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ASTM E388-72(1998)

(Test Method)

Standard Test Method for Spectral Bandwidth and Wavelength Accuracy of Fluorescence Spectrometers

Standard Test Method for Spectral Bandwidth and Wavelength Accuracy of Fluorescence Spectrometers

SCOPE
1.1 This test method covers the testing of the spectral bandwidth and wavelength accuracy of fluorescence spectrometers.

General Information

Status
Historical
Publication Date
09-Oct-1998
ICS
17.180.30 - Optical measuring instruments
Technical Committee
E13 - Molecular Spectroscopy and Separation Science
Current Stage
Ref Project

Relations

Replaced By
ASTM E388-04 - Standard Test Method for Wavelength Accuracy of Spectral Bandwidth Fluorescence Spectrometers
Effective Date
01-Nov-2004
Referred By
ASTM E2056-04(2016) - Standard Practice for Qualifying Spectrometers and Spectrophotometers for Use in Multivariate Analyses, Calibrated Using Surrogate Mixtures
Effective Date
10-Oct-1998
Referred By
ASTM E3029-15(2023) - Standard Practice for Determining Relative Spectral Correction Factors for Emission Signal of Fluorescence Spectrometers
Effective Date
10-Oct-1998
Referred By
ASTM D5412-93(2017)e1 - Standard Test Method for Quantification of Complex Polycyclic Aromatic Hydrocarbon Mixtures or Petroleum Oils in Water
Effective Date
10-Oct-1998
Referred By
ASTM E2143-01(2021) - Standard Test Method for Using Field-Portable Fiber Optics Synchronous Fluorescence Spectrometer for Quantification of Field Samples for Aromatic and Polycyclic Aromatic Hydrocarbons
Effective Date
10-Oct-1998
Referred By
ASTM E2719-09(2022) - Standard Guide for Fluorescence—Instrument Calibration and Qualification
Effective Date
10-Oct-1998

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ASTM E388-72(1998) - Standard Test Method for Spectral Bandwidth and Wavelength Accuracy of Fluorescence SpectrometersASTM E388-72(1998) - Standard Test Method for Spectral Bandwidth and Wavelength Accuracy of Fluorescence Spectrometers
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ASTM E388-72(1998) - Standard Test Method for Spectral Bandwidth and Wavelength Accuracy of Fluorescence Spectrometers
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NOTICE: This standard has either been superseded and replaced by a new version or withdrawn.
Contact ASTM International (www.astm.org) for the latest information
Designation:E388–72 (Reapproved 1998)
Standard Test Method for
Spectral Bandwidth and Wavelength Accuracy of
Fluorescence Spectrometers
This standard is issued under the fixed designation E 388; 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 (e) indicates an editorial change since the last revision or reapproval.
1. Scope 576.96, and 579.07 nm. These are listed in the Mercury Arc
Emission Spectrum in the Ultraviolet and Visible Regions fig.
1.1 This test method covers the testing of the spectral
of Practice E 275.
bandwidth and wavelength accuracy of fluorescence spectrom-
6.1.1 Other lines are suitable for calibration such as some of
eters.
the weaker lines included in the Mercury Arc Emission
2. Referenced Documents Spectrum in the Ultraviolet andVisible Regions fig. of Practice
E 275, but not included in the above list. The comparatively
2.1 ASTM Standards:
low resolution monochromators often used in fluorescence
E 275 Practice for Describing and Measuring Performance
equipment may not resolve pairs of lines such as at 404.66 and
of Ultraviolet, Visible, and Near Infrared Spectrophotom-
407.78, or at 576.96 and 579.07 nm.
eters
6.1.2 In instruments using grating monochromators, addi-
3. Summary of Test Method
tional weaker lines are found due to second order diffraction of
mercury lines. Thus, lines appear at 507.30, 593.46, 668.30
3.1 The difference between the apparent wavelength and the
nm, arising from the 253.65, 296.73, and 334.15-nm lines,
known wavelength for a series of mercury emission lines is
respectively.
used as a test for wavelength accuracy. The apparent width of
6.2 Calibration and Adjustment of Emission Monochroma-
some of these lines is used as a test for spectral bandwidth.
tor:
4. Apparatus
6.2.1 With the mercury arc source properly aligned in the
sample cell compartment, adjust the position of the wavelength
4.1 Fluorescence Spectrometer to be tested.
dial to give maximum signal for each of the mercury lines and
4.2 Mercury Arc, Low-pressure , sufficiently small to be
record the wavelength reading. The difference between the
placed in the sample cell holder of the instrument.
observed value and the corresponding value in 6.1 represents
5. Reagent
the correction that must be subtracted algebraically from the
reading on the dial. The corrections may be recorded or the
5.1 Glycogen Suspension—Dissolve1gof glycogen per
monochromator adjusted to give the proper values. Since there
litre of water, or use a Ludox suspension containing 1 mL of
is some backlash in the wavelength drive, always adjust the
Ludox per litre of water.
dial to the peak reading from the same direction.
6. Procedure
6.2.2 When calibrating scanning-type instruments, turn the
dial to give the peak reading in the same direction that the dial
6.1 Lines suitable for calibration are the nine mercury lines
at 253.65, 296.73, 334.15, 404.66, 407.78, 435.84, 546.07, is turned by the scan motor. Check the dial reading against the
value recorded while scanning and, if necessary, correct as in
6.2.1.
This test method is under the jurisdiction of ASTM Committee E-13 on
6.3 In cases where the monochromator is designed so that a
Molecular Spectroscopy and is the direct responsibility of Subcommittee E13.06 on
lateral displacement of the calibration source from a position
Molecular Luminescence.
directly in front of the entrance slit appears as a wavelength
Current edition approved June 29, 1972. Published September 1972. Originally
published as E 388 – 69 T. Last previous edition E 388 – 69 T.
shift, proceed as follows:
Annual Book of ASTM Standards, Vol 14.01.
The Pen Ray Quartz Lamp, manufactured by Ultraviolet Products, Inc., San
Gabriel, CA, and available from apparatus distributors, has been found satisfactory.
Ludox is an aqueous suspension of colloidal silica, manufactured by E. I.
DuPont De Nemours and Co., Industrial and Biochemicals Dept., Wilmington, DE.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
E388
6.3.1 Instead of placing the mercury lamp in front of the 6.4.6 The match of the excitation monochromator with the
entrance slit of the monochromator, fill a sample cell with a emission monochromator may
...

