This international standard was developed in accordance with internationally recognized principles on standardization established in the Decision on Principles

for the
Development of International Standards, Guides and Recommendations issued by the World Trade Organization Technical Barriers to Trade (TBT) Committee.

Designation: E925 − 09 (Reapproved 2022)

Standard Practice for

Monitoring the Calibration of Ultraviolet-Visible
Spectrophotometers whose Spectral Bandwidth does not
Exceed 2 nm1
This standard is issued under the fixed designation E925; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.

INTRODUCTION

In the application of spectrophotometric methods of analysis it is the responsibility of the analyst

to verify and validate that the instrument is functioning properly and is capable of providing
acceptable analytical results. It is preferable that the verification of instrument performance be
accomplished through the use of reference materials whose properties have been accurately
determined. Such materials are readily available, and their use in the tests and measurements described
in this practice is satisfactory for evaluating the performance of spectrophotometers whose spectral
bandwidth does not exceed the value for which the intrinsic or certified properties are valid. A
compromise maximum permissible spectral bandwidth of 2 nm is recommended for the reference
materials and error tolerances recommended here.

iTeh Standards
This practice covers some of the essential instrumental parameters that should be evaluated to
ensure the acceptability of the analytical data routinely obtained on the instrument. These parameters
include the accuracy of the wavelength and absorbance scales and stray radiant power levels.
(https://standards.iteh.ai)
The accuracy of the wavelength scale in both the ultraviolet (UV) and visible regions is determined
using the sharp absorption bands of a holmium oxide glass or solution filter. The absorbance scale

Document Preview
accuracy in the UV region (235 nm to 350 nm) is determined using acidic solutions of potassium
dichromate. In the visible region (440 nm to 635 nm) the absorbance accuracy is determined using
individually certified neutral density glass filters. The use of these reference materials provides a valid
and relatively simple means to test the errors in the wavelength and absorbance scales of small spectral
bandwidth spectrophotometers in ASTM E925-09(2022)
the spectral ranges indicated. A simplified version of the opaque
https://standards.iteh.ai/catalog/standards/sist/4e9723a2-8e57-4f46-ba59-8fc47e90ff79/astm-e925-092022
filter method is provided as a test for excessive stray radiant energy.

1. Scope 1.2 This practice may be used as a significant test of the

performance of instruments for which the spectral bandwidth
1.1 This practice covers the parameters of spectrophotomet-
does not exceed 2 nm and for which the manufacturer’s
ric performance that are critical for testing the adequacy of
specifications for wavelength and absorbance accuracy do not
instrumentation for most routine tests and methods2 within the
exceed the performance tolerances employed here. This prac-
wavelength range of 200 nm to 700 nm and the absorbance
tice employs an illustrative tolerance of 61 % relative for the
range of 0 to 2. The recommended tests provide a measurement
error of the absorbance scale over the range of 0.2 to 2.0, and
of the important parameters controlling results in spectropho-
of 61.0 nm for the error of the wavelength scale. A suggested
tometric methods, but it is specifically not to be inferred that all
maximum stray radiant power ratio of 4 × 10-4 yields <1 %
factors in instrument performance are measured.
absorbance bias at an absorbance of 2. These tolerances are
chosen to be compatible with many chemical applications
while comfortably exceeding the uncertainty of the certified
1
This practice is under the jurisdiction of ASTM Committee E13 on Molecular values for the reference materials and typical manufacturer’s
Spectroscopy and Separation Science and is the direct responsibility of Subcom-
mittee E13.01 on Ultra-Violet, Visible, and Luminescence Spectroscopy. specifications for error in the wavelength and absorbance
Current edition approved Nov. 1, 2022. Published November 2022. Originally scales of the instrument under test. The user is encouraged to
approved in 1983. Last previous edition approved in 2014 as E925 – 09 (2014). develop and use tolerance values more appropriate to the
DOI: 10.1520/E0925-09R22.
2
Routine tests are defined as those in which absorbance data obtained on a requirements of the end use application. This procedure is
sample are compared to those of a standard sample preparation. designed to verify quantitative performance on an ongoing

Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States

1
 E925 − 09 (2022)
basis and to compare one instrument’s performance with that 2.3 ISO Publications:5
of other similar units. Refer to Practice E275 to extensively ISO 17025 General Requirements for the Competence of
evaluate the performance of an instrument. Testing and Calibration Laboratories
1.3 This practice should be performed on a periodic basis, ISO Guide 34 General Requirements for the Competence of
the frequency of which depends on the physical environment Reference Material Producers
within which the instrumentation is used. Thus, units handled
3. Terminology
roughly or used under adverse conditions (exposed to dust,
chemical vapors, vibrations, or combinations thereof) should 3.1 Definitions:
be tested more frequently than those not exposed to such 3.1.1 For the definitions of terms used in this practice, refer
conditions. This practice should also be performed after any to Terminology E131.
significant repairs are made on a unit, such as those involving 3.1.2 For a description of the instrumental parameters
the optics, detector, or radiant energy source. evaluated in this practice, refer to Practice E275.
3.1.3 For a description of quantitative ultraviolet spectro-
1.4 The values stated in SI units are to be regarded as
photometric techniques, refer to Practices E169.
standard. No other units of measurement are included in this
standard. 4. Significance and Use
1.5 This standard does not purport to address all of the
4.1 This practice permits an analyst to compare the perfor-
safety concerns, if any, associated with its use. It is the
mance of an instrument to the manufacturer’s supplied perfor-
responsibility of the user of this standard to establish appro-
mance specifications and to verify its suitability for continued
priate safety, health, and environmental practices and deter-
routine use. It also provides generation of calibration monitor-
mine the applicability of regulatory limitations prior to use.
ing data on a periodic basis, forming a base from which any
1.6 This international standard was developed in accor-
changes in the performance of the instrument will be evident.
dance with internationally recognized principles on standard-
ization established in the Decision on Principles for the 5. Reference to this Calibration-Monitoring Procedure
Development of International Standards, Guides and Recom-
iTeh Standards
mendations issued by the World Trade Organization Technical
Barriers to Trade (TBT) Committee.
5.1 Reference to this practice in any spectrophotometric
calibration-monitoring scheme shall constitute due notification
that the adequacy of the spectrophotometer performance has
2. Referenced Documents (https://standards.iteh.ai) been evaluated by means of this practice. Performance is
3 considered to be adequate when the data obtained are within
2.1 ASTM Standards:
Document
E131 Terminology Relating to Molecular Spectroscopy
E169 Practices for General Techniques of Ultraviolet-Visible
Preview
the stated tolerances from the true values.

6. Instrument Operation
Quantitative Analysis
6.1 In obtaining spectrophotometric calibration data the
ASTM E925-09(2022)
E275 Practice for Describing and Measuring Performance of
Ultraviolet and Visible Spectrophotometers analyst must select the proper instrumental operating condi-
https://standards.iteh.ai/catalog/standards/sist/4e9723a2-8e57-4f46-ba59-8fc47e90ff79/astm-e925-092022
E387 Test Method for Estimating Stray Radiant Power Ratio tions to realize satisfactory instrument performance. Operating
of Dispersive Spectrophotometers by the Opaque Filter conditions for individual instruments are best obtained from
Method the manufacturer’s literature because of variations in instru-
E1866 Guide for Establishing Spectrophotometer Perfor- ment design.
mance Tests 6.2 When using reference materials, all the components of
2.2 NIST Publications:4 the spectrophotometer must be functioning properly. In
NIST Special Publication 260-54 Certification and Use of addition, the temperature of the specimen compartment should
Acidic Potassium Dichromate Solutions As An Ultraviolet be between 20 and 25°C. Matched solution cells should be
Absorbance Standard used for calibration purposes.
NIST Special Publication 260-102 Holmium Oxide Solution 6.3 Each of the above factors in instrument operation is
Wavelength Standard from 240 to 640 nm—SRM 2034 important in the determination of wavelength and absorbance
NIST Special Publication 260-116 Glass Filters as a Stan- accuracy.
dard Reference Material for Spectrophotometry—
Selection, Preparation, Certification, and Use of SRM 930 7. Determination of Wavelength Accuracy in the
and SRM 1930 Ultraviolet and Visible Spectral Regions
NIST Special Publication 260-140 Technical Specifications
for Certification of Spectrophotometric NTRMs 7.1 Discussion—The holmium oxide glass filter (1, 2)6 or
solution standard (NIST Special Publication 260-102) may be

