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Eugenio Aprea1, Franco Biasioli1, Luca Cappellin1,2, Christos Soukoulis1, Flavia Gasperi1, Filomena Morisco3, Giuseppe D'Argenio3, Vincenzo Lembo3,
Vincenzo Fogliano4, Nicola Caporaso3
1
IASMA Research and Innovation Centre, Fondazione Edmund Mach, !"#$%&$'#()*$+$,$-./+/*$0#1$'"()232$#33456"72*$89#3:$ 2
$81;9"9<9$=>?$8@121A):;"B$<16$5172C#1692$D):;"B*$E2@A@36,F?#1G21;$H1"I2?;"9J9$811;K?<(B*$$ Technikerstr. 25, A-6020, Innsbruck, Austria
3
$L3"1"(#3$#16$%MA2?"N219#3$'26"("12*$O#;9?@2192?@3@7:$H1"9*$H1"I2?;"9:$@=$P#A32;$QF262?"(@$88R*$89#3:$ 4
$S2A#?9N219$@=$F@@6$0("21(2*$H1"I2?;"9:$@=$P#A32;$QF262?"(@$88R*$89#3:$
Summary:! T)2$ A?2;219$ ;9<6:$ 62N@1;9?#92;$ 9)2$ A@;;"K"3"9:$ 9@$ N2#;<?2$ @13"12$ 9)2$ 2M)#326$ K?2#9)$ @=$ #C#B2$ ?#9;&$ T)2$ A@9219"#3"9:$ @=$ DTU,T@F,'0$ )#I2$ K221$ 92;926$ A?@I"6"17$ #1#3:9"(#3$ "1=@?N#9"@1$ 9)#9$ #33@C;$ 9)2$ 6";(?"N"1#9"@1$ @=$ ;<KV2(9;$ #((@?6"17$ 9)2$ 6"29$ ?27"N2$ 62N@1;9?#9"17$ 9)2$ A@;;"K"3"9:$ 9@$ N@1"9@?$9)2$N29#K@3"($;9#92$@=$;<KV2(9;$9)?@<7)$=#;9$1@1$"1I#;"I2$K?2#9)$#1#3:;";&$
Keywords: proton transfer reaction-mass spectrometry; breath analysis; diet. ?&@%$"#>=-$.#%&
Volatile compound found in exhaled breath N#:$ A?@I"62$ "1=@?N#9"@1$ @1$ 9)2$ A):;"@3@7:$ ;9#92$ @=$ ;<KV2(9;&$ U#9;$ #?2$ (@NN@13:$ <;26$ "1$ preclinical trials to investigate the effect of the diet on the animal health before test in humans. 51#3:;";$ @=$ 9)2$ (@NA@1219$ A?2;219$ "1$ 9)2$ breath of such animals can give important information on state of their metabolism and N#:$ K?"17$ 9@$ 9)2$ "6219"="(#9"@1$ @=$ N#?B2?;$ specific for the different metabolic conditions. U2(2193:$ DTU,'0$ )#;$ K221$ A?@A@;26$ =@?$ 9)2$ rapid screening of potential pathologies N#?B2?;$ W+X&$ 5"N$ @=$ 9)2$ A?2;219$ ;9<6:$ C#;$ 9@$ I2?"=:$ 9)2$ =2#;"K"3"9:$ @=$ DTU,T@F,'0$ =@?$ 9)2$ online in vivo #1#3:;";$ @=$ ?#9$ K?2#9)$ 9@$ demonstrate the potential of such technique for #1"N#3$ K?2#9)$ #1#3:;";$ #16$ 9@$ =@<16$ A@;;"K32$ markers associated at different diet regimes.
