ASTM D3016-97(2003)

Standard Practice for Use of Liquid Exclusion Chromatography Terms and Relationships

ABSTRACT
This practice covers the definitions of terms, symbols, and relationships most often used in liquid exclusion chromatography. Wherever possible, these terms and symbols are consistent with those used in other chromatographic techniques.
SCOPE
1.1 This practice covers the definitions of terms and symbols most often used in liquid exclusion chromatography. Wherever possible, these terms and symbols are consistent with those used in other chromatographic techniques. As additional terms and relationships are developed, they will be incorporated.
Note 1—There is no similar or equivalent ISO standard.

General Information

• Status: Historical
• Publication Date: 09-Mar-2003
• ICS: 01.040.71 - Chemical technology (Vocabularies)
• ICS: 71.040.50 - Physicochemical methods of analysis
• Technical Committee: D20 - Plastics
• Drafting Committee: D20.70 - Analytical Methods

Relations

• Replaces: ASTM D3016-97 - Standard Practice for Use of Liquid Exclusion Chromatography Terms and Relationships
• Effective Date: 10-Mar-2003
• Replaced By: ASTM D3016-97(2010) - Standard Practice for Use of Liquid Exclusion Chromatography Terms and Relationships
• Effective Date: 10-Mar-2003
• Referred By: ASTM E682-92(2019) - Standard Practice for Liquid Chromatography Terms and Relationships
• Effective Date: 10-Mar-2003
• Referred By: ASTM D6579-11(2020) - Standard Practice for Molecular Weight Averages and Molecular Weight Distribution of Hydrocarbon, Rosin and Terpene Resins by Size-Exclusion Chromatography
• Effective Date: 10-Mar-2003
• Referred By: ASTM D6474-20 - Standard Test Method for Determining Molecular Weight Distribution and Molecular Weight Averages of Polyolefins by High Temperature Gel Permeation Chromatography
• Effective Date: 10-Mar-2003
• Referred By: ASTM F2150-19 - Standard Guide for Characterization and Testing of Biomaterial Scaffolds Used in Regenerative Medicine and Tissue-Engineered Medical Products
• Effective Date: 10-Mar-2003
• Referred By: ASTM F561-19 - Standard Practice for Retrieval and Analysis of Medical Devices, and Associated Tissues and Fluids
• Effective Date: 10-Mar-2003
• Referred By: ASTM D5296-19 - Standard Test Method for Molecular Weight Averages and Molecular Weight Distribution of Polystyrene by High Performance Size-Exclusion Chromatography
• Effective Date: 10-Mar-2003

Standards Content (Sample)

