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R*    L Bh% ?J e!  T*    L H(  ?  % R*    L H첸 ?J e   T*   H L 0 eA ? ̙3380___PPT10.j@> 0!0 (    6o H_  6SOLFRIZZO M., PASCALE M., VISCONTI A., AVANTAGGIATO G..76   l  6A"`4J  Z ?"`  RTOXICITY AND ANALYSIS OF MYCOTOXINS AND THEIR OCCURRENCE IN SOME ORGANIC PRODUCTS $S}SgR  jB  BDf>?jB  BDf>?      B  3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  r  ZHvd޽h @ ? ̙33y___PPT10Y+D=' = @B +X w@ (  #  N  ? GPRINCIPAL MYCOTOXINS PRESENT IN CEREALS AND RELEVANT FUNGAL PRODUCERS .HxFff H x8  ,`B   0DfԔ`B   0DfԔ`B   0DfԔ8  Qy    H  ? Q^ ^FUNGAL PRODUCERS    H!  ? :y -F. graminearum, F. culmorum, F. crookwellense.Z.            @`*   H4  ? ? "Aspergillus flavus, A. parasiticus#Z#0     @`#  H86  ? X  %Penicillium verrucosum, A. ochraceus&Z&&     @`3  H?  ? h  F. graminearum, F. culmorumZ@       @`b  HI  ? l  4F. verticillioides (F. moniliforme), F. proliferatum5Z5V         @`  <S  ?>? $Aflatoxins (B1, B2, G1, G2)Z@       H\  ?sm U MYCOTOXIN     N(a  ?W  ~ Ochratoxin A, Z "     Ne  ? j  p Fumonisins, Z     Nk  ?w q Zearalenone, Z     No  ? g  tDeoxynivalenol,Z   Ns  ?^  TCEREAL    BPw  ? w \ corn, wheat  Z     Nz  ?"  *wheat, barley, corn, rye, oats, buckwheat++*   Bx  ?C +  %wheat, corn, barley, oats, rye, spelt.&Z%%   N  ?v   ucorn ,Z    B  ?b Tcorn Z     B Gly  3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`!^l/H  0޽h ? ̙33y___PPT10Y+D=' = @B +{ PE 0 (   : 1  Z xaxa1?+ 6SUMMARY OF IARC EVALUATIONS CONCERNING SOME MYCOTOXINSJ70 f(f 7  2   ` xaxa1?l  aGlobal evaluation 0   3   `d xaxa1?lK Z Mycotoxins 0    5   f xaxa1?1< ZGroup 1:" 0    6   `h xaxa1?: 2 jCarcinogenic to humans$0   7   ` xaxa1?:2 Aflatoxin B1, B2, G1, G20     9   f xaxa1?   Group 2B:F 0333   :   `ĵ xaxa1? n  sPossibly carcinogenic to humans$ 0 3  x ;   ` xaxa1?b.  $Aflatoxin M1 Ochratoxin A Fumonisins^%- 3333B         =   f$ xaxa1?& 1  ZGroup 3:" 0   2 >   ` xaxa1?&   4Not classifiable as to its carcinogenicity to humans$505&    @ ?   ` xaxa1?& 1 :Deoxynivalenol Nivalenol Zearalenone Fusarenone X T2 toxin8;-:/   F  @  %`B A  0DfԔ`B B  0DfԔ`B C  0DfԔ  D  B \ & 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f E  0A"`6DH   0޽h ? ̙33y___PPT10Y+D=' = @B +    `$ (  $z $ 08  C"DDD`0O It is slowly absorbed from the gastrointestinal tract and once reaches the blood it binds to serum albumin It is distributed via the blood, to kidneys, liver, muscle and fat It is transferred to milk in rats, rabbits and humans, minimal in ruminants Its major metabolite is the less toxic ochratoxin  It is excreted in the urine and faeces Its serum half-life varies widely among species, e.g. 24-39h in mice, 55-120h in rats, 72-120h in pigs, 510h in one macaque monkey, 840h in a human volunteer.xfn<<          $  $ HH  s s?8 @OCHRATOXIN A ABSORPTION, DISTRIBUTION, METABOLISM AND EXCRETION<A0h f3f A 8 X- $tIfB  $ 6DfԔXXfB  $ 6DfԔ--  $ B de  3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44 ()  f $ 0A"`/We=H $ 0޽h ? ̙33y___PPT10Y+D=' = @B +x    p,/ (  ,- , s *| ys ENephrotoxic in all mammalian species tested Cytotoxic and carcinogenic effects (mainly at the renal proximal tubule) Sensitivity to nephrotoxicity: pig > rat > mice The biological basis and significance of the renal tumours are unclear (National Toxicology Program, USA 1989) Genotoxic Genotoxicity (induce DNA damage and chromosomal aberrations) both in vitro and in vivo The mechanism of genotoxicity is unclear, and there is no evidence that it is mediated by direct interaction with DNA Embryotoxic and teratogenic in rats and mice Immunosuppressive in a number of species,0x fZ0x Z 0x ZF0x ZV0x ZF,{F' BvV  "   P  G  5  ]   Y      ;  F X-  , M.`B  , 0DfԔXX`B  , 0DfԔ--  , H  s s?K OCHRATOXIN A Toxicity^0 Z(f f(f $f     , B8 \ & 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f , 0A"`6sDH , 0޽h ? ̙33y___PPT10Y+D=' = @B +  *"0(  0: 0 H,  s s?I{ $OCHRATOXIN A OBSERVATIONS IN HUMANSn%0 (f f(f$ff %  0 s *6 $ BIOMARKERS OF EXPOSURE Blood and urine concentrations of OTA are considered a convenient biomarker of exposure during recent weeks EPIDEMIOLOGICAL STUDIES It has been found in human blood samples (mainly in cool temperate climatic areas of the northern hemisphere) and in human milk in Europe (up to 6.6 ng/ml) and Africa (up to 337 ng/ml) No cases of acute intoxication in humans It is a possible determinant of urinary tract tumors and Balkan endemic nephropathy (BEN) Epidemiological and clinical data available do not provide a basis for calculating the carcinogenicity in humans and the etiology of BEN may involve other nephrotoxic agentsx Pl8 P(8 F<x P((x Z((MK  ft0      5    F X- 0 M^`B 0 0DfԔXX`B 0 0DfԔ--H 0 0޽h ? ̙33y___PPT10Y+D=' = @B +l kk~(7k(  4@ 4 H B  s s? &OCHRATOXIN A OCCURRENCE IN HUMAN BLOODr 0 <0 (f f(ff ' j  ) (7 #"^R)  4 Tp_ __ ?E q1 - 100$x2   @` 4 Tha __ ? 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E s 1138/1222$ x2    @` ?4 T01__ ?  |1994  95$ x2    @` @4 T9__ ?  yCzech Republic $x2   @` A4 TA__ ?Ep |0.1  1.3$ x2    @` B4 TJ__ ? pE p35/143$x2   @` C4 TX__ ?p  n1990$x2   @` D4 T`__ ? p xCzechoslovakia$x2   @` E4 T\__ ?Ep |0.27  35$ x2    @` F4 Tj__ ? Ep q207/303$x2   @` G4 Tc__ ? p |1988  94$ x2    @` H4 T __ ? p pCanada$x2   @` I4 T܈___ ?E v 1  35$x2   @` J4 T___ ? E p82/576$x2   @` K4 T(___ ?  |1984  90$ x2    @` L4 T___ ?  rBulgaria$ x2    @` M4 T__ ?E)  Range (ng/ml)"F&    @` N4 T<__ ? )E Positive/analysed"F&     @` O4 T__ ?)  lYear"F   @` P4 T<__ ? ) oCountry"F   @`B Q4 Z__f1 ? ))B R4 Z__f1 ? B S4 Z__f1 ? ) B T4 Z__f1 ?)B =5 Z__f1 ? )E)B >5 Z__f1 ?) )B 5 Z__f1 ?  B 5 Z__f1 ?E))B 5 Z__f1 ?B 5 Z__f1 ?  pB 5 Z__f1 ?pB 5 Z__f1 ? p B 5 Z__f1 ?pB 5 Z__f1 ?  B 6 Z__f1 ?B 6 Z__f1 ?  B  6 Z__f1 ?B  6 Z__f1 ?  