International Journal of Polymer Analysis and

Characterization

ISSN: 1023-666X (Print) 1563-5341 (Online) Journal homepage: https://www.tandfonline.com/loi/gpac20

Universal calibration of gel permeation

chromatography using evaporative light scattering
detector coupled with viscometer

Adrian Boborodea & Stephen O’Donohue

To cite this article: Adrian Boborodea & Stephen O’Donohue (2020): Universal calibration
of gel permeation chromatography using evaporative light scattering detector coupled
with viscometer, International Journal of Polymer Analysis and Characterization, DOI:
10.1080/1023666X.2020.1770403

To link to this article: https://doi.org/10.1080/1023666X.2020.1770403

Published online: 02 Jun 2020.

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INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION
https://doi.org/10.1080/1023666X.2020.1770403

Universal calibration of gel permeation chromatography using

evaporative light scattering detector coupled with viscometer
Adrian Boborodeaa and Stephen O’Donohueb
a
Certech ASBL, Seneffe, Belgium; bAgilent Technologies LDA UK Ltd, Craven Arms Shropshire, UK

ABSTRACT ARTICLE HISTORY

The linearized evaporative light scattering detector (ELSD) allows the evalu- Received 1 August 2019
ation of molecular weight distributions (MWD) of a range of polymers Accepted 11 May 2020
using conventional gel permeation chromatography (GPC), and is a suit-
KEYWORDS
able replacement for the differential refractive index (DRI) detector in appli-
Evaporative light scattering;
cations which will benefit from fast equilibration, solvent gradients or HDPE; high-
when working with low polymer concentrations. Further, when DRI is temperature GPC
coupled with a viscometer, the Universal Calibration GPC method also
allows the evaluation of Mark–Houwink–Sakurada parameters K and alpha,
which provides valuable information on the chemical structure of polymer
chains. Therefore, for a wider acceptance of ELSD in GPC, we investigated
the possibility of coupling the ELSD with the viscometer.
Recommendations are provided for successfully coupling the ELSD and
viscometer, as well as on software adjustments to calculate K and alpha
parameters. For linear polyethylene, a good correlation was found between
the measured and the expected K and alpha values.

Introduction
Gel permeation chromatography (GPC) is the recommended analytical method to evaluate the
molecular weight distribution (MWD) of soluble polymers, the differential refractive index (DRI)
being the first choice as detector due to its linear response in concentration and negligible variation
in sensitivity with molecular weight.[1] However, being also sensitive to temperature and pressure
fluctuations, the DRI requires a relatively long time for equilibration. In addition, negative and posi-
tive peaks present in DRI chromatograms can be problematic, and often an intense solvent peak can
impede the evaluation of concentrations for the analysis of low molecular weight materials.
Being classified, like DRI, as a universal concentration detector for liquid chromatography, the
evaporative light scattering detector (ELSD) was considered for many decades as a possible
replacement for DRI. However, its application in GPC was restricted because of the loss of low
molecular weight species during eluent nebulization/evaporation, and because of power law func-
tion relationship of the signal vs concentration:
ELSD signal intensity ¼ kELSD  concentrationaELSD
The evaporation of low molecular species was suppressed by introducing a diffuser trapping
device,[2] which prevents large particles from reaching the evaporation chamber, and by introduc-
ing a dry evaporator gas which allows solvent evaporation to be achieved at lower temperatures.[3]
In the case of polyolefins analyzed in trichlorobenzene (TCB), these improvements allow the
reduction of the solvent evaporation temperature from 270  C to 120  C, thus producing an ELSD

CONTACT Adruan Boborodea adrian.boborodea@certech.be Certech ASBL, Rue Jules Bordet, Zone industrielle C,
Seneffe, B-7180, Belgium.
ß 2020 Taylor & Francis Group, LLC
2 A. BOBORODEA AND S. O’DONOHUE

signal which is less dependent on molecular weight and covering a broader range of molecular
weights. These enhancements are now into a dedicated GPC model of the ELSD, and combined
with the linearization of the ELSD signal intensity, the correct evaluation of the MWD of poly-
mers is now achievable.[4] The following power coefficients were generated for different solvents
using this GPC model of ELSD allowing the analysis of polyolefins:

