Topics in.. .

Chemical Instrumentation
Edited by GALEN W. EWING, Seton Hall University, So. Orange, N. J. 07079
I
These articles are intended to serve the readers o f ~ ~JOURNAL
l s
by calling attention lo new developments in the theory, d e d n , or availability of
chemical laborato~yinstrumentation, or by presenting useful insights and ez-
planations of topics that are o j practical importance lo those who use, or teach
the use of, modem instrumentation and instrumental techniques. The editor
invites correspondence from prospective contributors.

LII. Current Trends in Gel Permeation

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Chromatography.
Part One: Theory and Equipment
JACK CAZES, Mobil Research and Development Corp.,
Poulsboro, N . J. 0 8 0 6 6
I. Introduction
This report is presented as asupplement
t o an earlier paper entitled, "Gel P m e a -
Lion Chromatography," which was pub-
lished in 1966 as part of the "Topics in
Chemical Instmmntation" series ( 1 ) . The
past four years have seen many changes in
the nature of gel permeation chromatog-
raphy (GPC). Whereas it was slmost
exclusively a polymer fractionation
method, i t has now taken its place as a
generally applicable separation technique
along with other, more commonly known
liquid-phase chromstographio techniques,
e.g., liquid-solid adsorption, liquid-liquid
partition, ion exchange, etc. I n fact,
GPC is, for d l practical purposes, a special
type of liquid-liquid partitioning in which
the stationary phase is the solvent con-
tained within the gel pores and the moving
phase is the solvent outside of the column
substrate particles. The development of
new, more universally applicable column
substrates that are capable of "sorting"
molecules on the basis of molecular size
and shape have extended the scope of GPC
t o verv, hieh . sndverv low molecular weight
sulwtiurw, t o use with n uiae vkrirty of
s~,lvcr.t.~, horh aqutow 8l.d wynnir, and l o
i n h - t r n l - i w l e wl,.tralions xnd pwlira-
tions.
There is no universally accepted name
for t,he technique described in this paper.
The name "gel permeation," which was
coined by John C. Moore of the Dow
Chemical Co., has persisted in the polymer
literature, but is no longer wholly suitable,
since there Itre now substrates that are
capable of separating molecules on the
basis of size, which are not gels, e.g.! po-
rous glass, silica, etc. "Gel filtratlon,"
long used by the biochemists, is not com-
pletely acceptable either, because the pro-
cess is not really one involving filtration in
its usual sense. Other expressions that
have been used include "molecular sieve
chromatography," "restricted diffusion
chromatography,'' and others. All of
these terms are the result of attempts t o
choose a name based either on the sus-
pected meohanism of the separation or on
the type of column substrate being used.
"Gel permeation" has been retained in the
present report primarily t o avoid confu-
sion. I t appears in much of the literature
cited, and, for that matter, almost all of
the non-biomedical literature dealing with
this subject.
Several excellent reviews (2-4) have
been published on this versatile technique,
including a book entitled "Gel Chromatog-
raphy" (6). A short two-day course en-
titled "Fundamentals of Gel Permeation
Chromatogmphy," is offered several times
a year a t various locations in the United
States under the sponsorship of the Ameri-
can Chemical Society. The course is
aimed a t the individual who has had little
or no exposure to GPC and who is inter-
ested in actually performing size separa-
tions.
A series of International Seminars on
GPC have been and are continuing to be
held a t least once a year under the sponsor-
ship of Waters Associates, Inc., Framing-
ham, Mass. Those that have already
been held or have been scheduled are
summarized here:
1st-Jan., 196&Cleveland
2 n d S e p t . 1617,196.5-Boston
3rd-May 19-20,1966--Geneva
4th-May 22-24,1967-Miami
5th-Summer, 1968-London
6 t h - 4 e t . 7-9,196&Miami
7 t h - 4 c t . 12-15,1969--Monaco
8th-July 1-3,1970-Prague
9 t h 4 c t . 5-8,1970--Miami
Jack Cares currently holds the position
~f Supervising Chemist in charge of the
iepnrations llesenrch Group at Mobil
tesenrch and Development Corp.. Pnuls-
,om. N. J. He had previously spent
even years with Mobil Chemical Co..
vhere he was involved in many areas of
esenreh, including organic chemical and
dymer process research, design and np-
,lieation of andytionl instrumentation.
:el permention ehromatogmphy, and
omputeriaed data proees~ing. Other
nterests have inoluded organic synthesis.
cgnnio free-mdienl reaction mechanisms.
.nd electronic circuitry for chemical in-
trumentntion.
Dr. Cnees reoeived a B.S. in Chemistry
1955) at the City College of New York,
.nd both MS. and Ph.D. degrees in
srganio chemistry at New York Univer-
itu.
Dr. Cams hns taught at Queens College
New York) and a t l t u t g e r ~ t h eState
lniversity (New Brunswlck). He eur-
ently teaches :L Short Course on Gel
'ermeation Chromatography for the
Lmerienn Chema:al Society.
Preprints of the papers presented were
published by Waters and some of the more
recent ones are still available from them.
They have also prepared an extensive
bibliography on GPC containing refer-
ences t o several hundred papers.
The ASTM D-20 committee is currently
involved in formulating a standardized
terminology for GPC as well as an accept-
able recommended practice. A "round
robin" experiment will certainly follow
acceptance of a suitable procedure.
Liquid chromatography and, conse-
quently, GPC are currently experiencing a
renaissance similar t o that which gas chro-
matography underwent in the late 1950's.
The development of suitable instrumen-
tation and a desperate need for a iraction-
ation technique applicable to non-volatile,
(Continued a page A4891

