The Lippmann Equation and the Characterization

of Block Lipid Films

An optically black lipid film in aqueous media approximates closely to an ideally polarizable
system and, in this respect, can be described by a form of the Lippmann equation. This
equation is derived and integrated assuming the film capacitance to be independent of applied
potential. Measurements of the contact angle and of the bulk interfacial tension are used to
find accurate values of the film tension and it is shown that, for a range of lipid films, the
integrated equation is obeyed for potential differences up to approximately 100 mV.
Various ways are then described in which the Lippmann equation may be used to find
film properties not readily accessible by other methods. Thus, for phospholipid films the
accurate measurement of both film and bulk interface tensions presents difficulties, but these
can be overcome if the specific capaci tance is known. In this way the specific free energy of
thinning of the film may be obtained. Conversely, for films in media of low conductivity or in
systems in which film area measurements are difficult the determination of the specific
capacitance is not readily achieved. A knowledge of the tension of the Plateau-Gibbs border
together with the contact angle at different applied potentials, however, circumvents this
problem. Finally it is pointed out that the use of the Lippmann equation provides an indepen-
dent means of finding the interfacial tension between bulk oil and water phases and hence of
checking the accuracy of the nonabsolu te direct methods, such as the drop-volu me technique,
which have to be employed in such systems.

INTRODUCTION of these quantities can, however, present diffi-

Optically black films formed in aqueous
media from solutions of lipids in nonpolar
solvents have been extensively examined during
the past few years and their structure and
properties have been the subject of several
recent review articles (1-4). Properties of
special interest in physico-chemical studies are
the specific capacitance of the thin film and the
interfacial tensions of the film and its adjacent
bulk liquid, the Plateau-Gibbs border. The
specific capacitance is important because,
among other reasons, it is closely related to the
thickness of the nonpolar part of the black film
(4, 5-7), and the two tensions, because their
difference reflects the free energy of thinning
of the film (4, 8). The accurate measurement
* Present address: Centro de Biofisica, Instituto
Venezolano de Investigaciones Cientificas, Caracas,
Venezuela.
culties. For example, the specific capacitance
is not easily found in media of 10\" conductivity
(4, 5), or for black films in which residual sol-
vent is trapped as micro-lenses (-!, 9, 10).
Again, while both tensions may, in principle,
be measured directly, their difference is so
small ["'10-2 dyne/cm (11, 12)J that, in
practice, this approach is hopelessly imprecise.
The fact that the tensions are interrelated
through the contact angle (8, 13, 14) is of great
assistance since the latter may be measured
very accurately and only one tension is then
required. For films stabilized by reversibly ad-
sorbed molecules, such as the monoglycerides
(15, 16), the interfacial tension of the liquid in
the Plateau-Gibbs border may be obtained
with sufficient accuracy from conventional
tension measurements on samples of the ap-
propriate bulk lipid and aqueous solutions.

Copyright © 1975 by Academic Press, Inc.

All rights of reproduction in any form reserved.
 Plateau-Gibbs
border y
Figure 1 depicts the thin film, aqueous solu-
I
tions and electrodes. At constant temperature
e
and pressure, the Gibbs equation for adsorption
I
1( phase (") on to the thin film may be \\Titten (18, 19)
Cu',Ag,AgCI (gCI, Ag, Cu"

