mass hunter / target deconvolution (td) and the maps 5977 ... · • target deconvolution (td) is a...
TRANSCRIPT
Mass Hunter / Target Deconvolution (TD)
and the MAPS 5977 MSD Surface Water Analyzer
Introduction
• Deconvolution Reporting Software (DRS) has been an add-on to GC-MSD
• Chemstation SW since May 2004. Brain child of Mike Szelewski, LFS
• DRS correlates results from 3 separate programs (MS Chemstation, AMDIS
• and NIST MS Search) to provide additional confidence in target analyte identification
• Target Deconvolution (TD) is a new feature of Mass Hunter Quant B.06.00,
• introduced in March 2013. The entire TD process is contained within Mass Hunter Quant
• TD provides additional confidence in target analyte identification by comparing
• deconvoluted component mass spectra against a target MS database
• TD avoids false positives (found by Quant but not confirmed) or false negatives (not found by quant
but found by TD) and increases the level of confirmation of identity
• TD can be used with either full scan data or synchronous SIM / Scan data
• Whilst essentially performing the same function, DRS and TD do not use the same processes for the confirmation of deconvoluted spectra against the target MS library and TD does not search against the NIST11 MS database
• TD has been developed using complex, ‘real World’ sample data and the results
• bench-marked against those produced by DRS
How does Target Deconvolution work in Mass Hunter Quant?
Mass Hunter Data Analysis Software
Mass Hunter’s 2 main Data Analysis modules have different functions:
Quantitative (Quant): (Target Analysis - Quantitation)
Powerful quantitation engine
Review Batch results - Batch at a Glance
Visualize quality outliers
Flexible and fully customizable data review
- Compounds at a Glance
Flexible result and report presentation
Qualitative (Qual): (Discovery of non-target compounds or features, & Method Development)
Finds compounds
Identifies compounds
Uses chromatogram and spectra extraction tools, i.e. deconvolution
Searches El Libraries
Full scan TIC of Hazardous Waste Landfill Site Water Extract
ISTD
TIC of complex samples may have many overlapping / co-eluting components
Why the Need for Target Deconvolution?
Data courtesy of the National Laboratory Service, UK Environment Agency
Deconvolution A Single Chromatographic Peak may contain Multiple Components
TIC & Spectrum
TIC TIC
Component 1
Component 2
Component 3
Components and Mixed Spectra
Failing ion ratio
Failing library match
Deconvoluted components and spectra
Component 1
Component 2
Component 3
Deconvolution Pulls Out Individual Components and their Spectra
TIC
Component 1
Component 2
Component 3
Deconvolution
Components and Mixed Spectra
Target Deconvolution Objective
From complex full scan TIC
to easy-to-interpret
report in < 1 minute per data file
Combined quantitative and qualitative .pdf Summary report
Use deconvoluted full scan mass spectra to provide additional,
increased confidence in target analyte identification
Complex samples may have interfering matrix peaks that compromise target
analyte identification based on ion ratios
Deconvoluted component spectrum
Deconvoluted component spectrum
Deconvoluted component spectrum
Deconvoluted component spectrum
Deconvoluted component spectrum
ECC
285 290 295 300 305 310 315 RT : Seconds
- 10 seconds + 10 seconds
Target RT Window
The peak apex of the deconvoluted component must be
within the RT Range of the target peak identified by the Quant Engine in order to be used for library matching. For quantitation also Quant/Qual ion ratios
TIC
Target Deconvolution Workflow in Mass Hunter Quant SW
Quant Results
Targets are identified by comparison to locked RTs and 1 Quant + 3 Qualifier ion ratios,
then quantified using Quant ion area versus calibration table
Review results in Mass Hunter Batch-at-a-Glance and
Compounds-at-a-Glance
Combined quantitative and qualitative
Target Deconvolution .pdf Summary report
MH deconvolutes component spectra and performs spectral matching of deconvoluted spectra vs target MS
database using RT Window and Library Match Score as qualifiers
Qualitative Results
All data processing performed within
Mass Hunter Quant SW
TIC
How to Set up a Mass Hunter Quant Method to use Target Deconvolution
Target Retention Time and Extraction Window are set in MH Quant Method
Target RT Extraction Window
Time window for the Extracted Ion Peak
Target Analyte Recognition Window is set under Globals Setup
Recognition window is set symmetrically around peak expected retention time, so where a peak should be found (tolerance retention time window)
Where to tell Mass Hunter Quant to use Target Deconvolution?
Inside Method Editor : from ‘Advanced Tasks’
- Spectrum Extraction Setup - Deconvoluted Scan
This tells not to use the raw spectrum but the deconvoluted spectrum
How Do I Create the Quant Target Reference MS Library?
