619 compounds were purchased from IROA technologies . These compounds were injected into UHPLC-QTOF instrument and whoever ionized and yielded a good MSMS spectra were included in the MSMS library.
Compounds were in dried state in 96-well plates. After dissoliving them in Water:MeOH(50:50), they were pooled by rows into glass vials and each vial contained 8 compounds. Injection volume was 2 micorLiter.
An Agilent 1290 UHPLC and 6550 quadruple time-of-flight mass spectrometer (Agilent Inc, Santa Clara, USA) were used for the acquisition of raw data. Instrument was tuned and calibrated for small molecule analysis, as per instructions provided by the manufacturer. The mass spectrometer was operated in ESI (Positive and negative) and MS1 and targeted ion MS/MS modes separately. Scan speed was 2 scans per seconds in both MS1 and MS/MS modes. Collision induced dissociation was performed at 10,20,40 eV energy. Active exclusion of precursor was enabled. Data were acquired in centroid mode only. Mass range was 50-1200. Source parameters were : Gas Temperature - 175 degree celcius, Gas Flow- 12 l/min, Nebulizer – 45 psig, Sheath Gas Temperature – 350 degree celsius, Sheath Gas flow – 12 l/min, VCap – 3500, Nozzle voltage – 300, Fragmenter – 175. Mz 121.0508 and 922.0097 was used as reference ions to calibrate the mass axis. The reference ion solution was delivered using an ancillary HPLC pump with a flow rate of 0.010 mL/min. For the main gradient, solvent A was water (0.1% formic acid) and solvent B was methanol (0.1% formic acid). Flow rate was 0.4 ml/min. The gradient started at 5% B, reaching to 100% B at 6.0 min, then stay at 100% B until 10.50, then drops to 5% B at 10.51 and stay at the same until 12.50. Column temperature was 45 degree celsius. An UHPLC reversed phase column (Waters Acquity HSS T3 1.8 microM, 2.1X100 mm, part number 186003539) was used for separating compounds. Data acquisition for entire library was done in a single batch lasted for three days.
"Find compound by Targeted MSMS algorithm" in the Agilent Mass Hunter version 6.0 was used to extract MSMS spectra for the compounds. Batch processing was achieved by Agilent DA reprocessor software. Exported CEF files were imported into Agilent PCDL manager software for creation of the library. Proton, Water, Sodium and Potassium adducts (whenever present) were included in the library. Compounds eluting in first one minute were excluded from the final database, due to their poor retention on the RP column.
To use the library, untargeted metabolomics datasets should be acquired in Auto MSMS mode with the same chromatographic method. The method is provided in the Google Drive folder. Processed the data with find compounds by Auto MSMS algorithm in Mass hunter software. After that, use identify compound with accurate mass library algorithm for annotation and use the download library (.cdb format) as input. In a typical blood sample, extracted with a classical blood:methanol (1:5) protocol, around 70-80 compounds can be routinely monitored with this library. Then, export the CEF files and merge them in MPP to export a data table.
