ACCESSION: MSBNK-UFZ-WANA0449237762PH
RECORD_TITLE: Methotrexate; LC-ESI-ITFT; MS2; CE: 80%; R=15000; [M+H]+
DATE: 2023.08.13
AUTHORS: Tobias Schulze, Martin Krauss, Department of Effect-Directed Analysis, Helmholtz Centre for Environmental Research - UFZ, Leipzig, Germany
LICENSE: CC0
COPYRIGHT: Copyright (C) 2023
COMMENT: CONFIDENCE Reference Standard (Level 1)

CH$NAME: Methotrexate
CH$NAME: (2S)-2-[[4-[(2,4-diaminopteridin-6-yl)methyl-methylamino]benzoyl]amino]pentanedioic acid
CH$COMPOUND_CLASS: N/A; Environmental Standard
CH$FORMULA: C₂₀H₂₂N₈O₅
CH$EXACT_MASS: 454.171315804
CH$SMILES: CN(CC1=NC2=C(N=C1)N=C(N)N=C2N)C1=CC=C(C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O
CH$IUPAC: InChI=1S/C20H22N8O5/c1-28(9-11-8-23-17-15(24-11)16(21)26-20(22)27-17)12-4-2-10(3-5-12)18(31)25-13(19(32)33)6-7-14(29)30/h2-5,8,13H,6-7,9H2,1H3,(H,25,31)(H,29,30)(H,32,33)(H4,21,22,23,26,27)/t13-/m0/s1
CH$LINK: CAS 59-05-2
CH$LINK: CHEBI 44185
CH$LINK: KEGG C01937
CH$LINK: PUBCHEM CID:126941
CH$LINK: INCHIKEY FBOZXECLQNJBKD-ZDUSSCGKSA-N
CH$LINK: CHEMSPIDER 112728
CH$LINK: COMPTOX DTXSID4020822

AC$INSTRUMENT: LTQ Orbitrap XL Thermo Scientific
AC$INSTRUMENT_TYPE: LC-ESI-ITFT
AC$MASS_SPECTROMETRY: MS_TYPE MS2
AC$MASS_SPECTROMETRY: ION_MODE POSITIVE
AC$MASS_SPECTROMETRY: IONIZATION ESI
AC$MASS_SPECTROMETRY: FRAGMENTATION_MODE HCD
AC$MASS_SPECTROMETRY: COLLISION_ENERGY 80 % (nominal)
AC$MASS_SPECTROMETRY: RESOLUTION 15000
AC$MASS_SPECTROMETRY: MASS_RANGE_M/Z 50-470
AC$CHROMATOGRAPHY: COLUMN_NAME Kinetex Evo C18 2.6 um 50 x 2.1 mm
AC$CHROMATOGRAPHY: FLOW_GRADIENT 95/5 at 0 min, 95/5 at 1 min, 0/100 at 13 min, 0/100 at 24 min, 95/5 at 24.3 min, 95/5 at 32 min
AC$CHROMATOGRAPHY: FLOW_RATE 300 uL/min
AC$CHROMATOGRAPHY: RETENTION_TIME 5.132 min

MS$FOCUSED_ION: BASE_PEAK 455.1799
MS$FOCUSED_ION: PRECURSOR_M/Z 455.1786
MS$FOCUSED_ION: PRECURSOR_TYPE [M+H]+
MS$FOCUSED_ION: PRECURSOR_INTENSITY 4464565
MS$DATA_PROCESSING: RECALIBRATE loess on assigned fragments and MS1
MS$DATA_PROCESSING: REANALYZE Peaks with additional N2/O included
MS$DATA_PROCESSING: WHOLE RMassBank 3.11.2

PK$SPLASH: splash10-0059-0900000000-bbea2752a81b5f1b7cb3
PK$ANNOTATION: m/z tentative_formula formula_count mass error(ppm)
  79.029 C4H3N2+ 1 79.0291 -0.67
  79.0543 C6H7+ 1 79.0542 1
  80.0242 C3H2N3+ 1 80.0243 -1.09
  81.0449 C4H5N2+ 1 81.0447 2.58
  106.0402 C5H4N3+ 1 106.04 1.8
  106.0653 C7H8N+ 1 106.0651 1.89
  107.0481 C5H5N3+ 1 107.0478 2.85
  108.0558 C5H6N3+ 1 108.0556 1.62
  110.0592 C4H6N4+ 1 110.0587 4.73
  118.0399 C6H4N3+ 1 118.04 -0.25
  121.0511 C5H5N4+ 1 121.0509 1.66
  133.0511 C6H5N4+ 1 133.0509 2.08
  134.0602 C8H8NO+ 1 134.06 1.05
  135.054 C5H5N5+ 1 135.0539 0.16
  135.0662 C6H7N4+ 1 135.0665 -2.24
  136.0502 C6H6N3O+ 1 136.0505 -2.29
  146.0464 C6H4N5+ 1 146.0461 2.07
  148.0621 C6H6N5+ 1 148.0618 1.94
  149.0697 C6H7N5+ 1 149.0696 0.69
  150.0776 C6H8N5+ 1 150.0774 1.09
  151.0621 C6H7N4O+ 2 151.0614 4.35
  158.0463 C7H4N5+ 1 158.0461 1.06
  159.0543 C7H5N5+ 1 159.0539 2.1
  160.062 C7H6N5+ 1 160.0618 1.71
  163.0727 C6H7N6+ 1 163.0727 0.11
  175.073 C7H7N6+ 1 175.0727 1.71
  176.0807 C7H8N6+ 1 176.0805 1.09
  177.0885 C7H9N6+ 1 177.0883 1
PK$NUM_PEAK: 28
PK$PEAK: m/z int. rel.int.
  79.029 2193.5 3
  79.0543 8778.1 15
  80.0242 1434.7 2
  81.0449 1002.6 1
  106.0402 20408 35
  106.0653 28943 50
  107.0481 3120.3 5
  108.0558 7606.3 13
  110.0592 1233.7 2
  118.0399 1769.8 3
  121.0511 29314.9 50
  133.0511 165775.6 288
  134.0602 287979.1 500
  135.054 7394.5 12
  135.0662 13952.3 24
  136.0502 1603.3 2
  146.0464 4450.1 7
  148.0621 61155.7 106
  149.0697 7530.2 13
  150.0776 7761.5 13
  151.0621 1315.9 2
  158.0463 3973.1 6
  159.0543 19469.8 33
  160.062 43558.5 75
  163.0727 5757.7 10
  175.073 574280.6 999
  176.0807 164973.7 286
  177.0885 25816.4 44
//