The Many Forms of NIST 2020 MS Libraries

Steve SteinNIST Mass Spectrometry Data CenterBiomolecular Measurement Division

Key MS Library Qualities

• Coverage• How big is it – NO!• Contains spectra of interest – YES!

• at conditions of interest

• Quality• Can you rely on it?• All spectra curated, intercompared using best possible software

• Software• Find and confirm identification• Assists if compound not in library

Tandem Library

EI Library

Available Chemicals

Coverage: Compound Selection

N

N N

N

CH3O

O

CH3

CH3

RYYVLZVUVIJVGH-UHFFFAOYSA-N

InChI

Wikipedia

HMDB

KEGG

FoodDB

EPA/FDA Lists

Drug Bank

30+ Collections

...

Combine & Rank

Acquire

GC/MS AMDISCustomLibrary

2nd

Evaluator

Chemicals of Interest

Available Chemicals

Chemical Inventory

Hold

3nd

Evaluator

NIST 2020

Acquire

Spectra & RI

InChIKey

Structures, Names

Resolve Inconsistencies

AcceptReject

Select Best Replicate

QC Software

Final Evaluator

Assemble

For all evaluations:Structure Search

Hybrid SearchMS Interpreter

Quality: EI Evaluation

TandemYang MOD 2:15Liang WOB 2:30

MS INTERPRETER

CONNECT PEAKS TO

STRUCTURES

MAJOR UPDATE

NISTMS.EXE

USER INTERFACE

HYBRID SEARCH

FOR COMPOUNDS NOT

IN LIBRARY

NEW AI RETENTION INDEX

ESTIMATES

SOFTWARE

Hybrid SearchIdentify Compounds Not in Library

MOD pm 3:50: Cooper et al.

OcinaplonAnxiolytic

drug

1,000s of New Compounds of Analytical InterestHuman & Plant MetabolitesFlavor/Fragrance – FoodDrugs & their MetabolitesForensics, ToxinsPesticides –ContaminantsIndustrial Chemicals Petrochemicals, Surfactants, Lipids, …

Also TMS, Ac and Me Derivatives

UWA-101Parkinson’s

drug

MK-212Serotonin

Agonist

PretomanidAntibiotic

Gardenia amideFlavor/Fragrance

BenfuresateHerbicide

LoliolidePlant

Metabolite

NIST/EPA/NIH EI MS Library – NIST 20

306,869 Compounds, 43,774 Replicate Spectra

40 K More Compounds than NIST 17

NIST Tandem Mass Spectral Library2020 Release

31K Compounds, 2X More than 2017186K Precursor Ions - 1.3M Spectra

Compounds: Fragmentation Methods27,840 HRAM (High Res Accurate Mass)29,890 QTOF, HCD, IT-HRAM, QqQ29,444 Ion Trap (Low Res., up to MS4)246 APCI HRAM ‘Extractables and Leachables’

Precursor Ion Types26,575 Protonated12,589 Deprotonated10,032 Water/Ammonia Loss24,167 Other In-Source Generated

CAS# 4261-42-1 C21H20O11 (hr_msms_nist2020_v42) Isoorientin [M+H]+ IT-FT 35%

P=449.1

280 300 320 340 360 380 400 420 440 4600

50

100

C17H13O7=p-C4H8O4

C20H15O8=p-CH6O3

C21H19O10=p-H2O

OH

HO

O

O

O

OH

HO

HO

HO

OH

OH

CAS# 121-75-5 C10H19O6PS2 (hr_msms_nist2020_v42) Malathion [M+H]+ HCD

10% 6 P=331

90 120 150 180 210 240 270 300 3300

50

100

C6H7O3=p-C4H13O3PS2

C8H13O4=p-C2H7S2O2P

C8H14O5PS2=p-C2H6O

p

S

P

O

O

OO

S

O

O

CAS# 67-97-0 C27H44O (hr_msms_nist2020_v42) Cholecalciferol [M+H]+ HCD 35%

2 P=385.3

80 120 160 200 240 280 320 360 4000

50

100

C9H13=p-C18H32O

C19H31=p-C8H14O

p

H

H

HO

CAS# 115550-35-1 C17H19FN4O4 (hr_msms_nist2020_v42) Marbofloxacin [M+H]+ IT

-FT 35% P=363.1

230 240 250 260 270 280 290 300 310 320 330 340 350 360 3700

50

100

C14FH15N3O2=p-C3H5NO2

C15FH15N3O4=p-C2H5N

C17FH18N4O3=p-H2ON

N

F

O

N

NO

OH

O

Vitamin D3Metabolite

CAS# 91-20-3 C10H8 (apci_msms_nist2020_v12) Naphthalene [M]+. QTOF 30V

P=128.1

70 80 90 100 110 120 130 1400

50

100

C6H6=p-C4H2

C8H6=p-C2H2

p

CAS# 437-38-7 C22H28N2O (hr_msms_nist2020_v42) Fentanyl [M+H]+ HCD 50% 3

P=337.2

60 90 120 150 180 210 240 270 300 3300

50

100

C8H9=p-C14H20N2O

C13H18N=p-C9H11NO

C19H25N2=p-C3H4Op

N

O

N

FentanylDrug

MalathionInsecticide

Luteolin glucoside Flavone

Fuc-GM1(d18:1/16:0)Glycolipid

2020 Tandem LibraryWide Range of Compounds

LopinavirAntiviral

NaphthaleneE&L, APCI

Marbofloxacin Antibiotic

• Annotated Recurrent Unidentified Spectra (ARUS)• Good quality, RUS converted to ‘consensus’ spectra

• Annotated by Hybrid Search, then by Evaluator

• LC/MS – Milk Oligos, Urine and Plasma/Serum, Acylcarnitines

• GC/MS – RI in Essential Oils/Foods

• Available: http://chemdata.NIST.gov/• Documented in Multiple Papers

• Urine/Plasma Updated for NIST 2020 Tandem Library

Coverage:Recurring, Unidentified Spectra

From Multiple NIST Standard Reference Materials

Plasma/Serum, Urine, MilkPos/Neg, Chemdata.NIST.gov

LC/MS ARUSHybrid Identified HRAM Spectra

Simon et al. TP 392

Peptide Library – 2020 Version

85% increase in peptide ions 489,921 (2016) -> 911,783

25% to 36% proteome coverage

Quality – 7 quality filters1 spectrum/peptide ion

Peptide.NIST.gov

ThP 291: Sheetlin et al.