I
'"
thin lipid
\\'here IJ' is the tension of the (t\\'o-sided) film,
film r i is the number of moles of species i per uni t
area of the interfacial region and jJ.i is the elec-
FIG. 1. A schematic diagram of the black film,
aqueous solu tions, electrodes and potentiometer. trochemical potential of the species i. If the
ionization of the water is ignored, Eq. [lJ may
be expanded as follo\\'s:
For phospholipid systems, ho\.I'ever, where the
monolayers are formed irreversibly and in an -dlJ' = rNa+'djlxa+' + rCI-'djlcl-'
uncontrolled manner (2, 4, 12) the conditions + r1I20'dJ.'1l20' + r,,'dJ.'''' + r{djl{
in the Plateau-Gibbs border are not readily + rxa+/ldjlxa+/I + rc1-/ldjlcl-/I
reproduced in separate experiments, and the
tensions obtained in this way may bear little
+ rll20/ldJ.'JI20/l + r,."dJ.',,/I + r/'djl{', [2J
relation to those in the film system. Only in \I'here It and l denote respectively the hydro-
situ determinations of the tension "'ill suffice carbon solvent and polar lipid of the film.
and these are unattractive from the technical Questions concerning equilibrium and sym-
point of vie\\'. metry in the system \\'ill be discussed later.
In this paper it \"ill be shown that difficulties Since
such as those described above may be over-
djlxa+ = dJ.'XaCI - djlCI-, [3J
come by using a form of the Lippmann equa-
tion to relate the tension of the film (or the Eq. [2J may be re\\Titten,
product of the bulk interface tension and the
-dlJ' = rNa+'dJ.'i\act' - ri\a+'djlci-'
cosine of the contact angle) to its specific
capacitance and the electrical potential differ- + rcl-'djlcl- + rH20'dJ.'lJ20' + r,.'dJ.''''
ence bet\\'een the t\\'o aqueous phases (17). + r{djl{ + rXa+/ldJ.'XaCI/I
With this relationship, a knowledge of the - rxa+/ldjlci-/l + rcl-/ldjlcl-/I
specific capacitance of the film and its contact
angle, as a function of the applied potential,
+ rJI20/ldJ.'1l20/l + r,,/ldJ.',,/I + r{'djl/'. [4J
enables us to calculate the tension of the inter-
faces of the Plateau-Gibbs border. Alterna-
tively, if the tension in the Plateau-Gibbs
border is kno\\'n from separate experiments, the \\'here e denotes the electrons m the copper
variation of the contact angle \\'ith applied \I'ire, so that Eqs. [-l-J and [5J may be com-
potential yields the specific capacitance \\'hich, bined to give
moreover, is independent of film area measure-
ments and hence of the presence of micro-
-dlJ' = (rCI-' - ri\a+')djlecu' + rNa+'dJ.'NaCI'
lenses. Examples of such uses of the Lippmann + ru20'dJ.'ll20' + r,,'dJ.".' + r{djl{
equation \\'ill be described belo\\'. First, how- + (rCl-/I - rNa+/I)djleculi
ever, it is appropriate to consider the ap- + rXa+/ldJ.'NaCt" + rH20/ldJ.'H20/l
plicabilityof the equation to black lipid films. + r,,/ldJ.',,/I + r/,djl{'. [6J
 It is quite common for black films to be [l-1J give
formed from nonionic lipids. In such instances
jll = JL1 and the excess charge q', on one side
-du = (qde)(dj1ecu' - dj1/u"). [15J
of the film is given by Equations [10J and [15J differ only in the
definition of the surface charge q. Ho\yever,
provided the charged lipid molecules are unable
to cross the thin film at an appreciable rate

and, since the film as a whole (including its fe' = ft-II = constant [16J
associated electrical double layers) must be and, on applying a potential across the film,
electrically neutral, the polarization charge qp is the same in each
instance. Thus, both Eqs. [1OJ and [15J may
be \Hitten
]f the bulk concentrations of the various com-
ponents are initially equal on the t\yO sides of
the film, and remain so during the course of an
experiment

d!1~aCI' = dJLNaCI" = dJLJl20'