Volume 47, Number 7, July 1970 / A461

 Chemical Instrumentation
thermally labile substances have been
largely responsible for this rebirth.
11. Mechanism of the Separation
An excellent review of the theory and
mechanics of GPC separations was pub-
lished by Altgelt (6). A t least lhree
mechanisms have been proposed t o de-
scribe the GPC separation process, and all
of them probably operate simultaneously
with one or more dominating under a
given set of operating parameters. GPC
is a. liquid-liquid partition chromata-
graphic process in which solute molecules
distribute themselver between two liquid
phases-the liquid contained in the gel
pores and the liquid outside of the gel, i.e.,
in t,he interstit,ial volume.
According t o the steric aclusion meeha-
nism i t is assumed that
0 different fractions of the total pore
volume are accessible t o different sise
pores into which they can enter, whereas
small molecules can enter a relatively
large number of pores.
0 there is diffusional equilibrium; the
time required for solute molecules t o enter
into and emerge from gel particles is much
smaller than the residence time of the
solute aone.
The process would, therefore, be expec-
ted t o he flow-rate insunsit,ive (not diffu-
sion controlled) over a relatively wide
range of linear velocities, and this is indeed
what is most often observed experimen-
tally. A distribution coefficient is thus
defined:
....
Kd = Vi. /Vi
and the retention volume of a given species
becomes
V n = V. + KdVi
where
Vi. ....= accessible porevolume
Vi = total pore volume
V , = interstitidvolume
The cffects of steric exclusion should be
most evidenl where s. major portion of the
solute molecules is larger than many of the
gel pores.
In the rml7iclcd diffusion ?ncehanism the
process is assumed Lo be, to a significant
degree, 8, dinusion-controlled one, i.e.,
there is no diffusional equilibrium. Ob-
served retention volumes should then be
affected by changes in flow-rate; the
shapes of broad chromatographic bands
observed for polydispe~.scmixbulw should
also be flow-ralc depende~~t.The absence
of difiusiord equilibrium should he most
pronounced s t very high linear velocities,
and it is under such condiliona that the
restricted dilfusion mechanism would be
the dominant one.
Several t h c m c l p a m i c lheories have
been advanced for the separation. Mars-
den has distinguished between the en-
thnlpy and entropy contributions t o Kc,
DEGASSING RESERVOIR
FRACT IONA-
COLUMN
I I DETECTOR
RECYCLE
SYSTEM
---- ELECrRKAL I FRACTION
CONNEC~/ONS 1 COLLECTOR
Figure 1. Liquid chromatography system.
(7). deVries relates Vn t o the molar
volume of the salute and the pore sise
distribution of the gel (8). Casassa (9)
proposes a theory which accounts far the
separation on the basis of the existence of
different chain conformations within either
spherical, eylindriesl, or deb-shaped pores.
He concludes that distribut,ion of solute
between the two phases is governed en-
tirely by the loss of conformational en-
tropy upon entering t,he pares. Car-
michael (10) relates the behavior of
random-coil molecules in uniform pores t o
the probability that t,he root-mean-square
end-to-end distance of the molecules will be
less than the average pore radius.
A secondary czclusion effect has recently
been ~ r o ~ o s e( 1d1 ) to account for the ex-
ceptidnaily good separalions observed
'
with columns operated under apparently
overloaded conditions, i.e., with as much
as ten times the normally used sample
loading. Briefly, if a mixture of large and
small molecules is placed onto R. gel, the
smnll molecules will diffuse most rapidly
into the gel pares. The larger molecules
will, eansoquent,ly, find relatively few un-
occupied pores and the probability of their
dilfusing into an occupied pare will be
reduced. The larger molecules will most
probably move further down the column
until they find unoecupiod pares. The
net result is enhancement of the separa-
tion.
Ill. Resolution
Classiedly, resolution of two chromato-
graphicponkfi has been defined as
where
Rw = n o . of peak widthsresolution;
Vnl and Vn, = retention volumes, at the
peak maxima, for the two species; WI
and W 1 = extrapolated base widths for
the two species.
VALVE-LOOP
SAMPLE AUTOMATIC
SAMPLING
SYSTEM
k-------------4 I
COMPUTER I
.
-. !
I
I
I-----
r
Baseline resolution is observed when Rw
= 15. This equation, however, does not
tell us anything about the factors that
affect resolution. An equation can be de-
rived
in which the first factor represents seleetiv-
ity, t,he second, capacity, and the third,
random dispersion.
The srleelivily term is concerned with the
distribution eoeffieient,s of the two solute
species and with how different they are.
The greater t,he differenceb e h e e n K, and
K2, the greater will be t,he difference be-
tween VR, and VE,. Thus, resolution of
a given solute pair could be increased by
choosing a gel such t,hat the difforenee
bet,weenK, and R2is increased.
The capacity term is related t o the
capacit,y ratio, K', which includes terms
involving both dislribution coefficient and
