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