Réduction combinée en chlorure de sodium et en matière grasse animale lors de la fabrication du saucisson sec : effets sur les propriétés physicochimiques et les réactions biochimiques en lien avec la production aromatique et les attributs sensoriels Hassan Safa
To cite this version:
Hassan Safa. Réduction combinée en chlorure de sodium et en matière grasse animale lors de la fabrication du saucisson sec : effets sur les propriétés physicochimiques et les réactions biochimiques en lien avec la production aromatique et les attributs sensoriels. Alimentation et Nutrition. Université Blaise Pascal - Clermont-Ferrand II, 2016. Français. NNT : 2016CLF22668. tel-01312196
HAL Id: tel-01312196 https://tel.archives-ouvertes.fr/tel-01312196 Submitted on 4 May 2016
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Mémoire de thèse de Hassan SAFA Page XVII Liste des abréviations, des figures et des tableaux
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Mémoire de thèse de Hassan SAFA Page XVIII
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1 TeRiFiQ: Combining Technologies to achieve significant binary Reductions in Sodium, Fat and Sugar content in everyday foods whilst optimizing their nutritional Quality (Convention de subvention n °289397, www.terifiq.eu)
Mémoire de thèse de Hassan SAFA Page 1 Introduction
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Mémoire de thèse de Hassan SAFA Page 2 Introduction
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Mémoire de thèse de Hassan SAFA Page 3 Introduction
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Mémoire de thèse de Hassan SAFA Page 4 Introduction
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émoire de thèse de Hassan SAFA Page 76 Chapitre 3 : Résultats 60 50 a bc ab c 40 c de cd ef 30 f g 20 gh ij ghi j Experiment S7 (45.4% at day 29) Weight loss of dry-fermented (%) sausages loss of dry-fermented Weight 10 k Experiments S4-S5 (49.0% at day 29) Experiments S1-S2-S3-S6-S8 control (42.9% at day 29) m kl o mn 0 0 7 14 21 28 35 A Time (days) HA> #*6 -%0M#)1! %%% #$&%!#)"!0!1#)1!0*- "!#$%0260*$! %"-%")%)#&$#$". K:+#" 1# " -%!"$" 2%#%M"%,0*$"M&" -% #$!, 260*$!%"-%")0*- "!#$%,%$!#$)%#*# ","!!$%" -% $!."#$) **$%-,% #,!%#00%#)$#0# "$! 2@,Z997A >-!1*'!1%%- !%M $0#*.2!1 "$-P2!%!%@". KKA!1"! 1#)1 #)1!&21#)1 2%#)$#0# "$!00 !%0!#*'." 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" $ $!"!#$0%" !#$!!1*"!#?'"$!1-%!" "%#$*"$"M&" - +",, #!"M&" -% !!10*"!#$0!1 "%%%00*- "!#$%@>#)-K A1 "%% %,$#$)!!1 M%!*"$&" -%0"M'M#!1&"#"!#$%0*9N7 "!"2:! 9 "!"2 N'#%0*.2."! 1% K' V"$ '#!1!11#)1%!6%" !0*- "!#$% $!"#$#$)"! "%!:: W"$#*" 0"!1 "%% %,$#$)!!11#)1%!#$6,- !*"$ "M&" -%'M#!1&"#"!#$%0*9N K"!"2:!9N9:"!"2 N'#%0*.2."! 1% L' 7 Mémoire de thèse de Hassan SAFA Page 78 Chapitre 3 : Résultats "$ ' # !1 !M M%!6" 0*- "!#$% @ 7 "$ A "$ !1 M%!60"! 0*- "!#$ @ $!"#$#$);-%! LW"$#*" 0"!A@ LA1!1# "%%#%0*.2?,#*$!% :"$ M1# 1 $!"#$"*"!" $!$!@ LWA"$"0"# 21#)1@b:LVWA"$#*" 0"! $!$! 1#% "%%,%$!"$#$!*#"!,"!!$#$!*%0!#*6 -%0*"$#$6,- !"M'M#!1 &" -%"$)#$)0*9N 9"!"2:!9 NV"!"2 N!!1"!" !10#$" *"$#$6,- ! "M&" -%". M9N "$!1"!"M&" -%. M9N " $%#%"0#$!*%0 * )M!1 "," #!2@$)1"*!" 99LA 0.980,98 Experiments S1-S2 (0.899 at day 29) a w ab Experiments S4-S5-S6 (0.901 at day 29) 0.960,96 ab bc abc cd Experiments S3-S7-S8 control (0.886 at day 29) cd de e ef ef 0.940,94 f g h hi 0.920,92 ij j k l 0.900,90 Mean value of dry-fermented a sausage of dry-fermented Mean value lm m 0.880,88 0 7 14 21 28 35 B Time (days) HAC #*6 -%0*"$M"!" !#&#!2@"MA&" -% #$&%!#)"!0!1#)1!0*- "!#$% 0 260*$! %"-%")% )#&$ #$ ". K: +#" 1# " -%! "$" 2%#% M"% ,0* $ "M &" -% #$ ! , 260*$! %"-%") 0*- "!#$% ,%$!#$) %#*# " ,"!!$% " -% $! ."#$) **$ %-,% #,!% #00 %#)$#0# "$! 2@,Z997A $"%#*# "M"2!!1!#*6 -%0M#)1! %%'!1%!"!#%!# " "$" 2%%,0*$!1 *"$"M&" -%'# "$-P2!%!%'%1M&21#)1 2%#)$#0# "$!00 !%0!#*' %" ! $!$!"$0"! $!$!@,Z999:A.-!$%#)$#0# "$!00 !0!1#$!" !#$.!M$%" ! "$0"! $!$!@,b997A@". KKA#!"!-"!"$!100 !00"! $!$!$"M&" -% #% #$ $%#%!$! * "-!1% 1"& ,! !1"! - #$) 0"! $!$! -#$) # %"-%") *"$-0" !- !1#)10#$" "M&" -%@" !" 9:LH=*<"$$< 9:KA 1#)1"M&" -%"!!10*$!"!#$%!")@ #&"%!" 9:9AM1"%!1%#$!0#$"$2 %#)$#0# "$!#00$ #$"M&" -%.!M$ M"$1#)160"!#%"-%")%*"$-0" !-M#!1 Mémoire de thèse de Hassan SAFA Page 79 Chapitre 3 : Résultats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"2 VAH!1#% #% ,"$ 2,%#%!"% ,+&" -% #$ "%&!1$?!!MMP%0, %%'.0)M#$)% #)1! 2%!$)-#$)!1 "%! Mémoire de thèse de Hassan SAFA Page 80 Chapitre 3 : Résultats MP0, %%!,"P"!9:7,+-$#!%"!"2 N+M&'" !1%,+&" -%'%, #" 2 !1 M%!"$0#$" &" -%'"0- 2,%$!"!#&0M1"! "%%# " 21",,$%-#$)>$ 1 260*$!%"-%")*"$-0" !-@"%$!" 99VA!!1"!2#$), %%' M "M&" -% "$!1,+&" -%"!0*$!"!#$%!")@"2VA"$"!!1$0#,$#$)@"2 NA.!"#$#$ !1#% %!-2 " $%# $-)1 ! $%- !1 *# .#" %"0!2 0 260*$! %"-%")% @","#*""$ -P"%:NNNA 6,56.5 Experiments S5-S6 (5.25 at day 29) Experiments S1-S2-S3-S4-S7-S8 control (5.40 at day 29) 6,06.0 a 5,55.5 b c c e d Dry-fermented sasuage pH value pH sasuage Dry-fermented 5,05.0 f 4,54.5 0 7 14 21 28 35 C Time (days) HAH #*6 -% 0 *"$ ,+ &" -% #$&%!#)"! 0 !1 #)1! 0*- "!#$% 0 26 0*$!%"-%")%)#&$#$". K:+#" 1# " -%!"$" 2%#%M"%,0*$ "M &" -% #$ ! , 260*$! %"-%") 0*- "!#$% ,%$!#$) %#*# " ,"!!$%" -%$!."#$) **$%-,% #,!%#00%#)$#0# "$! 2@,Z997A "$-P2!%!@". 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"$ "$#*" 0"! - !#$% -#$) !1 *"$-0" !-0260*$!%"-%")% !"0- 2$*" !#*6 -%0,+&" -%'M#!1 %!$)" ##0# "!#$-#$)!10#%!MP0, %%0 M.2",)%%#&#$ "%#$,+ &" - & ' #" -#$) 260*$! %"-%") *"$-0" !-' "$ %, #" 2 -#$) !1 0*$!"!#$ "$ #,$#$) %!")%' !1 *"! ,!#$ "$ #,# *,$$!% -$) $<2*"!# "$ 1*# " *#0# "!#$% 1 $<2*"!# *#0# "!#$% #$& & " 1#0 2 ,! 2%#% "$ #, 2%#% 1%.# 1*# " " !#$%"01#)1#$!%!. "-%!12%1",!1& ,*$!0%$%2 "!!#.-!%0260*$!%"-%")%'%- 1"%!?!-';-# #$%%"$"*" )" ' # >#)-KL 1"!%!1!#*6 -%0,! 2%#%#$? #)1!0*- "!#$%,- !1%"* ,"!!$0.1"&#-#$!*%0!#*6 -% $!10#%!"20, %%',! 2!# $<2* " !#&#!2#%" "2&#%#. 'M1# 1#%3-#! )# " . "-%,! 2!# $<2*%"1#)1 2" !#& "!"!*,"!-0".-! L @+"P-%%!" 9:LA! 2%#%#$?@A"$).!M$ NWMKW"!"2:"$" 1L M7NW"!"2V,$#$)$0*- "!#$>*"2V'"! 0,! 2%#% "%$!# ". 2',.". 2-!"$$6,!#*" ,+&" -0,! 2!# $<2*" !#&#!2@=" #"6="#!" 999A-!!1#$!$% " !# " ##0# "!#$ -#$)"! !1"!,#$!'"%%1M$#$>#)-KK2$"2 :'&2%-,#%#$) 2'&" -% "%0 " 0*- "!#$ "%%%@>#)-KLA .%-!1#% "%M"%$!-!",. *M#!1!10 -% $!! 1$#3--%'M *"%-#$%*%"*, %M#!1!1 "%%# O; 1" *!1 $%#%!#$)#$ " - "!#$)!1 , $!") "!# 0 $$6,!#$ $#!)$ $!$! ! !!" $#!)$ $!$! 1#% "%%# " *"%-*$!*!1 $0#*!1 "%#$ &" -%&!1 "%! MP0!12#$) , %%@"!"$!%1M$A1#%- !#*"! 2!$%!,&!1"!%*$6,- !%0,! 2%#% 1"#%",,"' #P 2 $%-*.2*# )"$#%*%,%$!#$!1260*$!%"-%")%"! !1"!,#$!#$!#*'"$% - $!.! !.2!1*"%-*$!! 1$#3-%-% Mémoire de thèse de Hassan SAFA Page 82 Chapitre 3 : Résultats 8 Experiments S3-S7-S8 control (4.8% at day 29) Experiments S1-S2-S4 (5.2% at day 29) g f Experiments S5-S6 (5.8% at day 28) e 6 e d d c c b 4 a a a 2 Proteolysis index in dry-fermented sausages (%) sausages in dry-fermented index Proteolysis 0 1 7 21 29 Time (days) HAI #*6 -% 0 ,! 2%#% #$? #$&%!#)"! 0 !1 #)1! 0*- "!#$% 0 26 0*$!%"-%")%)#&$#$". 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"$ " % !1 #$!" !#$.!M$%" !"$0"! $!$!@,Z999:A+M&'$*"P00 !00"! $!$! #%&#%#. #$>#)-KL&!1 %%'M1$ *,"#$) ?,#*$!% 7"$ !1"!. $)! !1%"* "%%'0#$6)"#$"$" 2%#%0!1"M%- !%&" !1"!,! 2%#%M"%% #)1! 2 Mémoire de thèse de Hassan SAFA Page 83 Chapitre 3 : Résultats * #$!$% #$ 7 !1"$ ' M#!1 "26 : &" -% 0 W "$ W' %, !#& 2 1#% ,.". 2%- !0*" M0"! $!$!@:: W,:VNWA!1"!,&P"**"P%" ! # -!#$"$!1-%" M%" ! $ $!"!#$#$!1 "$,"!0!1*"!."!!!1"! $!"#$!1 )"!3-"$!#!20 "$,P*"!@#?,#*$! 7A1%%- !%"#$)")*$! M#!1*$!%!" @ 99N' 9: A 1 ,%$! %- !% @>#)- KLA $0#* !1"! ,! 2%#% #$ 260*$! %"-%")% M"% *"#$ 2 )&$ .2 %" ! $!$! .-! " % 1#)1 #)1! !1"! *.#$ %" ! "$ "$#*" 0"! - !#$% - "*, #02 ,! 2!# $<2* " !#&#!2 ' - #$) !1 %" ! "$ 0"! $!$! #$ "%%,! 2%#%'!1#% "$,&!#*$!" !!10#$" !?!-0!1$6,- !% *' # "' ,, 2#$)+$!1" ##!2&" -%@ #, 2%#%A!*#$"!"2 N0" 0*- "!#$% !!10*"!#$0!M#%!#$ ! "%%%"%"0-$ !#$0"$#*" 0"! $!$!)" %%0" $!$! $ "%%, !10-0*- "!#$%0M1# 10"! $!$!M"%"!*%!3-" ! :LVW"$!1!1 "%%, !1:VNW6"$#*" 0"!"$ :W6"$#*" 0"!0*- "!#$% "$ " ##!2&" -M"%%#)$#0# "$! 21#)10!10*- "!#$% $!"#$#$)&:VW"$#*" 0"!'"! N ,V *)O +,)0,- !0"! %1M !1"! %" ! $!$! %#)$#0# "$! 2 "00 ! @, Z 99:A "! 0 #, 2%#% #$ 26 0*$!%"-%")%"$!1"!0"! $!$!M"%"1#)1 2%#)$#0# "$!0" !%!$) 2#*," !#$)"! 0 #, 2%#%@,Z999:A'-$ #P!1#$!" !#$.!M$%" !"$0"! $!$!@,b997A #&"% !" @ 9::A" % #!"%!$)00 !00"! $!$!$ #, 2%#%"$$!1)$"!#$00 0"!!2" #%1%"-!1%,!!1"!% M60*$!%"-%")%#$#!#" 2 $!"#$#$)K9W,P ." P0"! %1M1#)1"!%0 #, 2%#%!1"$#,- !% *"M#!1 M,P." P0"! $!$!@:9W"$ 9WA>-!1*'%&" "-!1%1"&,!"*#$!$% #, 2%#%#$ %" !6- #%"-%")% @ !"1$P:NN7A $ -$ #, 2%#%'-"!"%-))%!%!1"!!1#$!$%#!20!1#%.# 1*# " ,1$*$$ #%%!$) 2,$$!$0"! $!$!"$" %#$0 -$ .2%" ! $!$!'.-!M#!1-!%#)$#0# "$! #$!" !#$.!M$!1%!M0" !%'-$ #P0,! 2%#% )" '' < #,#"$,!#$?#"!#$%!1"! -!1-)1-!!1*"$-0" !-0260*$!%"-%")% "" %#$& &#$!1 "."!#$0!1!"%!'0 "&-"$-0!1%P#$%0# - ,- !%@" !" 9:KA Mémoire de thèse de Hassan SAFA Page 84 Chapitre 3 : Résultats *' < >#)-K7 1"!%!1!#*6 -%0 #,#?#"!#$3-"$!#0#.2!*#$#$)12% -. 1#00."%@+ A&" -%+M&' #,#?#"!#$M"%0#%!*"%-.2!1!1#.".#!-# " #" !#&%-.%!"$ %*!1@ A@"!"$!%1M$A.#,1"%# -&M"%.!"#$0 " #)1! 0*- "!#$%' M#!1 &" - "%#$) M#!1 !#* @*) XP) 0 %"*, A .2$"2:'M1# 1#%$!" )# " ,"!!$"$#$# "!%!1"!!1 3-"$!#0# "!#$*!1 M"%# 6%-#!!" -"!"%%%%*$!0 #,#?#"!#$#$!1,%$!%!-21#%,*,!-% !"%%%% #,#?#"!#$."%$!10 -% $ 0+ 1#00."%%"0*#$*"! -#$) %!") "$ P#$) @="! # ! " 99NA 12 %,$ ! !1 .#$#$) 0 !1 " 12# ,- !%0 #,#,?#"!#$!0"*#$)-,%0,!#$%1, "#!20 1#00 ."%%,$%$!1# *,%#!#$!M"%.%&#$"M*"!!1"!!1"3--%0 -,1% M%,$%#. 0-,!V W0!!" 0 -% $ #$!$%#!2'M1"% P#$)&%!1 , $!")%!" 1K W#$!1, ",1"%"$ LW#$!1", ",1"%@="! #!" 99NA >#)-K7%1M%!1%- !%0+",, #!+ &" -%0" 0*- "!#$%$!"2!!1 *!1@"!"