Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 1 of 13
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315BETWEEN DG TAXATION AND CUSTOMS UNION (DG TAXUD) AND THE JOINT RESEARCH CENTRE (JRC)for fast recognition of New Psychoactive Substances (NPS) and identification of unknown chemicalsThis report was generated on 27/01/2016 based on data from the European Customs laboratories and the JointResearch Centre. This report includes sample and molecular information, spectral data and associated tables andfigures. The chemical structure(s) was/were identified by Bio-chemical interactions & metabolomics (BCIM) groupchemists on the basis of analytical data available. NMR assignments proposed below were performed by ACD labstools in agreement with the chemical structure identified by analytical experts. Reported data are related to thesample in the following table:
Eurodat number 16010001 Received on 08 January 2016PACKAGING 1 glass vial white powder (30 mg)Registration date 08 January 2016 Name of customer Service commun des laboratoires FranceCustomer's identification 2015-30772
The following structure(s) was/were identified in the sample:
O
N
N
Data of identified compound(s)Formula C24H32N2OFW 364.5237Monoisotopic Mass 364.2515Chemicalize IUPAC name N-phenyl-N-[1-(2-phenylethyl)piperidin-4-yl]pentanamideInChI (v.1.04) InChI=1S/C24H32N2O/c1-2-3-14-24(27)26(22-12-8-5-9-13-22)23-16-19-25(20-17-23)18-15-21-10-6-4-7-11-21/h4-13,23
H,2-3,14-20H2,1H3InChI Key (v.1.04) VCCPXHWAJYWQMR-UHFFFAOYSA-NSMILES (v.14.01) CCCCC(=O)N(C2CCN(CCc1ccccc1)CC2)c3ccccc3
Claude Guillou, Fabiano Reniero, Hubert Chassaigne, Joana Lobo Vicente, Veronica Holland, Salvatore Tirendi,Kamil Kolar
European Commission, Joint Research CentreInstitute for Health and Consumer Protection (IHCP)via E. Fermi, 2749TP 281 I-21027 Ispra (VA) - ItalyPhone: +39 0332 785678Fax: +39 0332 [email protected]://ihcp.jrc.ec.europa.eu
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 2 of 13
Processing and interpretation based on data provided by:Service commun des laboratoires France
Title 30772File Name \\139.191.6.82\ihcp∴I01\BIO_CHEMICAL_INTERACTION_METABONOMICS\CLEN2SAND\!PROPOSED-STRUCTURE\CLEN
_NAS\16010001_2015-30772\30772.SPADate Stamp 24 Nov 2015 13:51:25 Date 24 Nov 2015 14:51:38Technique Infrared Spectral Region IRX Axis Wavenumber (cm-1) Y Axis %ReflectanceSpectrum Range 525.0251 - 3500.1670 Points Count 6172Data Spacing 0.4821
Wavenumber (cm-1)3500 3000 2500 2000 1500
%R
efle
ctan
ce
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
3052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.283052.28
3031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.073031.07
2954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.892954.89
2858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472858.472756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.742756.74
2728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.32728.3
2704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.672704.67
2678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642678.642638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.142638.14
2508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.452508.45
2459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.282459.28
2415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.42415.4
2203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272203.272185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432185.432053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332053.332032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.62032.6
1988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251988.251949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21949.21942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451942.451770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.331770.33
1747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671747.671729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.831729.83
1714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411714.411704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281704.281688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.371688.37
1643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.541643.54
1594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.361594.36
1568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.811568.81
1492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.631492.63
1464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.671464.67
1423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.691423.69
1389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.941389.94
1373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.551373.55
1343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.661343.66
1326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.791326.79
1272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.791272.79
1245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.791245.79
1201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.921201.92
1159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.011159.01
1110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.81110.8
1076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.081076.08
1044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.261044.26
1025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.941025.94
952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18952.18
921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81921.81
877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45877.45 804.17
804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17804.17
785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37785.37
750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66750.66
701701701701701701701701701701701701701701701701701701701701701701
648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45648.45
614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7614.7
594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45594.45
545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27545.27
CH3
O
N
N
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 3 of 13
Retention Time: 6.352 Ion Mode: EI+
m/z360340320300280260240220200180160140120100806040
Rel
ativ
e In
tens
ity (%
)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
364.
2563
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
273.
2000
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
230.
1838
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
189.
1000
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
160.
0999
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
146.
1000
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
132.
0999
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
118.