amount of stationery phase in tho column.
The magnitude of K' actually determines
how long a fiolute will remain on the col-
umn. Small values of K ' result in small
retention volumes.
The random dispersion term is a fune-
tion of the number of theoretical plates,
N, in the column. Separation, per se, is
not achieved by a large N; a hlgh plate
count will only assure a minimum of
sprendiug of an olhereise narrow, well
shaped chromai.ographic band. llandom
dispersion includes the effects of flow in-
equalities within the column due t o in-
homogeneities in the packing density in
the column, nonequilihrium of the iolute
between the two phases, and longitudinal
diffusion, primarily in the moving phase.
IV. Equipment
Since GPC is a specific l,ype of liquid
chromatography, much of the equipment
t h a t is customarily used for the more
classical liquid-phase column chromato-
graphio techniques can be employed for
this fractionation method. I t is of value
here t o outline briefly some of the various
(Conlinued on page A4fi4)
 Toble 1. Some Manufacturers of Liquid Chromatogrophic Equipment
Chemical Instrumentotion Com- Fraction
plete Col- Deteo- collec-
kinds of apparatus that are applicable, but Manufacturer svstem Pumm umns tors tors Valves
no attempt will be made to provide a com-
plete listing of all available paraphernalia;
first we shall look at component types that
can be used in "do-it-yourself" approaches
and then we'll see what is available com-
mercially in the way of complete liquid
chromatographs.
Figure 1is a flow-sheet that outlines the
various components that can be included
in a. complete liquid chromatographic sys-
tem. The only part that is absolutely
essential, of course, is the fractionation
column, for it is in the column that the
actual separation of a. mixture into its
components takes place; furthermore, it
is merely the nature of the column suh-
strate that classifies a chromatograph as a
GPC instrument, i.e., when a substrate is
used that separates molecules on the basis
of molecular size one refers to the instru-
ment as s. gel permeation chromatograph.
Otherwise, it is simply a liquid chromato-
graph. All of the components, with the
exception of the column, are chosen either
t o improve reproducibility and/or reliabil-
ity or to render operation more convenient.
GPC can be performed, in its simplest
form, with an open glass chromatographic
column packed with a suitable aubstrat,e.
Solvent is generally supplied from a reser-
voir at the top of the column via gravity
feed. Fraction collection can involve no
more than periodically changing a con-
tainer situated under the bottom of the
column with the detection method being
either visuitl observation (in the case of
colored substances), or removal of solvent
from the collected fractions followed by oh-
servation and/or weighing of the sample
residue contained in each vessel. At the
other extreme one might include in his
instrument such "trimmings" as auto-
matic sample injection and fraction collec-
tion, multiple liquid chromatographic
detectors, provision for recycling all or
part of the sample through the same
column for improved resolution, and a
digital computer for automatic control of
various functions and operating param-
eters (temperature, pressure, flow-rate,
cycle control, etc.) as well as automatic
data processing. At least one manu-
facturer (Problematits, Inc., Waltham,
Mass.) bas described such an integrated
system designed around a. small dedicated
computer. Of course, increased com-
~ l e x i t vgenerally results in increased cost,
often beyond the limits that can be justi-
fied on the basis of the utility of the tech-
nique itself (better known as "payout" in
the jmgonaf the accountant).
Table 1 lists manufacturers of liquid
chromatography components. (2) Syringe pumps are available in hack to the large solvent reservoir in the
volume capacities from a few milliliters to Pump.
A. Pumps several liters (without refilling) for opera- (3) Reciprocating piston pumps, avail-
Three basic pump designs probably tion up to several thousand pounds pres- able for operation up to several thousand
account for most of the commercially sure. They are pulseless and therefore pounds pressure can be readily used for
avdable liquid metering pumps: ean be used with flow- and pressurebensi- conventiond as well as recycling chrome
(1) Peristallic pumps are useful, pri- tive detectors without pulsation damping tography.
marily, for low pressure applicetions for systems. Their chief drawbacks are the 0. Columns
flow-rates at or near those at grwity-feed need for periodic refilling and their large Many column designs are available, of
conditions. They are not positive dis- internd volume; in cases where recycle both metal and glass, of fixed length and
placement systems and feed rate for a operation is desired, syringe pumps cannot adjustable length. Variable length col-
given motor speed generally is not repro- be used, became fractionated sample com-
duciblefrom day to day. ponents would be remixed when directed (Continued a page A466)