1B 6 Z__f1 ?1B 6 Z__f1 ? 1 B 6 Z__f1 ?1B 6 Z__f1 ?   B !6 Z__f1 ? B #6 Z__f1 ?  B B )6 Z__f1 ? B B +6 Z__f1 ? B  B 16 Z__f1 ?B  B 36 Z__f1 ?   B 96 Z__f1 ?  B ;6 Z__f1 ?  R B A6 Z__f1 ? R B C6 Z__f1 ? R  B I6 Z__f1 ?R  B K6 Z__f1 ?   B Q6 Z__f1 ?  B S6 Z__f1 ?  {B Y6 Z__f1 ? {B [6 Z__f1 ? { 3B a6 Z__f1 ?{3B c6 Z__f1 ? 3 B i6 Z__f1 ?3B k6 Z__f1 ?  B q6 Z__f1 ?B s6 Z__f1 ? B u6 Z__f1 ? EB w6 Z__f1 ?EfB y6 6o ? H 4 0޽h ? ̙33y___PPT10Y+D=' = @B + 8e(  8 8 <  1JOINT FAO/WHO EXPERT COMMITTEE ON FOOD ADDITIVES(20 2f 2  8 N s s?"`j!$ OCHRATOXIN A Provisional Tolerable Weekly Intake PTWI = 0.1 g/kg body weight NOEL = 21 g/kg of body weight per day (rats) Safety factor = 1500< 0 A0 D0 ,  ,  D  F X- 8 (`B 8 0DfԔXX`B 8 0DfԔ--  8 B z 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f 8 0A"`H 8 0޽h ? ̙33y___PPT10Y+D=' = @B + NF D(  D D 0P "`]D<4___PPT9 TFPoor absorption from the digestive tract (d" 4% of the dose) and rapid distribution and elimination Accumulation (as unmetabolized fumonisins) in liver and kidney Little or no evidence of residues in food products derived from animals Little or no evidence of in vitro or in vivo metabolism cpxxx fHt       7 D HŸ s s?. 3FUMONISINS ABSORPTION, DISTRIBUTION AND METABOLISM\40 (f  ff(f 4 ^B D 6DfԔ}^B D 6DfԔ<<  D Bϸ\er & 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  D 0A"`6WeDH D 0޽h ? ̙33  M E  <(  < < H s s?B< iFUMONISINS TOXICITY&0  f   < 04& 3" =High acute and chronic toxicity towards animals Target organs: liver, kidney, heart or homologous organs Leukoencephalomalacia in equids and rabbits; pulmonary edema and hydrothorax in pigs; nephrotoxicity in rabbits and rats; cardiovascular effects in several species; liver and kidney cancer in rodents Biochemical modes of action Disruption of lipid metabolism (sphingolipid metabolism through inhibition of ceramide synthase and metabolism of fatty acids and glycero-phospholipids) both in vitro and in vivo Not mutagenic and not genotoxic both in vitro and in vivo.(:8<0 l    7     !  "              F X- < 6-`B  < 0DfԔXX`B  < 0DfԔ--   < B\;H & 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  < 0A"`6-;DH < 0޽h ? ̙33 @=(  @ @ < 1JOINT FAO/WHO EXPERT COMMITTEE ON FOOD ADDITIVES(20 n2f 2 a @ N48 s s?"`/  FUMONISINS Provisional Tolerable Daily Intake PTDI = 2 g/kg body weight NOEL = 200 g/kg of body weight per day (rodents) Safety factor = 100I0 0 F0     F  F X- @ TI`B @ 0DfԔXX`B @ 0DfԔ--  @ B C\ & 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f @ 0A"`6DH @ 0޽h ? ̙33  7 /  \(  \ \ 0K "` [UD<4___PPT9 rapid appearance and low bioavailability (d"7%) in sheep and cows; higher bioavailability (55%) in pigs main distribution to plasma, kidneys, and liver biotransformation (de-epoxidation and glucuronidation) to less toxic compounds elimination half-life at 3.9h in pigs pPfn<<2" >   !  ?           \ HR s s?|? 