$! %1M$A'+ &" -% #$ "%.%&". 2"% "0-$ !#$0!#*' *"$#$)!1"! #,#?#"!#$ )# " 2#$ "%M#!1!#*,, 2#$)+ ! "%%#02!1 0*- "!#$%#$!!1#%!#$ ! "%%%"%"0-$ !#$00"! $!$!H1#)10"! $!$!*"$% 1#)1 + &" -% "$ !1-% 1#)1 #,# ?#"!#$ 1 0#%! "%% #% 0* 0 !1 !1 0*- "!#$% $!"#$#$)"!*%!:: W"$#*" 0"!@?,#*$!% L' 7"$ VA'!1% $ "%% #% 0* 0 !1 !1 0*- "!#$% $!"#$#$) .!M$ :LVW "$ :VNW "$#*" 0"! @?,#*$!% :' K"$ A'"$!1!1# "%%)"!1!1!M :W6"$#*" 0"!0*- "!#$% @?,#*$!% "$ $! A>#)-K7%1M%!1"!!1%!1 "%%%#00#$!*%0 #,#?#"!#$'%, #" 2"!"2 :.-! %%%"!!1$0,- !2#$)@"2 NA"$$! " 2" 2$#$!1, %%@"2%:"$VA "$ -P2 !%!% @". KKA #$# "! &2 1#)1 2 %#)$#0# "$! 00 !% 0 !#* "$ "$#*" 0"! $!$!@,Z999:A$ #,#?#"!#$.-!$%#)$#0# "$!00 !0%" ! $!$!'$!1 #$!" !#$.!M$%" !"$0"! $!$!@,b997A1%%- !%"#$ #$M#!1#"%!" @ 99NA"$ 2!" @ 997AM1,!!1"! M60"!%"-%"),%$! M #,#?#"!#$ & %!1"$ $! %"-%")% #&"%!" @ 9::A" %,!1#)1 &" -%#$1#)16 0"!%"-%")% *,"! M60"!%"-%")%1 #!"!-#% %% "$!100 !0%" !$ #,#?#"!#$>?"*, '&2 $! 2'" !" @ 9:7A,!1#)1 &" -% "$1#)1".-$"$ 0& "!# *,-$%#&0* #,#?#"!#$#$%" !6- # %"-%")%'!1-% $0#*#$)!1#,&#-%%- !%@" !" 9:KA+M&'8#$!" @ 9:KA Mémoire de thèse de Hassan SAFA Page 85 Chapitre 3 : Résultats ,!!1"!1#)1%" ! & % !1#)1?#"!#$.2 "%#$)"$!#?#"$!$<2*" !#&#!2 "$ "%#$) #$ 0* 1*6.#$#$) ,!#$ >-!1*' " ! " @ 9:7A $! 2 ,!1#)1 #,#?#"!#$ & %#$%" !6"$60"!6- %"-%")% 6 Experiments S4-S5-S7 (4.8 μmolar/kg at day 29) f f Experiments S1-S3-S6 (5.2 μmolar/kg at day 29) e 5 Experiments S2-S8 control (5.2 μmolar/kg at day 29) e d 4 c b b b 3 a a a 2 Hydrosoluble Schiff base content Schiff base content Hydrosoluble (μ molar/kg dry-fermented sausage) (μ molar/kg dry-fermented 1 0 1 7 21 29 A Time (days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émoire de thèse de Hassan SAFA Page 86 Chapitre 3 : Résultats ,!#$?#"!#$M"%0-$0!1!10*- "!#$% $!"#$#$)"!*%!:: W"$#*" 0"! @?,#*$!% L' 7"$ VA$#$!*#"! "%%0?,#*$!% :' K"$ M"%0* 0*!10*- "!#$% $!"#$#$)#!1:LVW"$#*" 0"!:VNW"$#*" 0"!+M&'%#$ $ " 1"$)#$ ,!#$ ?#"!#$ ",,"%M#!1 !#*'.2$"2 : @>#)- K A',!#$ ?#"!#$%*%! -",# 2'*"2.# ! 2-#$)!1*"!."!!,,""!#$'M#!1-! %-.%3-$!#$!$%#0# "!#$ 40 Experiments S4-S5-S7 (28.4 nmol/mg at day 29) Experiments S1-S3-S6 (25.9 nmol/mg at day 29) Experiments S2-S8 control (21.0 nmol/mg at day 29) a b 30 b b b bc bc bc c d d d 20 10 Free-thiol-group content (nmol/mg protein) content Free-thiol-group 0 1 7 21 29 B Time (days) HA? #*6 -%0,!#$?#"!#$" !#$#$&%!#)"!0!1#)1!0*- "!#$%026 0*$!%"-%")%0". K:!#$?#"!#$M"%3-"$!#0#.2!*#$#$)06 !1# 6)-, $!$!+#" 1# " -%!"$" 2%#%M"%,0*$"M&" -%#$ !, 260*$!%"-%")%0*- "!#$%,%$!#$)%#*# ","!!$%>" 1 "%% 0 # %"-%")%' &" -% M *"$% / %!"$" &#"!#$ " - "! 0* " !1 #$,$$!*"%-*$!&" -%0" 1?,#*$! $%!#!-!#$)!1 "%%" -%$! ."#$) **$%-,% #,!%#00%#)$#0# "$! 2@,Z997A #P0 #,#?#"!#$' "$-P2!%!@". 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b997A' M1"%"!!1$02#$)',+&" -%M&2%#)$#0# "$! 2"00 !.20 "&-#$).-!" % .2 & 0"$#*" 0"!@,Z999:A1%0#$#$)%"#$ #$M#!1!1%- !%,!.2" !" @ 9:KA'= ".!!" @ 99KA'=#*$!" @ 99:.A'=-"#"!" @ 99 A'."<!" @:NN7A"$ #&"%!" @ 9:9A'.-!#&)0*!1%- !%0=" «"'*#$)-<'=" &<' "%"%'"$ )"%@ 99 A'#"%!" @ 99NA' "6=" )!" @ 9:KA' -)-<"!" @ 99:A"$","#*""$ -P"%@:NNNAM1#$!0#$%#)$#0# "$!#00$ %.!M$,+ "$X " !# ." !#" -$!%M1$ *,"#$) $! 2%"-%")%M#!1,- !% "."! M#!1,"!#" %-.%!#!-!#$0" M#!1O M60"!6 $!$!%"-%")% Mémoire de thèse de Hassan SAFA Page 109 Chapitre 3 : Résultats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ormulation RF (4.94% at day 29) Formulations Control, NF (4.65% at day 29) Formulations K-RS, SFO-RF (4.90% at day 29) Formulations K-RSF, K-SFO-RSF (4.88% at day 29) 6 h fgh gh def def def efg efg de cde de cd 4 a ab a bc 2 Proteolysis index in dry-fermented (%) sausages in dry-fermented index Proteolysis 0 1 7 21 29 Time (days) HAL #*6 -% 0 ,! 2%#% #$?' #$&%!#)"! #$ !1 %&$ 0*- "!#$% 0 26 0*$!%"-%")%+#" 1# " -%!"$" 2%#%M"%,0*$"M&" -%#$ ! , ,- ! 0*- "!#$% ,%$!#$) %#*# " ,"!!$% > " 1 "%% 0 # %"-%")%'&" -%"*"$%/%!"$"&#"!#$ " - "!0*" !1#$,$$! *"%- &" -% 0 " 1 0*- "!#$ #$ !1"! "%% " -% $! ."#$) **$ %-,% #,!%#00%#)$#0# "$! 2@,Z997A Mémoire de thèse de Hassan SAFA Page 110 Chapitre 3 : Résultats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b997A1%%- !%"#$")*$!M#!1" !" @ 9:KA'."<'F-#$!"$# "'#'%!#"%"$'"$ @:NNVA'"$ #&"%!" @ 9::AM1 0-$1#)1,! 2!# " !#&#!#%#$0"!6- 260*$!%"-%")%"$#$#,- !% *"M#!1,"!#" %-.%!#!-!#$0" M#!1O Mémoire de thèse de Hassan SAFA Page 111 Chapitre 3 : Résultats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b997A1%0#$#$)%"#$ #$M#!1!1%- !%,! .2 #&"%!" @ 9::A"$ 2' !"2'"$< * ) -@ 997A 14 c 12 b 10 8 a 6 4 2 Acidity value (mg KOH/g dry-fermented fat) sausage dry-fermented (mg KOH/g value Acidity 0 Formulations Control, NF, Formulation RF Formulation K-RS (b) K-RSF, SFO-RF, K-SFO-RSF HA>K "! 0 #, 2%#% 3-"$!#0#!1-)1"$ " ##!2&" - "!"2 N+#" 1# " -%! "$" 2%#%M"%,0*$"M&" -%#$!, ,- !0*- "!#$%,%$!#$) %#*# ","!!$%>" 1 "%%0#%"-%")%'&" -%"*"$%/%!"$"&#"!#$ " - "!0*" !1#$,$$!*"%-&" -%0" 10*- "!#$#$!1"! "%% " -%$!."#$) **$%-,% #,!%#00%#)$#0# "$! 2@,Z997A Mémoire de thèse de Hassan SAFA Page 112 Chapitre 3 : Résultats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ormulations RF, K-RSF (3.7 μmolar/kg at day 29) g Formulations Control, NF, K-RS, SFO-RF, K-SFO-RSF (5.1 μmolar/kg at day 29) 5 f 4 e 3 d c b 2 ab a Hydrosoluble Schiff base content Schiff base content Hydrosoluble (μ molar/kg dry-fermented sausage) (μ molar/kg dry-fermented 1 0 1 7 21 29 Time (days) HA>> #*6 -%0 #,#?#"!#$" !#$#$&%!#)"!#$!1%&$0*- "!#$%026 0*$!%"-%")%#,#?#"!#$M"%3-"$!#0#.2!*#$#$)12% -. 1#00 ."%@+ A $!$!+#" 1# " -%!"$" 2%#%M"%,0*$"M&" -%#$ ! , ,- ! 0*- "!#$% ,%$!#$) %#*# " ,"!!$% > " 1 "%% 0 # %"-%")%'&" -%"*"$%/%!"$"&#"!#$ " - "!0*" !1#$,$$! *"%-*$!% 0 " 1 0*- "!#$ #$ !1"! "%% " -% $! ."#$) **$ %-,% #,!%#00%#)$#0# "$! 2@,Z997A Mémoire de thèse de Hassan SAFA Page 113 Chapitre 3 : Résultats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b997A#"%!" @ 99NA0-$ M #,#?#"!#$#$ 0"!6- #%"-%")% !1"-!1%1"&,!%#*# " & %0 #,#?#"!#$M1$ "$#*" 0"!M"%,"!#" 2, " M#!1&)!". # %- 1"% 96K9W #&# @ -)-<"!" ' 99:AV W > @>-$!%' %! &<'$!"$"%'Q$!"$"%' 9:LA2 $!"%!'!1%!-#% ,!!1"!-%#$)&)!". # % - " "! #,#?#"!#$@ &#$#!" ' 99KA>#$" 2' 1M0-$$%#)$#0# "$!00 !0%" !%-.%!#!-!#$' $!"2!" !" @ 9:KA'> %' #!'>"$> %@ 997A"$n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émoire de thèse de Hassan SAFA Page 114 Chapitre 3 : Résultats # ! 2-#$)*"!."!!,,""!#$'"0!M1# 1!1M"%$%-.%3-$!#$!$%#0# "!#$' . "-%!1!1# 6)-,6 $!$!*"%-"!"2:M"%" "2 M!1"$!1"!*"%-#$!1 "M*"!+M&'!1#%,"!# - "!$*"2" %%- !0*+ ,- !#$0* 2%!#$ "$ 2%!#$ .2 *# )"$#%*% %- 1 "% " !# ." !#"' 2"%!% "$ *- % !1"! ?,#$ ?,$$!#" )M!1-#$)!10#%!MP0, %% 40 Formulations RF, K-RSF, SFO-RF, K-SFO-RSF (28.6 nmol/mg at day 29) Formulations Control, NF, K-RS (21.5 nmol/mg at day 29) d c 30 c c b a a a 20 10 Free-thiol-group content (nmol/mg protein) content Free-thiol-group 0 1 7 21 29 Time (days) HA>C #*6 -%0,!#$?#"!#$" !#$#$&%!#)"!#$!1%&$0*- "!#$%026 0*$!%"-%")%!#$?#"!#$M"%3-"$!#0#.2!*#$#$)06!1# 6)-, $!$!+#" 1# " -%!"$" 2%#%M"%,0*$"M&" -%#$!, ,- ! 0*- "!#$% ,%$!#$) %#*# " ,"!!$% > " 1 "%% 0 # %"-%")%' &" -% " *"$% / %!"$" &#"!#$ " - "! 0* " !1 #$,$$! *"%-*$!% 0 " 1 0*- "!#$ #$ !1"! "%% " -% $! ."#$) **$ %-,% #,!%#00%#)$#0# "$! 2@,Z997A "$-P2!%!%@". K A $0#*!1"!,!#$?#"!#$M"%&2%#)$#0# "$! 2 "00 ! .2 0"! $!$! "$ # > $!$! @, Z 999:A' %#)$#0# "$! 2 "00 ! .2 !#* @, Z 99:A' .-! -$"00 ! .2 0 "&-#$)' %" ! $!$! "$ !1 !M #$!" !#$% #$&%!#)"! @, b 997A 1% 0#$#$)% " #$ #$ M#!1 %- !% ,-. #%1 .2 >-$!% ! " @ 9:LA M1 ,!1#)1 #,#?#"!#$"$ M,!#$?#"!#$#$#%"-%")%*"M#!1 # > )"#$) %" ! $!$!' %* "-!1% 1"& ,! !1"! %" ! ! 1#)1 ?#"!#$ .2 #$ "%#$) !1,6?#"$!" !#&#!20#$"$!1%-% ,!#.# #!20*20#.# ! ".$2 "!#$ Mémoire de thèse de Hassan SAFA Page 115 Chapitre 3 : Résultats @<6" 1'=" "$6 &# "'$%,'Q #$" #' 997A'M1"%!1%!-#% 1"&0-$"$"$!#?#"$!00 !0%" !@1' *#!1'Q ':N KA $ # #" 5 %- !%0#$%!-*$!" -*"%-*$!%'# #)1!$%%@cA'$%%@"cA"$2 M$%% @.cA&" -%'",!#$>#)-K:K"!""$" 2%#%00#)-K:K"%1M%!1"!.!10"! & "$ 0 "&-#$) 1" &2 %#)$#0# "$! 00 !% $ #)1!$%% @, Z 999:A 1 :W6"$#*" 0"! 0*- "!#$%@$! '>'"$O6 A%1M1#)1c&" -%!1"$!1!1L0*- "!#$% @>'O6 >' > 6>"$O6 > 6 >A'!1-% $0#*#$),&#-%,!%#$# "!#$)1#)1c &" -%#$1#)160"!%"-%")%@ "6=" )!" ' 9:LA10" !!1"!!11#)1%!c&" -% M0-$#$1#)160"!%"-%")% - ."%%#)$!" M, $!")0M#)1! %%%"$ M$#!%*2) .#$,#)*$! $!$!#$!1%."! 1%@ -P"%!" 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" *"!# )"%% "$#*" ," R1-# !-$% #3- $R"00 !$! ,"% % , %%-% 0*$!"#% 6,56,5 Formulation NF Formulations Control, K-RS Formulations RF, K-RSF, SFO-RF, K-SFO-RSF f 6,06,0 e de de 5,55,5 d c c b Valeur de pH des saucissons Valeur a 5,05,0 4,54,5 0 7 14 21 28 35 Temps (jours) IA? #$ !#3-%,+*%- %"$% %0*- "!#$% "% $% #0".# "!#$% #%! %"$% !". "-K7@,:9 A $&"$ 1' R & -!#$-,+ "0*- "!#$>%"$%"*"!#%"!#$";-! @$#"# ' $#,#&A%!!!" *$!"$*" ,-%%"- #%%$%0*$! %'"& -$,1"%" ##0# "!#$ #$%-00#%"$!"- -% ",*# %*"#$@,+"!!#)$"$!7'7"-.-!-V *;-A!! &" -,+ "!#&*$! & ' "0#$ ",1"%0*$!"!#$',-!!, ;-# #". Mémoire de thèse de Hassan SAFA Page 138 Chapitre 4: Discussion générale 3-"$! " $%&"!#$! "% -#! *# .# )#3--,-#!',-#%3- "0*$!"!#$! " 1-!-,+ $%!#!-$!%."# % $! ", #0 "!#$%*# )"$#%*%R" ! 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"-? &" -% ,+ "%%#3-*$! *%- % "$% % %"- #%%$% % % 0*$! %@#")$ ' 99 A Mémoire de thèse de Hassan SAFA Page 139 Chapitre 4: Discussion générale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b997A% %- !"!%.!$-%%$!!-!0"#!$" "& -? Mémoire de thèse de Hassan SAFA Page 140 Chapitre 4: Discussion générale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émoire de thèse de Hassan SAFA Page 141 Chapitre 4: Discussion générale %-.%!#!-!#$,"!# -% ," O @,Z999:A@>#)-K:9',:: !". "-K ',:9 A! $$,"%," R"*"!#%"!#$' R";-!R1-# !-$% ! %-?#$!" !#$%#$&%!#)- % % %- !"!% $ $!"