A464 / Journal o f Chemicol Education


Chemical instrumentation
umns often include a plunger that passes
through the end-fitting of the column,
which can be moved in or out t o obtain a
given packing length inside the column.
A "column kit" is available from Lahora-
tory Data. Control, Inc., which includes
small-volume end-fittings together with
precision bore tubing for assembling a
variety of columns. Packed columns can
he purchased from some of the manufae-
turers of complete instmments.
Extremely high resolution is reportedly
attainable through the use of very narrow
columns (1-2 mm diam) packed with very
fine materials (200 mesh and smaller).
The high pressures often required t o
achieve practical flow-rates with narrow
columns of course makes instrument design
considerations more stringent (high pres-
sure pumps, low volume "plumbing" and
detectors, safety, etc.) and this must he
kept in mind when considering their use.
C. Detectors
Many kinds of liquid chromatographic
detectors have been used and/or are avail-
able commercially. These are based on a
variety of physical properties of molecules
in 8dution. Some rely a n the presence of
a specific chemical functionality. Others
do not, and are more universal in their
scope of applicability. Often, multiple
detectors are used, one, non-specific (e.g.,
a differential refractometer) t o indicate the
elution of all sample components, another,
a specific one (e.g., a U.V. photometer), t o
det,eet the presence of a specific chemical
type in the eluate. With suitably selected
dual detector systems one can, thus, ob-
tain information concerning bath the
molecular weight distribution and the re-
lationship between MWD and chemical
ential refractometer and an ultraviolet
spectrophotometer, to monitor the GPC
fractionation of styrene-hutadiene copoly-
mers; the former responds t o all polymer
components, regardless of chemical type,
while the latter is sensitive only t o the
aromatic styrene blocks in the copolymer.
A description of some liquid chromato-
graphic detectors follows:
(1) Differentiel rejraetomete7-extremely
sensitive, with some commercial units
capable of detecting a difference in refrac-
tive index of 1 0 ' or even 10-8 R.I. units.
Detectors based on refractive index are
almost universally applicable, since they
do not rely on the presence of specific
functional groups. Also, since many poly-
meric systems exhibit a constant refractive
index over a wide molecular weight range,
i t is often possible t o relate detector re-
sponse t o quantity of sample eluting from
the chramatogra,phic column. I t is far
this reason that the differentialrefreetom-
eter has found wide acceptranee in GPC
frclctionations where molecular weight
averages and distributions are t o he com-
puted.
These detectors are extremely flow- and
temperaturesensitive (many organic sol-
vents exhibit a, temperature coefficient of
R.I. unitsIoC).
A466 / Journal o f Chemical Education
(2) Ultraviolet and visible photometers are tial t o a static electrode pair; a supporting
available as both single and double beam electrolyte would be inbroduced into the
systems. Use of these detectors is limited flowing liquid stream just ahead of the
t o those applications where the solutes of deketor. A recording of both half-wave
interest absorb radiation of the wavelength potential and diffusion current would pro-
employed and the solvent does not. For vide both qualitative and quantitative in-
specific solutes that have high extinction formation concerning the species present.
coefficients, U.V. and visible photometers (8) Grauimetric detectors involving direct
offer extreme sensitivity with good tem- automatic continuous weighing of the
perature and flow stability. chromatographic effluent (to yield den-
(3) Flame ionization-column effluent is sity), or automatic evaporation of solvent
allowed to fall on and wet a moving and weighing of the resultant residue from
met~llicwire, chain, or band which then discrete micro-fractions would seem t o
er ters an oven maintained a t a temper* offer a most valuable detector for liquid
t. re h'qh emugh t o completely evaporate chromatography. I t might well represent
solvent f r a n the moving element, but not as important an advance in liquid chrom*
disturb the residual sample. This resid- tography as the katharometer (thermal
ual material is then either passed directly conductivity detector) did in gas chrorna-
through a flame ionization detector where tography. Both approaches mentioned
i t is pyrolyzed snd ionized or it is first above have been shown t o he feasible in
pyrolyaed in a high temperature furnace studies based on the use of an Electrobal-
