07 県産食品資源の市場流通を促進するための評価技術の確立

26
Quercetin
Quercetin
Quercetin
Quercetin 3,4’-­Glucoside
Quercetin 3-­Glucoside
Quercetin
Quercetin
1.
1
1,267
23
1,243
24
2
4), 5)
22%
3)
20%
Premier-­S
6) 8)
9), 10) PR
6
26
4 5ºC
80%
Methanol
-­20ºC
30
20,000 x g, 4ºC, 15
0.2 ȝm
ESI
1
500 g
Methanol
Quercetin
Acetonitrile
1
2.4 ESI
2.
2.1
ESI
2.2
Quercetin
Que
Que 3-­Glucoside Glc
Que 4’-­Glc
Que 3-­4’-­Glc
2.3
200 g
80ºC
1
30
1
Quercetin
Que, R1 = R2 = R3 = H;; Que 3-­Glucoside (Glc);; R1 = Glc & R2 = R3 = H;; Que 4’-­Glc, R2 = Glc & R1 = R3 = H;; Que 3,4’-­Glc, R1 = R2 = Glc & R3 = H Liquid Chromatograph-­ESI Ion Trap Mass Spectrometer;; LC-­IT/MS
Agilent 1100 Agilent Technologies
LC
IT/MS
HCT ultra Bruker Daltonics
LC
IT/MS
: , Cadenza CD-­C18
2 mm i.d. x 250 mm, Imtakt
;; , 40ºC;; , 0.2 mL/min;; , Acetonitrile/
/Formic acid = 10/89.9/0.1
A
Acetonitrile/
/Formic acid = 50/49.9/0.1
B ;; , B
0% 0 min
80% 20 min 100% 21 min
100%
5
;; , 368 nm;; , Negative;; . 200 m/z – 800 m/z;; , 8,100 (m/z)/sec;; , 45 psi;; N2
, 350ºC;; MS/MS
MS/MS/MS
, MS
MS/MS
2.5
Liquid Chromatograph-­quadrupole Time of Flight Mass Spectrometer;; LC-­qTOF/MS
Nexera X2
LC
qTOF/MS
impact II Bruker Daltonics
LC
26
2
Que
LC-­IT/MS
50ºC
2.4
qTOF/MS
: , ESI;; , Negative;; . 100 m/z – 1,500 m/z;; , 23.2 psi;; N2
, 180ºC;; MS/MS
, 462.00
464.00
624.00
626.00
LC
3.
3.1
Que
DataAnalysis
Formula
of Chemistry
ChemSpider12)
Que Que 3-­Glc Que 4’-­Glc
3,4’-­Glc
LC-­IT/MS
Total Ion Chromatogram;; TIC
Que 3,4’-­Glc
2
Que
[M-­H] 463 m/z
MetFrag11)
Royal Society [M-­H] 625 m/z
Extracted Ion Chromatogram;; EIC
3
LC
EIC
Que 3-­Glc
Que 3,4’-­Glc
Que 625 m/z EIC
2
TIC
Que Que 3-­Glc
3
Que
2
3-­Glc
qTOF/MS
ver. 4.3
Smart-­ MS
MS/MS
TIC 463 m/z
625 m/z
EIC 26
4 Que 3-­Glc
3.2 IT/MS
MS/MS
MSn
Que 4’-­Glc
MS/MS
301 m/z
MS/MS/MS
MS/MS/MS
4
MS
Que 3-­Glc
MS/MS MS/MS/MS
Que
Que 4’-­Glc
MSn
5
TIC
Collision-­ Induced Dissociation;; CID
[M-­H]
463 m/z MS
[M-­H]
MS/MS
m/z 301
Que [M-­H]
Glc
162 m/z
5
Glc
Que 3,4’-­Glc
MSn
Que 3,4’-­Glc [M-­H]
625 m/z
CID
MS/MS
Glc
1
463 m/z
Glc 2
301 m/z
MS/MS
463 m/z
MS/MS/MS
Glc 1
301 m/z
4
MS/MS
MSn
12.5 min
m/z
Que 3,4’-­Glc
12.5 min
3,4’-­Glc
3
Que 625 m/z
12.7 min;; 3,4’-­Glc
MS/MS
Que
5
12.7 min
14.8 min
5
MSn
: 14.8 min;; 5
2
Que MS/MS/MS
MS/MS
2
Que
2
3.3 qTOF/MS
6
MS
LC-­qTOF/MS
EIC 463.0000 m/z
MS/MS
TIC
625.1426 m/z
26
5 Que 3,4’-­Glc
TIC 12.5 min 12.7 min
0.5 m/z
625.1426 m/z
3
463.0000 m/z
4
1
MSn
14.8 min
9.7 min 10.9 min 12.3 min 12.7 min
4
625.1426 m/z
EIC
26
6
LC-­qTOF/MS
TIC
625.1426 m/z
463.0000 m/z
EIC 7
6
EIC 625.1426 m/z
MS
MS/MS
4
MS
7
0.0003
10.9 min
min
12.7 min
2
9.7 min 10.9 min 12.3 min
9.7 min
MS/MS
4
625.1425
MS
C27H29O17
9.7 min
12.3 [M-­H]
462.7639 -­ 463.7796 m/z
[M-­H]
10.9 min
625.1426 m/z
MS/MS
C27H29O17
4
300.0280 – 300.0284 m/z
12.7 min
MS
2
MS MS/MS
26
ChemSpider
85
1) 24
25
23
25
2
28
www.e-­stat.go.jp/SG1/ estat/List.do?lid=000001104918 Que
IT/MS
www.maff.go.jp/j/tokei/ 2) Que 3-­Glc
25
kouhyou/nougyou_sansyutu/ Que
Que 3,4’-­Glc
12
3) 162 m/z
24
www.pref.saga.lg.jp/web/kensei/_1366/toukei/t-­sougo
u/t-­nenpou/_72049.html Que
qTOF/MS
MS MS/MS
4) K. Hayashi, M. Yamanaka, K. Toko, and K. Yamafuji, Multichannel taste sensor using lipid membranes;; Sensors and Actuators B: Chemical, 2(3), Que
205-­213, 1990. 5) A. Legin, A. Rudnitskaya, Yu Vlasov, C. Di Natale, E. Mazzone, and A. D’Amico, Application of electric tongue for qualitative and quantitative analysis of complex liquid media;; Sensors and Actuators B: Chemical, 65(1-­3), 232-­ 234, 2000. 6) , 4.
, 52(8), 355-­365, 2005. 7) , 39(1), 49-­56, 2006. 8) K. Chikuni, M. Oe, K. Sasaki, M. Shibata, I. Nakajima, K. Ojima, and S. Muroya, Effects of muscle type on beef taste-­traits assessed by an electric CID
sensing system;; Animal Science Journal., 81(5), 600-­605, 2010. 9) V. Anand, M. Kataria, V. Kukkar, V. Saharan, and P.K. Choudhury, The latest trends in the taste assessment of pharmaceuticals;; Drug Discovery Today, ESI
12(5-­6), 257-­ 265, 2007. 10) K. Woertz, C. Tissen, P. Kleinebudde, and J. Breitkreutz, Taste sensing systems (electric tongues) for pharma-­ ceutical applications;; International Journal of Pharmaceutics, 417(1-­2), 256-­271, 2011. 11) MetFrag;; In silico fragmentation for computer 26
assisted identification of metabolite mass spectra;; http://msbi.ipb-­halle.de/MetFrag/. 12) ChemSpider;; Search and share chemistry;; Royal society of chemistry;; http://www.chemspider.com.