& -?",,! %," #&"% @ 9::A! 2 @ 997A " *,""#%$ $! % -? %!"! )#% - !#$ @ - !#$ # ! *.#$ ! %-.%!#!-!#$,"!# *.#$ A#$#3-3-' $!"#*$! " - !#$# ! *.#$ 3-# 0#$ % , %%-% #, 2!#3-%' , -$ %-.%!#!-!#$ ,"!# *.#$ ,*! R.!$#%$#&"-? #, 2%#$!#3-% -?*%- %"$% %0*- "!#$%Q $! R @>#)-LNA >C >K G ? 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' 3Z4 "JK "?K IA>H #$ !#3-%,!$,#%*%- %"$% "% "0*- "!#$Q$! R! " 0*- "!#$Q LR!$-% -#!%$% !$*"!# )"%%"$#*" -! !!,!,#%) ." '$-%"&$%, -$ !-%,0# %#$!$% !$-%$"-!$% "$% %-?0*- "!#$%%"- #%%$%- 0"#'!#%%"- #%%$% 1"3-0*- "!#$$! ! , & %L!*,%0".# "!#$#00 $!%8 '87'8:L!8 N $%-#!',"!# 1"3-%"- #%%$'%!"$ 1%$! ! -, %!"$%&%" *$!',-#% 1"3-!"$ 1" ! -, '% $ % 1 *"#$#3- %- "0#)-L:L'0" $0* %"$$"-? $ $!#3-% %,$"$!#00 $!%,0$-%,"",,! "%-0" - Mémoire de thèse de Hassan SAFA Page 155 Chapitre 4: Discussion générale ,-#!' :**' **' K**' **' :9**' :7** ! b:7** - 1"3- "$$"- $ $!#3-' %!$-%$"-!$% '"#$%#3- R"M$! ! *%- % IA>I %#!#$$*$!%, &*$!%% 1"$!# $%%- %!"$ 1%%"- #%%$%,- %#00 $!%*%-% 1#*#3-%!$-%$% !$"-'!*%-%R" !#&#! R"-"0#$!" $%-#! %,0# % 0#$!$#$ *,! ",%#!#$ $%# !%-!-!"0#$,$$ *,! " !" !"!#$-,-#!"- -%-% 1")' %#%!"$ %@9'#!$A,"!#- L--,-#! $! ! 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émoire de thèse de Hassan SAFA Page 177 Références bibliographiques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émoire de thèse de Hassan SAFA Page 178 Références bibliographiques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|#",1#'' % 1$"%'='="#!#%'>"$ !"?,- %'8 @ 997A1"" !#<"!#$0!1*# .#" 0 " 0* " !"#!#$" =P 0*$!%"-%") "! # N@ AK9V6K:V Mémoire de thèse de Hassan SAFA Page 179 Références bibliographiques D-' '> %' "$ '>@ 99LA 00 !0."2*2 %%,,$!1,! 2%#% 0260*$!%"-%")% "! # @ AK:N6K DM#&#'O"$ P*"$$' @:NV7A "!0 "&#!&> #7@LAL V6 7K7 E Ed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émoire de thèse de Hassan SAFA Page 180 Références bibliographiques F %' '#!''>'8 "$> %'8@ 997A 00 !0 " #-* 1 #$!1 & "!# ,"!!$"$%$%2" ,!"$ 0260*$!%"-%")% ->% 1$ :@7A L6 K9 F %' '-' ' " ''Q '>@ 99LA 00 !0."2*2 %%,,$ "*"0*"!#$"$%$%23-" #!20260*$!%"-%")% "! # @KALKN6LL F %' ' "$<'e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e'"&"6 "% ''%!#"%""$'"$$%$"'@ 9:9A $#-*'##$'*)"6K>"$$"!-" "$!#?#"$! 0* #%%" 00# #$" #% " *.#$"!#$ 0 *,$$!% 0* 1" !1# 2 0*$!%"-%")%0*- "!#$ "! # 7@ A VL6 VN G" "6="#'8'F-# %6n"0"''",#"'8'Q-3-"%!' @ 999A !#&#!2 0 "!1,%#$''+"$#$ ,"$#%12 -1"*0$*" "$0 !#&!?!- "! # 7 @:A:M G" «"' '*#$)-<''=" &<' '"%"%'"$ )"%' @ 99 A!# #<"!#$ 0 " "$0-#!0#.%#$ M0"!20*$!%"-%")% "! # 9@KA V6 K Mémoire de thèse de Hassan SAFA Page 181 Références bibliographiques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émoire de thèse de Hassan SAFA Page 182 Références bibliographiques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émoire de thèse de Hassan SAFA Page 183 Références bibliographiques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émoire de thèse de Hassan SAFA Page 184 Références bibliographiques I$)1"*' '-)'',,''Q%#$%P#'8@ 99LA -&#&" 0#%!#"*$ 2!)$% -#$) %!") 0 "26!6"! *"! ,- !% , %% .2 2#$)' 0*$!"!#$' "$X %*P#$)8>! V@: A N M V9 J J#* $<6 *$' >' "." ' 8 "$ 0"%' @ 99:A +" !1# *"! 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M#!1 ,-',6 *- %#0#$ ",%- "!0#%1# "! #N9@:A :6 K K"."P'"$.%$'U@ 9::A$$!- !#$!!1"#!#$" >*$!>% "$&")%0-P2#!&> #7:@KA L 6 9 K"$$'8'+" ' "$8"00'@:NN:A#,#,?#"!#$0*-% 0"%"00 !.2 " 8)# >1*KN:9:VM:9 : L L"P' > U' O*,' 8' >?'8 "$ "$) #' @:N VA 00 !% 0 .$#$) !#*' * 1"$# " !$#<"!#$ "$ ,"!#" , " *$! 0 %#-* 1 # $ !1 3-" #!2 "$ *# 0 "0.$ %%26 -1"*8> #7 K6 L' O' #'' P' '@:NNVA$0 -$ 0%#-* 1 #$"$!#?#"$!$<2* " !#&#!2"$ #,#?#"!#$#$0<$)-$,P "! #L @LAKLN6K77 Mémoire de thèse de Hassan SAFA Page 185 Références bibliographiques L$%,>'""$ "$ 1<'$"$8' !"$+#" >' "!" $ ' $! <6"3- "$ @:N 7A ! #%#% 2 #, #%#% $ " *"-" #$ 1#< #*$!"#": K7:67K L&#$'"$ !"!*"$' @ 99:A ?#"!#&*#0# "!#$0,!#$%-#$)")#$) ?,=$! 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" - "*"%%& -*#3-R-$%"- #%%$'$R#*,! 3- !*,%0".# "!#$',"!# "&" -#$#!#" !!*"%%& -*#3-@ A! "!$-$"-'"-*2$ " "!#$%-#&"$! ݑܽ݁ܥ ଵାಾೄ ߩൌߩιܯܵቌ ೌೠ ቍ ൌ 3@: A ಾೄ ܯܶ ଵାఘιெௌൈ ೌೠ ܺ݁ܽݑ ഐೌೠ &=&%% Food Bioprocess Technol (2015) 8:2109–2122 DOI 10.1007/s11947-015-1563-3 ORIGINAL PAPER Effect of Combined Salt and Animal Fat Reductions on Physicochemical and Biochemical Changes During the Manufacture of Dry-Fermented Sausages Hassan Safa1 & Philippe Gatellier 1 & André Lebert 2 & Laurent Picgirard3 & Pierre-Sylvain Mirade1 Received: 13 April 2015 /Accepted: 13 July 2015 /Published online: 23 July 2015 # Springer Science+Business Media New York 2015 Abstract Reducing salt and fat contents in dry-fermented Keywords Dry-fermented sausage . Combined salt and fat sausages could benefit consumer health. This study aimed to reductions . Physicochemical properties . Proteolysis . quantify, from an experimental design, the effects of salt and Lipolysis . Lipid and protein oxidations fat contents and combined salt and fat reductions on the time- course of several physicochemical (product weight loss, mean water activity and pH values) and biochemical (proteolysis, Introduction lipolysis and lipid and protein oxidations) parameters. Statistical analyses found that time, salt and fat contents had Meat and meat products are very important elements in a very significant impact on weight loss and aw and that time human diet as they are especially rich in proteins, vitamins and salt content (not fat content) had a significant impact on and minerals (Josquin et al. 2012). Dry fermentation and pH. Biochemical results indicated that proteolysis was salt- dry curing are centuries-old techniques of meat preserva- content-dependent and amplified by combined salt and fat tion, and dry-processed meats have become typical, authen- reductions. Intensity of lipolysis was mainly dependent on tic and very popular products across Europe. To illustrate, fat content. Lipid and protein oxidations were more intense Spain produces about 200,000 t of dry-fermented sausage in higher-fat formulations. Combined salt and fat reductions in per year (Martin-Sanchez et al. 2011) while France pro- dry-fermented sausages increased acidification, weight losses duces another 110,000 t (Hoz et al. 2004). Dry-fermented and aw, leading to more proteolysis, less lipolysis and less sausages are made to different recipes using different tech- oxidation. Sensory studies are now required to investigate nologies in different countries and local regions, but all are consumer acceptability of these healthier sausages. generally prepared from raw meats consisting of about two- However, the present results constitute a valuable set of data thirds lean meat and one-third fat. In general, the lean meat for helping professionals wishing to reduce salt and fat con- comes from different animal sources such as pork and beef, tents in dry-fermented sausages. whereas the fat is almost always pig backfat. The term Bdry-fermented sausage^ corresponds to raw meats pre- served under the effect of salting, lactic fermentation and drying. These products are manufactured by mincing raw meat and pork backfat added with 5 % of spices, curing * Pierre-Sylvain Mirade agents—chiefly sodium chloride—additives and starter cul- [email protected] ture. The meat batter thus obtained is pushed into artificial or natural casings before undergoing fermentation, steaming 1 INRA, UR370 Qualité des Produits Animaux, and drying processes (Olivares et al. 2009). 63122 Saint-Genès-Champanelle, France Excessive consumption of salt, especially sodium chloride, 2 Institut Pascal, UMR6602 UBP/CNRS/IFMA, 24 avenue des in human diet is strongly related to increased blood pressure Landais, BP80026, 63171 Aubière Cedex, France and consequently cardiovascular disease (Gelabert et al. 3 ADIV, 10 rue Jacqueline Auriol, ZAC Les Gravanches, 2003), and to certain cancers, including stomach cancer 63039 Clermont-Ferrand Cedex 2, France (Stollewerk et al. 2012). To limit the development of these 2110 Food Bioprocess Technol (2015) 8:2109–2122 human diseases, several health organizations in industrialized Rhee et al. (1983) reported that lipid oxidation is inhibited in countries worldwide recommend limiting salt consumption to ground pork meat prepared with high NaCl concentrations. 