and then the pyrolyzate is "blown" into ance as the sensor, and reported by Rad-
the flame ionization detector. I n both awski and Williams, but i t does not appear
ceses, the flame ionization cell is similar t o that a commercial instrument will he
that which is used with gas chromato- forthcoming in the foreseeable future (16).
graphic instruments and the measured "Necessity is the mother of inventionn-
ionization current shoukl be related t o the as the need dictates, there is no doubt that
amount of sample on the moving element specific detection systems will be forth-
and the relative proportion of carbon con- coming that are based on other molecular
. ~.
tained in the sample molecules. I n actual ~1onerties .
such as oolarizahilitv idielect.rir
. ~ ~
practice, secondary ionization effects com- co!.iluul , pie,oelrctriv eITe(.t, viwhily,
plicate the situation, and the observed Krlviu "elertric pvl" effwi ( ~ ~ ~ a . - u r e n ~ e r ~ t
signal t o noise ratio leaves much t o he de- o f deflecrmn of ihnrgnl liquid Jroplet:. iu a
sired. Consequently, the currently avail- field), and.. ?? .
able flame ionization detectors have not
gained wide acceptance by liquid chroma- D. Complete Systems
tographers. Also, these detectors are
limited t o those applications where solvent Commercially available instruments for
e m be removed from the moving element performing liquid-phase molecular size
without removing any of the solute (sam- separations can be classified, for our pur-
ple components). poses as
(4) Kcat of adsorption detectors gener-
1. gel permeation chromatographs
ally consist of s. "micro-column" contain- 2. general-purpose liquid chromato-
ing a small amount of an adsorbent in graphs
which a temperature sensing element (e.g.,
thermistor) is imbedded. The heat of ad- Although sepuralionr Imicd on molerulhr
sorption and desorption is detected and s i x difirrence> ( n u 1w performed will, d l
measured as a change in temperature of liquid chromatographs equipped with a
the adsorbent column. Thin detector is suitable column substrate, some are better
often very sensitive and useful for qualit* suited from the standpoint of convenience
tive detection; i t is not well suited for and separation efficiency attainable.
quantitative work, requiring frequent celi- Generally, maximum efficiency can be
hration. realized with systems containing a mini-
mum of dead volume with narrow bore
(5) Electrical conductivity detectors gen-
erally consist of a pair of metallic elec- columns containing finely divided packing
(substrate) with a narrow particle size di8-
trodes cmtained in a microcell together
with a Wheetstane bridge measuring tribution. Some of the newer instruments
circuit. Applications are limited t o those have been designed with this capability.
systems that conduct electricity. Future Some are modular, others are not; some
developments in the direction of high- have several detectors, automatic injec-
frequency eleotrodeless conductivity tion, fraction collection, high pressure
bridges might extend their usefulness t o pumping systems, etc. as options. Table
poor conductors. 2 i s s s u r n m a q of some of the commercially
available liquid chromatographs. The
(6) Infrared detection has, thus far, specialized bio-medical analyzers have
been limited t o homemade modifications been omitted primarily because they would
of conventional recording infrared spectro- have t o be extensively modified t o make
photometers (14, 16). Applications are them conveniently applicable t o GPC
limited t o samples containing functional separations.
groups that exhibit strong absorption
bands within the wavelength range of the
instrument. Sensitivity is relatively poor V. Column Substrates
except in cases where an extremely strong
absorption hand can be monitored (e.g., Many new column substrates have he-
come commercially available since the
carbonyl, C-H, etc.)
earlier report (1). They can be classified
(7) Polarographic detectors are not yet as being rigid, semi-rigid, or soft. The
commercially wailable. A practical de- rigid substrates, char.racterized by their
sign, however, might involve the applica- fixed, rather uniform, pore volume, high
tion of a fixed or rapidly varying (in the
case of oscillographic instruments) poten- (Continued on page A468)
?
01
Table 2. Commercial Liquid Chromatographic Systems
0
, Max.
col-
Diei-
ti&
2 Basic GPC Auto. . Detectors .
Auto Largest Max. u r n Sample &/or
z
%
Manufacturer Model
prm
(8)
eol-
umns
mn- Refr.
ject. index
Flame
U.V. ionis'n.
fract.
Other collect.
diam.
column
Pump
type
p r ~ s . temp. re- Re- eom-
(pslg) ("C) cycle carder pttter Remarks