A&BC6+"./+%$(1& Animals.$0"M9221$?#9;$6"I"626$"1$Y$7?@<A;$C2?2$ <;26$ =@?$ 9)2$ 2MA2?"N219&$ %#()$ 7?@<A$ C#;$ ;<KV2(926$9@$#$6"==2?219$6"29$?27"N2Z$QP@?&[#9R$ ?#9;$ =226$ C"9)$ ;9#16#?6$ A23329;$ #16$ C#92?\$ QP@?&L@=R$ ?#9;$ =226$ C"9)$ ;9#16#?6$ A23329;$ #16$ 62(#==2"1#926$(@==22\$QF#9&[#9! ?#9;$=226$C"9)$ =#9$ ;<AA32N219$ #16$ C#92?\$ QF#9&(@=R$ ?#9;$ =226$ C"9)$=#9$;<AA32N219$#16$62(#==2"1#926$(@==22&$
Breath collection. %M)#326$K?2#9)$C#;$;#NA326$
"19?@6<("17$9)2$N<GG32$?#9$"1$#$]/$N3$F#3(@1^$ (@1"(#3$9<K2$9)#9$C#;$6"?2(93:$(@112(926$9@$9)2$
DTU,'0$ "1329$ _F"7&$ +`&$ a?2#9)$ (@332(9"@1$ C#;$ achieved for at least 20 seconds. Breath of all 9)2$#1"N#3;$C#;$(@332(926$@1$9)2$;#N2$6#:$#16$ 9)2$2MA2?"N219$C#;$?2A3"(#926$#=92?$#$C22B&$
Food volatile compounds. Headspace of the
(@==22$#16$@=$;9#16#?6$#16$=#9$;<AA32N219$C#;$ 6292?N"1#926$#;$C233$9@$I2?"=:$9)2$A?2;21(2$#16$ the levels of the detected compounds and their A@;;"K32$ "192?=2?21(2$ C"9)$ 9)2$ (@1(219?#9"@1$ found in exhaled breath.
& Fig. 1. Sampling of rat exhaled breath. &
D&,+)=1$)&
F"7<?2$ b$ ?2A@?9;$ #$ D?"1("A#3$ L@NA@1219$ 51#3:;";$ _DL5`$ @=$ #33$ 9)2$ (@332(926$ 6#9#$ @I2?$ 9)2$ 9C@$ 2MA2?"N219$ 6#:;&$ T)2$ =@<?$ 7?@<A;$ #?2$ C233$ ;2A#?#926$ "16"(#9"17$ 9)#9$ 9)2$ 2M)#326$ breath composition is related to the different diet regimes. Both fat supplementation and
coffee influenced the metabolic state of rats.
!" !# !$ !% !& !' !( ) ( ' & % $ # " !* !+ !" !# !$ !% !& !' !( ) ( ' & % $ # " + * ,-' . ,-(. /0,1234 /0,150, 6371234 637150, /0,1234 /0,1234 /0,1234 /0,1234 /0,1234 /0,1234 /0,1234 /0,1234 /0,150, /0,150, /0,150, /0,150, /0,150, /0,150, /0,150, /0,150, /0,150, /0,150, 6371234 6371234 6371234 6371234 63712346371234 6371234 637150, 637150,637150, 637150, 637150, 637150, 637150, 637150, 89:2;!<=>('1)1(>!>')((!)"!'$>('?(%?$#>@AB2=(C>
Fig. 2. PCA score plot of first 2 components of the all collected data (two days replicates are included).
Different masses were identified as potential
markers. For example m/z 42.0336
corresponding to the protonated acetonitrile (theoretical mass 42.03382 Th) is high in the breath of rats fed with standard diet and even higher in rats drinking coffee (Fig. 3).
637150, 6371234 /0,150, /0,1234 )1) )1$ (1) (1$ '1) '1$ &1) &1$ 2 3 D E F D ,7 0 ,G 3 D >@ H H IJ C ;EF,3DG,7GKF !"#$% &$% '()*+,"*-".)% (+,(),*-/*"+,% ",% *0)% -/*12% exhaled breath. Boxes: 25-75 percentile; whiskers: outliers; small boxes: mean value.
Concentration of methanol (m/z 33.0335) is higher in the breath of rats under standard diet regime and is not influenced by the coffee (Fig. 4).
Acetonitrile and methanol were both present at different concentrations in the headspace of coffee and of solids food. From the comparison with these data (not shown), the measurement !"# $%&'# &!()!%*+$# ,*# -'.# /0-$1# 2/.0-'# $..($# not be influenced by supplemented foods.
637150, 6371234 /0,150, /0,1234 () ') &) %) $) #) 2 3 D E F D ,7 0 ,G 3 D >@ H H IJ C :F,L0D3K !"#$%3$%4)*0/,+.%(+,(),*-/*"+,%",%*0)%-/*12%)50/.)6% breath. !"#$%&'()*$%)"
This study demonstrated the possibility to analyze online the composition of exhaled breath of awake rats minimally affecting their metabolic state. Markers of diet regime can be identified by PTR-ToF-MS showing the
potentiality of this technique for the
monitoring of metabolic conditions of the mammals through the fast non-invasive breath analysis.
"
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1.> J. Rieder, P. Lirk, C. Ebenbichler, G. Gruber, P. Prazeller, W. Lindinger, A. Amann, Wiener Klinische Wochenschrift, 113 (2001), pp. 181-185.