| Compound Name | Formula | Mass | RT (min) | Num Spectra |
|---|---|---|---|---|
| 10-HYDROXYDECANOATE | C10H20O3 | 188.1416 | 5.144 | 5 |
| 1-2-DIDECANOYL-SN-GLYCERO-3-PHOSPHOCHOLINE | C28H56NO8P | 565.3748 | 7.105 | 3 |
| 1-2-DIPALMITOYL-SN-GLYCEROL | C35H68O5 | 568.5086 | 10.269 | 6 |
| 12-HYDROXYDODECANOIC ACID | C12H24O3 | 216.1733 | 5.824 | 9 |
| 17A-21-DIHYDROXY-4-PREGNENE-3-20-DIONE | C21H30O4 | 346.2154 | 5.551 | 9 |
| 1-NAPHTHYLAMINE | C10H9N | 143.0742 | 3.955 | 3 |
| 2'-4'-DIHYDROXYACETOPHENONE | C8H8O3 | 152.047 | 1.81 | 3 |
| 2-4-DIHYDROXYPTERIDINE | C6H4N4O2 | 164.0331 | 1.532 | 6 |
| 2-4-DIHYDROXYPYRIMIDINE-5-CARBOXYLIC ACID | C5H4N2O4 | 156.017 | 1.191 | 6 |
| 25-HYDROXYCHOLESTEROL | C27H46O2 | 402.3502 | 7.299 | 3 |
| 2'-DEOXYADENOSINE | C10H15N5O4 | 269.1122 | 1.674 | 3 |
| 2'-DEOXYGUANOSINE | C10H15N5O5 | 285.1072 | 1.766 | 3 |
| 2-HYDROXYPHENYLACETIC ACID | C8H8O3 | 152.0476 | 3.458 | 6 |
| 2-HYDROXYPYRIDINE | C5H5NO | 95.037 | 1.639 | 3 |
| 2-METHYLMALEATE | C5H6O4 | 130.0266 | 2.097 | 3 |
| 2-OXOADIPATE | C6H8O5 | 160.0361 | 1.332 | 3 |
| 2-QUINOLINECARBOXYLIC ACID | C10H7NO2 | 173.0477 | 3.152 | 9 |
| 3(2-HYDROXYPHENYL)PROPANOATE | C9H10O3 | 166.0624 | 4.036 | 3 |
| 3(4-HYDROXYPHENYL)LACTATE | C9H10O4 | 182.0577 | 2.714 | 6 |
| 3'-5'-CYCLIC AMP | C10H12N5O6P | 329.0523 | 1.785 | 6 |
| 3-5-DIIODO-L-THYRONINE | C15H13I2NO4 | 524.8935 | 4.584 | 3 |
| 3ALPHA-11BETA-17ALPHA-21-TRETRAHYDROXY-5ALPHA-PREGNAN-20-ONE | C21H34O5 | 366.2415 | 5.033 | 6 |
| 3-HYDROXY-3-METHYLGLUTARATE | C6H10O5 | 162.0524 | 1.574 | 6 |
| 3-HYDROXYANTHRANILATE | C7H7NO3 | 153.0426 | 2.495 | 6 |
| 3-HYDROXYKYNURENINE | C10H12N2O4 | 224.0794 | 1.105 | 3 |
| 3-HYDROXYPHENYLACETATE | C8H8O3 | 152.0473 | 3.286 | 3 |
| 3-METHOXY-4-HYDROXYMANDELATE | C9H10O5 | 198.0522 | 1.81 | 6 |
| 3-METHOXYTYRAMINE | C9H13NO2 | 167.0946 | 3.091 | 3 |
| 3-METHYL-2-OXINDOLE | C9H9NO | 147.0683 | 4.38 | 6 |
| 3-METHYLGLUTARIC ACID | C6H10O4 | 146.0577 | 2.692 | 3 |
| 4-ACETAMIDOBUTANOATE | C6H11NO3 | 145.0736 | 1.648 | 9 |
| 4-AMINOBENZOATE | C7H7NO2 | 137.0478 | 2.306 | 5 |
| 4-COUMARATE | C9H8O3 | 164.0479 | 3.808 | 6 |
| 4-HYDROXYPHENYLACETATE | C8H8O3 | 152.047 | 2.946 | 3 |
| 4-PYRIDOXATE | C8H9NO4 | 183.0529 | 1.741 | 9 |
| 5'-DEOXYADENOSINE | C10H13N5O3 | 251.1016 | 2.064 | 6 |
| 5-HYDROXYINDOLEACETATE | C10H9NO3 | 191.0577 | 2.816 | 6 |
| 5-HYDROXY-L-TRYPTOPHAN | C11H12N2O3 | 220.0846 | 1.666 | 6 |
| 5'-METHYLTHIOADENOSINE | C11H15N5O3S | 297.0896 | 2.742 | 6 |
| 5-OXO-D-PROLINE | C5H7NO3 | 129.0423 | 1.111 | 6 |
| 5-VALEROLACTONE | C5H8O2 | 100.0526 | 1.951 | 3 |
| 6-HYDROXYNICOTINATE | C6H5NO3 | 139.0273 | 1.735 | 3 |
| ADENOSINE | C10H13N5O4 | 267.0964 | 1.624 | 3 |