6 g per day (e.g., US Department of Health and Human Services Some authors also observed a significant increase of lipid oxida- 2005). These organizations also prone reducing fat intake to less tion in sodium-reduced dry-fermented sausage formulations than 30 % of total calories to limit the development of serious compared to the traditional one, when using calcium chloride health disorders like obesity, high blood cholesterol and coronary to partially replace sodium chloride (Flores et al. 2005; Zanardi heart diseases which are highly related to excessive intake of fat- et al. 2010). Reducing fat content logically reduces the extent of rich foods, especially saturated fatty acids (SFAs) and cholesterol lipolysis and the production of lipid oxidation products. Sanchez- (Ansorena and Astiasarán 2004; Ruiz-Capillas et al. 2012). Zapata et al. (2013) highlighted a positive impact of adding tiger Given that dry-fermented sausages contain high levels of sodium nut fibre as a source of fibre and walnut oil as a source of unsat- and SFA, from a health standpoint, it would be beneficial to urated fatty acids during the manufacture of a traditional Spanish reduce the salt and fat contents in this family of meat products. dry-cured fermented sausage. However, all the above cited stud- However, salt and fat play key functional ingredients in dry- ies focussed on the impact of separate reductions in salt or fat fermented sausage manufacture. For instance, salt ensures micro- content during dry-fermented sausage manufacture. To our biological safety by decreasing water activity (aw), as well as knowledge, few studies have addressed a combined reduction acting on the drying process by decreasing weight losses. It has in salt and fat. also been reported that salt affects biochemical reactions such as Therefore, the objective of this study was to use an exper- proteolysis, lipolysis, oxidation and fermentation that are in- imental design to quantify the effect of salt content, fat content volved in the development of the final sensory attributes of and combined salt and fat reductions on the time-course of dry-fermented sausages, such as texture, flavour and appearance. several physicochemical properties and on the biochemical On the other hand, fat in meat products is considered a valuable reactions that occur during the manufacture of dry-fermented source of energy, fatty acids and vitamins, and biochemical reac- sausages. tions affecting the lipid fraction are also involved in the develop- ment of the final sensory attributes of dry-fermented sausages, such as flavour, juiciness and texture. Several studies have fo- Materials and Methods cussed on the effect of reducing salt or fat content on physico- chemical properties (weight losses, aw and pH) and biochemical Manufacture of Dry-Fermented Sausages reactions (proteolysis, lipolysis and lipid and protein oxidations). For instance, Mora-Gallego et al. (2013) recently reported that To investigate the effects of time, fat content, salt content and salt slows the drying process by affecting the binding water ca- their interactions on physicochemical and biochemical chang- pacity of protein and that reducing fat content accelerates product es during the processing of dry-fermented sausages, a weight losses. Furthermore, Roseiro et al. (2008) showed that Doehlert design (Doehlert 1970) was established on the basis salt-reduced dry-fermented sausages showed higher aw values of two factors: initial animal fat content in the range 8.4–21 % than control batches. Likewise, Olivares et al. (2010)showed and initial salt content in the range 2.0–2.8 %. The first factor, that reducing fat content in dry-fermented sausages led to slightly i.e., fat content, was studied at five levels: 8.4, 11.6, 14.7, 17.9 higher end-of-process aw. The literature on pH is far less consis- and 21.0 %, while the second factor, i.e., salt content, was tent. Corral et al. (2013) did not find significant difference be- studied at three levels: 2.0, 2.4 and 2.8 %. In this study, setting tween pH values of salt-reduced and control dry-fermented sau- up a Doehlert design leads to seven formulations of dry- sages, whereas Roseiro et al. (2008) reported higher fermentation fermented sausages with different amounts of salt and fat, as rates in salt-reduced dry-fermented sausages. With respect to fat indicated in Table 1. We added a control fabrication numbered content, Olivares et al. (2010) found a faster pH decline in low- Beight^ in which dry-fermented sausages were manufactured fat sausages whereas Muguerza et al. (2002) did not find any from a 2.8 % initial salt content and a 21 % initial fat content to effect of fat level on pH value. Other authors have focussed on be used as baseline reference for the other seven fabrications. the effect of salt content and fat content on the biochemical For each formulation in Table 1, about 30 dry-fermented reactions in meat products. Harkouss et al. (2014), for example, sausages were manufactured as per the following procedure. reported higher proteolytic activity when salt is reduced in dry- Raw pork meat, i.e., pork shoulder and pork backfat, was cured ham. Several authors have found higher proteolysis rates in purchased from a local distributor (DISTRIPORC, salt-reduced dry-fermented sausages (Armenteros et al. 2012). Clermont-Ferrand, France). On receipt of the pork meat, we Likewise, the literature reports higher lipolysis rates in dried verified the pH, moisture and water activity (aw) of the pork salted meat products (Armenteros et al. 2012; Quintanilla et al. lean as pH=5.98, moisture=73.5 % and aw=0.97. Pork shoul- 1996; Stahnke 1995). Several studies have reported that NaCl is ders were defatted and cut into small parallelepipeds. For each able to accelerate the oxidation process (Jin et al. 2013), whereas formulation, the corresponding amount of defatted pork other authors find that salt plays an antioxidant role. For example, shoulder and backfat was weighed, ground to 6-mm diameter Food Bioprocess Technol (2015) 8:2109–2122 2111 Table 1 Details of experiments giving all the formulations of dry- Moisture content was expressed on a total matter basis fermented sausages performed in the present study (kg H2O/kg TM). All moisture measurements were performed Experiments Animal fat content (%) Sodium chloride in six replicates. content (%) S1 14.7 2.4 Salt Content S2 21 2.4 S3 17.9 2.8 Salt content was measured using 2 g of powdered-down sam- ple. The sample was homogenized (Ultra-Turrax system, Ika, S4 8.4 2.4 Germany) with 20 mL of ultrapure water. After a 3-h rest S5 11.6 2.0 period, the homogenate was centrifuged at 11,300g for S6 17.9 2.0 10 min at room temperature (MiniSpin Plus, Eppendorf, S7 11.6 2.8 France). The supernatant was recovered, diluted in ultrapure S8 control 21 2.8 water and run through an ion chromatography system (850 This list was built from a Doehlert design on the basis of two factors: professional IC, Metrohm France SAS, France) to systemati- initial animal fat content in the range 8.4–21 % total matter and initial salt cally measure chloride ion and sodium ion contents. The chlo- – content in the range 2.0 2.8 % total matter ride ion or sodium ion values were then used to calculate an equivalent NaCl content (%). All the details on this measure- and mixed with the corresponding amount of salt, a set of ment technique developed in the laboratory can be found in additives and a starter culture corresponding to a mid- Bombrun (2013). All salt content measurements were per- acidification kinetic starter. The starter culture (MF55, formed in six replicates. Biovitec, Lissieu, France) was prepared at 100 kg/L concen- tration and added to each formulation at 10 g/kg. In each meat batter, we added dextrose (5 g/kg), potassium nitrate Fat Content (0.3 g/kg), potassium erythorbate (0.5 g/kg), black pepper (2 g/kg), garlic powder (0.5 g/kg) and finally a solution of Fat content was determined on powdered-down samples starters (10 g/kg). The meat batter was then stuffed into 50- based on the method of Folch et al. (1957) but using mm-diameter collagen casings. The raw sausages, weighing dichloromethane/ethanol (2:1) instead of chloroform/ 450 g and about 20 cm in length, were then plunged in a methanol (2:1) as solvent. Total lipids from 0.5 g of sample Penicillium nalgiovensis solution to cover their surface during were extracted with 50 mL of solvent. The organic phase the drying stage. All products were steamed for 4 days at (dichloromethane) containing total lipids was separated using 24 °C and 70 % relative humidity (RH) and then dried for 10 mL of salt solution at 0.73 %, after centrifugation at 2000g 25 days at 13 °C and 70 % RH in the same ripening room. for 5 min, at 4 °C. The extract obtained was evaporated in a For the eight batches of Table 1, three sausages were taken at vacuum evaporator and weighed to determine total lipid con- days 1, 2, 5, 7, 21 and 29 of drying to evaluate the time-course tent. All fat content measurements were performed in six patterns of chemical composition, physicochemical parameters replicates. (aw, weight loss, pH) and biochemical parameters (proteolysis, fermentation, protein oxidation, lipid oxidation). As the experi- Physicochemical Analysis of Dry-Fermented Sausages ments were cumbersome to set up, the biochemical parameters were ultimately only assessed at four timepoints, i.e., days 1, 7, Weight Loss 21 and 29. In addition, for biochemical and basic chemical anal- yses, all the dried products were individually treated with liquid Throughout the dry-fermented sausage drying period, nine nitrogen, ground down into fine powder to minimize problems products from each batch arranged on the same bar were tied to heterogeneity of sampling in subsequent analysis and weighed together practically every day to determine the kinet- stored at −80 °C until analysis. ics of weight loss. Weight loss was expressed as percentage of initial weight. Chemical Composition of Dry-Fermented Sausages Water Activity (aw) Moisture Wateractivity(aw) was measured at 20 °C with a laboratory Moisture was determined by drying about 1.5 g of powdered- aw meter (aw Sprint TH-500, Novasina, Switzerland). down sample at 80±2 °C in a temperature-controlled chamber Preliminary tests performed to measure aw individually on (Model FT127U, Firlabo, France) until constant weight, the three sausages of each formulation showed that there which took at least 48 h (adapted from Norm NF V 04401). was no significant difference between the three values 2112 Food Bioprocess Technol (2015) 8:2109–2122 (<0.001 aw unit). Water activity values were therefore deter- nm excitation wavelength (excitation slit, 10 nm), a 470-nm mined on a mixture of the three sausages for each formulation. emission wavelength (emission slit, 10 nm) and a 3-s integra- tion time. A linear standard curve of quinine was plotted, and pH sample HSB levels were expressed