0,
n
3'
m
s.
5 A . Gel Permeation Chromatog~aphs
Problematics, Inc. 1000 -20,000 X* X X X N N N 5 mm i.d. S 10,000 200 N X A On-line still for
-%. 223 Crescent St.,
Waltham, Mass. 02154
solvent recovery

Waters Associates, Inc. GPC-200 16,500 X A X N N N A 1" 0.d. R 300 150 N X N

61 Fountain St. GPC/ALC-301 9,900 X N X A N N A 1'o.d. R 1,000 R.T. X A N See Noteb
Frarningham, Mass. 01701 GPC/ALCdOl 4,980 X N X N N N N a/8" a d . R 1,000 R.T. N A N See Notee
GPC/ALC-100 8,400 X N A A N N X ZL/,' a d . R 1,000 R.T. A A N
GPC/ALC-101 12,825 X N X A N N X 21/~X~.d. R 1,000 R.T. X X N SeeNote
Ana-Prep 29,500 X X X N N N X 2'/2 0.d. R 250 150 N X N See Noted
(X) (A) (XI (N) (N) (N) (A) (1"o.d.) (R) (250) (150) (A) (XI (N)

B. General Purpose Lipllid Chromatographs

1)uPont Instrument TJiv. 620 16,500 A N X(A) X(A) N N N 8 mm i.d. S 3,000 200 A X N See Notes*,'
Wilmil~grou,Del. 19\98
Nester-Faust, Ine. 1200-H 7,025 A N A A A A A 25 mm i.d. S 2,500 R.T. N A A See Notes'.'
2401 Ogletown Rd. 1240 4,600 A N A X A A N 8mmi.d. R 1,000 125 N A A
Newark, Del. 19711
Varian Aerograpb 4000-1 1,700A N A X N A N 0.d. G 750 R.T. N A A See Note'
2700 Mitchell Dr. 4000-2 2,600 A N X A N A N / 0.. G 750 R.T. N A A See Not@
Walnut Creek, Calif. 94598 400C-O 2,000A N A A N X N o.d. G 750 R.T. N A A See Notes6.G
4120-5 12,550 A N X X N A N a/8" 0.d. S 5,000 90 N A A Micro beat of sd-
sorpt. detector
available
Waters Associates, Inc. ALC-100 7,850 A N X A N N A 1" 0.d. R 1,000 R.T. X A N See Noteb
61 Fountain St. ALC-201 3,900A N X N N N N a/8" o.d. R 1,000 R.T. N A N See Notec
Framingham, Mass. 01701

,-.
a X = included in basic price; A = avdable as option at additional cost; N = notavdsble; R = reciprocating pump; S = syringe pump; G = gas pressurized solvent reservoir; R T =
j! room temperature (ambient).
2' An accessory will be wailable late in 1970 to convert to ooeration UD to 3000 mie and 200'C.
An accessory will be available late in 1970 t o convert to operation ub to 3000 bsii.
*The Ana-prep instrument incorporates both an analytical and a preparative instrument in a single cabinet. Both sections can operate simultaneously, but only one can be monitored a t any
given time with the refrectometer, via a refractameter selector velve. Deta in parentheses refer t o the analytical section of the instrument.
' Recycle accessory will be available late in 1970.
$ J The DuPont 820 is available with either the refraotametric or U,V. detector for the basic mice shown.

; 'Art awrssuty oven for operation up to L O O T is available as a n option.