The term (dj1ecu' - dj1ecu") is simply edE,
= dJLH20" = ... = 0, [9J "'here E is the difference in potential applied
and Eqs. [6J-[9J reduce to from the potentiometer to the copper connect-
ing ,,·ires. Hence Eq. [17J becomes
-du = (qI/e)(djlecu' - djl/u"). [10J
- (du/dE)I"l',p = qp, [19J
In order to treat the more general case in
"'hich the lipid molecules do ionize, it is as- i.e., the Lippmann equation.
sumed that the equilibrium In deriving Eq. [19J ithas been assumed that
the film surfaces are ahyays in equilibrium \yith
their adjacent aqueous phases. But for the
exists. Hence, as for the NaCl possibility of fluxes across the film there is no
reason to suppose that this \yould not be so.
djlt- = dJLNat - djlNa+ [12J
Fluxes of uncharged components should, from
and, together \yith Eqs. [3J, [5J, and [6J, \ye symmetry be zero. Fluxes of ions may be com-
find posed of the inorganic electrolyte and, if the
lipids ionize, of these molecules also crossing
-du = (fel' +f t-' - fNa+')djlecu'
from one film surface to the other. Applied
+ fNa+'dJLNaC/ + f 0'dJLlI 0' H2 2
potentials of 100 m V in systems containing
+ fh'dJLI.' + fe'dJLNat' 0.1 M NaCl produce steady state currents of
+ (f + ft-
CI-"
II
fNa+")dj1/u" - ca. 10-10 A cm-2, and it is readily inferred that
+ fNa+"dJLNact + fH201dJLJl20" the diffuse double layers at the film surfaces
+ fhldJLh" + ft-ldJLNat". [13J \,"ould not, under these circumstances, be
significantly perturbed. Thus, for films of non-
The excess charge q2 on one side of the film is ionic or zwitterionic lipids, disturbance of the
no\\' given by equilibrium by the electric field should not in-
fCI-' + ft-' - fxa+' validate Eq. [19]. For ionic lipids the validity
of Eq. [19J rests on the validity of Eq. [16J
= - (fCI-" + ft-" - fNa+") = qde [14J
and this depends on whether, in the time of an
If the system is again symmetrical with respect e:o..'[)eriment,an appreciable asymmetry in the
to the uncharged species Eqs. [9J, [13J, and surface concentrations of lipid ions on the t\\'o
sides of the film is built up, As to "'hether or
not this happens depends on the nature of the
lipid and, in general, therefore, it cannot be
assumed that Eq. [16J \\'ill be correct. HO\y-
ever, for phospholipids, there is evidence (20,
21) that the rate at which they cross lipid
bilayers is so small that the present type of
experiment ,,'auld normally be complete before
the held significantly affected the ion distribu-
tion, Certainly, for the phosphatidyl serine,
examined beIO\\', no time dependence of the
contact angle \\'as observed, so that for Eq.
[16J to be in error, the diffusion of this lipid
acros the film ,\"QuId have to be rather faster