as micromoles per kilo- gram of meat versus quinine equivalent. All lipid oxidation pH was determined using 1 g of powdered-down sample ho- measurements were performed in nine replicates. mogenized (Ultra-Turrax system, Ika, Germany) with 10 mL of ultrapure water. pH was measured conventionally with a pH Protein Oxidation meter (InLab427, Mettler Toledo, France) calibrated with standard solutions of pH 4 and pH 7. All pH measurements Protein oxidation was assessed in powdered-down samples of were performed in nine replicates. dry-fermented sausages by determining free thiol groups of cysteine residues from Ellman’s assay using 2,2′-dithiobis(5- Biochemical Analysis of Dry-Fermented Sausages nitropyridine) (DTNP) as reagent (Morzel et al. 2006). In al- kaline solution, DTNP binds to the anionic free thiol groups of Proteolysis cysteine residues to form a complex that absorbs at 386 nm. The results are expressed in nanomoles of bound DTNP per The proteolysis index (PI) of each powdered-down sample of milligram of protein. All protein oxidation measurements dry-fermented sausages was determined using a fluorescence were performed in nine replicates. method based on the reaction of fluorescamine with N termi- nal α-amino groups of peptides in TCA soluble fractions Statistical Analyses (Harkouss et al. 2012). In this method, PI is defined as the percentage of the ratio of amino group content to total protein To make the results easier to interpret and the figures easier to content. All PI measurements were performed in nine read, a specific statistical treatment called hierarchical cluster replicates. analysis (HCA) (Chrétien and Szymoniak 1987; Wishart 1969) was applied to all measured raw values. HCA consists Lipolysis in clustering samples that lead to similar results on a given parameter, thereby creating classes of dry-fermented sausages. The degradation of fat into fatty acids was quantified by de- HCA was calculated using Ward’smethodonSTATISTICA termining the acid value of fat (Norm NF T 60–204) in sam- 10-V2014 software. In addition, when a class of sausages is ples of dry-fermented sausages, at day 29. Briefly, total free formed, the values of the parameter corresponding to the class fatty acids from 25 g of sample were solubilized in a solvent are calculated, at each time point, by averaging the values of mix of ether/ethanol. Total free fatty acids were determined all the same-class sausages. quantitatively by potassium hydroxide (0.1 N) in the presence Statistical analyses of the results were completed by anal- of a colour indicator (phenolphthalein). Before neutralization, ysis of variance (ANOVA) using STATISTICA 10-V2014 phenolphthalein (acid medium) is colourless. Under basic software. The objective was to assess the effect of each factor conditions (beyond neutralization), it colours pink. The num- (time, fat content, salt content and salt content × fat content ber of equivalents of potassium hydroxide poured is equal to interaction) on each variable measured in this study. When the number of equivalents of acid present in the sample, and ANOVA found a significant effect (p<0.05), post hoc proce- acid value is the mass of potassium hydroxide, in milligrams, dures were used: Multiple comparisons among means were required to neutralize 1 g of fat. All these measurements were examined by the Tukey test to determine the level of signifi- performed in three replicates. cance between groups. Lipid Oxidation Results and Discussion Lipid oxidation was quantified in powdered-down samples of dry-fermented sausages by determining hydrosoluble Schiff Effects on Chemical Composition of Dry-Fermented bases (HSBs) using the procedure reported in Gatellier et al. Sausages (2009). For this purpose, the aqueous phase obtained to deter- mine fat content was collected to quantify HSB levels. A Drying dry-fermented sausages globally leads to a reduction standard curve of commercial Schiff bases (quinine) was pre- in in-product water content due to water evaporation from the pared in parallel. Fluorescence of each point of the standard product surface and, in turn, to a fat and salt concentration that curve and fluorescence of each sample were measured with a increases fat and salt contents, respectively. We therefore ran spectrofluorometer (FP 8300, Jasco France, France) at a 370- chemical analysis of moisture, salt content and fat content at Food Bioprocess Technol (2015) 8:2109–2122 2113 four timepoints, i.e., day 1, day 7, day 21 and day 29 values of all formulations were lower than the water content (end of drying) to track and trend the time-course of measured at reception of the fresh lean meat that was not dried these parameters. Table 2 shows the measured values (73.5 %), probably as a result of the pork backfat, curing of moisture, salt content and fat content in these dry- agents (NaCl, nitrite) and additives added to the pork lean fermented sausages. when preparing the meat batter. Indeed, at day 1, the higher moisture contents were measured on the three formulations containing less than 12 % animal fat, and the highest value Moisture (68.1 %) was even obtained for the lowest-fat formulation (S4). Analysis of Table 2 confirms that in-product moisture de- It can be concluded that the respective proportions of lean creases globally with drying, with mean values decreasing meat and pork backfat logically influence moisture values – from initial values of 59.9 68.1 % at day 1 to final values of during dry-fermented sausage manufacture (Table 2). For an – 36.9 43.6 % at day 29 (Table 2). The lowest final water con- identical drying process, the highest moisture values tent values were obtained for the two 21 % animal fat formu- expressed as percentage of total matter are obtained for the lations (S2 and S8, i.e., the control) and for S6 corresponding low-fat products. to a high-fat but low-salt formulation (17.9 % fat and 2.0 % salt, respectively). Note that at day 1, in-product water content Salt Content Table 2 Time-course evolution of chemical composition (moisture, sodium chloride and animal fat contents) measured in dry-fermented sausages for the eight formulations of Table 1 Analysis of Table 2 logically indicates a net increase in in- product salt content with drying. Note that fresh pork lean is Experiments Day 1 Day 7 Day 21 Day 29 known to naturally contain small amounts of sodium and chlo- Moisture content (% total matter) ride ions; here we measured a 0.17 % equivalent of NaCl S1 62.5±0.8c 57.7±0.7c 46.1±0.6bc 43.6±1.0a content in the fresh pork lean. At day 1, measured salt content S2 61.1±0.7b 54.8±0.6de 41.3±0.8e 37.4±0.8c value ranged from 2.34 to 3.13 % depending on formulation S3 59.9±0.5a 55.6±0.8d 43.4±1.2de 41.1±0.6b (Table 2), whereas intended salt content corresponded to the S4 68.1±0.6f 62.9±1.0a 49.8±0.9a 40.7±1.0b levels of the Doehlert design: 2.0, 2.4 and 2.8 %. This implies S5 66.0±0.5e 59.8±1.2b 48.1±0.3ab 39.7±0.7b that real salt content values are globally higher than intended bc f e c values. Accurate analysis between real and intended salt con- S6 61.7±0.5 51.7±0.7 42.0±0.5 38.2±0.5 tent shows that discrepancies ranged from 0.18 % (S8) to S7 64.8±0.3d 59.3±0.6b 45.4±0.3cd 40.4±0.7b 0.52 % (S2) depending on formulation, with a mean discrep- S8 control 60.8±0.6ab 54.1±0.5e 42.8±1.0e 36.9±0.7c ancy of 0.37 %. This underlines just how difficult it is to add Sodium chloride content (% total matter) exactly the intended amount of NaCl to meat batters with S1 2.86±0.50bcd 3.76±0.30c 5.77±0.41cd 6.54±0.60bcd varying proportions of lean meat and fat, even at pilot-scale S2 2.92±0.25abc 4.35±0.15ab 5.93±0.12c 6.56±0.23bcd level. At day 29, the final NaCl content values ranged from S3 3.13±0.30a 4.5±0.40a 6.76±0.40a 7.15±0.50ab 6.26 to 7.64 % depending on experiment and on the weight S4 2.84±0.23cd 3.77±0.29c 6.27±0.20b 6.86±0.20bc loss experienced by the dry-fermented sausages. The highest S5 2.34±0.17de 3.62±0.13d 5.61±0.18d 6.26±0.13d final NaCl content value was measured for the control (S8) S6 2.44±0.13cde 3.64±0.14d 5.66±0.14d 6.39±0.20cd that initially contained about 2.98 % NaCl (day 1) and that S7 3.08±0.14ab 4.23±0.16b 5.71±0.15cd 6.34±0.16cd ab ab ab a counted one of the lowest final water contents measured S8 control 2.98±0.16 4.33±0.20 6.57±0.17 7.64±0.23 (60.8 %, Table 2). Animal fat content (% total matter) Regarding in-product salt content, it can be concluded that c b c d S1 14.3±0.2 17.7±1.3 22.8±1.0 25.6±0.4 the measured salt content values at day 1 were still higher than ab a a ab S2 18.9±0.2 23.3±0.6 31.1±1.6 34.0±1.4 the intended values, probably as a result of the natural pres- b b b c S3 17.1±0.4 19.8±1.0 25.7±1.6 29.5±1.6 ence of sodium and chloride ions in lean pork meat and e d e f S4 8.0±0.3 10.8±1.3 14.7±0.2 16.5±0.4 very probably as a result of the real difficulty in per- d c d e S5 11.2±0.2 14.2±1.1 19.7±0.2 20.8±0.4 fectly adjusting the amount of salt added during the b a b bc S6 18.1±0.4 22.7±0.8 27.8±1.6 31.0±1.8 meat batter preparation. However, the formulations with d c d e S7 12.0±0.2 14.5±1.4 19.3±1.4 21.0±1.4 the higher intended salt content values nevertheless a a a a S8 control 20.6±0.5 25.2±1.1 31.9±1.3 35.0±1.3 contained more salt than the others once the meat bat- ters were prepared, thus a priori lowering the impact of Measured values were the means±standard deviation calculated from six independent determinations. Values not bearing common superscripts this observed discrepancy on the physicochemical and differed significantly (p<0.05) biochemical results subsequently obtained. 