Esso Research and Engineering Co., Brit. Patent 798,542 Shell International Research Maatschappiy N V , Brit.

(July 23, 1958). Patent 973,393 (Oct. 28, 1964).

Hollyday, W. C., Jr., Munsell, M. W. (to Esso Research
and Engineering Co.), U. S. Patent 3,076,791 (Feb. 5,
1963).
Moyer, R. C . (to Pure Oil C o . ) , U. S. Patent 3,143,877
(Aug. 11, 1964).
Rogers, D. T., et al., “Mechanism of Engine Sludge Forma-
tion and Additives Action,” SAE Golden Anniversary
Fuels and Lubricants Meeting, Philadelphia, Pa.,
November 1955.
Rohm & Haas G.m.b.H., Brit. Patent 937,734 (Sept. 25,
1963).
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Engine Deposits,” SAE Golden Anniversary Fuels and
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(July 17, 1962).
RECEIVED for review April 29, 1968
ACCEPTED August 23, 1968
Division of Petrochemistry, 155th Meeting, ACS, San
Francisco, Calif., April 1968. Permission to publish granted
by the British Petroleum Co., Ltd.

GEL PERMEATION CHROMATOGRAPHY

Calibration Curue from Polydisperse Standards

S . T . B A L K E , A . E . H A M I E L E C , A N D B . P . LECLAIR
Chemical Engineering Department, McMaster Uniuersity, Hamilton, Ontario, Canada

S . 1. P E A R C E
Shell Oil,Ltd., Montreal, P. Q., Canada

A search technique and computer program (Fortran IV) have been developed
to determine the calibration curve for a gel permeation chromatograph using
broad molecular weight distribution standards. Two average molecular weights
(number, weight, or viscosity) for one or two polydisperse standards are required.
A Rosenbrock search is used to find the calibration curve constants. This search
has been tested for a variety of cases on an IBM 7040 computer. An evaluation
of these results is presented.

THEuse of the gel permeation chromatograph (GPC)
for the determination of molecular weight distribution
(MWD) of polymers is developing rapidly. T o interpret
the GPC chromatogram it is necessary to establish a
quantitative relationship between molecular weight and
elution volume. The GPC is usually calibrated with narrow
MWD standards ( M , / M , 5 1.1) using the elution volume
a t the peak position. I t is not valid to use the peak
position for broad MWD standards. For many linear
homopolymers narrow MWD standards are not readily
available, thus presenting the problem of determining the
calibration curve with broad MWD standards. Rodriguez
et ai. (1966) suggested a graphical procedure, but with
little elaboration. Cantow et al. (1967) proposed a pro-
cedure for finding the calibration curve given the complete
MWD of one broad MWD standard. They are also able
to correct their calibration curve for the effect of concen-
tration.
More recently, Frank et al. (1968) developed a technique
for finding a calibration curve using a number- and a
weight-average molecular weight of one broad MWD
standard. Their technique involves a graphical approxima-
tion which makes it difficult to employ.
I n this paper a precise computer technique is presented,
tested, and evaluated for several cases. These include
searches for linear calibration curves given the following
data: one broad MWD standard and its number- and
weight-average molecular weights; two broad MWD stand-
ards, given two number-average molecular weights, two
weight-average molecular weights, two viscosity-average
molecular weights, and one number- and one weight-
average molecular weight. Searches for nonlinear calibra-
tions are also discussed.
Development
Linear Calibration Curve. A linear calibration curve may
be expressed as
VOL = C1 - C2 logm(MW)
where
VOL = elution volume in counts
MW = molecular weight of a monodisperse
sample
C l and C2 = constants to be found by search
program
The method involves the injection of one or two broad
MWD standards into the GPC under conditions where
the effect of correction for imperfect resolution on the
molecular weight averages of a polydisperse sample is
negligible. [If skewing and symmetrical axial dispersion
correction may be estimated, the true calibration curve

54 I & E C PRODUCT RESEARCH A N D DEVELOPMENT

 where
Table I. Test Samples
Sample FMAX2 = objective function
No. M" M, M,IMn MW(1) = experimental weight-average
7A 48,000 55,000 1.14
108 197,400 250,000 1.26
molecular weight for sample 1
3A 333,000 430,000 1.29 MN(1) = experimental number-average
17 64,000 125,800 1.95
103 48,000 107,100 2.21 molecular weight for sample 1
VMW (1) = experimental viscosity-average
molecular weight for sample 1
may be obtained even. under extreme conditions (Balke T E M P 1 = calculated number-average
and Hamielec, 1968)]. This usually means residence times
of about 100 minutes or greater, according to Duerksen
molecular weight
and Hamielec (1968). This assumption of negligible correc- T E M P 2 = calculated weight-average
tion for imperfect resolution can be checked by injecting molecular weight
other broad MWD standards to test the validity of the
GPC calibration curve over a range of molecular weights. T E M P 5 = calculated viscosity-average
I n this program valu'es of C1 and C2 are guessed by molecular weight
the search and the calibration curve is used to convert
the GPC chromatogram to a differential MWD, assuming The computer program uses a Rosenbrock search as
infinite resolution. This differential M W D is then used described by Wilde (1964). The search is restricted to
to calculate the appropriate molecular weight averages the desired range of possible C1 and C2 by a "high failure"
(M,,, M,, or M u ) . One of the following objective functions technique.
is minimized. Nonlinear Calibration Curve. A nonlinear calibration
curve might be expressed as

