of basal zinc-deficient diet

Table 2 Mineral compositions of oyster extracts
Minerals

NWZ

WZ

Ingredient%Egg albumin

20Sodium(g/100 g)

3.6

3.7

Dextrose

63.7Calcium(mg/100 g)

170

210

Corn oil

10Phosphorus(mg/100 g)

680

737

Cellulose powder

2

Potassium(g/100 g)

1.70

2.13

Magnesium(mg/100 g)

280

403

Iron(mg/100 g)

10.5

14.6
9.48

Mineral mix a

3.13

Vitamin mix b

1.17

a

Supplied per 100 g of diet : 17.75 g of NaCl, 34.16 g of K2HPO4,5.28 g of MgSO4, 7.96 g of CaHPO4, 31.79 g of CaCO3, 2.91 g of FeCitrate, 0.084 g of KI, 0.028 g of MnSO4#4H2O, 0.032 g of CuSO4#
5H2O, 0.006 g of CoCl2#6H2O.
bSupplied per 100 g of diet : 85,470 IU of Vitamin A, 10,680 IU ofVitamin D3, 940 mg of Vitamin E, 2.82 mg of Vitamin K3, 85.5 mgof Vitamin B1, 51.3 mg of Vitamin B2, 34.2 mg of Vitamin B6, 0.17
mg of Vitamin B12, 34.2 mg of Biotin, 4.27 mg of Folic acid, 136.8
mg of Calcium pantothenate, 213.7 mg of Nicotinic acid, 12.8 g ofCholine chloride with cellulose powder as a carrier.

! 30 !

Copper(mg/100 g)

8.72

Zinc(mg/100 g)

209

150Manganese(mg/100 g)

5.46

5.59
193Selenium( µg/100 g)

140

Chromium(mg/100 g)

0.08

0.07

Lithium( µg/100 g)

100

139

Vanadium( µg/100 g)

33

10.0

Cadmium(mg/100 g)

0.19

0.14

Oyster extract was manufactured by Japan Clinic Co., Ltd.

Y±"1 N EéÁlks\qéQA pH W 7.6 D&ë

86"5 mg E«ÛkspcxWM 10 mL EksdÁControlZinc sulfate

Þ$50 mM, pH 7.4%W-"37 (A5RD 3 Ç;ÒY±Eç¶ÜQFøP/Då6@ßìB(.DCU6<!
4!/0Zn (μg/mL)

59<!;61"%&$01

NWZ
WZ

42úIF>"BWÚ¡7T<N"Yekx&ãWé&YekxEØDïá6<!7CV="úIJ 0.1 g DïáW00:U;U10 mL-ª2æ_6"1ÇE!±"ÑB+,
+WØD¸><*B"A6@ÅZB6<!2BEö"¬ì"úI.QGç¶ÜQB:EßìF"8ÝÁWR+12Time (h)34Fig!
1 Serum zinc concentration after oral administration of different zinc sources.

@ÊÉ6",Î±"ou#q¯Ã ²²/©A÷*6<!C."Ex. 1 E¬ìy8/E÷*DF"5-Br-PAPS
FWR+< Zn-jdkwb#$^²×KÍí% WÂR6<!
7%

Table 3 The area under the curve (AUC4) of serum zinc concentration

Zinc added2"+.(-/0AUC4 (µg㹺h/mL)

None

5.4 㼳1.1 a<!÷*DFo]mtsxÁËFWR+< ALP `[nZinc sulfate

12.5 㼳2.5 b8%
d$`[ndÍí%WÂR6<!

NWZ

14.8 㼳1.2 bWZ

13.8 㼳2.1 b¬ìYt`so^dgoX#f$ALP%êW÷*6)'#*,

Values are means'SD (n = 6). Values in the same row not sharing
a common superscript differ significantly (p < 0.05).

Ex. 1 D.3TyEþ¨´0O±E¥rikE¬

B6@·-RU<!