2114 Food Bioprocess Technol (2015) 8:2109–2122 Total Fat Content significant effect of the interaction between salt and fat content (p>0.05). These results are in line with several studies de- Analysis of Table 2 confirms that animal fat concentrates with signed to assess the impact of reducing salt or fat content on drying. Like for salt content, discrepancies emerged between the sensory qualities of these kinds of meat products (Corral measured in-product total lipid content and the intended fat con- et al. 2014;Liarosetal.2009), but disagree with Corral et al. tent fixed by the Doehlert design, but the order is respected, (2013) who did not find any differences in weight losses for meaning that the formulations with the highest intended fat con- slow-fermented dried sausages prepared with different levels tents have the highest measured total lipid contents, and vice of salt (2.7 and 2.26 %). However, the salt effect observed here versa (Table 2). For example, formulations corresponding to S2 was for a higher level of salt reduction (−30 %) combined with and S8 for which intended fat content was 21 % really contained high fat reduction. Regarding the effect of fat content on prod- 18.9 and 20.6 % total lipids, respectively, at day 1. For the 17.9 % uct water loss, Olivares et al. (2010) failed to find differences fat formulations (S3 and S6), the discrepancy in total lipid con- in water loss between high and low-fat sausages, but they tent was −0.8 and +0.2 %, respectively. Moreover, 8.0 % total studied dried slow-fermented sausages slow-ripened for lipid content was measured in the specific case of formulation S4 63 days, whereas here where we applied high fermentation where 8.4 % fat content was planned. Finally, discrepancy in and faster drying. total lipid content ranged from −1.1 to +0.2 % depending on experiment investigated, again underlining the real difficulty in Time-Course of Mean Water Activity Values perfectly adjusting the amount of added fat during meat batter preparation. The formulations can be sorted as a function of their Figure 1b charts the kinetics of mean in-product aw values real total lipid content in the same order, from the beginning (day measured for the eight formulations of Table 1. Generally 1) until the end (day 29) of the drying process. However, fortu- speaking, the drying process led to a reduction in water con- nately, the formulations with the higher intended fat content tent and to a concentration of salt into the matrix, and thus to a values really contained more fat than the others, once the meat decrease in mean aw value. batters were prepared, thus a priori lowering the impact of this HCA applied to aw values led to the formation of three observed discrepancy on the results subsequently obtained. classes of formulations (Fig. 1b). The class corresponding to the lowest mean values of aw,withvariationsfrom0.958at Effect on Physicochemical Parameters in Dry-Fermented day 1 to 0.886 at day 29, is formed by batches S3, S7 and S8, Sausages i.e., the three highest-salt formulations containing at least 11.6 % animal fat. The class corresponding to the highest in- Time-Course of Weight Loss product mean aw values, with variations from 0.963 at day 1 to 0.901 at day 29, is formed by batches S4, S5 and S6, i.e., the Figure 1a charts weight loss kinetics during the ripening pro- two lowest-NaCl formulations (S5 and S6) and the lowest-fat cess. First of all, from day 1, all the dry-fermented sausages formulation (containing just 8.4 % animal fat) (S4). The third lose weight corresponding to the water lost through evapora- class is formed by experiments S1 and S2 which contain a tion during steaming and ripening. HCA applied to all weight moderate NaCl content (2.4 %) and a fairly high (>14.7 %) loss values led to the formation of three classes of formula- animal fat content. This class presented an intermediate pat- tions, as shown in Fig. 1a. The first class, corresponding to the tern in terms of time-course of mean in-product aw, with lowest weight loss (about 43 % at day 29), is composed of values ranging from 0.960 at day 1 to 0.897 at day 29. Note experiments S1, S2, S3, S6 and S8 (control). The two other that all the final mean in-product aw values are below 0.92 and classes strongly diverge from the first class from day 5, and that aw values below 0.92 are considered safe in terms of they pool the three lowest-fat formulations that generate Listeria monocytogenes growth capacity (Ingham et al. 2004). higher water losses from the sausages. These two classes stay In a similar way to the time-course of weight loss, the very close up to day 14 and thus separate the three lowest-fat statistical analyses performed on the mean aw values, i.e., formulations as a function of their relative NaCl content; the ANOVA and Tukey tests, showed very highly significant ef- highest-salt formulation (S7) led to lower weight loss (45.4 % fects of time, salt content and fat content (p<0.001) but no at day 29) than the two other formulations (S4 and S5) which significant effect of the interaction between salt and fat content are less salty (2.4 and 2.0 %, respectively) for which water loss (p>0.05) (Table 3). Literature data on the effect of fat content reached 49 % at the end of drying. Thus, Fig. 1a shows that on aw values is inconsistent. Some authors have reported that lean dry-fermented sausages globally lose more water than the reducing fat content during dried sausage manufacture led to fattiest products. higher final aw values (Corral et al. 2014; Gómez and Lorenzo Furthermore, these results were confirmed by the ANOVA 2013)orhigheraw values at the fermentation stage (Olivares and Tukey tests (Table 3) that highlighted very highly signif- et al. 2010) whereas others did not find any significant differ- icant effects of time, backfat and salt content (p<0.001) but no ence in aw values between low and high-fat dried sausages Food Bioprocess Technol (2015) 8:2109–2122 2115 Fig. 1 Time-course of three 60 physicochemical parameters investigated for the eight formulations of dry-fermented 50 a sausages given in Table 1: a bc ab weight losses, b mean water c 40 c activity (aw) values and c pH values. Hierarchical cluster de cd analysis was performed on raw ef 30 values in order to pool dry- f fermented sausage formulations presenting similar patterns. g 20 gh Values not bearing common ij ghi superscripts differed significantly (p<0.05) j Experiment S7 (45.4% at day 29) 10 k Experiments S4-S5 (49.0% at day 29) Experiments S1-S2-S3-S6-S8 control (42.9% at day 29) m kl Weight loss of dry-fermented sausages (%) o mn 0 0 7 14 21 28 35 A Time (days) 0.98 Experiments S1-S2 (0.899 at day 29) w a Experiments S4-S5-S6 (0.901 at day 29) 0.96 ab ab bc abc cd Experiments S3-S7-S8 control (0.886 at day 29) cd de e ef ef 0.94 f g h hi 0.92 ij j k l 0.90 lm Mean value of dry-fermented sausage a m 0.88 0 7 14 21 28 35 B Time (days) 6.5 Experiments S5-S6 (5.25 at day 29) Experiments S1-S2-S3-S4-S7-S8 control (5.40 at day 29) 6.0 a 5.5 b c c e d 5.0 f Dry-fermented sasuage pH value 4.5 0 7 14 21 28 35 C Time (days) 2116 Food Bioprocess Technol (2015) 8:2109–2122 Table 3 Details of statistical analyses: A – Analysis of variance and B – protein oxidations) parameters measured in the dry-fermented sausages Post hoc Tukey procedure, performed on the values of physicochemical for all the formulations of Table 1 (weight loss, aw, pH) and biochemical (proteolysis, lipolysis and lipid and A – Analysis of variance (ANOVA) Weight losses aw pH Proteolysis (proteolysis Lipolysis (acidity Lipid oxidation Protein oxidation index) value) (HSB) (thiol group) Time *** *** *** *** (−) *** *** Salt *** *** ns *** ** ns ns Fat *** *** *** *** *** *** *** Salt*Fat ns ns ns *** ns ns ns B – Post hoc procedure (Tukey test) S1 19.0a 0.940de 5.45±0.34a 4.80±1.25b 8.1±0.33ab 3.70±1.07b 26.64±1.19b S2 18.9a 0.939cd 5.43±0.35 a 4.75±1.22b 10.4±0.26cd 3.87±1.11cd 21.80±1.21a S3 18.7a 0.935bc 5.41±0.34a 4.39±0.92a 9.0±0.13b 3.82±1.10bc 26.67±0.83b S4 22.3b 0.943de 5.41±0.31a 4.89±1.28b 7.4±0.30a 3.51±0.86a 30.16±0.53d S5 21.6b 0.944e 5.33±0.37b 5.45±1.51d 7.9±0.25ab 3.52±0.89a 28.60±0.62c S6 20.5ab 0.944e 5.31±0.36b 5.17±1.38c 9.7±0.35c 3.83±1.08bc 26.32±0.6b S7 21.0b 0.935bc 5.41±0.33a 4.34±0.96a 7.5±0.45a 3.50±0.94a 28.72±1.46c S8 control 18.6a 0.933a 5.40±0.34a 4.25±0.81a 10.0±0.38c 3.96±1.26d 21.81±0.64a ANOVAwas run to test the effects of time, salt content, fat content and the interaction salt content × fat content on all the parameters studied. Significance is noted as follows: ns (p>0.05), * (p<0.05), ** (p<0.01), *** (p<0.001), and (−) corresponds to a factor not studied. ATukey test was applied on the means±standard deviation calculated from all independent determinations and for all timepoints. Values not bearing common superscripts differed significantly (p<0.05) manufactured with cereal and fruit fibres (García et al. 2002). corresponding to intense acidification due to the action of the Concerning salt content, several studies have mentioned that lactic acid bacteria (LAB) added during sausage manufacture. reducing salt logically led to higher aw values (Olesen et al. This period is characterized by an exponential growth phase of 2004; Roseiro et al. 2008). In contrast, other authors did not LAB that fermented sugar substrate into lactic acid. Beyond find any difference in final aw values of dried sausages made day 7, pH values increased progressively until the end of the with different levels of salt. For example, Corral et al. (2013) drying process