ACCESSION: MSBNK-UFZ-WANA044925AF82PH
RECORD_TITLE: Methotrexate; LC-ESI-ITFT; MS2; CE: 90%; 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$MASS_SPECTROMETRY: COLLISION_ENERGY 90 % (nominal)
AC$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$SPLASH: splash10-0059-0900000000-48e3f2cf253bbc7081dc
PK$ANNOTATION: m/z tentative_formula formula_count mass error(ppm)
  79.0293 C4H3N2+ 1 79.0291 3.19
  79.0544 C6H7+ 1 79.0542 1.67
  80.0244 C3H2N3+ 1 80.0243 1.39
  81.0448 C4H5N2+ 1 81.0447 0.7
  106.0403 C5H4N3+ 1 106.04 2.66
  106.0654 C7H8N+ 1 106.0651 2.61
  107.0481 C5H5N3+ 1 107.0478 2.49
  108.0559 C5H6N3+ 1 108.0556 2.46
  110.0589 C4H6N4+ 1 110.0587 1.82
  118.0402 C6H4N3+ 1 118.04 2.27
  121.0511 C5H5N4+ 1 121.0509 2.23
  133.0513 C6H5N4+ 1 133.0509 2.88
  134.0602 C8H8NO+ 1 134.06 1.5
  135.0542 C5H5N5+ 1 135.0539 1.97
  135.0668 C6H7N4+ 1 135.0665 2.06
  136.0508 C6H6N3O+ 1 136.0505 2.2
  146.0465 C6H4N5+ 1 146.0461 2.8
  148.0622 C6H6N5+ 1 148.0618 2.66
  149.0699 C6H7N5+ 1 149.0696 1.82
  150.0778 C6H8N5+ 1 150.0774 2.51
  151.0615 C6H7N4O+ 1 151.0614 0.71
  158.0465 C7H4N5+ 1 158.0461 2.5
  159.0544 C7H5N5+ 1 159.0539 2.58
  160.0622 C7H6N5+ 1 160.0618 2.47
  163.0731 C6H7N6+ 1 163.0727 2.92
  175.0731 C7H7N6+ 1 175.0727 2.49
  176.0809 C7H8N6+ 1 176.0805 2.22
  177.0888 C7H9N6+ 2 177.0883 2.64
PK$NUM_PEAK: 28
PK$PEAK: m/z int. rel.int.
  79.0293 4296.4 12
  79.0544 17374.1 51
  80.0244 5655.9 16
  81.0448 1342.9 3
  106.0403 47882.7 141
  106.0654 49437.7 146
  107.0481 8404.9 24
  108.0559 9728.1 28
  110.0589 2020.4 5
  118.0402 3105.5 9
  121.0511 43074.8 127
  133.0513 210354.1 621
  134.0602 173062.4 511
  135.0542 11162.6 32
  135.0668 11342.6 33
  136.0508 2297.7 6
  146.0465 5542.3 16
  148.0622 71022.3 209
  149.0699 9044.3 26
  150.0778 10632.6 31
  151.0615 1363 4
  158.0465 7094.5 20
  159.0544 28083.8 82
  160.0622 32555.3 96
  163.0731 5369.2 15
  175.0731 338207.5 999
  176.0809 100409 296
  177.0888 9260.7 27
//