Quantification - Calibration

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In This Section, We Will Discuss:

The steps necessary to build a calibration table.

How to select calibration settings.

Calibration table options.

Peak identification tools.

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1. Inject standards with concentrations that bracket yourunknowns.

2. Set-up appropriate integration events for the low standard.Integrate to obtain the peak height or area. Save the integration events to the method.

3. Begin a new calibration table with level one, the lowestlevel. Fill in compound names and amounts.

4. Integrate each additional standard level and add to the calibration table.

5. Examine the calibration curve and save it to the method.

Building a Calibration Table

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Example Calibration

1. Standard 30.0 ISTD 50.02. Standard 50.0 ISTD 50.03. Standard 75.0 ISTD 50.0

ng/L ng/L

3 Levelsmin1 2 3 4 5 6

mAU

0

20

40

60

80

100

DAD1 A, Sig=254,4 Ref=550,100 (DEMO\005-0101.D)

0.7

47

1.0

21

2.5

65

5.8

37

StandardISTD

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Define signals to be evaluated during a method.

Signal Details

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Calibration Settings

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Load the Signal(s) for the Low Level Standard and Integrate

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Create a Calibration Table: Level One

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Fill in:Compound NameGroupAmount

Check if:Reference PeakISTDIndicate #

Calibration Table Overview: Fill in the Dialog

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Load the Signal - Add Level Two

Fill in Amount for Level 2

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Add Additional Levels

Add Level 3 and fill in the

amount

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Compound, Amt, Area, Rsp Factor,Ref, ISTD#

Compound, Grp, Amt, Low LimitHigh Limit

Compound, Amt, Area, Def, CurveType, Origin, Weight

Compound, Area, Rsp%, Def, +-, Pk Usage

Calibration Table Options

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Use Low and High Limits as qualifiers to confirm peak identification.

Compound Details

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Check Def to apply thedefault curve type and origin from the Calibration Settingsdialog box.

Curve TypesPiece wise ExponentLinear QuadraticLog CubicPower Average Rsp/Amt

Origin TreatmentsIgnoreIncludeForceConnect straight segment

Peak Details

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By default, the largest peak of a compound is the Main peak. Others are set to Ignore.

Identification Details

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Qualifiers

The ratio of 230 to 254 is 38%for Biphenyl

The ratio of 230 to 254 is 224%.The peak would be rejected

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Define the columns included in your Calibration Table

Edit Calibration Table Options

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Calibration TaskTool

New CalibrationTable

Recalibrate withCurrent Chromatogram

Add New Level

Signal DetailsAdd New Peaks

Select Calibrated peak(s) fromChromatogram and delete fromcalibration table.

Select peak(s) from chromatogram and addto calibration table

Select compoundsfrom chromatogram and recalibrate

Edit CalibrationTable Options

Quantification Tools

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Save Table to Method

To Save the Calibration Table as Part of a Method...