as a result of an array of phenomena including found significant differences in aw values after 9 days but, a strong decrease in LAB acidifying action due to the deple- surprisingly, not at the end of processing. However, Corral tion of sugar substrate, the transformation of lactic acid into et al. (2014) reported that a combined reduction of salt and other chemical substances, consumption of lactic acid by fat contents affected sausage quality by producing an increase moulds (Flores et al. 2004) and/or the production of alkaline in final aw values. molecules due to proteolytic mechanisms (Ordóñez et al. Concerning the time-course of mean aw values, the HCA- 1999). based results presented here logically showed a strong impact HCA applied to the pH values of all the formulations led to of salt content and also an effect of fat content. Indeed, mod- the formation of two distinct classes differentiated according ifying the fat content of the meat batter modifies the salt con- to NaCl content of the formulations (Fig. 1c). Lower pH centration in the lean part of the batter, and thus the water values were obtained for the two lowest-salt formulations, activity value. From a water activity perspective, reducing i.e., S5 and S6 that contain only 2.0 % NaCl, probably due fat content in dry-fermented sausages provokes the same in- to more intense LAB activity. The second class pooled all the crease on aw as reducing salt content. So, binary reductions in other formulations that contain at least 2.4 % NaCl. The dif- fat and salt contents may prove detrimental from a safety ference between the two classes resided in the fact that a more standpoint if the products are not sufficiently dried afterwards. intense acidification was observed for the two lowest-salt for- mulations during the first week of process (with 0.10 pH unit Time-Course of pH Values less at day 7); this discrepancy persisted as pH values in- creased over the next 2 weeks of process, before growing Figure 1c charts the time-course of pH values. All formula- slightly stronger during the last week of process to peak at tions followed a normal time-course of pH values with a 0.15 pH units at day 29. However, all these pH values, espe- strong decrease in pH during the first week of process, from cially the lowest and final values, are fully representative of 5.91 to 5.93 at day 1 to a minimum of about 5.0 at day 7, what classically happens during French dry-fermented Food Bioprocess Technol (2015) 8:2109–2122 2117 sausage manufacture (Rason et al. 2007). Note that in the dry- day 1 and reached 4.6–5.9 % at day 7 depending on ing process, low aw values and the pH values at fermentation formulation. From day 7, rate of proteolysis decreased stage (day 7) and at the end of ripening (day 29) obtained in this noticeably, probably due to a non-optimal pH value for study are considered enough to ensure the microbial safety of proteolytic enzyme activity (García-Garrido et al. 2000) dry-fermented sausages (Papadima and Bloukas 1999). due to the intense lactic acidification occurring at that ANOVA and Tukey test (Table 3) showed very highly sig- point, as shown in Fig. 1c. Beyond day 21, very surpris- nificant effects of time and fat content (p<0.001). On the ingly, PI values decreased for all formulation classes other hand, salt content and the interaction between salt and (Fig. 2). To be sure that this decrease was not due to a fat content appeared non-significant (p>0.05). There is exten- problem with the fluorescent technique used, we measured sive research on the effect of reducing salt or fat content on pH PI in some samples with the classic Kjeldhal method values in dry-fermented sausages. For example, Olesen et al. consisting in calculating the percentage ratio of non- (2004)andStahnke(1995) reported lower pH values at fer- protein nitrogen content to total nitrogen content. This mentation stage in sausages made with lower amounts of classical PI measurement method confirmed the decrease NaCl. In contrast, Corral et al. (2013) did not find a significant in PI values over the last week of the drying process (data effect on pH evolution nor on final pH value when NaCl was not shown). This ultimately tends to prove that some end reduced by 16 % or substituted by 16 % KCl in slow- products of proteolysis had disappeared, likely consumed fermented sausages. The literature reports conflicting data on by microorganisms present in the dry-fermented sausages the effect of fat content on pH values. For instance, some at that point in time, and so could not be detected by the studies did not find differences in pH values when comparing measurement techniques used. high- and low-fat dried sausages (García et al. 2002;Liaros Figure 2 shows that HCA formed three classes of formula- et al. 2009; Papadima and Bloukas 1999). In contrast, Olivares tions. In-depth analysis of results highlighted that the formu- et al. (2010) highlighted a faster pH decline in the case of fat- lations are perfectly classified as a function of their respective reduced dried sausages. Finally, Corral et al. (2014) did not NaCl content, thus further confirming the inhibitory effect of find any significant difference in pH values between control salt content on proteolytic enzyme activity. Indeed, the first sausages and sausages made with combined reduced-fat and class that led to the lowest PI values is formed by the three reduced-salt formulations. 2.8%NaClformulations (S3, S7 and control S8), the second Our results (Fig 1c) show that combined salt and animal fat class by the 2.4 % NaCl formulations (S1, S2 and S4) and the reductions during the manufacture of dry-fermented sausages third class by the 2.0 % NaCl formulations (S5 and S6). At led to a fully normal time-course of pH values, with strong day 21, PI values were 5.2, 6.3 and 6.7 %, respectively, ac- acidification during the first week of process followed by a cording to class/salt content considered. progressive increase in pH value. Moreover, ANOVA and Tukey test (Table 3)highlighted, for the first time, very highly significant effects of all the Effect on Biochemical Parameters in Dry-Fermented factors tested, i.e., time, salt content, fat content and also the Sausages interaction between salt and fat content (p<0.001). However, no marked effect of fat content is visible in During dry-fermented sausage manufacture, and especially Fig. 2. Nevertheless, when comparing experiments S5 during the fermentation and ripening stages, the meat protein and S6 that belong to the same class, finer-grained anal- and lipid components undergo enzymatic and chemical mod- ysis of the raw results revealed that proteolysis was ifications. The enzymatic modifications involved are chiefly slightlymoreintenseinS5thaninS6,withday21PI proteolysis and lipolysis. These biochemical reactions are of values of 6.8 and 6.6 %, respectively. This probably high interest because they shape the development of sensory resulted from a lower fat content (11.6 vs. 17.9 %) that attributes of dry-fermented sausages, such as texture, juiciness provoked a more marked salt dilution and thus a lower and aroma. salt concentration in the lean part of the meat batter that contained the greater quantity of lean pork meat (i.e., Time-Course of Proteolysis experiment S5). These results are in good agreement with Armenteros et al. (2009, 2012). Figure 2 charts the time-course of proteolysis index. All eight The present results (Fig. 2) confirmed that proteolysis formulations produced the same pattern of behaviour in in dry-fermented sausages was mainly governed by salt terms of PI time-course. On the first day of process, pro- content but also highlighted that combined salt and an- teolytic enzyme activity is already visible, which is quite imal fat reductions could amplify proteolytic enzyme logical because proteolytic enzymes are highly active at a activity. So, reducing the salt and fat content increases temperature of about 24 °C (Harkouss et al. 2014). proteolysis; this can prove detrimental to the final tex- Proteolysis index (PI) ranged between 2.9 and 3 % at ture of the end products. 2118 Food Bioprocess Technol (2015) 8:2109–2122 Fig. 2 Time-course of 8 proteolysis index investigated for Experiments S3-S7-S8 control (4.8% at day 29) the eight formulations of dry- Experiments S1-S2-S4 (5.2% at day 29) g f fermented sausages given in Experiments S5-S6 (5.8% at day 28) e Table 1. Hierarchical cluster 6 e analysis was performed on raw d d values in order to pool dry- c c fermented sausage formulations b presenting similar patterns. For each class of dried sausages, 4 values were means±standard a deviation calculated from all the a a independent measurement values of each experiment constituting 2 the class. Values not bearing common superscripts differed significantly (p<0.05) Proteolysis index in dry-fermented sausages (%) 0 1 7 21 29 Time (days) Lipolysis of End Products elaboration of the taste, flavour and odour of these kinds of dried cured products (Corral et al. 2013). Applying HCA on the acidity values (lipolysis) determined at day 29 for all eights formulations led to the formation of two Lipid Oxidation distinct classes as a function of animal fat content regardless of NaCl content. One class pooled the four formulations for Figure 3a charts the time-course of lipid oxidation quantified which fat content was at most equal to 14.7 % and the other by determining hydrosoluble Schiff base (HSB) values. class pooled the 17.9 % animal fat and 21 % animal fat for- However, lipid oxidation was first measured