0957
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
105.
0996
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
96.0
998
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
783
77.0
78357
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
857
.099
8
EI+
Processing and interpretation based on data provided by:Service commun des laboratoires France
Combine <852-860> Count 299 Data Source Name 2015-30772_Centroid_EI+Data Type Condensed Date 24 Nov 2015 18:54:48Date Stamp 24 Nov 15 06:42 pmFile Name 2015-30772_Centroid_EI+ Ion Mode EI+Mass Spec Model GCMS Plot Type Stick Retention Time 6.352Sample Ds CH2Cl2/MeOH : 2015-30772 Scan 858Scan Mode Centroid Spectrum Assigned 0.0% [35-503/0-100]TIC 529.31 Total Signal 7190323.5
CH3
O
N
N
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 4 of 13
Table of fragmentsNo. Fragment Structure Formula Label m/z Calc. TIC Calc.
(%)Difference
(Da)m/z Exp. RI Exp.
(%)TIC Exp.
(%)Origin
1 8-12,20-21(-H)N11
10
9
CH8
21
20
CH2+
12
C6H10N M - C18H22NO 96.081 2.604 0.019 96.100 12.837 2.604 Manual
2 12-19
CH2+
1213
14
15
1617
18
19 C8H9 M - C16H23N2O 105.070 5.012 0.030 105.100 24.260 5.012 Manual
3 4-5,7,22-27(+H) N7
C5
22
23 24
25
2627
CH2+
4
C8H8N M - C16H24NO 118.065 1.702 0.031 118.096 8.209 1.702 Manual
4 7-8,20-27
CH+
821
20 N7 22
2324
25
2627
C9H10N M - C15H22NO 132.081 2.613 0.019 132.100 12.464 2.613 Manual
5 9-19(-H)N+
11 109
1213
14
15
1617
18
19
C10H12N M - C14H20NO 146.096 13.718 0.004 146.100 64.697 13.718 Manual
6 5-8,20-27
CH+
821
20N7
5
O6
22
23 24
25
2627 C10H10NO M - C14H22N 160.076 0.819 0.024 160.100 3.854 0.819 Manual
7 5-8,11-12,20-27(+H)N11
CH+
821
20
CH2 12
N7
C5
O6
22
23 24
25
2627
C11H13N2O M - C13H19 189.102 10.133 -0.002 189.100 46.973 10.133 Manual
8 5-13,20-27
N11
10
9 8
21
20
12
13
N7
C+
5O6
22
23 24
25
2627 C14H18N2O M - C10H14 230.141 0.774 0.042 230.184 3.471 0.774 Manual
9 1-12,20-27
N11 10
98
21
20
CH2+
12
N75
O6 22
2324
25
2627
43
2
CH31
C17H25N2O M - C7H7 273.196 23.081 0.004 273.200 100.000 23.081 Manual
10 MN
+11
10
9
821
20
1213
1415
16
17 18
19
N7
5O6
22
23 24
25
2627
4
3
2
CH31
C24H32N2O M 364.251 0.012 0.005 364.256 0.047 0.012 Manual
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 5 of 13
Data from BCIM group at JRC, IHCP IspraNMR spectraAcquisition Time (sec) 2.7263 Comment 2015-30772 10 mg in 600 uL DMSO-d6 Paris 01/ 2016Date 08 Jan 2016 15:17:51 Date Stamp 08 Jan 2016
15:17:51File Name \\139.191.6.82\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCIM_NAS\BCIM_6
00\16010001\1\PDATA\1\1rFrequency (MHz) 600.13 Nucleus 1H Number of Transients 128Origin spect Original Points Count 32768 Owner nmrsuPoints Count 65536 Pulse Sequence zg30 Receiver Gain 1.91SW(cyclical) (Hz) 12019.23 Solvent DMSO-d6 Spectrum Offset (Hz) 3706.0515Spectrum Type standard Sweep Width (Hz) 12019.05 Temperature (degree C) 26.998
16010001.001.001.1r.esp
Chemical Shift (ppm)10 9 8 7 6 5 4 3 2
Nor
mal
ized
Inte
nsity
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
2.93
2.03
2.01
1.96
2.03
2.02
2.03
2.08
1.98
1.97
2.66
0.98
4.86
2.01
1.02
1.97
0.95
DMSO
M14(m, 25)
M17(m, 26,24)
M13(m, 18,16)
M16(m, 12)