3
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
5
contact ASTM Customer Service at [email protected]. For Annual Book of ASTM Available from International Organization for Standardization (ISO), ISO
Standards volume information, refer to the standard’s Document Summary page on Central Secretariat, Chemin de Blandonnet 8, CP 401, 1214 Vernier, Geneva,
the ASTM website. Switzerland, https://www.iso.org.
4 6
Available from National Technical Information Service (NTIS), 5301 Shawnee The boldface numbers in parentheses refer to a list of references at the end of
Rd., Alexandria, VA 22312, http://www.ntis.gov. this standard.

2
 E925 − 09 (2022)
used for evaluating wavelength accuracy. The glass and solu- that digital filter widths should be smaller than the full-width-
tion standards are both available commercially from reference half-maximum recommendation of that guide.
material producers, in the sealed cuvette format (a cuvette- 7.1.4 In the absence of drift or slippage in the wavelength
shaped metal holder is used for the glass) or as a bottled drive train, repeatability of the band positions should be on the
solution, prepared from high purity Holmium Oxide (> order of 60.1 nm for a given instrument, especially with the
99.99 %), where value assignment is by self assertion (Note 1). use of a computer based peak location algorithm.
A purchaser should require certification by the supplier that the 7.2 Procedure:
wavelengths of the absorption bands are within 0.2-nm of the 7.2.1 Examine the holmium oxide reference material and
values given in Ref. (2), and reported below. The appropriate remove any surface contamination using a soft brush or
solution standard is 4 % (mass fraction) holmium oxide in lint-free cloth. Measure the temperature of the sample com-
10 % (volume fraction) perchloric acid, contained in a 10-mm partment by placing an appropriate sensor into the cell com-
path length cuvette. For this material, the transmittance minima partment of a stabilized instrument and replacing the compart-
of 18 absorption bands have been certified by a multi- ment cover securely. Place the sensor as close as possible to the
laboratory inter-comparison, at the highest level, allowing the actual position that will be occupied by the standard. After a
peak value assignments as an intrinsic wavelength standard (3). suitable period of time record the temperature reading, remove
Absorbance maxima or transmittance minima must be located the sensor, and resume normal operations.
within 61 nm of the wavelengths given below: 7.2.2 Record the blank absorbance or transmittance (air
Glass FilterA Dilute Acidic SolutionB versus air) spectrum at the desired resolution and at the
241.5 nmC 241.1 nm
... 249.9 nm
appropriate wavelength intervals and scan speeds, in order to
279.3 nm 278.1 nm perform any necessary baseline adjustments. The wavelength
287.6 nm 287.2 nm intervals should be no greater than the spectral bandwidth used.
333.8 nm 333.5 nm
... 345.4 nm Acquire the appropriate spectrum of the holmium oxide
360.8 nm 361.3 nm reference material with respect to air and baseline correct if
385.8 nm 385.6 nm necessary using the blank spectrum. Record the wavelengths of
418.5 nm 416.3 nm
the positions of the relevant bands, and compare these values to
453.4 nm
459.9 nm
...
iTeh Standards ...D
467.8 nm
485.3 nm
the expected values. If large discrepancies (>1 nm) exist
between the true and measured wavelengths, repeat the proce-
(https://standards.iteh.ai)
536.4 nm 536.6 nm
637.5 nm 640.5 nm dure at a slower scan speed and smaller spectral bandwidth, if
possible, to verify the nonconformity.