than has, so far, seemed likely.
For practical purposes it is convenien t to
integrate Eq. [19J making use of the expression
for the integral capacitance, C, of the film,
C = ijp/ f!.,
From Eqs, [19J and [20J,
f
-
u 1E
drT = C lid H,
0"0 Eo
\\'here it is assumed that C is independent of E.
[The origin of C has been discussed in terms of
the film structure in an earlier publication (7).J
The relationship bet\\"een the film tension, rT,
and that of the Plateau-Gibbs border, 1', may
be \\TitLen (8, 13, 1-1)
rTo= 21' cosOo at f~ = Eo,
and
rT= 21' cosO at f~ = E,
where 0 is the contact angle. Integrating Eq.
[21J and substituting for rTand rTofrom Eqs.
[22J we obtain
cosOo - COSOE= (C/-1-y)(E2 - E02). [23J
The assumption that C is independent of E
is not generally valid, as has been sho\1'll by
several investigators (15, 22-2-1). For applied
potentials of less than 100 m V, ho\\"ever, the
specific capacitance of many types of black
lipid film is constant to ,\"ithin experimental
error. The use of Eqs. [22J requires that the
film be in mechanical equilibrium \\"ith its
border. That this is normally so will be made
clear in due course. It should also be clear that
I' is not affected by the applied potential since
the field produced across the interfacial region
bet\\"een the bulk oil and aqueous phases "'ill
be negligibly small compared ,\"ith that across
the thin film.
Equation [23J is the basis of the approach
outlined in the Introduction. The main experi-
mental task i to measure the contact angle a
a function of the applied potential.
EXPERIJ\IEKT AL
Apparatus. The apparatus employed is il-
lustrated in Fig. 2. The formation of the black
films and the measurement of their contact
angles has been described in detail else\\'here
(-1, 12, 25). In brief, the films \\'ere formed by
the micrometer injection method (1) (-1, 26)
in the 0,7 mm diameter hole (A) in the base of
the inner PTFE vessel (B), For setting-up
purposes the film could be illuminated from
belO\\' but, for the determination of the contact
angle, incident illumination from a high pres-
sure mercury lamp, \\'ith an interference filter,
,,'as used. The mercury green ligh t \\"as trans-
mitted through a metallurgical microscope, the
objective (0) of \\'hich is shOll'll in the diagram.
\Yhen this light is reflected, either from the
Plateau-Gibbs border or from a lens in the
film, interference fringes may be seen and can
be photographed (see Fig. 3). The film and
[22J aqueous phases were surrounded by a '''<lter
jacket (D) by means of which the temperature
FIG. 2, The apparatus for black film formation and
associated electrical measurements, A detailed descrip-
tion is given in the text.
 E4-1-51

