Library Matching

Alyssa Loo
Sales Applications Chemist, Waters Pacific
 Library Creation/ Library Matching

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Spectral Libraries in Empower
 A spectral library is a collection of spectra that you extract from known compounds
under known chromatographic conditions
 You create libraries in the Review window
 Empower 3 software uses the Spectral Contrast algorithm for library matching
 Spectral Contrast Angle is called Match Angle
 Threshold Angle is called Match Threshold
 You perform library matching to:
− Confirm that peaks are spectrally the same as known library peaks
− Provide evidence to detect co-elution

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Guidelines for Creating Spectral Libraries
 Use pure standards to create libraries
 If you do not have a standard, use a representative batch of material
 Collect the library and sample spectra with the same conditions:
− Use the same instrument method parameters, such as the wavelength range, sampling rate,
resolution and filter response
− Use the same mobile phase, including organic modifiers and pH
− Use the same column and column temperature
− Use the same sample diluent
− Use the same detector calibration
 Perform Peak Purity testing on the peaks before using the spectra in a library
 Keep absorbance under 1 AU across the entire collected wavelength range

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Creating a Library

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Bringing Standard into Review

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PDA Review Window, Creating a New Library

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Naming the Library

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Adding Spectra to the Library

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Adding Spectra to the Library

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Adding Spectra to the Library

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Spectra Added to Library

7 Spectra added to Library

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Spectral Table Properties

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Viewing Spectral Information

Library information added to table

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Setting Library Search Parameters in the Processing Method

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Library Search Parameters

Set the angle difference in degrees for when the search should stop

Select how
many matches
to display

Select Libraries
To search

Set the Threshold Criteria

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Library Search Parameters

Select if spectra should be derived

Select if smoothing
should be applied
to the spectra

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Library Search Parameters

Select to apply a
retention time
pre-search, and set
the RT window

Select to apply a
wavelength
pre-search, and set
the λmax window

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Saving the Changes to the Processing Method

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Testing the Library Search Parameters
Results for Std5, Inj #1, The Library Standard

To apply the
library search
parameters

Analgesics Std5, inj #1

Match angle is 0 since we

are matching spectra from
the standard that was used
to create the library
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Results for Std5, Inj #2

Analgesics Std5, inj #2

3 peaks has
Match Angle > Match Threshold

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Results for Std5, inj #3-6

Analgesics Std5, inj #3 Analgesics Std5, inj #4

Analgesics Std5, inj #5 Analgesics Std5, inj #6

Largest Match Angle = 0.158 ~ 0.16

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Adjusting Search Parameters

Threshold Criteria set to Noise+Solvent

Solvent Angle
set to highest
Match Angle

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Re-applying the Method Set

Match Threshold
w. Criteria set to
Noise

Match Threshold
w. Criteria set to
Noise+Solvent,
Solvent Angle = 0.16
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New Results for Std5, inj #2

Analgesics Std5, inj #2

All matches now OK

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New Results for Std5, inj #3-6

Analgesics Std5, inj #3 Analgesics Std5, inj #4

Analgesics Std5, inj #5 Analgesics Std5, inj #6

Matches for all injections of standard 5 now OK

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Saving the Changes

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Processing the Sample Set

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Viewing the Result Set

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Reviewing the Result Set

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Results for Standard 1

All peaks
Match

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Results for Sample 1

All peaks
Match

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Results for Sample 2
Caffeine contains impurity at 20%

Caffeine peak does not give good match

but we also know it is not pure

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View Results

General Results Table

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Match Plot - Properties

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Normalized Spectra in Match Plot

Library Match results table

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Triple Plot

Spectrum from Acetaminophen peak

Best match from Library Search

Difference between the two spectra

Observe the scale

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Match Plot for Caffeine Peak

Change in Spectra caused by presence of impurity

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Triple Plot for Caffeine Peak

An indication of that the Impurity has a spectrum with λmax at ~240nm and 294nm

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Match Plot for Salicylic Acid Peak

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Spectrum Index Plot w. Library Match Spectra
Library Match icon

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 Manual Library Search

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Performing Manual Library Search

Spectrum Extracted at
Max Impurity

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Opening a Library in Review

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Loading a Library in Review

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Library Spectra Loaded

Library Spectra Loaded

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Removing Spectra from the Spectrum Review Window

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Selecting the Extracted Spectrum

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Manual Library Search

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Modifying the Library Search Parameters

90°is the largest

angle difference possible,
So this will always give a
match result

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New Library Search Result

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Library Match Table

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Results for Sample 3
Caffeine contains impurity at 10% level

Caffeine peak still does not give good

match but we also know it is not pure

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Results for Sample 4
Caffeine contains impurity at 5% level

Caffeine peak still does not give good

match but we also know it is not pure

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Results for Sample 5
Caffeine contains impurity at 2% level

Caffeine peak now gives a good

match but we still see it is not pure

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Overview of Libraries in Configuration Manager

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Opening the Library

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Editing the View Filter

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Saving the View Filter

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Viewing Library Spectra Details

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Export Libraries/Spectra

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Library Creation/ Matching Summary
 Build libraries in Review Window
 Library searching can be automated in Processing method
 Use match parameters to narrow search
 A good match is when Match Angle < Match Threshold
 Library matching in conjunction with peak purity testing can reveal perfect co-elution
of compounds that cannot be detected by peak purity testing alone

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