Accurate Mass for GC/MSD & LC/MSD

Transcription

1 Accurate Mass for GC/MSD & LC/MSD A Presentation at Agilent Hospitality VIP Event Yongdong Wang, PhD Cerno Bioscience Norwalk, CT, USA June 6, 2018

2 Agenda Cerno s Novel MS Calibration: Why & How Application to GC/MSD Application to LC/MSD

3 Cerno/Agilent Startup in Norwalk, CT, USA MS calibration & analysis technology 14 granted/pending patents world-wide MassWorks software product: Standalone post acquisition application software V1.0, PittCon Bronze Medal (now V5.0) Supports nearly all MS systems but works the best on Agilent MSD Over 1,200 customers in US, Canada, Europe, Japan, China etc Agilent partnership Key function: the only approach for accurate mass formula determination / compound ID with a quadrupole MS Real key technical advance: Spectral Accuracy

4 Rel. Abundance Actual MS (Profile/Raw): C 25 H 23 N 2 OS Quadrupole, R = m/z

5 Rel. Abundance After MS Centroiding: C 25 H 23 N 2 OS Quadrupole, R = m/z

6 Rel. Abundance True Mass Spectrum: C 25 H 23 N 2 OS True Mass R = Infinity m/z

7 Rel. Abundance Enough Details Even at Low Res Mass R= C 33 S 15 N 2 H C25H23N2OS C4H19N16OF2NaP C20H27N2FNaP2 C9H33N6O3S2P2 C10H27N10OS m/z

8 True Raw True m/z-only & Profile Fully Mass MS Mass Calibrated Calibration Spectrum and True MS MS Mass Accuracy Plus Spectral Accuracy m/z 8

9 counts Mass Accuracy Average of all RT Windows - C:\TestData\T42Desktop\BobStrife\MS-MS\101306_1.raw\_FUNC001.crn Raw: mass accuracy ±0.2~0.5Da Calibrated: mass accuracy ±0.002~0.005Da x better mass accuracy m/z

10 ±5mDa/400Da = ±12ppm?!

11 ±5ppm Tough Even for TOF Row Formula Mono Isotope Mass Error (mda) Mass Error (ppm) Spectral Accuracy RMSE DBE 1 C19H15N10O E C19H23N6S E C6H23N8O E C11H19N12O3S E C14H27N2O9S E C11H27N8O2S E C26H23O2S E C22H23O E C18H19N6O E C15H31N2O4S E C18H27N2O4S E C7H19N12O E C10H23N8O7S E C15H23N6O5S E C23H27O2S E C23H19N4O E C17H23N2O E C8H23N12O3S E C16H27N6S E C8H15N16O E C17H27N4O3S E

12 Enters Spectral Accuracy

13 counts Spectral Accuracy Average of Scans 14 thru 49 (0.25 to 0.87) Calibrated MS Theoretical MS Unparalleled Spectral Accuracy m/z

14 Technology: Patented, Peer-Reviewed & Published Wang, Y.; Gu, M. Anal. Chem. 2010, 82, (17).

15 Agilent/Cerno Partnership MassWorks for GC/MSD, Agilent Part # G6851AA MassWorks for LC/MSD, Agilent Part # G6887AA Specially configured version with highly attractive pricing when bundled with new MSD purchase Unique ease of use: GC/MSD-only AutoCal Unique spec among all SQ systems: Mass accuracy: ±0.005 Da Spectral accuracy: 99%

16 GC/MSD: Hardware Advantages Heated gold-coated ceramic quads Superb stability and reproducibility /how to quantify? High mass accuracy demonstratable w/ RIGHT calibration Raw Scan (Profile) mode readily available! None of detrimental on-board processing/filtering Compromised spectral accuracy Wide (5-6 orders of) linear dynamic range High spectral accuracy

17 counts relative abundance PFTBA Valve On PFTBA Valve Off GC/MS: Readily Available Cal Gas E:\Mulligan\ \TETSR3.D\DATA.MS GC Peak of Unknown Average of Scans 1398 thru 1439 (12.23 to 12.47) minutes CF C 9 F 20 N m/z Available on all GC/MS systems Computer-controllable & user-programmable Fragment ions covering Da Even internal to a GC/MS run

