Presenter: Stephan Baumann

Agilent Technologies, Inc.; Santa Clara, CA

ASTS Talk

November, 2016

5977 SQ 7000 TQ 7200 QTOF

6560 IM QTOF

6550 QTOF6540 QTOF6420 QQQ 6470 QQQ 6490 QQQ

6120 SQ

6130 SQ

6150 SQ

8800 ICO-QQQ7700 ICP-MS

When is it Time to Move to MassHunter?

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Our Goal is to provide you a basic introduction and understanding of the MassHunter Workflow and MSD Operating System

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MassHunter SoftwareOne software for all Agilent Mass Specs

Minimize the learning and optimize the use of software in yourlab across different mass spec instrument platforms• Control of Agilent 6000 Series QQQ, TOF and Q-TOF• LC/MS Data Processing for QQQ, TOF, Q-TOF, SQ• Control and data processing for GC/MS including GC-QQQ, GC Single

Quad and GC-QTOF• Control & data processing for ICP/MS

LC/MS GC/MS ICP-MS

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MassHunter – A Software SuiteMassHunter software is modular: Different modules that focus on these specific tasks:

A common reporting engine

Suite of accessory applications: Unknown Analysis: batch versions of deconvolution and library

searching Library editor Method development assistants Data Translators

Acquisition (Instrument Specific) Quantitative Analysis (Quant) Qualitative Analysis (Qual)

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GC/MS SoftwareMassHunter Qual

MassHunter QuantMH

CS

MassHunter GC/MS Acquisition

And Use EitherMassHunter Data Analysis

ORMSD ChemStation Data AnalysisMSD ChemStation DA

G1701FA

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GC/MS MassHunter EnviroQuantConcerns relative to ChemStation EnviroQuant

Analyze Button• The analyze button was inexplicably removed by MassHunter Quant

software team. The feature can be restored by changing setting within MassHunter.

Changing File Name• ChemStation EnviroQuant has the capacity to change sample information.

Any changes are tracked. MassHunter does not have this flexibility. However, the file name can be changed in the sample directory and this name is displayed in the Quant results.

Printing an Injection Log as opposed to a Sequence Log• ChemStation EnviroQuant provides a true Sequence Log. MassHunter

EnviroQuant can print a log of files in a directory. These are identical if all the data (blanks, calibrators, continuing calibrators, etc. are stored together.

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GC/MS ChemStation DrugQuantDifferent than ChemStation DrugQuant

Resolution, Fronting and Tailing parameters • These tools are built into MassHunter but they are not in the report. Tune Time Stamp (Fixed in B.07.05)• MassHunter can’t read the time stamp directly. There are five report types in ChemStations• MassHunter has a several templates and custom templates can be

created. This provides us with additional flexibility.

Note that there is a good MassHunter DrugQuant workflow guide (G6845-90007) that is very helpful in setting up the software correctly. There is also the Sequence_Decisions.Log that helps with intelligent sequencing troubleshooting.

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GC/MS Acquisition MethodsMSD Productivity ChemStation MassHunter GC/MS Acquisition

GC/MS Acquisition Methods are Transparentfrom MSD ChemStation to MassHunter GC/MS

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MassHunter Data Analysis SoftwareMassHunter’s main data analysis modules have different functions:

Quantitative (Quant):(Target Analysis - Quantitation) Review Batch results Visualize quality outliers Powerful quantitation engine Compounds-at-a-Glance Library Editor Unknowns Analysis

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

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Migrate Data to MassHunter Format

Data

Data

Data

MassHunter Quant

MassHunter Unknowns Analysis

MassHunter Qual

ChemStation (.D)

WorkStation (.SMS)

Generic (.CDF)

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Migrate Quantitative Methods to MassHunter Format

Method

Method

ChemStation (QDB.MTH)

WorkStation (.mth)

MassHunter Quant

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MassHunter WorkStation Qualitative AnalysisData Navigator

Method Explorer

Chromatogram Results

Method Editor

Spectrum Results

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Extract Chromatograms

Types of Chromatograms

TIC – Total Ion Chromatogram

BPC – Base Peak Chromatogram

EIC – Extracted Ion Chromatogram

SIM – Selected Ion Monitor

MRM – Multiple Reaction Monitor (Triple Quad)

pNLC - Precursor Neutral Loss Chromatogram (Triple Quad)

Other Chromatograms – GC, ADC, DAD, VWD

Instrument Curve - %Comp., Temps, etc.

Extract multiple signal types to create Chromatograms

Definition of signals can be stored in the method and used with automation

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Extract Spectra

•Manual selection or based on integrated peaks

•Background subtraction based on defined regions or peak definition

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Find Compound by Chromatogram Deconvolution

•Agilent deconvolution algorithm, similar to AMDIS, integrated into MassHunter

• Creates Compounds with chromatographic and spectral components

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Identify Compounds with Library Search

•Library search User Spectra or Compounds.•Library search parameters are part of a method.•May use Retention Time or Retention Index in search

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Utilize NIST library searches• Export spectra to NIST

• Requires NIST (not included in Qual)

• Information does not flow back to Qualitative MassHunter reports

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A dedicated application, the Library Editor, is used to create, edit, and modify user libraries.