ìy8/E¨ÇDW Fig. 1 DÈ6<!bxkv#t¥

Table 4 D"Ex. 2 D.3TÆ|ÒY±E¥riErikE¬ìy8/F"0O'ñ/R 4 Ç±LAE

kEúÓBúÓ 100 g 1<SEóäÓ[WÈ6<!¥AV8/Dß6<0"JK}*A*><!y0O 1

rikEúÓ"úÓõ["ÆZ³V"úÓ 100 g 1<S.QG 2 Ç±D"y0O¥E¬ìy8/W<7TEóäÓ[F7I@E¥D¿F6NRU8"ÆZHEB"NWZ 0O¥B WZ 0O¥0XÁy0O¥QS¹ÿ

oZhxÁ-.QGy°DQT~¢W6NC/><!

WÈ7§µD*><!

L<"ò¥ErikDÆ|#EPÃPGÙ­D.+@{y0O 4 Ç±LAE±E¥rikE¬ìy8/

àW6NC/><!

EDWîC6<£ðHî AUC4 W Table 3 DÈ6<!

NWZ Wy°B6<B1"ÆZ#oZhxÁlks\¬ìyE AUC4 F"XÁy0O¥"WZ 0O¥"NWZ

q8/ 0"0.2"0.5) EB1EyE­/3E0O¥EØDý1+§µ0­RU":U;Ubxkv#t

U;U 43.4"37.0"34.4) A*><!3PD"XÁyW¥E 2.3"2.5"2.7 :A*><!L<"XÁy0O¥D<y°B6<B1F"43.0"38.8"33.3) A*><!y°6@"NWZ 0O¥F 1.2 :"WZ 0O¥F 1.1 :A*S"

EÐ\D¦C2"ÆZ#oZhxÁlks\q8/0¹

NWZ 0O¥.QG WZ 0O¥FXÁy0O¥D<62CTD6<0>@"yE­/3E@ý1+§µD*><!

µ0­RU<!

AUC FúÖ¬#D5><BË[D<]7TEA "ÑVF":ÑVF)7T§

Table 5 D"Ex. 2 D.3T¥rikEóäy8/9%

ú4DÏS¼LU<y[WÈ7Ä=B6@·-RUT!

WLBN<!ö"èö"`»"¬ì"%Ey8/F"

6<0>@"NWZ #EyF WZ #EyPXÁyDy°EÐ\DVR8"ÆZ#oZhxÁlks\q8<6@"ÜD.+@/0¹2CTD6<0>@")7T§µ0­RU<!B

Ñ5UP7+9ê0*TB·-RUT!

2D"ö"èö"`»Ey8/FÆZHEoZhxÁ

WZ F`aE7é$ÕBBy8ÔBA*T ZRF BElks\q-DQTLzC~¢0­RU<!ÆZ#oZ¾ºBA*T!ZRF #EyF#ê>¤E pH AFQhxÁlks\q8/EõBBOD"yE­/3Eê0)2"[_xEâüAùÀ6@+C+4B0Nô5UÑV0PP)6"}@Eóäy8/D~¢0­RU<T10%!4E<ND"WZ #EyE

4B/R"oZhxÁE~¢DQSyESL³ê0)ÑêF NWZ #Ey

D<6@)2C>@+T9ê0·-RUT!;ûD"7T4B06A1<!

NWZ F#ê>¤E pH AQCCA*T<N"NWZ#Ey0ÜD.+@

Ñ5UP72CTTME}?

ÆZ#oZhxÁlks\q8/B`»Ey8/F?
E'ð¦D*TB+VU@+T11%!½EÌ®AO"+! 31 !