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:D3016–97 (Reapproved 2003)
Standard Practice for
Use of Liquid Exclusion Chromatography Terms and
Relationships
This standard is issued under the fixed designation D3016; 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
Liquid exclusion chromatography (LEC) began as “gel filtration chromatography” which is
2 3
attributed to Porath and Flodin. With the invention of new column packings by Moore for
organic-phaseworkanewformofLECdevelopedwhichcommonlybecameknownasgelpermeation
chromatography or GPC. Liquid exclusion chromatography is a form of liquid chromatography (some
other forms being partition, ion-exchange, and adsorption) and as such is the preferred name for the
technique; however, the reader must be aware that other names are common in the literature, the most
prevalent being those cited above. LEC differs from all other chromatographic techniques in that only
the exclusion mechanism may be operative if meaningful data are to result. Most other chromato-
graphic mechanisms operate in essentially the opposite way, that is, with small molecules exiting first.
Any combination of mechanisms causes confusion and is misleading.
Liquidexclusionchromatographyasusedfortheanalysisofpolymershasgrownandmaturedsince
the first issuance of this practice in 1972. Therefore, some infrequently used or “outdated” terms have
beendeletedandsomemodernpractices(orterms)havebeenincluded.Moderndevelopmentsinclude
the use of constant-volume pumps, use of “microparticle” column packings and much smaller
columns, and automated data-handling procedures. In addition, SI units as recommended in ASTM
Standard IEEE/ASTM SI 102.1 for Metric Practice are now used.
1. Scope* IEEE/ASTM SI 10 Standard for Use of the International
System of Units (SI): The Modern Metric System
1.1 This practice covers the definitions of terms and sym-
bols most often used in liquid exclusion chromatography.
3. Terminology Apparatus Definitions
Wherever possible, these terms and symbols are consistent
3.1 absolute detectors—devices that sense and measure the
with those used in other chromatographic techniques. As
absolute concentration or other physical property of solute
additional terms and relationships are developed, they will be
components contained in the eluate.
incorporated.
3.2 by-pass or loop injector—the injector most common in
NOTE 1—There is no similar or equivalent ISO standard.
liquid exclusion chromatography and which utilizes a sample
chamber that can be filled with sample while it is temporarily
2. Referenced Documents
external to the flowing liquid stream. It can be manipulated by
2.1 ASTM Standards:
means of a valving device to sweep the sample with eluent into
the column.
3.3 collection devices—devices used to collect discrete
ThispracticeisunderthejurisdictionofASTMCommitteeD20onPlasticsand
is the direct responsibility of Subcommittee D20.70 on Analytical Methods. portions of an eluate according to a preset cycle (for example,
Current edition approved March 10, 2003. Published April 2003. Originally
times, volume, etc.).
approved in 1972. Last previous edition approved in 1997 as D3016 – 97. DOI:
3.4 column end-fittings—devices that prevent the column
10.1520/D3016-97R03.
packingfrompassingthroughthembutwhicharepermeableto
Porath, J., and Flodin, P., Nature, NTRWA, Vol 183, 1959, p. 1657.
Moore, J. C., Journal of Polymer Science, JPYAA, PartA, Vol 2, 1964, p. 835.
the eluent (solvent or solution).
Journal of Gas Chromatography, JCHSB, Vol 66, 1968.
3.5 columns—tubes that contain the column packing.
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at [e-mail]. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
*A Summary of Changes section appears at the end of this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
D3016–97 (2003)
3.6 detectors—devices that sense and measure the concen- comparison with other methods. For example, the molecular-
tration or other physical property of solute components in the weight distribution can be compared with that obtained from
solution (eluate) passing through. equilibrium ultracentrifugation. More commonly the weight-
3.7 differential detectors—devices that sense and measure and number-average molecular weights computed from the
the difference in a physical or chemical property between a LEC (GPC) trace are compared with those measured by light
solution (solvent containing solute components) and a refer- scattering, (both static and on-line), osmometry, and on-line
ence liquid (for example, solvent alone). viscometry.
3.8 direct injector—a device for introducing a sample from
6. Readout Definitions
a source external to the column directly onto the column (for
example, septum-syringe injector).
6.1 chromatogram—a plot of detector response against
3.9 pump—any device that causes mobile phase to flow
volume of eluate emerging from the system. An idealized
through the columns.
chromatogram obtained with a detector providing differential
3.10 sample inlet system—a means of introducing samples
response is shown in Fig. 1.
onto the column.
6.2 The definitions that follow apply to chromatograms
obtained directly by means of differential detectors:
4. Reagent Definitions
6.2.1 baseline—the portion of the chromatogram recording
the detector response when only eluent emerges from the
4.1 column packing—the stationary phase which consists of
column.
microporous material and the stationary liquid phase contained
6.2.2 half width, GH—the retention volume interval of a
in the pores.
line parallel to the peak base, bisecting the peak height, and
4.2 eluate—the liquid emerging from the column.
terminating at the sides of the peak.
4.3 eluent—the mobile phase or solvent used to sweep or
6.2.3 peak—the portion of the chromatogram recording the
elute the sample (solute) components into, through, and out of
detector response while a single component or a single
the column. Its composition is the same as the stationary liquid
distribution (two or more sample components that emerge
phase.
together as a single peak) is eluted from the column.
4.4 solutes—dissolved substances that, in LEC, are caused
6.2.4 peak area, BGDHCAB—the area enclosed between
to pass through the column and to influence the detector
the peak and the peak base.
response.
6.2.5 peak base, BC—an interpolation of the baseline be-
5. Performance in Terms of Resolution, Column tween extremities of the peak.
6.2.6 peak height, AD—the dimension from the peak maxi-
Efficiency, and Precision
mum to the base measured in the direction of detector
5.1 Resolution is the resultant of two effects, the separating
response.
power of the column packing and the efficiency or peak
6.2.7 peak width, EF—the retention volume interval of the
broadening. The separating power of the column packing is
segment of peak base intercepted by tangents to the inflection
dependent on pore size and pore volume. Peak broadening
points on either side of the peak.
depends on the nature of the column packings, on how well the
columns are packed, and on instrumental components external
7. Volume Parameters
to the columns. The equations used in LEC (GPC) are similar
7.1 Volume parameters expected to be involved in LEC are
to those used in other chromatographic techniques. Reference
listed in the glossary of terms found in Tables 1 and 2. These
may be made to any standard chromatography text. Resolution
terms are intended for use where the primary mechanism of
for any two samples is defined by the following equation:
separation depends on the size of the solute molecules. The
R 5 2[ V 2 V / W 1 W (1)
~ ! ~ !
1,2 R R 1 2
1 2
hydrodynamic volume of polymers usually may be related to
wheresubscripts1and2refertosamples1and2.(Seetables
V independentofmolecularweightandstructure,forexample,
R
for symbols used in this equation.) For complete separation,
in describing the exclusion limit. Hydrodynamic volume is
R must be 1.25 or greater.
1,2
5.2 Column efficiency is a measure of peak spreading or the
rate of generation of variance with column length. For a
monodisperse material, efficiency is the number of theoretical
plates, N, for the entire system defined as follows: (See tables
for symbols used in this equation.)
N 5 16~V /W! ors5 N/V (2)
R R
This expression includes all contributions to peak broaden-
ing.
5.3 Precision and accuracy are used according to their
accepted definitions. Precision is inherent to the system. Both
precision and accuracy are dependent on the method of
calibration and treatment of the data as well as on the resolving
power of the columns. The accuracy must be determined by FIG. 1 Typical Chromatogram
D3016–97 (2003)
TABLE 1 Recommended List of Terms and Symbols for Use in LEC—Part I
PART I—LEC PARAMETERS
A
Units
No. Parameter Symbol Definitions
SI Common
1 Peak width W cm ml The distance between the baseline intercepts of lines
drawn tangent to the points of inflection of the trace
2 Peak width at half height W cm ml Peak width measured parallel to baseline at 50 % of its
1/2
height
2 2
3 Peak area A cm cm The area enclosed between the peak and its base,
ABGDHCA in Fig. 1
4 Retention volume V cm ml Volume of liquid that has passed through the system
R
from middle of sample injection period to peak
maximum
5 Volume of mobile phase in interstitial V cm ml Volume of mobile p
...