5011

FIG. 3. A photograph of a lens trapped in a black film under reflected monochromatic light. Interference
fringes are visible in both the lens and in the Plateau-Gibbs border. The film was formed from a solution
of monoolein in n-octane under 0.1 M NaCl at 20°C.
in the system could be controlled to ±0.2°C.
The aqueous phases in the outer and inner
vessels were electrically isolated from each
other and potentials \yere applied from a
Tinsley potentiometer (f) through the Ag/
AgCl electrodes (E). Also connected to the
electrodes \yas an fET low bias current opera-
tional amplifier (OPA:\IP) (input resistance
'" 1013 £1, :\Iodel -12K, Analog Devices, ~Iass.,
U.S.A.), \yhich \yas used as an inverting ampli-
fIer to measure the current flowing or the
potential across the film. The closed loop gain
of the amplifier \\"as determined by a precision
resistor inserted in the feedback loop of the
amplifier. Its value could be selected \\"ith a
PTfE insulated \Yafer rotary switch from a
bank of resistors ranging from 10c1010 £1. An-
other PTFE insulated wafer rotary s\\"itch
connected the feedback resistor in parallel to
a silver mica precision capacitor chosen from
a bank of capacitors ranging from 10-1000 pI',
or to open contacts. The electrode in the inner
compartment could be connected to the nega-
tive input (virtual earth) of the amplifier
either directly or via a 10~ £1precision resistor
in parallel \yith a 100 pI' capacitor. The former
arrangement allo\\"ed the measurement of the
current \yhile the latter yielded the potential
bet""een the electrode .
The capacitance of the film was determined
by means of a Wayne Kerr Universal Bridge
(UB) \yith an external variable frequency
signal generator (G) (\Yayne Kerr, :Model
S121) and an oscilloscope (CRO) ,,"ith a sensi-
tivity of 100 }J'v / em as the detector (5).
Interfacial tensions were determined by the
drop-volume method (27, 28).
All the experiments to be reported ,,"ere
carried out at 20°C.
Methods. Reproducible results ,,"ere ob-
tained only by paying great attention to the
cleanliness of the apparatus. The PTFE com-
ponents \yere especially important in this re-
spect and \yere subjected in succession to a
gentle scrubbing in detergent solution, rinsing
in distilled water, immersion in fresh chromic-
sulphuric acid mixture, further rinsing in water
and, finally, refluxing in twice-distilled \yater
for some hours. The treatment of other COI11-
ponents differed only in the degree of rigor
required to give satisfactory resul ts.
Lenses, such as the one illustrated in fig. 3,
were formed either spontaneously or by apply-
ing suddenly a potential C,,180 mY) just as
the film began to drain. The lens \yas centered
in the field of vie'" and focused. The light in-
tensity \YaSadjusted to give maximum contrast
and the negative exposed. for a given lens,
this procedure was repeated for each increment
in the applied potential, the sequence ending
"'ith a repeat of the zero potential condition.
The response of the fringe pattern to a change
in the applied potential "'as almost instanta-
neous, but an interval of two minutes was
nevertheless allo\\"ed before a photograph \yas
taken. Alternate sequences on a lens \\"ere con-
ducted \\"ith the polarity of the potential
reversed.
The contact angle, 0, \\"as calculated from
the expression (-1, 12, 25)
sinO = (-
>..) --po
211 r III fiX ,
where>.. is the wavelength of the monochro-
matic illumination and 11 is the refractive index
of the lipid solution. rmax is the radius of the
lens and po is the order of the fringe of de-
structive interference at its center. Both these
quantities can be obtained by extrapolation of
the linear plots of fringe order p against fringe
radius r2•
The change in the cosine of the con tact angle
at a potential, E, with respect to its value at
zero potential for each sequence (tl COSO)E,
was obtained by averaging the 1\\"0 zero po-
tential results-the one at the beginning and
the one at the end-and subtracting from it
the cosine of the angle at E, thus,
The experimental data which appear m the
figures and tables are the means of (tl COSO)E
taken over all sequences on a given system.
The regression lines shown in the figures were
computed from the individual (~cosO) E values.
The behavior of all the systems to be de-
scribed was very sensitive to contamination.
The criteria for acceptability of a given experi-
ment were (a) that the film area could be con-
trolled by the addition or withdrawal of lipid
solution from the hole \yith the micrometer
syringe; (b) that the lenses survived at least
sufficiently long for a complete sequence of
potentials to be applied; (c) that movement
of the lenses was minimal, i.e., that an exposure
of up to 2 sec did not produce a blurred photo-