6DEOXYNIVALENOL ABSORPTION, DISTRIBUTION AND METABOLISM870  f(f 7 F X- \ `B \ 0DfԔXX`B  \ 0DfԔ--   \ B|M%2 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  \ 0A"`%H \ 0޽h ? ̙3380___PPT10.cp~+#   X#(  XF X- X ]`B X 0DfԔXX`B X 0DfԔ-- X Hl s s?g Deoxynivalenol Toxicity:0 Z(f $f      X Bq  3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  X 0A"` 5  X s *t]h Animal Toxicity reduced growth; decreased feed consumption (anorexia) and emesis (vomiting) are related to increased central serotoninergic activity Inhibition of protein synthesis primary toxic effect Genotoxic Genotoxicity (chromosomal aberrations) both in vitro and in vivo The mechanism of genotoxicity is unclear, and the results are considered equivocal Immunosuppressive in a number of species Embryotoxic and teratogenic in mice^0x fZ0x Z60x fZ0x Z  ,TM~  @  G   _         H X 0޽h ? ̙3380___PPT10.cpl&C  TC(  T6 T s *؆fp BInternational Agency for Research on Cancer (1993) not classifiable as to its carcinogenicity to humans (Group 3). Clinical observations acute toxic effects including nausea, vomiting, abdominal pain, headache, dizziness have been attributed to consumption of contaminated grains (3-93 mg/kg) in Asia no deaths in humans.@3h P@h Ph Ph Ph P34f&N   Pq& T H s s? c &Deoxynivalenol OBSERVATIONS IN HUMANSJ'0 (f$ff  F X- T ]!`B T 0DfԔXX`B T 0DfԔ--   T B{_ ) 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  T 0A"`9{GH T 0޽h ? ̙3380___PPT10.cp`)"N  PN(  P P <` 1JOINT FAO/WHO EXPERT COMMITTEE ON FOOD ADDITIVES(20 2f 2 r P N s s?"`j!$ Deoxynivalenol Provisional Tolerable Daily Intake PTDI = 1 g/kg body weight NOEL = 100 g/kg of body weight per day (mice) Safety factor = 1000 >0 D0 ,  D  F X- P (`B P 0DfԔXX`B P 0DfԔ--   P B z 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  P 0A"`H P 0޽h ? ̙3380___PPT10.bp (  :  6  The use of validated analytical methods is essential to ensure that results of surveys provide a reliable assessment of intake. $g&    B8c  Analytical methods of mycotoxins$! !g"f$ F X-   0Q,`B   0DfԔXX`B   0DfԔ--    B z 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`H  0޽h ? ̙33  ((    6"] vThe working group CEN/TC 275/WG 5   Biotoxins selects and elaborates methods of analysis for mycotoxins that are to become European Standards. The Standards can be used for mycotoxins which are subject to legal limits.r|'''f'L'&%    e  B  ACEN Report CR 13505 Criteria for analytical methods of mycotoxins:B xg$f.efT  F X-  `B   0DfԔXX`B   0DfԔ--    B z 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`H  0޽h ? ̙33* 0j(    6d jPERFORMANCE CHARACTERISTICS OF REPEATABILITY (RSDr), REPRODUCIBILITY (RSDR) AND RECOVERY FOR FUMONISNS k i2ememee efeV-    %   Hd  ?q   LEVEL (g/g)J GgG    H ?g ea  FB1 or FB2` GOGO    T& 33?e Z   < 0.5 0.5  5.04GG 2  T& 33?p *   30 20` GG  2  T & 33?\ s Z   60 30` GG    T|&& 33?