 Trichlorobenzene, aELSD ¼ 1.61[4]

 Dibutoxymethane, aELSD ¼ 1.35[5]
 Xylene, aELSD ¼ 1.40 [6]

The conventional GPC experiment with a single concentration detector is used in a large
majority of GPC applications, the MWD being calculated using Mark–Houwink–Sakurada (MHS)
parameters of the standards and sample provided by literature. However, with the separation
mechanism in GPC being driven by the polymer coil dimensions in solution, it is becoming more
common to combine the concentration detector with a viscometer, to evaluate the intrinsic vis-
cosity of eluted species. Accordingly, the universal GPC method is based on the direct evaluation
of K and alpha parameters of unknown samples, using the following relationships:[7,8]
Vh ¼ ½gM ¼ KMaþ1
where Vh, [g] and M are respectively the hydrodynamic volume, the intrinsic viscosity and the
molecular weight of a monodisperse fraction of the polymer.
Whilst combining a DRI with a viscometer is relatively straightforward, both parallel and serial
connections being possible, coupling the ELSD with the viscometer is slightly more challenging,
mainly because ideally both detectors should be the last one in the flow path, but also requiring a
minimum flow rate for an adequate signal to noise.
In this study we provide the necessary information for successfully coupling the ELSD with
the viscometer in a parallel configuration, as well as the software modifications to correctly evalu-
ate the MWD and the MHS parameters. The universal calibration method was evaluated on a lin-
ear polyethylene (PE) reference, for which the average molecular weights are given by its
certificate and the expected MHS parameters are provided in the literature.[1]

Experimental
Solvent
1,2,4-Trichlorobenzene (TCB Spectropure dry, Biosolve, Netherlands, CAS 120-82-1).

Samples
The method was evaluated using a high-density polyethylene (HDPE) reference (SRM 1475a), from
National Institute of Standards and Technology, having the following certified average molecular
weights: Mn ¼ 18,310 ± 360 g/mol, Mw ¼ 53,070 ± 620 g/mol, Mz ¼ 138,000 ± 3,700 g/mol. The column
calibration was created with polystyrene (PS) EasiVials standards from Agilent Technologies described
in Table 1.

GPC apparatus and method

All GPC data were recorded using an Agilent PL-GPC 220 High-Temperature GPC System
equipped with ELSD in parallel with a DRI detector followed by a viscometer (VS). The SRM
1475a was dissolved to obtain concentrations of 1 mg/ml in TCB at 160  C. The injection volume
 INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION 3

- M), peak molecular weight and the quantity for PS

Table 1. Average molecular weights (Mn, Mw), molar mass dispersity (D
standards (batch 0006339313) provided in 4 mL vials.
EasiVial Code Mn (g/mol) Mw (g/mol) -M
D Peak MW (g/mol) Mass (mg)
PS Red PS 6035k 5,405,000 5,680,000 1.05 6,035,000 0.8
PS 436k 419,300 430,900 1.03 436,200 1.6
PS 20k 19,740 20,260 1.03 19,920 2.4
PS 1150 1,080 1,170 1.08 1,150 3.2
PS Yellow PS 3187k 2,777,000 2,996,000 1.08 3,187,000 0.8
PS 202k 197,900 204,000 1.03 202,100 1.6
PS 10k 9,370 9,580 1.02 9,590 2.4
PS 580 580 655 1.12 580 3.2
PS Green PS 1074k 1,013,000 1,059,000 1.05 1,074,000 0.8
PS 70k 68,100 69,350 1.02 70,500 1.6
PS 4730 4,550 4,690 1.03 4,730 2.4
PS 162 – – 1.0 162 3.2

45
40
35
VS
30
25
signal (mV)

20
ELSD
15
10
5
DRI
0
-5
-10
0 5 10 15 20 25
me (min)
Figure 1. Representative DRI, linearized ELSD and viscometer (VS) chromatograms for EasiVial PS Red.

was 200 mL and the chromatographic separation was performed using two PLgel 10 mm Mixed-B
(300  7.5 mm) columns at a flow rate of 1.2 mL/min and a temperature of 160  C.
Agilent GPC software (v1.2) was used for data acquisition. The raw data (DRI, ELSD and vis-
cosity) were exported to Excel template files allowing the linearization of ELSD chromatograms
using a power exponent of aELSD ¼ 1.61.