graph, and (d) that the contact angles did not
show hysteresis on the application and removal
of a potential.
 Materials. All the 11l0noglycerides \yere
purchased from Ku-Chek Prep. Ine. (Min-
nesota, U.S.A.), stored under vacuum at
-20°C and used \yithout further purification.
They \yere claimed to be greater than 99%
pure but did not necessarily consist of more
than 95% i-isomer. The fatty acid composition
,,-as checked by GLC to be better than 99%
of the theoretical value. The 111onoglycerides
from micelles in alkanes (15) and, to obtain
stable films, concentrations above the CMC
are necessary. 3 mglml of the monoglyceride
in the alkane proved suitable in all the present
systems.
The egg yolk lecithin was supplied by N.
:Miller (Agricultural Research Council Institute
of Animal Physiology, Babraham), and the 1,2-
dioleyl phosphatidyl choline by Dr. J. C.
;'1etcalfe of the Medical Research Council,
Kational Institute for Medical Research, Mill
Hill, London. Bacterial phosphatidyl ethanol-
amine ,,-as obtained from Supelco \\"hile the
brain phosphatidyl serine \\"as purchased in its
acid form from Lipid Products. All the phos-
pholipids "-ere shO\m by TLC to be > 98%
pure. The principal fatty acids of the natural
compounds \yere shO\m by GLC to be in %
(\\"1\\-) as follo,,-s: egg yolk lecithin: oleic, 32%;
palmitic, 26%; stearic, 15%; linoleic, 12% and
arachidonic 6%; bacterial phosphatidyl etha-
nolamine: palmitic, 49%; palmitoleic, 20%;
oleic, 18%; bisdihydrosterculoic, 13%; brain
phosphatidyl serine: oleic, 41%; stearic, 36%;
CIS polyunsaturated, 12%.
All the phospholipids were stored in chloro-
form-methanol (2: 1 v Iv) at - 20°C. In the
preparation of the dispersions in the alkanes it
was extremely important to exclude water.
This may be achieved in a variety of ways, one
of which is by repeated slo\Y evaporation at
reduced pressure, of the phospholipid in dry
solvents (12). A stoichiometric concentration
of 1% (w/v) phospholipid in decane \\"as used
in each instance.
The alkanes \yere obtained from Koch-Light
Laboratories and \\"ere all "puriss" grade
(> 99%). Before use they \\"ere passed through
an alumina column in order to remove polar
impurities. The glycerol and the sodium chlo-
ride \yere both AR grade; the latter \\"as roasted
for 6 hr at 700°C to remove organic impurities.
The ,,-ater \yas t\yice-distilled, the second
distillation being carried out in a Pyrex still
fitted \yith a fused silica column and condenser.
RES LTS ND DISCUSSIO~
Tests of the theory. The effect of the applied
potential on the diameter and fringe pattern
of a lens in a black film is shown in Fig. 4.
Some two minutes elapsed bet\\"een each
photograph. From the final observation at zero
potential it can be shown that the influence of
the field is completely reversible_ The calcula-
tion of the contact angle from the fringes is a
straightforward geometrical exercise and has
been described elsewhere (11, 12, 25). It has
also been shown that, to \yithin experimental
error, the same value of the contact angle is
found from a lens as from the Gibbs-Plateau
border (12, 25).
In Table I are given the contact angles for
a range of applied potentials for black films
formed from glyceryl monooleate-n-decane
solutions under 0.1 M KaCI. Figure 5 sho\ys
plots of the change in cosO against the square
of the applied potential. As can be seen, the
relationship bet\yeen the t\\"o quantities is
Iinear below abou t 100111V. The specific capaci-
tance of the film and the tension of the bulk
oil/aqueous solution interface are given in
Table II. In the last two columns of this table
are given the slope of the plot in Fig. 5 (b) and
the theoretical slope (C/h) according to Eq_
[23]. These two quantities are in excellent
agreement.l
I An equation essentially the same as Eq_ [23J has
been tested by Rovin, Kryglyakov and Koretskii (3-1)-
These authors worked with films formed from a 0_3%
solution of "xylane" (Co4H4005) in undecane_ They
found a minimum in the plot of () against 1:."'2 at lo\\" J~
and, although the plot became linear at higher values
of E, the specific capacitance calculated from the
equation differed substantially from that obtained by
direct measurement.
It should also be mentioned that the variations in
tension of a lecithin and an oxidized cholesterol film
with applied potential have been reported by Tien (35),
but no details of the experiments or interpretation of
the data is given.
 The above system was chosen to test the
Lippmann equation largely because it has been
repeatedly examined (15, 16, 26) and is prob-
60 /
ably the most precisely characterized of all
black lipid films. In other respects, however,
the choice was arbitrary and it was felt neces-
sary to test the equation with other systems
~::~
[J
~