'
-
a
(:oIumt.s rre jacketed; n tmqwraturr-vorrrrolld circulatmg bath may he uued for operation at higher ramp
' TIM mstr!rrue!rt inrludes a r~ricrohenr-of-ndsorprion detector.
 Table 3. Commercially Available Column Substrates
Pore Moleculhr weight
Designation diameter. 1 exoluaion limit Distributor

A. Porous glass
Bio-Glas 200
Bio-Glas 500
Rio-Gisr 1000
Bio-Glas 1500
Bio-Glas 2500
CPG-10-75
CPG-10-125
CPG-10-175
CPG-10-240
CPG-10-370
CPG-10-700
CPG-10-1250
cPG-10-*000
B. Porous Szizco
Porasil-60
Porasil-250
Porasil-400
Poraail-1000
Porasil-1500
Poreail-2000
Merek-0-gel Si-150
Merok-o-gel SI-500
Merok-o-gel Si-1000
I Semi-Rigid Gala
A. Polystyrene Gels
Styragel 39720 ... 00 A'
Stvrad39721

~<yragel39731
Bio-Beads S-X8
Aquapak A-440
B. Polyu%nylocclotcQel3
Merok-o-gel-OR-750
Merok-0-gel-OR-1500
Merek-o-gel-OR-5000
Merok-0-gel-OR-20.000
Merok-o- el-OR-100,000
Merek-0-gel-OR-l.000.000

A. Dcztran Qels
Sephadsx G-10
Sephadex G-15
Sephadex 6 2 5
Sephsdex G-50
Sephsdex G-75
Sephhdex G I 0 0
Sephsdex G-150
Sephader G-200
Sephadeli L H 2 0
B. ~ ~ i asla ~ ~ ~ ~ ~ l ~ ~ ~ a ~
Bia-Gel P-2
n'io-Gel P-4
11io-Gel P-0
Bio-Gel P-10
Bio-Gel P-30
Bio-Gel P-60
Bio-Gel P-100
Tho-Gel P-150
Hio-Gel P-200
Ilio-Gel P-300
C . Agarose Gels
Ria-Gel A-0.5
nio-Gel A-1.5
Ilio-Gel A-5
Bio-Gel A-15
Ilio-Gel A-50
Hia-Gel A-150
Sepharose B
S e ~ h h r o s e2B
Sag (Ago-Gell-10
Saz (Ipo-Gel)-8
Sag (Ago-Gell-0
Sag (Ago-Gell-4
Sag (Am-Gel)-%
D. Polyslyrene Gels
no-Reads S-XI
Bio-Beads S-X2
Hio-Reads 5-X3
nio-Beads S-X4
'Determined with soluble dertrans.
6 Determined with polystyrene.
~ ~ t ~ ~chain
d e lengths
d , of polystyrene. ~ u l t i p l yby 41 t o convert these values t o approximate
moleoulsr w a g h t of polystyrene.
Determined with po1y?thylene glycols (+qusoual.
' Determined with peptldes and/or protems (aweous).
J Exclusion limit depends upon solvent mnployed a n d resultant degree of swelling. Some solvents
t h a t can be used are ~ a t e r methanol,
, ethanol, ohlorofarm, r h u t a n ~ l dmrsne.
, THF. D M F , acetone.
ethyl aoathte, a n d toluene.,
(1) Rio-Rad Laboratorms, Richmond. Calif. 84802.
(2) Cornin& Glass Works. Cornlog. N. Y. 14830.
(3) Whtera Associates, 61 Fountain St., Framlngham. Mass. 01701.
(4) Merek AG, Dhrmstad? \Vest Germany.
(5) Pharmaoia Fine chemlbala Inc. Uppaala, Sweden: also Pisoataway, N. J. 08854.
(6) Seravho Lahoratoriea ( ~ t y ~ j t d Holyport
: Mmden Head, Berkahire, England.
(7) Mhnn Reaearoh ~ ~ l , ~ IOC:, ~ ~ New ~ i N.&Y..10000.
t ~York,

A470 / Journal of Chemical Education

 egarose, and polyacrylamide gels. K. H.. A N D h l o o i l ~J.
2. ALTOELT. , C . in "Polu-
mor Fioctionolion," M. J. R. c a n t o r , e l l .
Academic Press, N e w Tork, 1967; pp
fractionation depends primarily upon the 123-180.
restrictions imposed by the solute/solvent , F.. P o n ~ ~R.
3. J o a r s a ~J. n ,S . , AXD C w ~ o r .
column permeability, and ability t o be
"wet" water and organic "Ivents
include porous silica and porous glass.
These substances are not affected by most
packillg must
be wettable in the syst,em of interest and
should not be degraded by it; if
temperature is indicated
4. ,J,,,
(1966).