Inside Method Editor : from ‘Method’ - Setup Reference Library…
Conversion to MH format
Where is the MS Target Reference Library specified in Mass Hunter Quant?
Inside Method Editor : from ‘Globals Setup’ - Reference Library
Inside Method Editor : from ‘Outlier Setup Tasks’ - Library Match Score
Where is the Minimum LibraryMatchS specified in Mass Hunter Quant?
If LMS < setting, than component will not be reported
Inside Method Editor : from ‘Outlier Setup Tasks’ - Peak Purity
Where is the Purity specified in Mass Hunter Quant?
Purity: Peak is corrected for interfering ions from neighbor peaks
Data file review - Mass Hunter Quant Batch-at-a-Glance
Batch Information
Compound Information
TIC
Quant ion EIC
Overlayed Quant EIC and Qualifier EICs
‘Head to Tail’ Deconvoluted component spectrum (top)
and reference library spectrum (bottom)
Expected RT
Compound Information Mass Hunter Quant Batch-at-a-Glance with TD
Recognition Window
EIC Window
Library Match Score from TD
Data file review - Mass Hunter Quant Compounds-at-a-Glance
Review / compare multiple samples / compounds in a fully customizable UI
Data file review - Mass Hunter Quant Compounds at a Glance
Data file review - Mass Hunter Quant Compounds-at-a-Glance No Outliers Applied
Target Deconvolution
Data file review - Mass Hunter Quant Compounds-at-a-Glance
Applying outliers to speed data review
Outliers passed Outlier(s) failed
Data file review - Mass Hunter Quant Compounds-at-a-Glance
Applying outliers to speed data review Targets with outliers are high-lighted
All white picture meet requirements; so well confirmed and well quantified components
MH Quant B.07 New Feature : Outlier Filtering
Outlier Filtering Only Display Target Compounds without Outliers
DRS versus TD : Summary Comparison
Function DRS Target Deconvolution
SW Platform MS Chemstation Mass Hunter Quant B.06.00
Quantitation 1 target ion + up to 3 qualifying ions 1 target ion + up to 3 qualifying ions
Full MS Deconvolution Yes Yes
Identification RT match + Library search vs
target MS library RT Match + Library match vs
target MS library
Additional Confirmation with NIST Library search
Yes No
Interactive data review QEDIT Batch at a Glance
Compounds at a Glance
Quant results using raw and deconvoluted data
Yes Yes
Reporting HTML .pdf
Comparison of TD #Hits and LMS versus DRS-AMDIS 21 Environmental Water Extracts
#Compounds confirmed Average LMS for the same confirmed compound
Data File DRS TD_Agile TD_Agile2 * DRS TD_Agile TD_Agile2 *
32701401.d 2 3 3 81 85 88
32701402.d 7 9 10 79 76 78
32701403.d 13 17 21 76 77 81
32701404.d 25 27 33 74 71 78
32701405.d 35 44 48 78 75 80
32701406.d 25 25 32 75 67 75
32701407.d 16 18 23 80 77 82
32701408.d 25 30 31 76 71 79
32701409.d 32 34 43 74 69 76
32701410.d 26 29 37 73 69 76
32701411.d 38 39 44 75 70 74
32701412.d 21 23 24 78 76 78
32701413.d 23 21 27 73 68 75
32701414.d 23 27 35 71 74 79
32701415.d 29 30 36 68 72 77
32701416.d 24 27 29 69 71 74
32701417.d 39 44 46 78 73 78
32701418.d 63 64 67 75 69 73
32701419.d 43 48 46 77 72 76
32701420.d 26 26 33 76 73 76
32701421.d 71 80 87 79 74 77
* Agile2 integrator will be available with release of MH Quant B.07 (Feb / Mar 2014)
Mass Hunter Target Deconvolution
• Legacy MSD Chemstation Methods and data files can be converted for use in MH
• Provides analytical data ‘the same as’ (if not ‘better than’) DRS
• Data processing faster than DRS
(~ 1 minute / data file for 971 targets)
Interactive data review using MH Batch at a Glance (BAG) Sorting / displaying results by Concentration, Library Match Factor etc in BAG Rapid data review using Compounds at a Glance and Outlier flagging
• Very fast .pdf reporting of individual samples or entire batch
Setting up the MAPS L5 5977 MSD
GC/MS Environmental Water Screening Analyzer
Environment Agency Requirements
• Screening method to detect presence / absence of organic pollutants in a given water body under WFD
• To identify VOCs and SVOCs from a single analysis
• Include provision to easily add new substances to method
• Typical Limit of Detection (LOD) 0.1 mg/L
• Low cost analytical solution for validating the pressures and risks to water bodies
• Agilent UK have been working with the EA on this application since 2006
Sample Types
• Surface freshwater bodies - Lakes - Rivers - Streams - Canals - Ground waters - Transitional water bodies (Estuaries)
• Sewage & trade effluent discharge