Table 4 Body and relative tissue wet weights of rats fed diets supplemented with different zinc sources and sodium phytate levels inEx. 2Tissue

Zinc sulfate

NWZ

Sodium phytate

Sodium phytate

0%292 㼳 15Whole body (g)

0.2 %

0.5 %

0%

0.2 %

0.5 %

308 㼳 3310 㼳 7314 㼳 18302 㼳 9304 㼳 19Liver(g/100 g WB)

3.16 㼳 0.17

3.60 㼳 0.30

3.56 㼳 0.19

3.46 㼳 0.33

3.49 㼳 0.25

3.52 㼳 0.24

Thymus(g/100 g WB)

0.23 㼳 0.03

0.25 㼳 0.03

0.26 㼳 0.04

0.22 㼳 0.02

0.26 㼳 0.02

0.26 㼳 0.04

Spleen(g/100 g WB)

0.16 㼳 0.01

0.16 㼳 0.01

0.18 㼳 0.01

0.17 㼳 0.02

0.16 㼳 0.01

0.17 㼳 0.01

Kidney(g/100 g WB)

0.69 㼳 0.01

0.69 㼳 0.05

0.67 㼳 0.04

0.66 㼳 0.01

0.71 㼳 0.06

0.71 㼳 0.04

Testis(g/100 g WB)

1.01 㼳 0.07

0.99 㼳 0.04

1.01 㼳 0.07

0.94 㼳 0.03

1.00 㼳 0.07

0.98 㼳 0.05

Cecum

(g/100 g WB)

0.21 㼳 0.01

0.21 㼳 0.01

0.21 㼳 0.02

0.20 㼳 0.02

0.22 㼳 0.02

0.21 㼳 0.01

Table 5 Distribution of zinc (µg/g wet weight) in several tissues of rats supplemented with different zinc sources and sodium phytate
levels in Ex. 2Tissue

Zinc sulfate

NWZ

Sodium phytate

Sodium phytate

0%

0.2 %

0.5 %

0%

0.2 %

0.5 %

Liver28.9 㼳 1.9 b

26.8 㼳 1.8 ab

26.0 㼳 1.7 a

28.7 㼳 1.8 ab26.7 㼳 2.4 ab

25.0 㼳 2.8 ab

Thymus20.4 㼳 0.5 a

20.0 㼳 1.0 a

19.8 㼳 0.3 a

20.3 㼳 0.9 a

20.1 㼳 0.4 a

19.7 㼳 0.5 a

Spleen21.1 㼳 1.3 a

20.0 㼳 0.8 a

20.9 㼳 0.7 a

20.7 㼳 1.1 a

20.4 㼳 0.6 a

20.9 㼳 0.7 a

Kidney27.0 㼳 1.0 b

27.4 㼳 1.6 b

23.6 㼳 1.3 a

27.2 㼳 1.6 b

26.9 㼳 1.3 b

28.7 㼳 1.0 a

Tibia

143 㼳 11 b

138 㼳 7 b

104 㼳 4 a

136 㼳 9 b

134 㼳 6 b

116 㼳 6 a

Testis24.6 㼳 0.6 a

24.5 㼳 0.7 a

24.2 㼳 0.6 a

24.4 㼳 1.3 a

23.1 㼳 1.9 a

23.4 㼳 2.4 aSerum *

1.77 㼳 0.21 a

1.74 㼳 0.14 a

1.64 㼳 0.17 a

1.81 㼳 0.14 a

1.73 㼳 0.12 a

1.63 㼳 0.12 aHair **

168 㼳 9 a

169 㼳 5 a

167 㼳 2 a

170 㼳 9 a

172 㼳 5 a

170 㼳 8 a

Duodenum

22.9 㼳 0.7 a

25.7 㼳 1.1 b

23.4 㼳 0.7 a

23.6 㼳 1.2 a

24.4 㼳 1.1 ab23.1 㼳 1.7 a

Jejunum

21.2 㼳 2.3 ab23.2 㼳 2.4 b

19.7 㼳 1.1 a

20.9 㼳 1.7 ab23.1 㼳 1.4 b

20.3 㼳 0.5 ab

Ileum

49.5 㼳 6.7 abc

61.3 㼳 10.4 c

40.8 㼳 6.7 a

50.5 㼳 7.4 bc

53.2 㼳 8.0 ab

43.7 㼳 4.4 a

Cecum

21.8 㼳 1.1 a

22.1 㼳 2.1 a

20.9 㼳 2.7 a

20.5 㼳 2.9 a

22.6 㼳 0.4 a

22.5 㼳 2.0 a

Values are means%SD (n= 6). Values in the same row not sharing a common superscript differ significantly (p < 0.05).
* unit of serum zinc µg/mL dry weight, ** unit of hair zinc µg/g dry weight.