M12(m, 19,15,17,27,23)
M01(t, 1)
M07(m, 13)
M09(br d, 20,10)
M08(m, 20,10)
M05(t, 4)
M11(m, 8)
M02(m, 2)
M03(quin, 3)
M04(qd, 21,9)
M06(br d, 21,9)
M15(br s, 11a) M10(br s)
δH (DMSO-d6): 10.39 (1H, br s, M15), 7.49-7.54 (2H, m, M17), 7.44-7.49 (1H, m, M14), 7.31-7.36 (2H, m,M13), 7.22-7.28 (5H, m, M12), 4.69-4.77 (1H, m, M11), 4.39 (3H, br s, M10), 3.53 (2H, br d, J=11 Hz,M09), 3.16-3.21 (2H, m, M16), 3.08-3.16 (2H, m, M08), 2.96-3.03 (2H, m, M07), 1.94 (2H, br d, J=13 Hz,M06), 1.84 (2H, t, J=7 Hz, M05), 1.65 (2H, qd, J=13 and 3 Hz, M04), 1.41 (2H, quin, J=7 Hz, M03),1.08-1.17 (2H, m, M02), 0.74 (3H, t, J=7 Hz, M01)
CH3 1
2
34
ON8
910
N+
12
13
15
1617
18
19
2021
23
2425
26
27
H11a
Joint Research CentreThe European Commission's in-house science service
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Page 6 of 13
Table of assignments and zoomed parts:
Chemical Shift (ppm)7.7 7.6 7.5 7.4 7.3 7.2 7.1
Nor
mal
ized
Inte
nsity
0
0.25
0.50
4.86
2.01
1.02
1.97
M14(m, 25)
M17(m, 26,24)M13(m, 18,16)
M12(m, 19,15,17,27,23)
No. Atom Exp. Shift(ppm)
Multiplet
1 1 0.74 M012 2 1.12 M023 3 1.41 M034 21 1.65 M045 9 1.65 M046 4 1.84 M057 21 1.94 M068 9 1.94 M069 13 3.00 M0710 20 3.12 M0811 10 3.12 M0812 12 3.18 M1613 20 3.53 M09
No. Atom Exp. Shift(ppm)
14 10 3.5315 8 4.7316 19 7.2517 15 7.2518 17 7.2519 27 7.2520 23 7.2521 18 7.3322 16 7.3323 25 7.4724 26 7.5125 24 7.5126 11a 10.39
Chemical Shift (ppm)4.5 4.0 3.5 3.0 2.5 2.0 1.5
Nor
mal
ized
Inte
nsity
0
0.1
0.2
0.3
0.4
0.5
2.93
2.03
2.01
1.96
2.03
2.02
2.03
2.08
1.98
1.97
2.66
0.98
DMSOM16(m, 12)
M01(t, 1)
M07(m, 13)M09(br d, 20,10)
M08(m, 20,10)
M05(t, 4)
M11(m, 8)
M02(m, 2)
M03(quin, 3)
M04(qd, 21,9)
M06(br d, 21,9)
M10(br s)
CH3 1
2
34
ON8
910
N+
12
13
15
1617
18
19
2021
23
2425
26
27
H11a
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 7 of 13
16010001.002.001.1r.esp
Chemical Shift (ppm)170 160 150 140 130 120 110 100 90 80 70 60 50 40
Nor
mal
ized
Inte
nsity
-1.0
-0.9
-0.8
-0.7
-0.6
-0.5
-0.4
-0.3
-0.2
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
DMSO
M01(s, 1)
M02(s, 2)
M03(s, 3)
M04(s, 21,9)
M05(s, 13)
M11(s, 8)
M12(s, 20,10)
M15(s, 25)
M16(s, 17)
M17(s, 18,16)
M18(s, 26,24)
M19(s, 27,23)
M20(s, 14)M21(s, 22)
M22(s, 5)M06(s, 4)
M13(s, 12)
Acquisition Time (sec) 0.9044 Comment 5 mm CPQCI 1H/19F-13C/15N/D Z-GRD Z114073/0012Date 08 Jan 2016 15:30:45 Date Stamp 08 Jan 2016 15:30:45File Name \\139.191.6.82\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCIM_NAS\BCIM_6
00\16010001_2015-30772\2\PDATA\1\1rFrequency (MHz) 150.90 Nucleus 13CNumber of Transients 256 Origin spectOriginal Points Count 32768 Owner nmrsuPoints Count 32768 Pulse Sequence jmodReceiver Gain 183.05 SW(cyclical) (Hz) 36231.88Solvent DMSO-d6 Spectrum Offset (Hz) 18108.3359Spectrum Type APT Sweep Width (Hz) 36230.78Temperature (degree C) 27.002
δC (DMSO-d6): 171.9 (C-5), 138.7 (C-22), 137.6 (C-14), 130.8 (C-23, 27), 129.9 (C-24, 26), 129.1 (C-16,18), 129.1 (C-17), 129.0 (C-25), 127.2 (C-15, 19), 56.9 (C-12), 51.4 (C-10, 20), 49.5 (C-8), 34.3 (C-4),29.9 (C-13), 27.8 (C-9, 21), 27.4 (C-3), 22.1 (C-2), 14.1 (C-1)
CH3 1
2
34
5ON
89
10
N+
12
13
14
15
1617
18
19
2021
2223