Document Preview
A
Wavelengths taken from Ref. (2) for Corning Glass Works Code 3130 glass, 7.2.3 Report the wavelength calibration data in the manner
superceded by Corning Glass Works Code 3131 glass and Kopp Glass Code 3131 of Table 1, given as an example for the holmium oxide glass
glass, for which the wavelengths are also valid.
B
Wavelengths rounded to 0.1 nm for a 1 nm spectral bandwidth taken from Ref. reference material.
(3).
C
D
ASTM E925-09(2022)
May not be usable, depending on the base glass of the filter.
Peak omitted because it resolves into a doublet at spectral bandwidth values
TABLE 1 UV-VIS Spectrophotometer Wavelength and Stray
https://standards.iteh.ai/catalog/standards/sist/4e9723a2-8e57-4f46-ba59-8fc47e90ff79/astm-e925-092022
less than 1 nm. Radiant Power Ratio Calibration
NOTE 1—‘Self assertion’ may take the form of value assignment and Instrument
certification in many forms. Some specific examples are: Date
(1) By a national metrology institute (NMI), Temperature
Analyst
(2) By an ISO 17025 and ISO Guide 34 accredited Reference Mate-
Wavelength Calibration: Holmium Oxide Filter
rial producer, and
(3) By a laboratory claiming ‘traceability’ to an NMI.
In all cases, the user should be satisfied that the quality of the value True Observed Conformance
Difference
assignment data meets the laboratory requirements. Wavelength Wavelength
(nm) Does Does Not
(nm) (nm)
7.1.1 If the observed absorption bands of the holmium oxide 241.5 ± 1
279.3 ± 1
glass or solution deviate by more than 61 nm from the values 287.6 ± 1
stated, then corrective service must by performed on the 333.8 ± 1
instrument by qualified personnel. If the user performs this 360.8 ± 1
385.8 ± 1
service, the manufacturer’s recommended procedure should be 418.5 ± 1
followed carefully. 453.4 ± 1
7.1.2 The wavelength accuracy is dependent on the spectral 459.9 ± 1
536.4 ± 1
bandwidth and thus on the physical bandwidth. Spectral 637.5 ± 1
bandwidths may be determined from the manufacturer’s speci-
fications. Stray Radiant Power Ratio
7.1.3 Computer based peak location algorithms that may be Wavelength Transmittance
Conforms
Does Not
used to assign absorbance maxima or transmittance minima are (nm) or Absorbance Conform
220
discussed in 7.6 of Guide E1866. It should be noted that peak 340
asymmetries in the holmium oxide reference materials are such

3
 E925 − 09 (2022)
8. Evaluation of Stray Radiant Power Ratio (SRPR) tified absorbances should be traceable to the regular transmit-
8.1 Discussion—A portion of the unwanted stray radiant tance scale maintained by an NMI.
power detected by the photodetector can be measured using the 9.1.1 If the blank-corrected absorbances (Acorr) of the stan-
following sharp cut-off solution filters in 1 cm cells: dards are outside the acceptable range, then corrective service
Solution Wavelength
must be performed on the instrument by qualified personnel. If
KI or NaL, 10.0 g/L in H2O 220 nm the user performs this service, the manufacturer’s recom-
NaNo2, 50.0 g/L in H2O 370 nm mended procedure should be followed carefully.
8.1.1 Reagent grade materials should be used for these 9.1.2 An acceptable absorbance range for each standard for
solutions. They are essentially opaque at the indicated wave- any instrument must be determined based on the instrument
lengths; any observed transmittance is equivalent to the effec- manufacturer’s specifications and on the analytical demands of
tive SRPR. the end-use application of the instrument. As a guide to the
8.1.2 An acceptable level of SRPR depends on the spectral acceptability of photometric accuracy data a tolerance of
character and absorbance level of the sample under investiga- 61.0 % relative (0.2 ≤ A ≤ 2.0) is employed in this practice.
tion. However, an upper limit of 4 × 10-4 is consistent with a The user is encouraged to establish tolerance limits more
worst-case absorbance bias of ~1 % at the upper limit of the appropriate to the application in question, and use the tables of
absorbance range (0 < A ≤ 2) covered by this practice, and is this practice as templates for custom tables that reflect the
suggested in the absence of other criteria. appropriate tolerances. One approach often used in defining
8.1.3 While the stray radiant power ratio is equivalent to the these limits is to linearly add the certified expanded uncertainty
transmittance described previously, it is often more convenient budget (k = 2) for a given reference material, to the manufac-
to make the measurement in the absorbance mode and math- turer quoted instrument photometric accuracy specification.
ematically convert absorbance to transmittance. The value 9.1.3 Rigorous treatment of the construction and use of an
quoted in 8.1.2 (4 × 10-4) equates to an absorbance value of absorbance correction curve for high accuracy work is beyond
3.4A. the scope of this practice.
8.1.4 An excessive SRPR usually arises from dust, 9.1.4 Studies by NIST and other ISO 17025 and
iTeh Standards
scratches, or corrosion on the collimator or disperser, or both.
Qualified personnel should correct this problem. Care should
be taken to discriminate between SRPR and light leaks. The
ISO Guide 34 accredited organizations have indicated that
solutions of acidic potassium dichromate are stable for at least

