MassHunter Quantitative Software Review and Quant Method ...

MassHunter Quantitation4/30/2015

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Agilent 6400 Series Triple QuadrupoleUsers Workshop

David Presser John HughesAnabel Fandino David Weil Doug McIntyre Shane TichyPeter Stone Laura Phillips André Szczesnieski

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MH QQQ Users workshop

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MassHunter Quantitative SoftwareReview and Quant Method Optimization

Topics • Creating a batch of processed results

• Creating a quantitation method

• Applying a previously saved method

• Updating a method

• Outlier setup and flagging

• Data review and manual integration

• Generating a report, modifying graphic output

• Exporting data

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MassHunter Quantitative SoftwareBatch-at-a-Glance

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Important MassHunter Quant concepts and rules

A Batch is a file which contains all the Quant results from a set of data files and the Quant method used.

All the data files in a Batch must reside in a single directory, so put them all together before creating a New Batch.

Select the data directory before creating the Batch

I would not recommend using the Browse to Copy Samples push. You will have two copies of the same files. Preferred solution is to create the batch in the directory which contains the data files.

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Important MassHunter Quant concepts and rules:Quant methods

The most common method problem: Calibration/QC level names in Batch and Quant method do not match, e.g.

- Method: levels are 1,2,3,4,5

- Batch: levels are L1,L2,L3,L4,L5

Second most common problem: a hidden column in the Method with a key parameter, e.g. Ion Polarity if method created manually.

Many Quant parameters can be copied between compounds with Apply to All button.

When in doubt, Right-Click to look for convenient features and shortcuts, like Fill Down or Fill Column.

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Creating a batch of processed resultsBatch-at-a-Glance

1) File | New Batch…

2) Create the batch in the directory in which the data files reside

3) Add Samples

4) Method | Open and Apply from existing file…

Or Method | Open method from existing Batch

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Creating a quantitation method for QQQMethod | New Method from acquired MRM data…

Select a high concentration standard

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Insure there are no zero RT’s

You may work from top downwards

Creating a quantitation methodAssign internal standards and provide an ISTD concentration

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Creating a quantitation method

Create a csv file which contains compound names and concentrations

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Creating a calibration table for B.05.02 and later

Note: columns B and C are blank (CAS number and purity)

Creating a quantitation methodConcentration Setup – import calibration levels

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Creating a quantitation methodAlthough other steps remain, one may analyze at this point

Insure that the level names match before pressing “Analyze Batch”

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Creating a quantitation method – Analyze Batch

A

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MassHunter Quantitation

Set Peak Filter Area Threshold will set the integration area reject to 50% of the average response of the lowest calibration level

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Setting a reporting threshold:

Creating a quantitation methodEdit and then Save method

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Creating a batch of processed resultsUse an existing method

1) File | New Batch…

2) Create the batch in the directory in which the data files reside

3) Add Samples

4) Method | Open and Apply from existing file…

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Creating a batch of processed resultsUse existing method• Method | Open and Apply from existing file…select saved method

• Analyze batch – note calibration curve recalled from method

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Report Generation

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Report Method EditAdd a report template. Scroll to the right to modify options such as creating a .csv file, opening the result file, and selecting a printer.

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Report Method EditAdd a report template. Scroll to the right to modify options such as creating a .csv file, opening the result file, and selecting a printer.

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Report Method EditSelect the Results tab. Click “Yes - Always generate a results file

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Report Method EditSelect the Graphics settings tab. If the report has no graphics, disable graphics generation to save time. Delete graphics after report generation.

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Report Method Edit Change graphics options for desired output, such as disabling normalization of qualifiers and labeling TIC with compound names.

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Report Method Edit – Quantifier/Qualifier

Example: uncheck normalize qualifiers.

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Report Method Edit – Sample chromatogramExample: labeling TIC with compound names.

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Report Method Edit – Sample chromatogramExample: labeling TIC with compound names.

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Report Method EditExample: Fixed range graphics settings.

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Report GenerationExample: Fixed range graphics settings.

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Report Method Edit

Save the report method.

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Report Generation Dialogue

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Changing report graphics prior to B.06.00Right-click in the compound graphics window and select “Properties…”

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Changing report graphics prior to B.06.00

Select desired attributes such as no gridlines

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Changing report graphics prior to B.06.00Select desired attributes for qualifier – turn off normalize

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Changing report graphics prior to B.06.00On B.05.00 and later one can change the sample chromatogram labels. View | Sample Info. Then Right-click the chromatogram window, select Properties…

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Changing report graphics prior to B.06.00On B.05.00 and later one can change the sample chromatogram labels

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Creating a quantitation method – TOF or Q-TOFCreate a library

• Copy and paste from Excel to Library Editor

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Creating a quantitation method – TOF or Q-TOFCreate a library

• Synthesize spectra (adds isotopic cluster information)

• Save library

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Creating a quantitation batch –TOF or Q-TOFCreate a batch

• File | New Batch

• Create the batch in the directory in which the data files reside

• Add samples

• Method Edit

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Creating a quantitation method –TOF or Q-TOFFrom Library• Method | New Method from File…

• Select the library

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Creating a quantitation method – TOF or Q-TOFFrom Library• Select “No RT calibration”

• Click Ok

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Creating a quantitation method – TOF or Q-TOFFrom Library• Compounds and retention times are imported

• Continue adding concentrations, etc.

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Creating a quantitation method – TOF or Q-TOFFrom Library• Optional: in Globals, designate Reference Pattern Library

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Creating a quantitation method – TOF or Q-TOFFrom Library• “red” Exit, Apply to Batch, Analyze Batch, and save.

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Creating a quantitation method – TOF or Q-TOFFrom .cef• Find the compounds in Qualitative Data Analysis, such as Find by Formula

• Export .cef file

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Creating a quantitation method – TOF or Q-TOFFrom .cef• In Quant – Method | New Method from File…

• Select .cef file – compounds are imported

• Continue adding concentrations, etc.

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Creating a quantitation method – TOF or Q-TOFFrom .cef• “red” Exit, Analyze Batch, and save

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MassHunter Quantitative Software Review and Quant Method ...

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