by the thiobarbi- mulations. Mean acidity value was significantly higher for the turic acid reactive substances (TBARS) method (data not formulations containing over 17 % animal fat, at 9.8 vs. shown). A biphasic curve was obtained for all eight formula- 7.6 mg KOH per gram of product fat. tions, with TBARS value decreasing with time (mg MDA/kg ANOVA showed that salt content significantly affected of sample) beyond day 1, which is not a logical pattern and (p<0.01) rate of lipolysis in dry-fermented sausages and that indicates that the TBARS quantification method was ill-suited fat content was a highly significant factor strongly impacting to accurate assessment of lipid oxidation in the present study. rate of lipolysis (p<0.001), unlike the interaction between salt This prompted us to assess lipid oxidation based on the fluo- and fat content (p>0.05). Olivares et al. (2011) also cited a rescence of HSB. Schiff bases are formed in meat during strong effect of fat content on lipolysis and on the generation storage and cooking (Gatellier et al. 2009). They correspond of free fatty acids. These authors reported that slow-fermented to the binding of the aldehydic products of lipid peroxidation sausages initially containing 30 % pork backfat showed higher to free amino groups of proteins. The polarity of Schiff bases rates of lipolysis than dried products made with lower pork depends on their composition. It was observed in raw meat backfat content (10 and 20 %). Furthermore, several authors that the aqueous fluorophores were responsible for up to 72 % have reported a more intense lipolysis in salt-reduced dried of total fluorescence intensity, whereas cooking reversed the sausages (Stahnke 1995). percentages to reach 36 % in the polar phase and 64 % in the To conclude on lipolysis, our data suggests that the inten- apolar phase (Gatellier et al. 2009). sity of this biochemical phenomenon is strongly dependent on Figure 3a shows the results of HCA applied to HSB values fat content and also influenced by salt content, but without of all formulations. Contrary to the TBARS method (data not significant interaction between these two factors, unlike for shown), HSB values increased observably as a function of proteolysis. time, meaning that lipid oxidation logically increased with time. Applying HCA led to classify the formulations into three distinct classes as a function of fat content; higher fat content Time-Course of Lipid and Protein Oxidations means higher HSB values and thus higher lipid oxidation. The first class is formed of the three formulations containing at Lipid and protein oxidations that occur throughout the manu- most 11.6 % animal fat (experiments S4, S5 and S7), the facture of dry-fermented sausages are also involved in the second class is formed of the three formulations containing Food Bioprocess Technol (2015) 8:2109–2122 2119 Fig. 3 Time-course of two 6 oxidation reactions investigated Experiments S4-S5-S7 (4.8 μmolar/kg at day 29) f f Experiments S1-S3-S6 (5.2 μmolar/kg at day 29) for the eight formulations of dry- e fermented sausages of Table 1: a 5 Experiments S2-S8 control (5.2 μmolar/kg at day 29) e lipid oxidation quantified by d determining hydrosoluble Schiff base (HSB) content and b protein 4 c oxidation quantified by determining free thiol group b b b content. Hierarchical cluster 3 a analysis was performed on raw a a values in order to pool dry- fermented sausage formulations 2 presenting similar patterns. For Hydrosoluble Schiff base content each class of dried sausages, (μ molar/kg dry-fermented sausage) values were means±standard 1 deviation calculated from all the independent measurement values of each experiment constituting 0 the class. Values not bearing 1 7 21 29 A Time (days) common superscripts differed significantly (p<0.05) 40 Experiments S4-S5-S7 (28.4 nmol/mg at day 29) Experiments S1-S3-S6 (25.9 nmol/mg at day 29) Experiments S2-S8 control (21.0 nmol/mg at day 29) a b 30 b b b bc bc bc c d d d 20 10 Free-thiol-group content (nmol/mg protein) 0 1 7 21 29 B Time (days) between 14.7 and 17.9 % animal fat (experiments S1, S3 and reduced dried sausages, thus confirming their previous results S6), and the third class gathered the two 21 % animal fat (Corral et al. 2013). However, Jin et al. (2013)reportedthat formulations (experiments S2 and control S8). Figure 3a high salt levels led to higher oxidation by decreasing antiox- shows that these three classes differ in terms of lipid oxidation, idant enzyme activity and releasing iron from heme-binding especially at day 21 but less so at the end of product drying protein. Furthermore, Corral et al. (2015) recently reported (day 29) and not really early on in the process (days 1 and 7) higher lipid oxidation levels in salt-and-fat-reduced sausages. ANOVA and Tukey tests (Table 3) indicated very highly significant effects of time and animal fat content (p<0.001) on Protein Oxidation lipid oxidation but no significant effect of salt content, nor the interaction between salt and fat content (p>0.05). These re- Protein oxidation was assessed by quantifying free-thiol- sults are in line with Liaros et al. (2009) and Soyer et al. (2005) group content. Indeed, thiol groups of cysteine are very sen- who reported that low-fat sausage presented lower lipid oxi- sitive to the oxidative process and their oxidation leads to the dation levels than control sausages. Olivares et al. (2011)also formation of disulfide bridges. Note that protein oxidation is reported higher TBARS values in high-fat sausages compared maximal when free thiol group content is minimal. to low-fat sausages. The literature is less clear on the effect of HCA-based results, shown in Fig. 3b, led to the formation salt on lipid oxidation. For example, very recently, Corral et al. of three classes of formulations, once again classified as a (2015) reported higher TBARS values and higher abundance function of their respective fat content. Maximal protein oxi- of volatile compounds derived from lipid oxidation in salt- dation was observed for the two 21 % animal fat formulations 2120 Food Bioprocess Technol (2015) 8:2109–2122 (experiments S2 and S8). Minimal protein oxidation was in fat and salt content may prove detrimental from a safety found for the three formulations containing at most 11.6 % standpoint if the products are not sufficiently dried. The results animal fat (experiments S4, S5 and S7). An intermediate class on proteolysis confirmed that this biochemical phenomenon of experiments S1, S3 and S6 was formed from the formula- in dry-fermented sausages was mainly governed by salt con- tions containing either 14.7 % animal fat or 17.9 % animal fat. tent and highlighted that it can be amplified by a combined However, since no clear change in protein oxidation appears reduction in fat and salt contents. Our experimental data also with time, beyond day 1 (Fig. 3b), protein oxidation seems to indicated that intensity of lipolysis was mainly dependent on occur rapidly, maybe directly during the meat batter prepara- fat content and that lipid and protein oxidations were more tion, without subsequent intensification. intense in high-fat formulations. Like for lipid oxidation, ANOVA and Tukey test (Table 3) Combined salt and fat reductions during the manufacture of showed very highly significant effects of time and fat content dry-fermented sausages globally affect end product quality by (p<0.001) on protein oxidation, but without any significant increasing acidification, weight losses and aw values. These effect of salt content, nor the interaction between salt and fat combined reductions also accelerate proteolysis and deceler- content (p>0.05). Protein and lipid oxidations are linked by ate lipolysis and oxidative mechanisms, which can negatively the fact that lipid oxidation produces free radicals that, in turn, affect the final sensory attributes of dry-fermented sausages. drive protein oxidation. Therefore, maximal protein oxidation Additional, sensory studies are thus necessary to investigate occurred in high-fat formulations. The present results are in the final product quality and consumer acceptability of low- good agreement with Fuentes et al. (2014) who recently re- salt and low-fat dry-fermented sausages. Furthermore, it will ported a strong impact of fat level on protein oxidation. also be important from an industrial point of view to obtain Regarding the effect of salt content on protein oxidation, accurate data on the potential application of new technologies Kanner et al. (1991) concluded that salt level increased the in dried sausage production, such as partial substitution of susceptibility of myofibril to carbonylation and the pro- NaCl by potassium chloride and adding vegetable oil rich in oxidantactivityofiron. mono-unsaturated fatty acids, in an effort to reduce end prod- The present results indicate that lipid and protein oxida- uct content of SFA by 60 % and salt content by 30 % in line tions in dry-fermented sausages are mainly affected by fat with nutritional health recommendations. level but not by salt level in the range studied here (2.0– 2.8 % NaCl) nor by the combined salt and animal fat reduc- tions. The fact that reducing fat level decreases both these Acknowledgments The research leading to these results received oxidative phenomena can potentially affect the final aroma funding from the European Union Seventh Framework Programme of these products. (FP7/2007-2013) under grant agreement No. 289397 (TeRiFiQ project). 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