FMAX2 = (MW(1) - T E M P 2 ) 2+ (MN(1) -

TEMP1)' (1)
FMAX2 = (MN(1:I - T E M P l ) ' + (MN(2) -
The search program can be easily modified to search for
TEMP1)' (2a) N constants given N bits of information. This involves
N / 2 to N broad MWD standards. An alternative approach
FMAX2 = (MW(1) - T E M P 2 ) *+ (MW(2) - might involve the search for straight-line segments of
TEMP2)' (2b) the nonlinear calibration using the existing program. No
attempt has been made here to evaluate this procedure.
FMAX2 = (VMW(1) - T E M P 5 ) ' + (VMW(2) - Effective Calibration Curves. An effective calibration
TEMP5)' (212) curve may be obtained where correction for imperfect
resolution is not negligible. This involves using the calibra-
tion curve constants obtained by search on the imperfectly
FMAX2 = (MW(1) - TEMP2)" (MiX(2) - resolved chromatogram of a broad standard only for
TEMP1)' (2d) unknowns with chromatograms similar to that of the

Table II. Input Data for Program Test

Search Parameters Initial
~ Estimate
Run Sample- First Sample Sample Second Sample Iststep, Accel, Deccel, ~

No. No. M, MI' ML No. M" Ma M, CC . A BE CI C2

1 17 64,430 125,800 0.005 2.5 -1 20 15
2 3A 333,000 430,000 0.005 1.5 -1 20 15
3 7A 48,000 3A 333,000 0.005 1.5 -1 50 6
4 7A 48,000 3A 333,000 0.002 2.5 -1 50 6
5 108 197,000 3A 333,000 0.005 1.5 -1 20 15
6 103 107,100 17 125,800 0.005 2.5 -1 80 5
7 103 107,100 17 125,800 0.1 2.5 -1 50 6
8 103 107,100 17 125,800 0.005 2.5 -1 50 6
9 103 107,100 17 125,800 0.005 1.5 -1 50 6
10 103 107,100 17 125,800 0.005 2.5 -1 20 15
11 103 107,100 17 125,800 0.05 2.5 -1 20 15
12 103 107.100 17 125,800 0.05 3.5 -1 20 15
13 7A 53,000" 3A 413,000" 0.005 1.5 -1 20 15
14 3A 333,000 17 125,800 0.005 1.5 -1 50 6
15 3A 333,000 17 125,800 0.005 1.5 -1 20 15
' a of 0.70 in Mark-Houwink-Sakurada relation used.

VOL. 8 NO. 1 M A R C H 1 9 6 9 55
 standard. The effective calibration curve constants
Table 111. Results of Program Test automatically accomplish necessary resolution correction.
output Another possibility is to use a series of broad standards
Cl c2 over the range of elution volumes desired and combine
(actual (actual the linear segments so obtained to obtain a complete
Run value value No. of APP~X effective calibration curve.
No. = 37.55) = 3.62) iterations time Remarks
A nonlinear calibration curve search would provide even
1 37.54 3.62 166 2 min. more flexibility in the above procedures. None of the
28 sec. above are evaluated in this paper.
2 37.56 3.62 409 4 min.
8 sec. Evaluation
3 37.52 3.61 209 3 min.
36 sec. I n this evaluation samples of high and low polydispersity
4 41.34 4.32 195 3 min. Slowb were used (Table I). Maximum computer time permitted
27 sec. per run was approximately 6 minutes. Previous programs
5 37.43 3.60 348 4 min.
0 sec. using a Hooke and Jeeves pattern search had required
6 72.31 9.42 198 4 min. Very slowb 7 to 30 minutes to reach an optimum. Tables I1 and
21 sec. I11 show that, in general, convergence to the correct
7 47.71 5.59 136 4 min. Very slowb optimum (&0.7%) occurred after about 200 iterations
44 sec.
8 49.79 6.00 58 5 min. Very slow* within approximately 3% minutes using an initial step
11 sec. size of 0.005, an acceleration factor of 2.5 or 1.5, and
9 49.79 6.00 219 5 min. Very slow6 a deceleration factor of -1.
1 2 sec. I n the only exceptional case slow convergence occurred
10 42.11 4.50 250 6 min. Very slowb
15 sec.
when two very broad molecular weight samples, each of
11 40.66 4.22 219 4 min. Lax criterion" known but of almost equal weight-average molecular
6 sec. weights, were used. These samples were examined in runs
12 36.74 3.47 260 5 min. Slow6 6 to 13. In none of these instances was the search criterion
6 sec. for compfetion, Y1 - Y2 = 0.1, reached although values
13 37.53 3.62 470 5 min.
30 sec. as low as Y1 - Y2 = 2.0 were obtained a t values far
14 37.35 3.58 234 3 min. from the correct C1 and C2-that is, the criterion was
39 sec. almost satisfied far from the optimum of these runs. I t
15 37.33 3.59 403 5 min. was evident from examination of the calculated molecular
16 sec.
weights that the objective function was not satisfied. This
situation was attributed to the automatic step size reduc-
tion in the search, which usually resulted in very rapid
"Search criterion was 20 rather than 0.1. bSearch criterion of
objective function variation (Y1 - Y2) = 0.1 not reached. convergence. Attempts to remedy this situation by
changing the three search parameters, the search criterion