~
3.0

20
also. The results for some of these are given in 1.0

Fig. 6 and Table II. It is again found that up 1 I ! !

o 4() 120 200

to 50 mV, at least, the change in cosOis a linear (E2-F6)l103 voltZ
function of E2 and that, as shown in Table II,
the observed and theoretical slopes are in very FIG. 5. Change in the cosine of the contact angle as a
function of the square of the applied potential for a
good agreement.
In order to test the equations still more gen-
monoolein +decane black film in 0.1 M NaC!. (a)
In the range 0-150 mV, showing the deviation from
erally, black films of glyceryl monooleate linearity at high potentials. (b) Showing the accuracy
+ decane in solutions of NaCI (1.0 M) in of the linearity at lower potentials; the slope of the
glycerol were examined. The accurate deter- line is 0.248 ± 0.003 V-2.
mination of the specific capacitance for these
films is made difficult by the low conductivity
of the glycerol solutions. O\ying partly to this,
and partly to the desire to see whether such
THE CONTACT ANGLE OF MONOOLEIN (3 mg/ml)-n-
films behaved as ideally as those in aqueous
DECANE BLACK FILMS IN AQUEOUS 0.1 At! NaCI
AT 20°C AS A FUNCTIO, OF THE
solutions, their impedance was measured as a
ApPLIED POTENTIAL function of frequency and the data displayed
in the complex plane as described in (5).
Applied Contact anglea cos Oa
Potential 8 (degrees) Figure 7 shows the resulting Cole-Cole plot of
(mV)
the complex capacitance C" (= G/w, where G
0 1.96 0.999415
is the parallel conductance of the system and
±0.000004 w the angular frequency) versus the real part
20 2.13 0.999311 C' of the capacitance of the system. The curve
±0.000005 is, within experimental error, a perfect semi-
30 2.32 0.999183
circle with its center on the abscissa showing
±0.000013
40 2.55 0.999013
that, as in aqueous solutions, the film imped-
±O.OOOOl1 ance is determined effectively by its hydro-
50 2.81 0.998801 carbon interior (5-7). The specific capacitance
±0.00001O obtained by extrapolation of the semicircle to
60 3.15 0.998486
the abscissa (zero frequency) is given in Table
±0.000029
80 3.80 0.997796
II. The change in the cosine of the contact
±0.OOO035 angle is again a linear function of E and the
100 4.56 0.996825 observed and calculated slopes ~~e again in
±O.OOOO90 close agreement.
120 5.37 0.995605
±0.000108
Applications: The Determination of the
140 6.18 0.994177
±0.000177
Free Energy of Thinning of a Black
150 6.60 0.993360 Phospholipid Film
±0.OOOI93
If the above evidence may be accepted as
• For 0-60 mV the values are the mean of 14 measure- establishing the validity of Eq. [23J, it is clear
ments while from 80-150 mV they are the mean of four that this equation may be used to find either
measuremen ts. Cor'Y if the other is knO\\"ll.It was pointed out
 THE PREDICTEDANDOBSERVEDSLOPESOF (COS 00 - cos OE) vs E2 (EQ. [23J) FORVARIOUSBLACK
LIPID FILMSAT20°C. THE MONOGLYCERIDES WERE3 mg/ml IN THEALKANES

System Bulk phase inter- Specific capacitance Predicted slope Observed slope
facial tension (/LF em-') (V-') (V-')
(dyne em-I)

Monoolein + 11-octane 4.38 ± 0.05 0.385 ± 0.002 0.220 ± 0.003 0.224 ± 0.009
Monoolein + 2,2,4-
trimethyl-pentane 4.25 ± 0.05 0.397 ± 0.005 0.234 ± 0.005 0.230 ± 0.008
aqueous
Monoolein + n-decane
O.IM
3.84 ± 0.03 0.386 ± 0.002 0.251 ± 0.004 0.248 ± 0.003
Monoolein + n-hexadecane
NaCI
2.51 ± 0.03 0.585 ± 0.003 0.583 ± 0.006 0.589 ± 0.011
Monolinolein +n-decane 3.30 ± 0.10 0.390 ± 0.006 0.295 ± 0.006 0.295 ± 0.007
Monolinolenin +n-decane 2.86 ± 0.06 0.554 ± 0.002 0.484 ± 0.004 0.484 ± 0.013
Monomyristolein +
n-decane 1.68 ± 0.03 0.415 ± 0.004 0.618 ± 0.006 0.615 ± 0.007
Monoolein + n-decane glycerol 1.0 M 4.06 ± 0.04 0.390 ± 0.005 0.238 ± 0.005 0.233 ± 0.003
NaCI

m the Introduction that')' for the Plateau-
Gibbs border of a phospholipid film is not ac-
curately assessable by ort.hodox methods and
that, for this reason, the free energy of thinning
of the films ,\"as difficult to determine. The
specific capacitance is, ho\\"ever, accurately
measurable. Hence, using Eq. [23J, ')' may be
found and, with the contact angle at zero
applied field, the free energy of thinning
(eTo - 2')') may be calculated.
The changes in the cosine of the contact
angle against E2 for four phospholipid films are
shown in Fig. 8. The systems satisfied the con-
ditions for reversibility described earlier and,
although the reproducibility was poorer than
for the monoglycerides, the plots are satis-
factorily linear. The slopes of the lines, to-
gether with the specific capacitances of the
films, are given in Table III. The tensions
deduced for the Plateau-Gibbs border are also
shown and, with cosO, yield the specific free
energy of thinning 6.A[ = 2')'(cosOo - 1)].2
These free energies have been shown elsewhere
(11, 12, 29) to originate almost entirely from
the van der Waals forces acting across the
films.