2425
26
27
H
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 8 of 13
Zoomed parts:
Chemical Shift (ppm)141 140 139 138 137 136 135 134 133 132 131 130 129
Nor
mal
ized
Inte
nsity
-0.2
-0.1
0
0.1
0.2
0.3
0.4
0.5
0.6
M14(s, 19,15)M15(s, 25)
M20(s, 14)M21(s, 22)
M16(s, 17)
M17(s, 18,16)
M18(s, 26,24)
M19(s, 27,23)
CH3 1
2
34
5ON
89
10
N+
12
13
14
15
1617
18
19
2021
2223
2425
26
27
H
Chemical Shift (ppm)60 55 50 45 40 35 30 25 20
Nor
mal
ized
Inte
nsity
-0.45
-0.40
-0.35
-0.30
-0.25
-0.20
-0.15
-0.10
-0.05
0
DMSO
M11(s, 8)M01(s, 1)
M02(s, 2)
M03(s, 3)
M04(s, 21,9)
M05(s, 13)
M12(s, 20,10)
M06(s, 4)
M13(s, 12)
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Page 9 of 13
MS spectraCombine <2-54> Count 351Data Source Name 16010001_2015-30772_MS_50-400_1_Centroided Mass Spectrum_ES+Data Type Centroided Mass Spectrum Date 19 Jan 2016 17:26:39Date Stamp 19 Jan 2016 16:45:45 Estimated Molecular Ion 365.198File Name \\139.191.6.82\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCIM_NAS\BCIM_
QTOF\16010001_2015-30772\16010001_2015-30772_MS_50-400.CDFInlet Model Electrospray Inlet Ion Mode ES+Plot Type Stick Retention Time 0.405Scan 22 Separation Type No ChromatographySpectrum Assigned 27.5% [50-400/0-100] TIC 479.63Total Signal 11909.3965
CH3
O
N
N
Retention Time: 0.405 Ion Mode: ES+
m/z3603403203002802602402202001801601401201008060
Rel
ativ
e In
tens
ity (%
)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
60.0
274
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
74.0
688
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
83.0
361
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
119.
0489
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
130.
1120
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
144.
9533
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
189.
9538
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
190.
9458
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
223.
0452
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
365.
1975
ES+
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 10 of 13
Combine <5-54> Count 351Data Source Name 16010001_2015-30772_MSMS_365_20EV_50-400_1_Centroided Mass Spectrum_ES+Data Type Centroided Mass Spectrum Date 19 Jan 2016 17:26:40Date Stamp 19 Jan 2016 16:47:22 Estimated Molecular Ion 366.195File Name \\139.191.6.82\ihcp∴I01\Bio_Chemical_Interaction_Metabonomics\CLEN2SAND\!proposed-structure\BCIM_NAS\BCIM_
QTOF\16010001_2015-30772\16010001_2015-30772_MSMS_365_20EV_50-400.CDFInlet Model Electrospray Inlet Ion Mode ES+Plot Type Stick Retention Time 0.829Scan 45 Separation Type No ChromatographySpectrum Assigned 87.5% [50-400/0-100] TIC 225.77Total Signal 18373.9004
CH3
O
N
N
Retention Time: 0.829 Ion Mode: ES+
m/z3603403203002802602402202001801601401201008060
Rel
ativ
e In
tens
ity (%
)
0
5
10
15
20
25
30
35
40
45
50
55
60
65
70
75
80
85
90
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379
85.0
379 10
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
2510
5.04
25
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
134.
0619
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
146.
