MOLECULAR SPECTROSCOPY

INSTRUMENT PERFORMANCE
VALIDATION KIT

User’s Guide
Release History

Part Number Release Publication Date

L1050096 B February 2011

Any comments about the documentation for this product should be addressed to:

User Assistance
PerkinElmer Ltd
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BUCKS HP9 2FX
United Kingdom

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Copyright © 2011 PerkinElmer, Inc.

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Table of Contents
Performance Validation Kits ............................................................................... 4
MIR Kit ...................................................................................................... 4
NIR Kit ....................................................................................................... 6
Validating Instrument Calibration Using the TRM ................................................. 7
Using Macros to Collect Reference Spectra ................................................... 7
Possible Reasons for Validation Failure ......................................................... 8
When Should I Recalibrate My Instrument? .................................................. 8
Routine Instrument Validation ...........................................................................10
Routine Validation in MIR ...........................................................................10
What Peaks Should I Use? .........................................................................10
4 . Instrument Performance Validation Kit User's Guide

Performance Validation Kits

The Food and Drug Administration (FDA), the United Kingdom Accreditation Service (UKAS)
and the Good Laboratory Practice (GLP) regulations all require that calibration of instrument
accuracy can be traced to a certified reference material (CRM).

The MIR Performance Validation Kit (L1250404) contains a polystyrene Traceable Reference
Material (TRM), and two types of reference material for routine use in the sample
compartment of your spectrometer: three polystyrene cards for routine validation of
wavenumber precision, and a glass transmission filter for recording ordinate repeatability.

The NIR Performance Validation Kit (L1250405) contains a polystyrene Traceable Reference
Material (TRM).

The use of both kits is described here.

MIR Kit
The Instrument Performance Validation (IPV) kit enables you to generate a record of
wavenumber calibration that is traceable to a standard reference material (SRM) calibrated
by the National Institute of Standards and Technology (NIST). It also enables you to confirm
that your instrument is performing consistently in day-to-day operation

In addition to the IPV kit, your instrument may have an internal APV (Automatic Precision
Validator) kit. Two reference materials are installed in the optical module of the instrument
and these internal reference materials can be used to provide automatic validation routines
when used with Spectrum software.

NOTE: If you have a Spectrum 65 or Spectrum Two instrument that does not have internal
reference materials, reference cards are provided that can be inserted in the sample
slide.

Polystyrene film

The polystyrene cards in the IPV kit are used to test that the instrument is performing
consistently, rather than for calibrating the accuracy of the wavenumber (abscissa) scale.

Calibration of wavenumber accuracy is performed using the TRM supplied in the IPV kit.

Traceability of the wavenumber axis comes from a certified polystyrene standard

(SRM 1921b), measured by the National Institute of Standards and Technology (NIST),
Gaithersburg, MD 20899, USA. The NIST certificate of measurement provides wavenumber
values for fourteen peaks, of which thirteen are recommended for certification using a
center-of-gravity (also known as centroid) algorithm. Of these, we have selected five that
show the least dependence on temperature and water vapor. These are nominally at
3082.22, 3060.14, 1601.38, 1583.04 and 1028.42 cm−1. The calibration is valid for
resolutions of 4 cm−1 or better.
 Instrument Performance Validation Kit User’s Guide . 5

NG11 Schott glass

The sample provided for testing the ordinate repeatability is an NG11 Schott glass filter with
a nominal thickness of 1 mm. This is a very stable material, and has a series of broad
spectral features with transmission between 70% and 0%. It can be used for testing that the
instrument is performing consistently, but not for testing the accuracy of the transmittance
scale. Accurate values for the transmittance of the glass sample are not available. Errors in
transmittance caused by inter-reflections between this sample and other optical components
vary from instrument to instrument, depending upon the precise location of the reference
material.

Limitations of traceable wavenumber calibration

Wavenumber calibration using polystyrene is derived from the NIST SRM 1921b. Certified
values for the most precisely known peak positions used have quoted uncertainties of about
±0.1 cm−1 for four of the peaks, and ±0.27 cm−1 for the peak at 1028.42 cm−1.