18 AutoCal : Out-of-Box Accurate Mass Mass accurate to 0.00x Da

19 Unique Specifications for 5977 EI SIM IDL (Helium Carrier gas with Auto Liquid Sampler) 10 fg or less IDL for Extractor ion source, turbo molecular pump system 24 fg or less for Inert EI source, turbo molecular pump system 30 fg or less for Inert EI source, diffusion pump system IDL statistically derived at 99% confidence level from the area precision of eight sequential splitless injections of 100 fg OFN,monitoring m/z 272. EI scan S/N (Helium carrier gas manual injection) 1500:1 or higher S/N for Extractor ion source, turbo molecular pump system 600:1 or higher for Inert EI Ion Source, turbo molecular pump system 300:1 or higher for Inert EI Ion Source, diffusion pump system These numbers will be given by 1-μL injection of 1 pg/μl OFN standard scanning from 50 to 300 u at nominal u ion. PCI scan S/N (Methane) 125:1 S/N will be given by 1-μL injection of 100-pg/μL BZP2 standard scanning from 80 to 230 u at nominal 183 u ion NCI scan S/N (Methane) 600:1 S/N will be given by 2-μL injection of 100 fg/μl OFN standard scanning from 50 to 300 u at nominal 272 u ion Mass Accuracy Spectral Accuracy 1-uL injection of a 100-pg/uL OFN standard scanning from u will give its monoisotope at m/z ± uL injection of a 100-pg/uL OFN standard scanning from u will give 99.0% spectral accuracy

20 Unique Specifications for LC/MSD EN: EN: InfinityLab LC/MSD Series Agilent LC/MSD XT For For OpenLAB CDS CDS and and OpenLAB CDS CDS ChemStation Edition

21 Applications Elemental composition determination (research/education/teaching) Forensics chemistry Pharma impurity ID Spectral accuracy for mixture analysis M+ and (M-H)+ in EI Labeled isotopes LC or MS peak purity: deamidation of biologics Metal organics, catalyst etc

22 Elemental Composition Determination Rank Formula Spectral Accuracy (%) C9H2N2O6F2 C2HN2O3F9 C10N2O3F4 C7HN4O7F C15N2O4 C10N4O6 C4N8O7 C20O2 C10F

23 counts relative abundance PFTBA Valve On PFTBA Valve Off Forensics Chemistry Unknown E:\Mulligan\ \TETSR3.D\DATA.MS GC Peak of Unknown Average of Scans 1398 thru 1439 (12.23 to 12.47) minutes CF C 9 F 20 N m/z

24 c ount s c ount s GC/MS Calibration A v er age of S c ans 785 t hr u 799 ( t o 8. 74) A v er age of S c ans 785 t hr u 799 ( t o 8. 74) m / z m / z MassWorks calibrates {m/z, peak shape} to improve mass accuracy by up to 100x

25 GC/MS Formula ID Correct identification w/ Spectral Accuracy

26 NIST Library Search w/same Data Acquisition

27 A Highly Toxic Compound Not Found in NIST Library

28 A Highly Toxic Compound Not Found in NIST Library

29 LC/MSD MassWorks Calibration Agilent tune solution: Universal standard (wide m/z range, ESI+ and ESI-) Stable and readily available External acquisition preferred Calibration drift: not significant (~5mDa/day) Known internal standards in sample, e.g., API Conveniently found within same sample Internal within same run No extra experimental work/acquisition Limited m/z range coverage: effective ±100Da

30 Agilent Tune Solution Calibration 10uL loop injection

31 Drug Impurity ID: Parent Drug as Calibration Standards Parent Impurity #1 Impurity #2 RT Window Name Formula Entry Net Formula Charge Mono Isotope Centroid Left Edge Right Edge 1 0: Scans Simvastatin C25H39O5 C25H39O : Scans NH4 Adduct C25H42NO5 C25H42NO : Scans K Adduct C25H38O5K C25H38O5K All

32 Drug Impurity ID #1

33 Drug Impurity ID #2 Impurity in impurity

34 [C14] Ring Labeled Compounds: Diclofenac Experimental * Diclofenac Sodium C14H10Cl2NNaO2 [14C] labeled SA=62.7 mci/mmol 7 possible isotopes Native, 1[14C] - 6[14C]

35 Results of Spectral Accuracy Mixture Analysis

36 GC/MSD RCM Paper: Flavor

37 GC/MSD RCM Paper: Forensics

38 LC/MS Paper: HDX Kinetics

39 LC/MS: Environmenal

40 Application Areas Pharmaceutical impurity ID: must have Small molecule or large biologics Leachables / Extractables Food / Beverages / Beer / Wine / Liquor Flavor / Fragrances / Cosmetics / Natural products Academic / Government Labs: Organic synthesis or other research support/acs journal accepted Novel tool to teach MS and accurate mass concept Forensics / Crime labs / Anti-doping / Homeland security / Environmental / Fine chemical / Petrochemical / Drug testing / Art museums / Paint / Automobile / Energy / Materials etc QA / QC labs Any GC/MSD with NIST library - need for compound ID

41 Cerno web Collaterals/Support/Contact Info Tech demo: Publications: App Notes: Agilent web Dedicated page: (another one for GC/MSD) Flyer: EN, EN App Notes: EN, EN, EN (polymer), EN (food) Support and Contacts Data acquisition setup: Training video: live demos / visits, anyone?