Results and discussion

Figure 1 shows the DRI, the linearized ELSD and the viscometer chromatograms of a mixture of
four PS standards (EasiVial Red). Several important observations are to be made:

 Although the flow is divided between the ELSD path and RI-viscometer path, using a flow rate of
1.2 mL/min provides a reasonable resolution of PS peaks, as recorded by the viscometer, for data
manipulation.
4 A. BOBORODEA AND S. O’DONOHUE

7.0
DRI ELSD
6.5
Linear (DRI) Linear (ELSD)
6.0
y = -0.7696x + 12.972 y = -0.7659x + 13.013
5.5 R² = 0.9998 R² = 0.9999

5.0
LOG (MW)

4.5

4.0

3.5

3.0

2.5

2.0
8.0 9.0 10.0 11.0 12.0 13.0 14.0
me (min)
Figure 2. Comparison of the calibration curves obtained with DRI and ELSD by injecting the EasiVial PS standards.

 The DRI and linearized ELSD chromatograms present similar features (same peak elution
times, peak shapes and separation), implying that ELSD could replace DRI as a concentra-
tion detector.
 For all peaks, the linearized ELSD signal to noise is about 5 times higher than the one given
by the DRI detector.
 When compared to the DRI chromatogram, no solvent peak was recorded in the ELSD chro-
matogram after 16 minutes.
 The DRI, ELSD and viscometer chromatograms allow measurement of the inter-detector vol-
ume between DRI and viscometer (0.4 mL), and between ELSD and viscometer (0.3 mL).
These two parameters are essential to evaluate the intrinsic viscosity of unknown samples.
 Most important, the peak areas of four PS standards evaluated based on the linearized ELSD
signal (0.09 mg/mL, 0.21 mg/mL, 0.31 mg/mL, 0.37 mg/mL) follow more closely the concentra-
tion series (0.10 mg/mL, 0.20 mg/mL, 0.30 mg/mL, 0.40 mg/mL) compared to the peak areas
measured with DRI (0.09 mg/mL, 0.20 mg/mL, 0.32 mg/mL, 0.25 mg/mL).

Therefore, we find that the linearized ELSD is sufficiently sensitive and responds linearly to
solute concentration, independently of molecular weight, thus qualifying ELSD as a GPC detector
conforming with ASTM 6474 criteria.[1]
Conventional GPC using only a concentration detector is a relative method in which the sam-
ples are differentiated based on the peak elution time. In this application, average molecular
weights are calculated based on calibrations with narrow distribution PS standards, which are
overlaid in Figure 2 for the ELSD and DRI detector. The plotted parallel lines have a time differ-
ence of 0.1 min, due to the slightly longer heated tubing to reach the external ELSD detector as
compared to the DRI detector located in the GPC oven immediately near the columns.
Representative chromatograms of SRM 1475a are presented in Figure 3. The DRI and ELSD
traces can be converted to MWD shown in Figure 4, using the established calibration curves and
the following K and alpha parameters:[1]

 For polystyrene: KPS ¼ 19  103 mL/g; aPS ¼ 0.655

 For linear polyethylene: KPE ¼ 39  103 mL/g; aPE ¼ 0.725
 INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION 5

65
60
55
50
45
VS
40
35
signal (mV)

30
25
ELSD
20
15
10
5
DRI
0
-5
-10
5 10 15 20
me (min)
Figure 3. Representative DRI, linearized ELSD and viscometer (VS) chromatograms for SRM 1475a.

1.0
RI
0.9

0.8 ELSD

0.7

0.6
dw/dlogM

0.5

0.4

0.3

0.2

0.1

0.0
2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0
logM
Figure 4. Comparison of conventional GPC molecular weight distributions given by DRI and linearized ELSD chromatograms for
SRM 1475a.