| ADIPIC ACID | C6H10O4 | 146.0578 | 2.626 | 3 |
| ANILINE-2-SULFONIC ACID | C6H7NO3S | 173.0144 | 1.623 | 3 |
| ANTHRANILATE | C7H7NO2 | 137.0478 | 2.306 | 5 |
| BENZALDEHYDE | C7H6O | 106.0421 | 10.141 | 3 |
| BIOTIN | C10H16N2O3S | 244.0888 | 3.537 | 6 |
| BUTANAL | C4H8O | 72.0577 | 3.469 | 3 |
| CAFFEIC ACID | C9H8O4 | 180.0414 | 3.24 | 3 |
| CAFFEINE | C8H10N4O2 | 194.0804 | 3.204 | 6 |
| CHENODEOXYCHOLATE | C24H40O4 | 392.2933 | 6.775 | 3 |
| CHOLIC ACID | C24H40O5 | 408.2881 | 6.464 | 6 |
| CORTICOSTERONE | C21H30O4 | 346.216 | 5.53 | 6 |
| CORTISOL | C21H30O5 | 362.21 | 5.17 | 9 |
| CORTISOL 21-ACETATE | C23H32O6 | 404.2206 | 5.553 | 9 |
| CORTISONE | C21H28O5 | 360.1951 | 5.011 | 3 |
| DEOXYADENOSINE MONOPHOSPHATE | C10H14N5O6P | 331.0679 | 1.134 | 3 |
| DEOXYCHOLIC ACID | C24H40O4 | 392.2933 | 6.775 | 3 |
| DEOXYCORTICOSTERONE | C23H32O4 | 372.2309 | 6.223 | 6 |
| DEOXYURIDINE | C9H12N2O5 | 228.0744 | 1.536 | 6 |
| DESMOSTEROL | C27H44O | 384.3391 | 7.299 | 3 |
| DETHIOBIOTIN | C10H18N2O3 | 214.1321 | 3.834 | 9 |
| DIACETYL | C4H6O2 | 86.0373 | 5.206 | 3 |
| D-PANTOTHENIC ACID | C9H17NO5 | 219.1107 | 2.354 | 6 |
| DTMP | C10H15N2O8P | 322.0561 | 1.594 | 6 |
| ESTRADIOL-17ALPHA | C18H24O2 | 272.1778 | 5.825 | 3 |
| ETHYLMALONIC ACID | C5H8O4 | 132.0415 | 2.39 | 6 |
| FERULATE | C10H10O4 | 194.0582 | 3.877 | 3 |
| FOLIC ACID | C19H19N7O6 | 441.1393 | 2.983 | 3 |
| FORMYL-L-METHIONYL | C6H11NO3S | 177.046 | 2.539 | 6 |
| GLUTARATE | C5H8O4 | 132.0414 | 1.91 | 3 |
| GLYCOCHOLATE | C26H43NO6 | 465.3092 | 6.162 | 9 |
| GUANOSINE | C10H13N5O5 | 283.0913 | 1.634 | 6 |
| GUANOSINE 3'-5'-CYCLIC | C10H11N5NaO7P | 367.0283 | 1.866 | 3 |
| HIPPURATE | C9H9NO3 | 179.0584 | 3.091 | 6 |
| HOMOGENTISATE | C8H8O4 | 168.0423 | 2.016 | 6 |
| HOMOVANILLATE | C9H10O4 | 182.0579 | 3.323 | 6 |
| HYPOXANTHINE | C5H4N4O | 136.0381 | 1.065 | 9 |
| INDOLE-3-ACETALDEHYDE | C10H9NO | 159.0684 | 4.413 | 6 |
| INDOLE-3-ACETAMIDE | C10H10N2O | 174.0795 | 3.369 | 9 |
| INDOLE-3-ACETATE | C10H9NO2 | 175.0636 | 4.145 | 6 |
| INDOLE-3-ETHANOL | C10H11NO | 161.0843 | 4.151 | 6 |
| INDOLE-3-PYRUVIC ACID | C11H9NO3 | 203.0563 | 3.068 | 3 |
| INOSINE | C10H12N4O5 | 268.0804 | 1.63 | 6 |
| INOSINE 5'-PHOSPHATE | C10H13N4O8P | 348.0466 | 1.414 | 3 |
| ISOLEUCINE | C6H13NO2 | 131.0944 | 1.346 | 6 |
| ITACONATE | C5H6O4 | 130.0266 | 2.097 | 3 |
| KYNURENIC ACID | C10H7NO3 | 189.0427 | 3.124 | 6 |
| KYNURENINE | C10H12N2O3 | 208.0846 | 1.841 | 6 |
| LEUCINE | C6H13NO2 | 131.0944 | 1.443 | 6 |
| LIPOAMIDE | C8H15NOS2 | 205.06 | 4.558 | 6 |
| L-METHYLADENOSINE | C11H15N5O4 | 281.1121 | 2.247 | 3 |
| L-OLEOYL-RAC-GLYCEROL | C21H40O4 | 356.2929 | 7.249 | 3 |
| LUMICHROME | C12H10N4O2 | 242.0812 | 4.602 | 3 |
| MALEIC ACID | C4H4O4 | 116.0103 | 1.204 | 6 |
| MANDELIC ACID | C8H8O3 | 152.047 | 2.946 | 3 |
| MELATONIN | C13H16N2O2 | 232.1215 | 4.038 | 9 |
| METHYL VANILLATE | C9H10O4 | 182.0575 | 2.739 | 3 |
| METHYLMALONATE | C4H6O4 | 118.0259 | 1.551 | 3 |