" A
J. F.,
~ ~ GSL
...
M. J. R., Reu. 111ociomo1ec. Chem.. 1, 393
porn,,, R, s,, .,I,,,
I ~ e ~~m e~o t i o~ nc h~r o m
rapt,y." J . ~~l~~~~s c i , C Z I , ( 1 9 ~ 1 .
I o~o~.
at an 5. D E T E R M ~ NH..
. 'V*l C h r ~ m a m ' ~ ~ h l i ~ "
solvents and, therefore, permit separations (to dissolve the sample, or to improve Springer-Verlag, N e w York. 1968.
to be achieved in that degrade molecular diffusion) then the column sub- 6. ALTO.LT, K. H . , in ,.arioonces i n chromdoe-
many organic substrates. strate must be stable a t that temperature. ropi,ll," ~ i d d i n q sJ, . c..and Keller. It. \ . .
Semi-rigid substrates also exhibit high The soft gels, because of their inherent eds., Mlareel Dekker. N e w Y o r k , 1968; V o l .
cO1umll permeahiIities, permitting Opera 7, pp. 3-16.
compressibility, cannot be used a t high 7, ht ,, ,,. ,,N. v . n., an-..v. Y. SC~.,
tion at hhih flow-1.ates, are mostly corn- pressures, and thus limited to opera- 125, 128 (1965).
patihle with organic solvents (although a t tion at relatively low flow-rates, ~h~ 8. J., at .I., Anal. Chrm., 39, 936
D E V ~ I E S ,A.
least one has been modified so that it is (1967): D E Y ~ E SA. . J.. et oi., .I. Polymai
Corning porous glasses, because of their Sci., CZ1. 119 (1968).
wettable aqueous solvent 'ysterns), and relatively narrow pore-size distributions, 9, cnrAss~, E. I?., J . ~ o l v m e rSei., B5, 773
are available in a broad spectrum of per- exhibit compsratively sharp cut-oKs a t (1967).
meabilities for high resolution applications their exclusion limits, often a desirable 10. C A R ~ C H A E1. L U.,
, J . Polymer Sci., AZ, 517
involving both monomers and high p"ly- (1968).
property for achieving specific separations. 11, I(, H., preprints, D ~ Vor. petroleum
mem. Included in this class are many of illany other have been Cbemistry, Am. C b e m . Soe., 15 (No. 2).
the commercially available cross-linked ,,fiered by but the fact 8.115 (Felr.. 1970); a paper presented at
polystyrenes (Styragel, POrqel,Aqua~ak)t tile S y m ~ o s i u m on Gel permeation
of life remains that, after compatibility C h r o m a t o g r a ~ l l y n, o u s t o n , Ter., I'eb. 22-
some ion exchange resins, and a considerations have been taken care of, the 27, 1970.
developed polyvin~lacetategel marketed choice 01 the "best type for the 12. cnaro-, H. J., et al.. K d . Gummi Kunald..
under the name, "Merck-0-gel-OR." job" is still made on the basis of trial and 13. R U21,
NYO 609 R., el
(1908).
N , J. J . A p p ~Potymrr
. SC;.,
This last substrate complements the poly- 13, 2359 (1969).
styrene gels since it can be used with many 'rable 3 lists many of the commercially 14. R ~ F.. ct a l . , ~I n d . ~ n g Cham..
~. Prod.~ ~
that be used with the available column substrates that can be I ? ~ s Iht,el,,
. 5, 121 (1960).
polystyrene gels, e.g., lower alcohols. J, H,, C*aTo, sf.E,,J , PolVme,
used for molecular size separations to- SC;., c z l , 143 (1968).
Soft gels are primarily applieable to gether with some of their reported proper- 16. R . , D * w ~ . ~ .J. L.. A X D W ~ L L I I M S , C . J.. , , A
aqueous systems and exhibit low column ties, ~ ~ ~~ ~ ~f o i ~ i rt~~ u ichroma-
d~~ t ~~ it
permeabilities (useful only a t low flow- to~raphy.''paper presented at the Pitts-
rates). Their permeability ranges are burg Conference on i n a l y t i e s i Chemistry
and Applied Speotroseopy. Cleveland.
variable and depend upon the extent of March, 1970.
"solvent swelling" when immersed in a.
given solvent. Included in this group are References "LII. Current Trends i n Gel Permeation
crosslinked dextrans (Sephadex), starch, c,,,,, J., J. A567, A625 Chrolnatogravhy-Part Two--Jfe(hodo(oguU
rubber, lightly crosslinked polystyrenes, (1966). wdl appear in Lhe August issue.

Volume 47, Number 7, July 1970 / A471