Agilent provides over 15 El Libraries such as:

NIST14Wiley11th+NIST14 Fiehn Metabolite Library Japanese Pesticide list Toxicology LibraryControlled Substances

Library Editor & Available Libraries

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Annotate and Document Annotate chromatograms and spectra using text, graphics (.jpg) and chemical structures (.mol)

Mass Caliper to document fragmentation and losses

Export or copy & paste graphics into presentations

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Quantitative Analysis Screen

Compound Information

Results Information

Batch Information(i.e., Calibration Curve)

Results Informationfor the batch can be displayed. Visual guides highlight specific data that fall outside specific, predefined conditions.

Batch Informationprovides easy visualization and customization of relevant desired data.

The Quant Batch screen looks like this.

Compound Information displays graphical representation of the peak, qualifier information, spectral information, and the ISTD. Visual guides help identify associated data problems.

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Batch Screen – Sample & Results Information

These icons filter outliers in the display.

In Results Information, the Quantitation Message is a human-readable string that contains a list of error messages that result from a compound's quantitation.

Blue = low / Red = high

“Outliers” define and specify results of known problem samples/substances that fall outside predefined conditions.

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Results Information: Outlier Options

>40 quality checks can be specified to highlight outliers in results.

Can set high and low limits.

User defined quality check can be added to predefined calculations.

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Batch Screen - Compound Information

CompoundIntegration

QualifierInformation

SpectrumInformation

ISTDIntegration(Quantitation: Part of Agilent’s ParameterlessIntegration)

ISTD QualifierInformation

ISTD SpectrumInformation

Compound information displays all relevant information in one view.

• Uncertainty Bands• Actual Ratio• Manual Integration

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Batch Screen - Library Reference SpectraSupports correct identification of compound• Visual comparison of Sample and Library Spectra• Seen in Batch-at-a-Glance and on Reports• Head to Tail Style, Head to Head not yet available• Extracts Spectra from Library by matching on CAS number based in Quant

method and creates a small reference library (reflibrary.xml)

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Batch Screen – Calibration Curve

View ISTD responses or display QC samples - click either the ISD or QC button.

Change Curve Fit

R2.

Zoom in/out button (right click/drag)

Concentration can be set as relative (to ISTD) or actual.

An example of Batch Information is this view of the Calibration Curve. The Curve fit can be changed and data can be updated instantaneously.

After editing the calibration curve, you must reanalyze the batch.

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Batch Screen - Curve Fit Assistant

Calculates all possible calibration curve combinations.

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Compounds-at-a-Glance

View up to 10 x 10 chromatograms Overlay Target with ISTD Overlay Quantifier with Qualifiers View chromatograms across 100 samples

5 x 55 x 5 10 x 1010 x 10

Compounds-at-a-Glance allows you to view multiple traces of compounds at a single glance.

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Compounds-at-a-Glance

Shaded automatically

You can view compound outliers at-a-glance too.

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Compounds-at-a-GlanceYou can view the manual integration of compounds at a glance too.

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Quantitation Method CreationEasy way to create a target method from acquired data (MRM, SIM or Scan).Using information from a data file, MassHunter Quant can create a target compound list - generating Compound Name, Retention Time, Quant and Qual ions, and ratios automatically.

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Automatic Compound Detection – Scan DataAutomatic compound detection works with both MRM and Scan data.For scan data, Library information can be used to add compound name and CAS#; The Quant and Qual ions and ratios are taken from the data file information.

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Overview of Unknowns AnalysisQuant addresses target compound list

What else is present in the samples?

Ion Extraction

Unknowns Analysis

Deconvolution

and

Library Search

Quantitation

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Quant & Unknowns Analysis WorkflowMS Scan data

Chromatogram Chromatogram

Extract

Integrate

Deconvolute

Library Match

Spectra

Target Match

Integrate

Quantitation

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Unknowns Analysis

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Chromatogram

Chromatogram displays selected component(s) in Blue

All non-selected ions (EIC) displayed in Green

TIC displayed in BlackClicking in the Chromatogram display selects the nearest peak and selects it in Components table

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Spectrum & Ion PeaksQuant Method

Qualifier Target Default number ions displayed in Ions Peaks is 5, limit is 10

Click on ion in Spectrum to make it add / remove in Ion Peaks

Click on label in Spectrum or Ion Peaks to remove

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Training Resource - Familiarization Guide

Familiarization guide covers basic operations

Examples use example data on media

Familiarization Guide in English Japanese Chinese

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Training Resource – Videos and Help

R&D Videos –• Good introduction• Lacking in details

Help

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MassHunter Excel Reporting

• Excel Based (Excel 2010/2013 supported)

• MassHunter application exports data in XML and graphics files

• Templates define report formatting

• Over 100 shipping report templates in Excel and PDF format

• Reports can be altered for desired information & appearance

• Customization using Excel features (function, formatting, code)

• Computer Based Training for report customization by users

• Some customization available through Agilent

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Report Template Design WorksheetsCustomize the template by placing data and graphics in the desired locations

Templates may have multiple Design worksheets with different layouts

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Excel XML Source Window• Reporting uses XML mapping to import results into templates

• Drag and drop to add data to existing reports

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