| MEVALOLACTONE | C6H10O3 | 130.0627 | 1.679 | 6 |
| MONO-ETHYL MALONATE | C5H8O4 | 132.0401 | 2.651 | 3 |
| MONO-METHYL GLUTARATE | C6H10O4 | 146.0578 | 2.767 | 3 |
| N-ACETYL-DL-GLUTAMIC ACID | C7H11NO5 | 189.0634 | 1.227 | 6 |
| N-ACETYL-DL-METHIONINE | C7H13NO3S | 191.0615 | 2.79 | 3 |
| N-ACETYL-D-TRYPTOPHAN | C13H14N2O3 | 246.1004 | 3.897 | 6 |
| N-ACETYL-L-ALANINE | C5H9NO3 | 131.0579 | 1.424 | 8 |
| N-ACETYL-L-CYSTEINE | C5H9NO3S | 163.0299 | 1.725 | 0 |
| N-ACETYL-L-LEUCINE | C8H15NO3 | 173.1055 | 3.78 | 9 |
| N-ACETYL-L-PHENYLALANINE | C11H13NO3 | 207.0899 | 3.911 | 6 |
| N-ACETYLSEROTONIN | C12H14N2O2 | 218.1058 | 2.919 | 9 |
| NAD | C21H27N7O14P2 | 663.109 | 1.232 | 3 |
| NICOTINAMIDE | C6H6N2O | 122.0479 | 1.129 | 3 |
| NITRO-L-TYROSINE | C9H10N2O5 | 226.059 | 2.204 | 3 |
| NORLEUCINE | C6H13NO2 | 131.0945 | 1.502 | 6 |
| OPHTHALMIC ACID | C11H19N3O6 | 289.127 | 1.046 | 6 |
| PARAXANTHINE | C7H8N4O2 | 180.0647 | 2.706 | 6 |
| PHENETHYLAMINE | C8H11N | 121.0894 | 2.067 | 3 |
| PHENYLALANINE | C9H11NO2 | 165.0788 | 2.05 | 5 |
| PHENYLETHANOLAMINE | C8H11NO | 137.0842 | 1.48 | 3 |
| PIMELIC ACID | C7H12O4 | 160.0736 | 3.342 | 6 |
| PRENOL | C5H10O | 86.0736 | 6.999 | 3 |
| PTERINE | C6H5N5O | 163.0489 | 1.466 | 3 |
| PYRIDINE-2-3-DICARBOXYLATE | C7H5NO4 | 167.0214 | 1.304 | 3 |
| PYRIDOXAL 5'-PHOSPHATE | C8H12NO7P | 265.0345 | 1.515 | 3 |
| RAC-GLYCEROL-L-MYRISTATE | C17H34O4 | 302.246 | 6.944 | 3 |
| RIBOFLAVIN | C17H20N4O6 | 376.1386 | 3.516 | 6 |
| S-(5'-ADENOSYL)-L-HOMOCYSTEINE | C14H20N6O5S | 384.1212 | 1.236 | 3 |
| SALICYLAMIDE | C7H7NO2 | 137.0479 | 3.425 | 3 |
| S-HEXYL-GLUTATHIONE | C16H29N3O6S | 391.1783 | 4.787 | 6 |
| SN-GLYCEROL 3-PHOSPHATE BIS (CYCLOHEXYLAMMONIUM) | C15H35N2O6P | 370.2204 | 3.442 | 3 |
| SPHINGANINE | C18H39NO2 | 301.2984 | 6.15 | 3 |
| SUBERIC ACID | C8H14O4 | 174.0891 | 2.626 | 3 |
| THEOBROMINE | C7H8N4O2 | 180.0646 | 2.377 | 6 |
| THEOPHYLLINE | C7H8N4O2 | 180.0647 | 2.815 | 6 |
| THIOPURINE S-METHYLESTER | C6H6N4S | 166.0317 | 3.369 | 6 |
| THYMIDINE | C10H14N2O5 | 242.0904 | 2.082 | 6 |
| THYMINE | C5H6N2O2 | 126.0427 | 1.596 | 3 |
| THYROXINE | C15H11I4NO4 | 776.6876 | 5.51 | 6 |
| TRIIODOTHYRONINE | C15H12I3NO4 | 650.7907 | 5.144 | 3 |
| TRYPTAMINE | C10H12N2 | 160.1005 | 2.428 | 6 |
| TRYPTOPHAN | C11H12N2O2 | 204.0899 | 2.511 | 6 |
| TYRAMINE | C8H11NO | 137.0843 | 1.046 | 3 |
| TYROSINE | C9H11NO3 | 181.0734 | 1.174 | 3 |
| URATE | C5H4N4O3 | 168.0282 | 1.069 | 3 |
| URIDINE | C9H12N2O6 | 244.0691 | 1.23 | 6 |
| VITAMIN D2 | C28H44O | 396.3392 | 8.38 | 3 |
| Xanthine | C5H4N4O2 | 152.0334 | 1.22 | 0 |
| XANTHOSINE | C10H12N4O6 | 284.0755 | 1.891 | 6 |
| XANTHURENIC ACID | C10H7NO4 | 205.0384 | 4.286 | 3 |
The data for developing the library were acquired at the International Agency for Research on Cancer (IARC) Lyon France.
Send an email to Dinesh Barupal at barupal@gmail.com for any questions and comments.