[ B  60 - 120 70 - 1104GG    B"    RSDr %P gfofgf&    B/   RSDR %P gfofgf    B84 4  \ Recovery %"  gf  8 Pj `B   0DԔP\ZB   s *D1^ j `B  0DԔP\ZB  s *D1b  B49Dyn (CEN Report CR 13505)$ }gf*     BD= z 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`H  0޽h ? ̙33@ @ (    6pFLE iCEN standard methods $f   NN"` >Ochratoxin A in corn, barley, wheat bran, rye Ochratoxin A in wheat whole meal Aflatoxins in cereals, shell fruits and derived products28n2828P282.8<2   .~  LF X-   0Q#`B   0DfԔXX`B   0DfԔ--    B   1PH     B$|qz ; 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`KlzYF X-   0Q`B   0DfԔXX`B   0DfԔ--H  0޽h ? ̙33G `(    60J mDevelop and validate an analytical method to determine FUMONISINS (FB1 and FB2) in maize and maize based foods at levels of interest for future EU legislation with good recoveries, repeatability and reproducibility.i7'! '!f'!/!'!/!'!A      ,     B [ OBJECTIVE"  g,f      Bqz ; 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`KlzYF X-   H0Qg`B  0DfԔXX`B  0DfԔ--H  0޽h ? ̙33 bZp(    B̞ (The project was designed in two phases:") )'f'    <8  PRELIMINARY STUDY (Ruggedness test) Maize flour, corn flakes, extruded corn, maize-based infant formula, and muffin COLLABORATIVE STUDY Maize flour, corn flakes$ Q 4 4  ''S%'%           q|   Bz e 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f  0A"`ulzF X-  p0Q9`B  0DfԔXX`B  0DfԔ--H  0޽h ? ̙33| ,$(    6*0Jj w}RUGGEDNESS TEST (involving 4 laboratories) Identify critical factors in the procedure of fumonisin analysis by testing different: Extraction solvent mixture compositions Ratios of test portion size/extraction solvent volume Extraction modes Types of clean-up COLLABORATIVE STUDY (involving 24 laboratories) Validation of the optimized method for maize flour and corn flakes<+ 4xY 4} Ff 4s 4x%f%'V'%'%%f%'C'      '/         ( p    B(z  3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`lzF X-  `B  0DfԔXX`B  0DfԔ--H  0޽h ? ̙33   S(    T ?z [RESULTS OF THE RUGGEDNESS-TEST x f  6Lv 'Extraction solvent composition and clean-up were critical factors in the analysis of fumonisins Test portion size, extraction solvent volume and extraction mode had minor or no effect Immunoaffinity column was essential for the purification of corn flakes, muffins and infant formula CH3CN-water (1:1, v/v) gave higher fumonisin recovery than CH3OH-water (3:1, v/v), but gave rise to separation of two phases in several cases# fn fn fn&>P       8XX   Blz ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`nlz|F X-   Y`B   0DfԔXX`B   0DfԔ--H  0޽h ? ̙33[   (  P   (B$CDE\Ff5%$#p YC,       /D[r$04@` _ :<L z z  #  P    B>CDEF&5%=> @`< J z b ;N ? z   ? z B  BCDETF^5% 0@Rbro`P@ 0 " ,0@` J ? d B  BZCDETF^5%  #/:FPZWMC9/%  ,0@`f M d 8N r z   r z "   BCDEDFN5%          $(@`. 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BB 8 3 o  BB 9 3 o  BB : 3 o: ; BB ; 3 o  BB < 3 o  BB = 3 o. / BB > 3 oBB ? 