0613
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
178.
0826
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
188.
0990
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080
218.
1080 24
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
1424
4.12
14
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
281.
1484
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
365.
1968
ES+
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 11 of 13
Table of fragmentsNo. Fragment Structure Formula Label m/z Calc. TIC Calc.
(%)Difference
(Da)m/z Exp. RI Exp.
(%)TIC Exp.
(%)Origin
1 1-6 C+
5
O6
43
2CH3 1
C5H9O M - C19H23N2 85.065 0.540 -0.027 85.038 1.149 0.540 Manual
2 12-19
CH2+
1213
14
15
1617
18
19 C8H9 M - C16H23N2O 105.070 3.350 -0.027 105.043 6.917 3.350 Manual
3 5-8,22-27(+H2)
CH+
8 N75
O6
22
23 24
25
2627
C8H8NO M - C16H24N 134.060 1.165 0.002 134.062 2.392 1.165 Manual
4 7-10,21-27CH2
+
10 98
21
N7 22
2324
25
2627
C10H12N M - C14H20NO 146.096 0.819 -0.035 146.061 1.648 0.819 Manual
5 1-7,22-27(+H2) C11H16NO M - C13H16N 178.123 0.312 -0.040 178.083 0.619 0.312 Manual
6 8-21N11
109
CH+
82120
12
1314
1516
17
1819
C13H18N M - C11H14NO 188.143 51.427 -0.044 188.099 100.000 51.427 Manual
7 1-10,22-27(+H)
CH2 109 CH+
8N7
5
O6
22
23 24
25
2627
4
32
CH3 1
C14H20NO M - C10H12N 218.154 0.316 -0.046 218.108 0.605 0.316 Manual
8 1-10,20-27(-H)
10
9 8
21CH2
+
20
N7
5
O6
22
23 24
25
26274
32
CH3 1
C16H22NO M - C8H10N 244.170 2.672 -0.048 244.121 5.013 2.672 Manual
9 7-27(+H2) C19H25N2 M - C5H7O 281.201 2.943 -0.053 281.148 5.331 2.943 Manual
10 M(+H)NH
+11
10
9
821
20
1213
1415
16
17 18
19
N7
5O6
22
23 24
25
2627
4
3
2
CH31
C24H33N2O M + H 365.259 23.922 -0.062 365.197 40.879 23.922 Manual
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 12 of 13
Fragmentation of the observed molecule and the virtual MS (EI+) spectrum predicted byThermo Scientific Mass Frontier
5075
100
125
150
175
200
225
250
275
300
325
350
375
400
425
013253850637588100
273.
20
146.
10
189.
10
105.
10
274.
2057
.10
96.1
013
2.10
91.1
077
.10
42.0
011
8.10
147.
1019
0.10
70.1
023
0.20
172.
1024
4.20
335.
2036
3.20
307.
2032
1.20
215.
2026
4.20
415.
1040
1.00
N
N O
•
N
N O
•ON
N
ON
N
N
NH
NH
N
ON
Joint Research CentreThe European Commission's in-house science service
ADMINISTRATIVE ARRANGEMENT JRC-Nr 33619-CLEN2SAND-DG TAXUD-Nr TAXUD/2014/DE/315
Page 13 of 13
Fragmentation of the observed molecule and the virtual MS and MSMS spectra predictedby Thermo Scientific Mass Frontier
5063
7588
100
113
125
138
150
163
175
188
200
213
225
238
250
263
275
288
300
313
325
338
350
363
375
388
400
013253850637588100
365.
20 366.
2160
.02
119.
0583
.03
144.
9518
9.95
130.
1274
.07
223.
05
367.
2110
2.01
208.
9526
2.02
171.
0615
7.98
244.
0829
3.96
311.
9634
4.86
278.
9333
1.02
387.
1835
7.91
96.0
2
N
N O
H
5063
7588
100
113
125
138
150
163
175
188
200
213
225
238
250
263
275
288
300
313
325
338
350
363
375
388
400
013253850637588100
188.
10
365.
20
189.
10
366.
2110
5.04
281.
1524
4.12
134.
0685
.04
160.
0717
8.08
218.
1157
.05
272.
1512
0.05
232.
1269
.04
96.0
519
7.61
374.
4935
7.56
307.
0825
7.91
393.
1833
4.99
321.
1029
0.07
N
N O
H
N 2H
N
N O
N O
NN 2
H
O
O
N O
NH