• Marine environment monitoring
• Potable raw water
• Landfill leachates
• Sediment / solids
• Pollution incidents
EA NLS Instrumentation for GC/MS Screening
2006 - 2013 Hardware
7890A GC System
7683B Series Injector
5975C inert XL MSD with Triple-Axis detector (TAD)
Programmed Temperature Vaporization Inlet (PTV)
Software
GC/MSD Chemstation
Retention Time Locked (RTL) Method
Deconvolution Reporting Software (DRS)
Automated Mass Spectral Deconvolution and Identification Software (AMDIS)
NIST08 Mass Spectral compound library
Hazardous Industrial Chemicals (HIC, 731 entries) MS Database extended to > 1000 compounds
http://natlabs.co.uk/our-services/gc-ms-multi-residue-screening/ More Information :
The EA have analyzed thousands of water samples with this system since 2006
November 2013 - Environment Agency System
November 2013 7890B / 5977A Extractor MSD System installed at EA Laboratory in Starcross
App. optimized
consumables
Agilent Pre-Configured Analyzers Increasing the Value Proposition
Application
report
System with
application setup
w/o Analyzer
Familiarization
Common
Components Installed & Configured with Chemical Testing
and Field Verification
Std. HW, SW Customer
Configured Analyzer Solution
36
Sample Preparation
1 litre water sample + ISTD + 50mL Dichloromethane (DCM)
Extract 15 minutes
Remove DCM, Acidify water,
Add 50mL fresh DCM
Extract 15 minutes
Combine DCM fractions
Reduce DCM to 1 mL
Transfer DCM to auto sampler vial,
Inject 1.5 mL Dry DCM extract with anhydrous
Sodium sulphate
MAPS 5977 MSD Surface Water Analyzer
8 page Solution Note 5991-4127EN
Consists of : • 7693A ALS + 150 position tray • 7890B GC with CO2 cooled MMI inlet • Standard 30m x 0.25mm ID x 0.25um HP5-MSUI Column • Retention time locked acquisition method • 5977 Extractor MSD Bundle • MH Quant data processing with Target Deconvolution • Application specific Reference MS Database • Customized .pdf TD reporting • Installation / check-out Test mix • Application Consultancy for installation of the screening
method and database, checkout and training using the checkout sample.
MAPS 5977 MSD Surface Water Analyzer
Analyzer Test Mix: For RTL set-up and TD Test Report Generation
Part Number : 5190-0473 (3 x 1mL ampoules) Dilute x10 with Dichloromethane for method re-lock and TD test report generation Fluorene is the RTL locking compound, locked to 15.577 minutes
Not all components (particular the deuterated IS) are in the MH Quant method nor Reference MS database so will not appear in TD test report
Quantitation
Based on Single point calibration for 1 mg/L equivalent Provided methods include calibration curves.
Quant Results are estimated amounts when customer has no standards
Full scan TIC of Hazardous Waste Landfill Site Water Extract
Mass Hunter TD Report : Hazardous Waste Landfill Site Water Extract Standard report with • Retention Time of identified target analyte • CAS number • Quant result Corrected for peak purity • % Peak purity • Time difference from locked RT • Library Match Score
Full scan TIC of Grape Extract
Analyzed using the Agilent RTL (1x) Pesticide Method and the RTLPEST3.L Pesticide library (converted to Mass Hunter .reflibrary.xml format)
Mass Hunter TD Report : Grape Extract
MH Target Deconvolution : Data Processing Times*
0.0
5.0
10.0
15.0
20.0
25.0
1 5 10 22
Min
ute
s
# Data Files in Batch
Processing time
* HP8530 laptop, 40 minute TICs, 971 targets in Quant method / Reference MS Library
DRS vs MH TD : Batch Reporting Time after Data Review*
* 22 data files, HP8530 laptop, 40 minute TICs, 971 targets in Quant method / Reference MS Library
0
1
2
3
4
5
6
7
Reporting Time (minutes)
DRS (html)
TD (.pdf)
TD .pdf Report Template
MAPS( level 5) 5977 MSD Turn-key Surface Water Analyzer
• Simple and complete GC/MS hardware configuration with dedicated column
• Retention Time Locked acquisition method
• Unique application specific MS Reference Library
• MassHunter Quant data processing with Target Deconvolution
• Data review using MassHunter Batch-at-a-Glance, Compounds-at-a-Glance , Outlier flagging
and filtering
• Very fast .pdf reporting of individual samples or entire batch
• Installation of software and libraries
• System check-out with special reference sample
• Standard Operating Procedure with description of analytical procedure
• Complete method of analysis incl sample prep, recommended consumables and materials
• On site training