These values were measured at 0.5 cm−1 resolution using an instrument at 23 ± 1 °C, but
the expanded uncertainty values are quoted for 23 ± 5 °C.

We suggest that the uncertainty associated with routine measurement of the five
recommended peaks in the spectrum of the traceable polystyrene sample, is realistically set
at ±0.5 cm−1. This level of precision is satisfactory for general analytical applications
involving solid and liquid samples.

Measurement precision with a polystyrene film

Spectra collected using the reference materials in the IPV give peaks 10 cm−1 or more wide,
and the peak positions can be consistently measured to ±0.1 cm−1, provided that the noise
level is low enough.

Peaks in the spectrum of polystyrene above 1800 cm−1 are broader than most of the peaks
below 1800 cm−1 and may, therefore, show more variation for a given noise level. The peaks
at 1943 and 1802 cm−1 are subject to interference by water vapor and should not be used.

NOTE: The recommended measurement conditions are 16 scans at 4 cm−1. Under these
conditions, the reported peak positions are unlikely to vary by more than ±0.1 cm−1
unless affected by water vapor.
6 . Instrument Performance Validation Kit User's Guide

NIR Kit
Instrument Performance Validation (IPV) enables you to routinely generate a record of
wavenumber calibration that is traceable to a National Physical Laboratory (NPL) CRM. Using
the NIR Performance Validation Macro, you can compare the result for a polystyrene
Traceable Reference Material (TRM) and your internal polystyrene Automatic Precision
Validator (APV) against a certified polystyrene standard (serial number: JR95), measured by
the NPL, Teddington, TW1 0LW, UK.

The NPL certificate of measurement provides wavenumber values for two peaks,
5669.3 cm−1 and 4571.6 cm−1, for resolutions of 4 cm−1 or better. The polystyrene TRM
supplied with your instrument has been calibrated against this CRM, and a Certificate of
Calibration printed at the factory is included with it. This certificate provides a record of
calibration and traceability for the TRM to the NPL CRM.

Limitations of traceable wavenumber calibration

Certified values for the peak positions used have quoted uncertainties of ±1.5 cm−1 for the
peak at 5669.3 cm−1 and ±1.0 cm−1 for the peak at 4571.6 cm−1. We suggest that the
uncertainty associated with routine measurement of the two recommended peaks, in the
spectrum of a traceable polystyrene sample, is realistically set at these limits. This level of
precision is satisfactory for general analytical applications involving solid and liquid samples.
 Instrument Performance Validation Kit User’s Guide . 7

Validating Instrument Calibration Using the TRM

To validate instrument calibration, tests must be performed under the same measurement
conditions as those used to calibrate the TRM (Traceable Reference Material) at the factory.
The Spectrum macros provided will use the appropriate instrument settings.

We also recommend that before starting the calibration you make sure that the instrument is
optimally aligned.

Using Macros to Collect Reference Spectra

NOTE: To run the macro, you must be connected to your instrument. No sampling accessory
other than the Slide Holder should be installed in the sample compartment.

1. Start Spectrum software and connect to your instrument.

2. Select Macro from the Setup menu.

The Setup Macros tab is displayed.

3. Select Add, and then select Import from the drop-down list.

4. Browse to and select the macro appropriate for your instrument.

The IPV macros are located in:
C:\Program Files\PerkinElmer\Spectrum\IPV Kit\Macros\NIST1921b
for Windows XP or Windows 7 (32-bit), or in
C:\Program Files\PerkinElmer (x86)\Spectrum\IPV Kit\Macros\NIST1921b
for Windows 7 (64-bit), where C: is the drive where Spectrum has been installed.

5. To run the macro, select Macros from the Process menu, and then select the
appropriate macro from the sub-menu.

OR

Click on the Process bar, and select the appropriate macro from the drop-down
list.

The macro runs. Follow the instructions on-screen. Spectra will be collected using the
instrument settings contained in the macro.
If you did not select the appropriate macro for your instrument, an error message will
appear.