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This test method covers the testing of the spectral bandwidth and wavelength accuracy of fluorescence spectrometers that use a monochromator for emission wavelength selection and photomultiplier tube detection. The method can be applied to instruments that use multi-element detectors, such as diode arrays, but results must be interpreted carefully. Atomic lines between 250 nm and 1000 nm are used in the method. The difference between the apparent wavelength and the known wavelength for a series of atomic emission lines is used as a test for wavelength accuracy. The apparent width of some of these lines is used as a test for spectral bandwidth.
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4.1 This practice permits an analyst to compare the performance of an instrument to the manufacturer's supplied performance specifications and to verify its suitability for continued routine use. It also provides generation of calibration monitoring data on a periodic basis, forming a base from which any changes in the performance of the instrument will be evident.
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4.1 This practice permits an analyst to compare the general performance of an instrument on any given day with the prior performance of an instrument. This practice is not necessarily meant for comparison of different instruments with each other even if the instruments are of the same type and model. This practice is not meant for comparison of the performance of one instrument operated under differing conditions.
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1.1 This practice describes two levels of tests to measure the performance of laboratory Fourier transform mid-infrared (FT-MIR) spectrometers equipped with a standard sample holder used for transmission measurements.  
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1.2.5 For FT-IR spectrometers run in a step scan mode, variations on this practice and information provided by the instrument vendor should be used.  
1.3 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.3.1 Exception—Informational inch-pound units are provided in 5.4.  
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Standard Practice for Estimation of the Spectral Bandwidth of Ultraviolet-Visible Spectrophotometers

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4.1 These practices should be used by a person who develops an analytical method to ensure that the spectral bandwidths cited in the practice are actually the ones used.
Note 2: The method developer should establish the spectral bandwidths that can be used to obtain satisfactory results.  
4.2 These practices should be used to determine whether a spectral bandwidth specified in a method can be realized with a given spectrophotometer and thus whether the instrument is suitable for use in this application. If accurate absorbance measurements are to be made on compounds with sharp absorption bands (natural half band widths of less than 15 nm) the spectral bandwidth of the spectrometer used should be better than 1/8th of the natural half band width of the compound’s absorption.  
4.3 These practices allow the user of a spectrophotometer to estimate the actual spectral bandwidth of the instrument under a given set of conditions and to compare the result to the spectral bandwidth calculated from data given in the manufacturer's literature or indicated by the instrument.
SCOPE
1.1 This practice describes procedures for estimating the spectral bandwidth of a spectrophotometer in the wavelength region of 185 nm to 820 nm.  
1.2 These practices are applicable to all modern spectrophotometer designs utilizing computer control and data handling. This includes conventional optical designs, where the sample is irradiated by monochromatic light, and ‘reverse’ optic designs coupled to photodiode arrays, where the light is separated by a polychromator after passing through the sample. For spectrophotometers that utilize servo-operated slits and maintain a constant period and a constant signal-to-noise ratio as the wavelength is automatically scanned, and/or utilize fixed slits and maintain a constant servo loop gain by automatically varying gain or dynode voltage, refer to the procedure described in Annex A1. This procedure is identical to that described in earlier versions of this practice.  
1.3 This practice does not cover the measurement of limiting spectral bandwidth, defined as the minimum spectral bandwidth achievable under optimum experimental conditions.  
1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
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ASTM E578-07(2021)(Test Method)
Standard Test Method for Linearity of Fluorescence Measuring Systems

SIGNIFICANCE AND USE
3.1 The range of concentration of a fluorescing substance in solution over which the fluorescence varies linearly with the concentration is the range most useful for quantitative analysis. This range is affected by properties of the solution under analysis and by features of the measuring system. This test method provides a means of testing the performance of a fluorescence measuring system and of determining the concentration range over which the system is suitable for making a given quantitative analysis.  
3.2 This test method is not meant for comparing the performance of different fluorescence measuring instruments.
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1.5 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard.  
1.6 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.  
1.7 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.

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4.1 Although it is possible to observe and measure each of the several characteristics of a detector under different and unique conditions, it is the intent of this practice that a complete set of detector specifications should be obtained under the same operating conditions. It should also be noted that to completely specify a detector’s capability, its performance should be measured at several sets of conditions within the useful range of the detector. The terms and tests described in this practice are sufficiently general that they may be used regardless of the ultimate operating parameters.  
4.2 Linearity and response time of the recorder or other readout device used should be such that they do not distort or otherwise interfere with the performance of the detector. This requires adjusting the gain, damping, and calibration in accordance with the manufacturer's directions. If additional electronic filters or amplifiers are used between the detector and the final readout device, their characteristics should also first be established.
SCOPE
1.1 This practice is intended to serve as a guide for the testing of the performance of a photometric detector (PD) used as the detection component of a liquid-chromatographic (LC) system operating at one or more fixed wavelengths in the range 210 nm to 800 nm. Measurements are made at 254 nm, if possible, and are optional at other wavelengths.  
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