The following average molecular weights on six separately prepared injections were obtained
with DRI detector:

 Mn, DRI ¼ 18,040 ± 190 g/mol

 Mw, DRI ¼ 54,630 ± 980 g/mol
 Mz, DRI ¼ 132,100 ± 2,700 g/mol

These values compare favorably with those obtained on the same six analyses with the
ELSD detector:

 Mn, ELSD ¼ 18,410 ± 240 g/mol

6 A. BOBORODEA AND S. O’DONOHUE

0.35
y = 1.007x
R² = 0.995
0.3

measured concentraon (mg/mL)

0.25

0.2

0.15

0.1

0.05

0
0 0.05 0.1 0.15 0.2 0.25 0.3 0.35
concentraon (mg/mL)
Figure 5. Correlation between the measured and the real concentration of PS standards (molecular weights between 4,000 and
6,000,000 g/mol) obtained with the optimized kELSD parameter.

 Mw, ELSD ¼ 55,050 ± 1,230 g/mol

 Mz, ELSD ¼ 131,700 ± 2,300

All measured average molecular weights are in good agreement with the values provided by
SRM 1475a Certificate.
Due to the fact that the peak is normalized to unit area, conventional GPC does not require the
evaluation of the concentration of an injected sample. However, to evaluate the intrinsic viscosity of
each eluted fraction (point of chromatogram) it is necessary to establish the relationship between
the DRI peak area and the injected concentration. This operation is known as “DRI detector cali-
bration” and it is usually implemented in the dedicated software for advanced GPC analysis. It
requires the injection of a PS standard solution with a molecular weight around 100,000 g/mol for
which the dn/dc parameter and the concentration are known. The relatively high molecular weight
is chosen to avoid the variations of dn/dc due to the presence of butyl end chain group.
For ELSD detector, no operation is implemented in the commercial software. Therefore, the
kELSD parameter was evaluated by an optimization method. Figure 5 presents the correlation
between the measured and the real concentration of different PS standards given by the opti-
mized kELSD parameter. We have previously shown[4] that in TCB, this parameter is the same for
both PE and PS samples.
Finally, the viscometer is also calibrated using a PS standard solution with a molecular weight
around 100,000 g/mol with known intrinsic viscosity and concentration.
Once the detectors are calibrated, the MHS plot (log(viscosity) vs log(molecular weight)) for
PS can be constructed based on the DRI, ELSD and viscosity peak areas generated by a series of
narrow PS standards, as shown in Figure 6. The following K and alpha were measured by cou-
pling DRI with viscometer:

 KPS, DRI-viscometer ¼ 14 mL/g

 aPS, DRI-viscometer ¼ 0.685

Similar values were evaluated by coupling ELSD with viscometer:

 KPS, ELSD-viscometer ¼ 13 mL/g

 aPS, ELSD-viscometer ¼ 0.691
 INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION 7

1.0
DRI ELSD

Linear (DRI) Linear (ELSD)

0.5
y = 0.6853x - 3.8502 y = 0.6907x - 3.8767
R² = 0.9997 R² = 0.9990

0.0
LOG([η])

-0.5

-1.0

-1.5
3.0 3.5 4.0 4.5 5.0 5.5 6.0 6.5 7.0
LOG(Mpeak)
Figure 6. Mark–Houwink–Sakurada plot for PS standards when DRI is coupled with viscometer compared with the same plot
given by coupling ELSD and viscometer.

The measured K and alpha values with both detectors for PS standards are in good agreement
with the values provided by ASTM D 6474 for PS.
For SRM 1475a analyses, the DRI, ELSD and viscosity peaks can be divided into narrow
domains. After compensating for the inter-detector delay volumes, each narrow elution domain
allows to calculate an intrinsic viscosity, and thus an accurate molecular weight of PE using the
relationship:
Vh ¼ ½gPE MPE ¼ ½gPS  MPS
where [g]PE and [g]PS are the intrinsic viscosities of PE and PS fractions eluting at the same
moment of time and having the molecular weights MPE and MPS.
Thus, by applying this universal calibration it is possible to evaluate also the MHS parameters
of a broad sample, as shown in Figure 7. The following K and alpha were measured by coupling
DRI with viscometer:

 KPE, DRI-viscometer ¼ 40 ± 2 mL/g

 aPE, DRI-viscometer ¼ 0.717 ± 0.005

Similar values were evaluated by coupling ELSD with viscometer:

 KPE, ELSD-viscometer ¼ 41 ± 2 mL/g

 aPE, ELSD-viscometer ¼ 0.720 ± 0.005

The measured K and alpha values with both detectors for SRM 1475a are in excellent agree-
ment with the values provided by ASTM D 6474 for linear PE.
Figure 8 shows a comparison of the MWD obtained by universal calibration when using as
concentration detector the DRI, or the ELSD.
The following average molecular weights on six separately prepared injections were obtained
with DRI detector coupled with viscometer:

 Mn, DRI, viscometer ¼ 18,630 ± 330 g/mol

 Mw, DRI, viscometer ¼ 59,430 ± 1,610 g/mol
8 A. BOBORODEA AND S. O’DONOHUE

0.6
DRI ELSD

0.4 Linear (DRI) Linear (ELSD)

y = 0.7206x - 3.4014 y = 0.7155x - 3.375
R² = 0.9958 R² = 0.9983
0.2

0.0
LOG([η])

-0.2

-0.4

-0.6

-0.8
4.0 4.2 4.4 4.6 4.8 5.0 5.2 5.4
LOG(Mpeak)
Figure 7. Mark–Houwink–Sakurada plot for SRM 1475a when DRI is coupled with viscometer compared with the plot given by
coupling ELSD and viscometer.

1.0

0.9 RI

0.8 ELSD

0.7

0.6
dw/dlogM

0.5

0.4

0.3

0.2

0.1

0.0
2.5 3.0 3.5 4.0 4.5 5.0 5.5 6.0
logM
Figure 8. Comparison of molecular weight distributions from universal calibrations for SRM 1475a.

 Mz, DRI, viscometer ¼ 149,100 ± 2,400 g/mol

These values are similar to those obtained on the same six analyses with the ELSD detector
coupled with viscometer:

 Mn, ELSD, viscometer ¼ 18,240 ± 220 g/mol

 Mw, ELSD, viscometer ¼ 57,570 ± 640 g/mol
 Mz, ELSD, viscometer ¼ 143,000 ± 1,500 g/mol

All measured average molecular weights by universal calibration method are in good agree-
ment with the values provided by SRM 1475a Certificate.
 INTERNATIONAL JOURNAL OF POLYMER ANALYSIS AND CHARACTERIZATION 9

Conclusions
For the first time we proved that for linear polyethylene samples the universal calibration method
using an evaporative light scattering detector (ELSD) coupled with a viscometer provides similar
molecular weight distributions and similar Mark Houwink Sakurada parameters to those obtained
by the differential refractive index (DRI) detector.
The main advantages of coupling ELSD with viscometer are a much faster equilibration of the
ELSD compared with DRI, especially for high-temperature GPC applications, no baseline drift
and the absence of solvent peak.
The main difficulty of using the universal calibration method with ELSD is the selection of the
adapted capillaries to split the flow between the ELSD and the viscometer. We found that slightly
increasing the flow rate to 1.2 mL reduces spiking in the ELSD and retains good peak resolution.
At the time we conducted this work the linearization of the ELSD was performed off-line.
However, this function is now implemented directly in the hardware, thus simplifying the soft-
ware required for universal calibration method. Work is in progress to test the universal calibra-
tion method with the ELSD and the viscometer, on low-density polyethylene samples to evaluate
the possibility of measuring the long-chain branching. It is our hope that other numerous appli-
cations of ELSD detector,[9,10] in which the linearization of the signal is not critical, could also
benefit from the introduction of this new firmware.

References
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Molecular Weight Averages of Polyolefins by High Temperature Gel Permeation.
[2] O’Donohue, S. J., and F. P. Warner. 1997. ELSD diffuser; US Patent 6151113.
[3] O’Donohue, S. J., N. J. Wrench, and P. E. Claes. 2003. Evaporative light scattering detector; US Patent
7561268.
[4] Boborodea, A., and S. O’Donohue. 2017. Linearization of evaporative light scattering detector signal. Int. J.
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[5] Boborodea, A., and A. Brookes. 2017. Polyolefin characterization in dibutoxymethane by high temperature
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[7] Grubisic, Z., P. Rempp, and H. Benoit. 1967. A universal calibration for gel permeation chromatography.
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