Table IV. Input Data

Polydisperse PVC sample 1
GPC response
Counts 23.0 23.5 24.0 24.5 25.0 25.5 26.0 26.5 27.0 27.5 28.0 28.5 29.0
Height 0.0 0.6 1.9 4.8 11.9 24.2 43.9 66.1 74.2 111.1 118.8 117.8 108.8
Counts 29.5 30.0 30.5 31.0 31.5 32.0 32.5 33.0 33.5 34.0 34.5 35.0 35.5
Height 90.9 70.8 51.6 38.7 27.2 18.0 11.5 7.9 4.7 2.9 1.6 0.3 0.0
Average molecular weight
M," = 67,800
M.b = 28,200
Search parameters
CC = 0.005
A = 1.5
BE = -1
First guess of C1 = 20
First guess of C2 = 15
Output data
C1 = 48.87
C2 = 4.36
Time = 2 min. 52 sec.
No. of iterations = 305
a Measured by light scattering. Measured by osmometry.

56 I & E C PRODUCT RESEARCH A N D DEVELOPMENT

 Table V. Verification of GPC Calibration Curve
P vc (MJos (MnJcPc[ ( M n ) c P C- (Mn)as] (MY)LS ( M ~ ) [ ( ~M s I~c p c~- ( M ~ L s ]loo
Sample No. (Osmometry) GPC ( M d O S (Light Scattering) (GPCI (Mi,
2 38,000 43,000 +13.2 128,000 115,000 -10.2
3 49,000 50,000 +2.1 148,000 133,000 -10.2
4 31,000 37,000 +19.4 99,000 98,000 -1.0
5 60,000 57,000 -5.0 176,000 141,000 -19.9

for convergence and particularly the initial guesses of C1
and C2, met with considerable success. Run 12, although
slow, was very near convergence when the time limit
expired.
Using the values of C 1 and C2 obtained with the search
technique and PVC sample 1 (Table IV) the GPC results
calculated assuming infinite resolution (Table V) differ in
M , from -5.0 to +19.4’% compared with osmometer values,
and in M, from -19.9 to -1.070, compared to SOFICA
light scattering measurements. Considering the sources
of uncertainty in all three methods of measurement as
well as the fact that the search technique was used only
for sample 1 (no attempt was made to optimize a C1
and C2 for the range of molecular weights of interest),
these results were comidered satisfactory.
Recommendations for Use crf Search Program
Use search parameter values of
CC = 0.005
A = 2.5
BB = -1
Use a t least two different sets of initial guesses of C1
and C2 to test each case.
Use a search criterion of Y1 - Y2 = 0.1 and a computer
time limit of 6 minutes.
Adjust initial guesses of C1 and C2 to speed convergence.
Altering search parameters, or criterion for convergence,
are less effective solutions to the problem.
Conclusions
A program to determine a linear calibration curve from
polymer samples of known weight number- or viscosity-
average molecular we:ights utilizing a Rosenbrock search
technique has been developed and used to determine a
linear calibration curve for a wide number of cases (Tables
I1 and 111).
The program may be very easily modified to search
for parameters of a nonlinear calibration curve. Results
of program evaluation and specific recommendations for
its use are presented. A source program listing is available
on request.
Acknowledgment
The authors appreciate financial support from the Chi-
nook Chemicals Corp., Ltd., and the National Research
Council of Canada. Data on PVC from Imperial Oil Enter-
prises, Sarnia, Ontario, and the original Rosenbrock search
program from John D. Sheel of McMaster University
are also gratefully acknowledged.
literature Cited
Balke, S. T., Hamielec, A. E., Reprints, Sixth International
Seminar, Gel Permeation Chromatography, Miami
Beach, Fla., 1968.
Cantow, M. J. R., Porter, R. S., Johnson, J. F., J .
Polymer Sci. 5 , Part A-1, 1391 (1967).
Duerksen, J. H., Hamielec, A. E., J . Polymer Sci. 6,
Part C, 83 (1968).
Frank, F. C., Ward, I. M., Williams, T., Reprints, Fifth
International Seminar, Gel Permeation Chromatog-
raphy, London, 1968.
Rodriguez, F., Kulakowski, R., Clark, 0. K., IND. ENG.
CHEM.PROD. RES.DEVELOP. 5,121 (1966).
Wilde, D. J., “Optimum Seeking Methods,” p. 151,
Prentice-Hall, Englewood Cliffs, N . J., 1964.
RECEIVED
for review May 20, 1968
ACCEPTED November 4, 1968

VOL. 8 NO. 1 M A R C H 1969 57