You will be prompted during the macro to insert the TRM from the IPV kit into the
sample slide in the sample compartment. If you do not have an internal filter wheel,
during the macro run you will also be prompted to insert the calibrated polystyrene card
(L1202057) supplied with your instrument into the sample slide.
Spectra generated during the procedure will be addded to the Samples View in the Data
Explorer and saved to C:\pel_data\Spectra.

6. Replace the TRM in its box and store it in a safe place.

8 . Instrument Performance Validation Kit User's Guide

After running the macro, we recommend that you use the Send To Word function to output
your results to Word:

 Select Send To Word from the File menu, and then select New Word Document from
the Word sub-menu.
You can send a graph of your spectra and the contents of the Results Table.

Possible Reasons for Validation Failure

If the peaks were not found or lie outside the tolerance, the failure may be because:

• the wrong spectra were used;

• the wrong reference material was used to collect one of the spectra;

• the instrument was not aligned correctly;

• the noise level is too high;

• the beam was obstructed during validation, for example by an accessory in the beam
path. The effect is shown most at high wavenumbers;

If you need more help, contact your PerkinElmer agent or office.

When Should I Recalibrate My Instrument?

You must check the calibration of your instrument if a new source, detector or power supply
has been fitted to your instrument, and recalibrate if necessary.

You should also consider recalibrating your instrument if the peak positions in a spectrum
collected from your TRM are not within ±0.3 cm−1 of those given in the column labeled TRM
on the Certificate of Performance Validation.

NOTE: If you recalibrate your instrument, you must collect new ASTM reference spectra.

For recalibration purposes, we recommend that you use the peak at nominally 3060.14 cm−1
for the MIR. It is a symmetrical band, at high wavenumber and therefore sensitive to
calibration errors. Any change in calibration will affect each peak by an amount proportional
to its wavenumber. The change in position of peaks around 3000 cm−1 will be approximately
double the change seen for peaks around 1600 cm−1.

NOTE: The reference laser controls the wavenumber scale of the instrument and the scale is
calibrated by measuring the effective wavenumber of the laser. This differs from the
absolute value because the infrared radiation in the interferometer is not all exactly
parallel to the laser beam. Slightly different calibrations are required for the different
J-stops used for 2 cm−1 and 1 cm−1 resolution. Any calibration error will be
proportional to the wavenumber value.
 Instrument Performance Validation Kit User’s Guide . 9

For recalibration purposes, we recommend that you use the peak at nominally 5669.3 cm−1,
for the NIR.

NOTE: The reference laser controls the wavenumber scale of the instrument and the scale is
calibrated by measuring the effective wavenumber of the laser. This differs from the
absolute value because the infrared radiation in the interferometer is not all exactly
parallel to the laser beam. A different calibration is required for the Near Infrared
Reflectance Accessory (NIRA) and remote accessories. Any calibration error will be
proportional to the wavenumber value.

We recommend that, before starting the test, you make sure that the instrument is optimally
aligned.
10 . Instrument Performance Validation Kit User's Guide

Routine Instrument Validation

For route instrument validation, we recommend that you use the Instrument Verification
functionality in Spectrum software. This enables you to use both the Internal APV reference
and an external sample card in the sample compartment (such as the TRM).

For more information see the Spectrum on-screen help which can be accessed by selecting
Contents from the Help menu in Spectrum.

Routine Validation in MIR

What Peaks Should I Use?

Polystyrene film

There are five NIST certified peak positions for the polystyrene film. We recommend that you
use the three peaks nominally at 3060.14, 1601.38 and 1028.42 cm−1 for testing
wavenumber repeatability.

NG11 Schott glass filter

The most useful region for measurements with the glass filter is 4000 to 2000 cm−1. Because
the spectral features are very broad, there is no significant variation in the spectrum for
resolutions of 8 cm−1 or worse. Those regions subject to atmospheric interference from water
vapor around 3600 cm−1 and carbon dioxide around 2350 cm−1 should not be used. We
recommend measuring the transmittance at 3990, 3030 and 2000 cm−1.

For information, see the Spectrum on-screen Help.