Chris Sandy GC/MS Applications Chemist

EMEA Technical Marketing Development Team Agilent Technologies UK

NACRW July 2013

An Introduction to ‘Target Deconvolution’ –

A Powerful New Feature of Agilent’s Mass Hunter Quantitative Software

Agilent Technologies Vendor Seminar

NACRW July 2013

Introduction

• Deconvolution Reporting Software (DRS) has been an add-on for 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) • Target Deconvolution (TD) is a new feature of Mass Hunter Quant SW B.06.00, introduced in March 2013

• The entire TD process is contained within Mass Hunter Quant SW

AMDIS

DRS Workflow : 3 Integrated Processes

Combined quantitative and qualitative HTML Summary report

Targets are identified by comparison to locked RTs and 1 Quant and 3 Qualifier ion ratios, then quantified using target ion

area versus cal table

Quant Results

QEdit

MS Chemstation

Deconvoluted target spectra found by AMDIS, searched against NIST MS database

NIST hits

NIST MS Search

AMDIS deconvolutes component spectra and searches target MS database using locked RT and

Library match as qualifiers

AMDIS Quant Results

AMDIS

AMDIS hits

MS Chemstation

NIST MS Search

DRS Data file review - Chemstation QEDIT

EICs

Spectra

Target List

MSD Quant EIC AMDIS Quant EIC

MSD & AMDIS peak areas and quant

results

DRS Report - River Water Extract

Typical processing + reporting time of 1 -2 minutes per data file

Chemstation (ion ratio) false positive

DRS Report - River Water Extract

Typical processing + reporting time of 1 -2 minutes per data file

Chemstation (ion ratio) false positive Chemstation (ion ratio) false negative, identified by AMDIS

DRS Report - River Water Extract

Typical processing + reporting time of 1 -2 minutes per data file

DRS Report - River Water Extract

Typical processing + reporting time of 1 -2 minutes per data file

Chemstation (ion ratio) positive, identified by AMDIS

Chemstation (ion ratio) false positive Chemstation (ion ratio) false negative, identified by AMDIS

New Agilent 7890B / 5977 Series GC/MSD What’s new with the 5977 MSD?

Better Performance • Higher Sensitivity Extractor Ion Source • Improved Agilent Vacuum system

More Productivity • Direct Communication 7890B5977A

• Better performance with H2 carrier gas • Vent the system faster • Power save (Sleep/Wake) • Helium conservation module

• New software revision G1701FA • Mass Hunter Data Acquisition • Both ChemStation and MH Data Analysis

5977A MSD : Mass Hunter Data Acquisition

Familiar Look & Feel to

MS Chemstation

Created data files (.d) may be processed by either Chemstation Data Analysis or Mass Hunter Qual / Quant software

Configure Mass Hunter Data Acquisition for MSD ChemStation Automated Data Processing

Convert MS ChemStation Quant Database to MH Format

Chemstation : qdb.mth MH : nnnnn.quantmethod.xml

Convert MS ChemStation Data Files to MH Format - Off-line Batch Mode

Once translated , data files may be processed in MS Chemstation or Mass Hunter SW

Target Deconvolution in Mass Hunter Quant B.06.00

Mass Hunter Data Analysis Software Mass Hunter’s 2 main Data Analysis modules have different functions:

MH Quantitative SW (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

MH Qualitative SW (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 English River Water Extract*

ISTD

* Data courtesy of the National Laboratory Service, UK Environment Agency

Would My Application Benefit from MS Deconvolution?

Full scan TIC of Former Hazardous Landfill Site Water Extract

TIC of complex samples may have many overlapping / co-eluting compounds

ISTD

A Single Chromatographic Peak may contain Multiple Components TIC & Spectrum

TIC TIC

Component 1 Component 2

Component 3

Components and Mixed Spectra

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 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

mike_szelewski@agilent.com

Mass Hunter Quant SW - Data Review

Batch-at-a-Glance

Compounds-at-a-Glance

Data file review - Mass Hunter Quant Batch-at-a-Glance

Batch Information

Compound Information

TIC

mike_szelewski@agilent.com

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

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

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

Mass Hunter Quant Target Deconvolution SUMMARY

• New feature fully integrated into Mass Hunter Quant B.06.00

• Provides additional confidence in the identification of target analytes by the additional use of deconvoluted full scan mass spectra

• Legacy MSD Chemstation Methods and data files can be converted for use in MH

• Interactive data review using Mass Hunter Batch-at-a-Glance (BAG)

• Rapid data review using Compounds-at- a-Glance (CAG) and Outlier flagging

• Very fast .pdf TD reporting of individual samples or entire batch

Mass Hunter Quant Target Deconvolution

Thank-you for your time and attention chris_sandy@agilent.com