.9+?¥;@"¥\c¸°@Ë«WMR[UTZNX¸°Lºa2"Ì]¦¯>®d3I0<,·CGJ5!NWZ ±Åx<Ê\c±Åx>¥\c¸°?Ä^Table 6 Percentage of soluble zinc in pepsin-trypsin digests ofexperimental diets containing zinc sulfate or oyster
zinc in vitro

Sodium phytate=@·CGJ=+85,"Ë«WMR[UTZNX¸°<Î?\c¸°?&'6?hs­¤?z-@"NWZ

Zinc added

0%

0.25 %

0.50 %±Åx@Ê\c±Åx>»k2:ÃG+>oE+;&H"

Zinc sulfate

51.4

29.0

11.9

NWZ «?\c@WMR[?`wL/>.'f¹¢,

NWZ

67.6

38.1

20.1&I<)GJ5!

WZ56.2

31.5

16.5}£ ALP m¢@\c|Á?¼>=H³I<'(¾,&I,"}£ ALP m¢@ixp>Ä^@·CG

12, 13$J4"\c?ÍEË«WMR[UTZNX¸°>

Percentage of soluble zinc is calculated by dividing supernatantzinc amount by total zinc amount.

?lD0?>'<)GJ5!

`wL/:='0<,j·1J5!

Âv;@"¡e?Æb?£>rBJI\c"3=K7fÆ

t?OP¬½;&I NWZ «?\c?

Ä¢L"YST>*/Iªh±Å"29 µp?±Å

e1J5\c@"¨´;ÉÇf¹=u1JI\c;&I

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L2"Table 6 >25!NWZ «?\c@"Ë5!

«WMR[UTZNX¸°>qy=."WZ «?\c*Ex. 1 >*':"NWZ L{±Å25YST?}£\FAÊ\c>»k2:"fÆ¢\c?l,'z>c¸°?V#Q,D8<D©-."AUC4 D©-+85!

&85!3=K7"fÆ¢\c?l@"NWZ ,D©

9BH"_g¢>25}£\c¸°,D8<D."-."9'; WZ"Ê\c?;&H"uf¹=\c¨n}b«>¶85\cÌD§+850<+G"NWZ! 32 !

¥?\c@uÐ,';)6!

zinc using water-soluble pyridylazo dye. Clin Chim

_Â"XVYZ,fo:u1KJ>@"Ìj29'=/KA=H=' !Ex. 2 >*'9"29 ´pYTU>\14%cMÔActa 120 : 127135.
8%Kind PR, King EJ (1954) Estimation of plasma phos-®Ë3J0;@"¤k}®Ë:&J Ex. 1 M½

phatase by determination of hydrolysed phenol withkzIÁ260;;¯s:&J;)HKJ!0?;-amino-antipyrine. J Clin Pathol 7 : 322326.

Ö?\cµ­, NWZ ®Ë{:'|>&76?@"

9%Matthews JN, Altman DG, Campbell MJ, Royston P

Ex. 3 ?~h+H¿+JG(>"\cMqDÒ,^F(1990) Analysis of serial measurements in medical re-§:f1K"§o:u1KJC:?iª>*'9"gsearch. BMJ 300 : 230235.

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19 : 4346.

(>"Åv:E"PQ¦¾¥?\c;Ñ\c@11%Zhou JR, Fordyce EJ, Raboy V, Dickinson DB, Wong

3B9WNS[>GJul?`yM/6,"rMS, Burns RA, Erdman JW (1992) Reduction of phytic?PQ¦¾:&J NWZ ¥?\c@"Î?PQ¦¾acid in soybean products improves zinc bioavailability

15, 16%¥?\cFÑ\c>·m29"WNS[?`yM/>.."u,."É,'g¶,&J;)in rats. J Nutr 122 : 24662473.
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