ASTM E2719-09(2022)

Standard Guide for Fluorescence—Instrument Calibration and Qualification

SIGNIFICANCE AND USE
4.1 By following the general guidelines (Section 5) and instrument calibration methods (Sections 6 – 16) in this guide, users should be able to more easily conform to good laboratory and manufacturing practices (GXP) and comply with regulatory and QA/QC requirements, related to fluorescence measurements.  
4.2 Each instrument parameter needing calibration (for example, wavelength, spectral responsivity) is treated in a separate section. A list of different calibration methods is given for each instrument parameter with a brief usage procedure. Precautions, achievable precision and accuracy, and other useful information are also given for each method to allow users to make a more informed decision as to which method is the best choice for their calibration needs. Additional details for each method can be found in the references given.
SCOPE
1.1 This guide (1)2 lists the available materials and methods for each type of calibration or correction for fluorescence instruments (spectral emission correction, wavelength accuracy, and so forth) with a general description, the level of quality, precision and accuracy attainable, limitations, and useful references given for each entry.  
1.2 The listed materials and methods are intended for the qualification of fluorometers as part of complying with regulatory and other quality assurance/quality control (QA/QC) requirements.  
1.3 Precision and accuracy or uncertainty are given at a 1 σ confidence level and are approximated in cases where these values have not been well established.3  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety, health, and environmental practices and determine the applicability of regulatory limitations prior to use.  
1.6 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.

General Information

• Status: Published
• Publication Date: 31-Oct-2022
• ICS: 71.040.50 - Physicochemical methods of analysis
• Technical Committee: E13 - Molecular Spectroscopy and Separation Science
• Drafting Committee: E13.01 - Ultra-Violet, Visible, and Luminescence Spectroscopy

Standards Content (Sample)

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: E2719 − 09 (Reapproved 2022)
Standard Guide for
1
Fluorescence—Instrument Calibration and Qualification
This standard is issued under the fixed designation E2719; 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.
1. Scope 2. Referenced Documents
2 4
1.1 This guide (1) lists the available materials and methods 2.1 ASTM Standards:
for each type of calibration or correction for fluorescence E131 Terminology Relating to Molecular Spectroscopy
instruments (spectral emission correction, wavelength E388 Test Method for Wavelength Accuracy and Spectral
accuracy, and so forth) with a general description, the level of Bandwidth of Fluorescence Spectrometers
quality, precision and accuracy attainable, limitations, and E578 Test Method for Linearity of Fluorescence Measuring
useful references given for each entry. Systems
E579 Test Method for Limit of Detection of Fluorescence of
1.2 The listed materials and methods are intended for the
Quinine Sulfate in Solution
qualification of fluorometers as part of complying with regu-
latory and other quality assurance/quality control (QA/QC)
3. Terminology
requirements.
3.1 Definitions (2):
1.3 Precision and accuracy or uncertainty are given at a 1 σ
3.1.1 absorption coeffıcient (α), n—a measure of absorption
confidence level and are approximated in cases where these
of radiant energy from an incident beam as it traverses an
3
values have not been well established.
-αb
absorbing medium according to Bouguer’s law, I/I = e ,
o
1.4 The values stated in SI units are to be regarded as
where I and I are the transmitted and incident intensities,
o
standard. No other units of measurement are included in this
respectively, and b is the path length of the beam through the
standard.
sample. E131
3.1.1.1 Discussion—Note that transmittance T = I/I and
1.5 This standard does not purport to address all of the
o
absorbance A = –log T.
safety concerns, if any, associated with its use. It is the
responsibility of the user of this standard to establish appro-
3.1.2 absorptivity (a), n—the absorbance divided by the
priate safety, health, and environmental practices and deter-
product of the concentration of the substance and the sample
mine the applicability of regulatory limitations prior to use.
pathlength, a = A/bc. E131
1.6 This international standard was developed in accor-
3.1.3 Beer-Lambert law, n—relates the dependence of the
dance with internationally recognized principles on standard-
absorbance (A) of a sample on its path length (see absorption
ization established in the Decision on Principles for the
coeffıcient, α) and concentration (c), such that A =abc.
Development of International Standards, Guides and Recom-
3.1.3.1 Discussion—Also called Beer’s law or Beer-
mendations issued by the World Trade Organization Technical
Lambert-Bouquer law. E131
Barriers to Trade (TBT) Committee.
3.1.4 calibrated detector (CD), n—opticalradiationdetector
whose responsivity as a function of wavelength has been
1
This guide is under the jurisdiction of ASTM Committee E13 on Molecular
determined along with corresponding uncertainties (3).
Spectroscopy and Separation Science and is the direct responsibility of Subcom-
3.1.5 calibrated diffuse reflector (CR), n—Lambertian re-
mittee E13.01 on Ultra-Violet, Visible, and Luminescence Spectroscopy.
Current edition approved Nov. 1, 2022. Published November 2022. Originally
flector whose reflectance as a function of wavelength has been
approved in 2009. Last previous edition approved in 2014 as E2719–09 (2014).
determined along with corresponding uncertainties (4).
DOI: 10.1520/E2719-09R22.
2
The boldface numbers in parentheses refer to the list of references at the end of
this standard.
3 4
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