3 oBB @ 3 o"#B A 3 VcPF B <@Ve% W*&   B C 3 VFy  D <PEVOG  W*&   B E 3 V  /B F 3 VA   G <\IV   W*&   B H 3 V` I <hNVTL W*&   BB JB 3 V. . 5 zB K <DV?"D zB L <DV?"Kp zB M <DV?"& zB N <DV?"= p= r zB O <DV?" zB PB <DV?"y" y zB QB <DV?"q q > zB R <DV?"   S H@T ?" R  =50 ml T TVKW ?" J  <10 g U T\[ ?"7Tk YShaking V TV ?"N=" ZBlending   W T7æc ?" T| _CH3OH> X Tdg ?"R 5 >100 ml Y TĦj ?" Tw  <25 g Z TԤ@n ?"VU* ;SAX [ Tq ?""R ;IMA \ H:s ?"G>t _CH3CN>[ ]  `1|x ?":/  FB1 + FB2 recovery (%)`  ^ <[5 c *p < 0.05*    zB _ <Do?"27  c BЃz ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f d 0A"`nlz|H  0޽h ? ̙33y___PPT10Y+D=' = @B +      (  U  C xm m fv @1PS?I SANALYTICAL METHOD FOR THE DETERMINATION OF FUMONISINS IN FIVE MAIZE-BASED MATRICES TxTfooNN  Hؖ ?aR* LSAMPLE PREPARATION(8 -Z  ]$<lB  <DfԔ?-lB  <DfԔ?1ZZw  ZDm m 1? $ mEXTRACT 20g corn material with 50mL acetonitrile:methanol:water (25:25:50, v/v/v) SHAKE for 20 min CENTRIFUGE for 10 min at 4000 rpm FILTER the supernatant through filter paper EXTRACT again the remaining solid material, SHAKE, CENTRIFUGE and FILTER COLLECT the extracts DILUTE extract with PBS solution (10 mL extract + 40 mL PBS) FILTER through microfibre filterRPn2Xn2K  &%    (NN    B8z ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`nlz|H  0޽h ? ̙33y___PPT10Y+D=' = @B +S  zr (    N0 ?  aIMMUNOAFFINITY CLEAN-UP.@8  mG>pxB  HDf>?xB  HDf>?~   `hm m 1?n kELUTE 10 mL of diluted extract through the FumoniTestTM column at 1-2 drop/second WASH with 10 mL PBS at 1-2 drops/second ELUTE fumonisins with 1.5 mL CH3OHH03$  NN  T ? SANALYTICAL METHOD FOR THE DETERMINATION OF FUMONISINS IN FIVE MAIZE-BASED MATRICES TxTf    Bz ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`nlz|H  0޽h ? ̙33y___PPT10Y+D=' = @B +? f ^   (    HX ?n G HPLC ANALYSIS48 1*  I:rB  BDf>?31*1rB  BDf>?2  ZH1?"` EVAPORATE to dryness under nitrogen stream at ca 60C and reconstitute with 200 l of HPLC grade acetonitrile:water (1:1, v/v) DERIVATIZE 50 L extract with 50 L OPA reagent MIX and INJECT at exactly 3 min of adding OPA reagent Column: RP C18 Mobile phase: methanol:0.1M NaH2PO4 (75:25, v/v) adjusted to pH 3.35 with H3PO4 Flow rate: 1.0 mL/min Detector: fluorescence (ex. = 335 nm, em. = 440 nm) Injected amount: 20 L (equivalent to 20 mg of matrix)*Px2 x ((  (    (   &  &  ooNN  B ?v4M RANALYTICAL METHOD FOR THE DETERMINATION OF FUMONISINS IN FIVE MAIZE-BASED MATRICES SxSf    Bt)z ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f   0A"`nlz|H  0޽h ? ̙33y___PPT10Y+D=' = @B +. "..8<-(    C x  11?"`o XRECOVERY AND REPEATABILITY RESULTS OBTAINED IN DIFFERENT FUMONISINS SPIKED CORN MATRICESDYxf>X   B ?a] 1C#RSDr = Relative Standard Deviation (n= 6) * Spiking level 2 g/g zD%P     N0 ?4 u Spiking level(   N4 ?A  ` CORN FLOUR     N9 ?   a CORN FLAKES     N> ?   