(https://standards.iteh.ai)
six months when prepared in the manner described in 9.3.2.1
latter most often originate in the sample compartment and can and stored in the dark in well-stoppered 1 L volumetric flasks,
be detected by blocking the sample beam alternately at the and for at least two years when permanently sealed in
Document Preview
ports on the source and detector sides of the sample compart-
ment. Any difference in the detected signals indicates a light
ampoules or far UV quartz cuvettes, by heat fusion. Neutral
density glass filters are certified by different sources for periods
leak.
of from two to five years, with appropriately adjusted uncer-
8.2 Procedure: ASTM E925-09(2022)
tainties.
8.2.1 Use the visible light source lamp in the 340 nm region
https://standards.iteh.ai/catalog/standards/sist/4e9723a2-8e57-4f46-ba59-8fc47e90ff79/astm-e925-092022
and the ultraviolet light source lamp in the 220 nm region.
9.2 Visible Region—The absorbance scale in the visible
region is tested using filters of a proprietary neutral glass. The
8.2.2 Determine the transmittance or absorbance of each
construction and certification of such filters is described in
solution at the appropriate wavelength using the indicated
some detail in NIST Special Publication 260-116 and
solvents for reference.
NIST Special Publication . Traceability of the certified absor-
8.2.3 Refer to Test Method E387 if the dynamic range of the bance values to the transmittance scale maintained by the
readout electronics of the instrument is not adequate for the Analytical Chemistry Division (ACD) of NIST is supported by
direct measurement of SRPR as described here. NIST for commercial participants in the NIST Traceable
8.2.4 In the manner of Table 1, report the transmittance or Reference Materials (NTRM) program, or by self assertion
absorbance of these solutions. Note whether the effective stray (Note 1) for other commercial sources. Traceability of these
radiant power ratio exceeds the suggested tolerance of 4 × 10-4 filters is normally maintained through NIST SRM 930 filters
or the user-defined tolerance. with nominal absorbances of 0.5, 0.7, and 1.0 and SRM 1930
filters with absorbances of 0.3, 1.5, and 2.0. (A letter series
9. Determination of the Absorbance Scale Accuracy in designation for SRM 930 is periodically adjusted without
the Ultraviolet and Visible Spectral Regions significant effect for this practice.) The wavelengths for which
9.1 Discussion—The accuracy of the absorbance scale is certified absorbance values are reported for individual filters
determined using reference materials with known absorbances. are close to local extrema in the nearly-neutral glass to
The absorbance scale accuracy in the ultraviolet region minimize the effect of wavelength error on the measured
(235 nm to 350 nm) is determined using acidic solutions of transmittance:
potassium dichromate as described in NIST Special Publica- 440.0 nm
tion 260-54. In the visible region (440 nm to 635 nm) the 465.0 nm
absorbance accuracy is determined using certified neutral 546.1 nm
density glass filters as described in NIST Special Publica- 590.0 nm
tion 260-116 and NIST Special Publication 260-140. The cer- 635.0 nm