c EXTRUDED CORN     N? ? jMean (%)  o  )   NhD ?R   RSDr# (%)l ooooo   NI ?O rx jMean (%)  s  )  NM ?  RSDr# (%)l sssss  NR ? jMean (%)      N@< ? |RSD# (%)X   N\ ?0p  <80o  N_ ?   <80s  Nhc ?;{  <94  NHg ?  ;3o  Nj ?A  ;4s  Ne ?e  ;4  Nq ?   ]MUFFINS   Nm ?   dINFANT FORMULA   Nx ?u  `0.5 g/g  o   N| ?  `1.5 g/g  s   N\ ?%_ `4.2 g/g     N ? ^Recovery    B ?   =88 o  N ?   <18o  N ? 0p  <94o   N ?   ;6o ! N ? 0p  Z80o  " N| ?   <10o # NĚ ? t-  Y-o  $ NH ?   ;-o48 |<  <| xB  HD?|<xB   HD?| < rB   BDjJ?|<rB % BDjJ?|< & NĢ ? c  >103 s ' NT ? A  ;3s ( N ?   =109 ) N0 ? e  ;5 * N| ?   <93s + N  ? N  <10s , NX ? |  =79*s - N ? A  ;8s . N ?   <70s / NH ? N  <12s 0 N ? ;{  Z86  1 N  ? e  ;3 2 Np ? ;{  Z92  3 N ? d  ;8 4 NH ? 8  Y-  5 Nt ? T  ;-  : Btz ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  f ; 0A"`nlz|H  0޽h ? ̙33y___PPT10Y+D=' = @B +9 88H@ 7(  H8 ln | Hnl | H BXz ^ 3Consiglio Nazionale delle Ricerche, ISPA-Bari-ITALY44()  n H 0A"`lnz| H <ld^ XEFFECT OF EXTRACTION SOLVENT COMPOSITION AND REPEATED EXTRACTIONS ON FUMONISIN RECOVERY"Y Y7.8 * H*< H # @ T|9 H4\9 "H 6D 9 K0.    #H 6} [  M200.    $H 6x} [  M400.    %H 6}p [  M600.    &H 6}W [  M800.    'H 6T8| N1000.    (H 6T| N1200.    )H 6 T| N1400.    *H 6T|j N1600.    +H 6T|P N1800.   @    }H N 4H S  RT b 8H# :j BB 9H 3 obBB :H 3 obZB ;H s *DovvN H S 5 HB H C oRSHB H C jJ  `B BH 0DoT N H S ̙ =e HB H C o m HB H C o m`B GH 0Do b@    ~Hi N H S  ;  HB H 0DoZ Zu N H S A* HB H C oHB H C ojk`B CH 0DolN  H S ̙ 2 HB H C o aHB H C o aEF`B HH 0Do {{Fb@ L ?  H, ? N  H S L n HB H C oy HB H C jJy `B @H 0Do N ,H S  t r HB .H C o I HB /H C o g I h `B EHB 0Do0 0 j HB H C o  ? @N -H S ̙ x N HB 0H C o  b c`B IH 0Do  A bb@   HN H S   HB  H C oA p HB !H C jJAm pn `B AH 0DoW Wn N 5H S  4 HB 6H C o  HB 7H C o Y Z `B FHB 0Do Z N 1H S ̙>  HB 2H C omRSHB 3H C om  `B JH 0DoU b@  a  Ht " N  H S  ; HB H C o HB H C jJ `B ?H 0Do N  H S B S HB H C oq o pHB H C or 4 5`B DH 0Do r 4N  H S ̙  a HB H C o  3 HB H C o  3 `B KH 0Do   BB NH 3 oBB OH 3 orsBB PH 3 o  BB QH 3 oY Z BB RH 3 o  BB SH 3 o? @ BB TH 3 o  BB UH 3 o' ( BB VH 3 o  BB WH 3 o  BB XH 3 o{ | BB YH 3 oBB ZH 3 oabBB [H 3 oBB \H 3 oIJBB ]H 3 oBB ^H 3 o/0BB _H 3 oBB `H 3 oBB aH 3 o~BB bH 3 o~  BB cH 3 o~  BB dH 3 o~  BB eH 3 o~  BB fH 3 o~{ | BB gH 3 o~abBB hH 3 o~IJBB iH 3 o~/0BB jH 3 o~BB kH 3 oZB lH s *Do/; nH B'  FB1 + FB2 content (g/kg)j    H Z.1Ȝ9? n Corn flour  $ H Z31Ȝ9?N  o Corn flakes  $ H Z$71Ȝ9?N I * s Extruded corn $ H Z$=1Ȝ9?3 . 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F23(F    B?x  F24(F  @  C ?d @  C ?]   B$?t  F24(F    B?q  F24(F    B$C(DEF?#$($ @`D.X  BRC$DEF?"R$R @`L^  BRC%DEF?#R%R @`I[L   c $?HY ! BSC$DEF?"S$S @`FX " B`?6R E0(F  @ # C ?1>o@ $ C ?17h % Bd?9N~ E0(F   & B?6K{ E0(F   ' B؜?2 G500(F  @ ( C ?-q@ ) C ?-k * B?5 G500(F   + B?2~ G500(F   , B4?1 H1000(F  @ - C ?,s@ . C ?,l / B?4 H1000(F   0 Bx?1 H1000(F   1 B?. H1500(F  @ 2 C ?)s@ 3 C