The Determination of the Specific Capacitance

::~ of Black Films in Media of

Low Cond~tctivity
For films in glycerol + 1.0 M NaCI it is
12~ possible, as sho\vn in Fig. 7, to obt.ain a fairly
Q 10
2 Moran and Ilani (36) have reported the specific
free energy of thinning of a lecithin-cholesterol-methyl
oleate black film to be ca. 1.2 erg cm-2• The reasons for
this value being very large compared with the present
results for phospholipid films are not clear. It may be
that the presence of the cholesterol and (or) the use of
methyl oleate instead of an alkane solvent is responsible.
However, Moran and Ilani's data were obtained by
subtracting the bulk intcrface tension (X2), measured
o 0.4 0.8 12 16 20 2.4 by the drop-volume method, from the film tension,
(E2_E6)~I03 1,10112 measured by bulging the film under a hydrostatic
pressure. For phospholipid systems, the drop-volume
FIG. 6. Change in cos 0 as a function of E;2 for black method is very inaccurate and for any system the film
films of monoolein +
hexadccane (6), monolinolein bulging method is also inaccurate and hence, as empha-
+ decane (0) and monoolein +
octane (.), all in sized in the Introduction, there must be some reser-
0.1 M NaC!. vation over the reliability of their very high free energy.
THE SPECIFIC FREE ENERGY OF THINNING OF SOME BLACK PHOSPHOLIPID n-DECANE +
FILMS IN AQUEOUS 0.1 "AI
NaCI AT 20°C. THE PHOSPHOLIPIDS WERE USED AT 1% (w/v) IN THE DECANE

Phospholipid Specific capacitance Obsen·ed slope Bulk phase inter- cos 00 M

u.F em-') of EQ. [23J facial tension (lO' erg em -')
(v-') (dyne em-I)

1,2 dioleyl phosphatidyl

choline 0.390 ± 0.005 0.306 ± 0.011 3.18 ± 0.15 0.999228 ± 0.000010 -4.90 ± 0.30
Egg phosphatidyl
choline 0.385 ± 0.008 0.399 ± 0.014 2.41 ± 0.13 0.999138 ± 0.000010 -4.15 ± 0.25
Bacterial phosphatidyl
ethanolamine 0.445 ± 0.002 0.282 ± 0.003 3.95 ± 0.06 0.999215 ± 0.000017 -6.20 ± 0.24
Brain phosphatidyl
serine 0.376 ± 0.003 0.464 ± 0.016 2.03 ± 0.09 0.999036 ± 0.000045 -3.91 ± 0.35

accurate value of the capacitance in the limit
of low frequency by means of the standard ac
bridge circuitry referred to under Apparatus
md, in more detail, elsewhere (4,5). Even so,
ming to the high impedance of the system, a
,hart extrapolation of the data was required
(Fig. 7). In glycerol +
0.1 M r aCI, the imped-
wce is some tenfold higher still and the zero
:requency capacitance can be obtained only by
neans of a long extrapolation \\"hich, nece -
;arily, yields inaccurate results. A dc transient
nethod could be used (6,30-32), but again, the
Jrecision is seriously limited by the low con-
luctivity of the glycerol solution.
The use of Eq. [23J, although indirect and
nvolving complex apparatus, yields specific
:apacitances which seem likely to be more ac-
:urate than those obtained by the orthodox
160
"-
Cl.
,~
120
~ in decane, whereas in aqueous systems the
15Cl.
~
"
.~
."
f;
0 80 I 0
Reol pori Ie; pF
FIG. 7. Cole-Cole plot for a black monoolein
11m in glycerol-Mil aCl. The relaxation frequency,
·0, of the system is 2.12 kHz, and the center of the
:ircle lies, within experimental error, on the real axis.
[he area of the film was 0.625 X 10-3 cm2•
methods. Table IV lists t.he consequences of
employing this approach for a range of mono-
glyceride black fIlms in glycerol +
0.1 M NaCI.
As for the phospholipid systems, these data
are relevant to the understanding of the
London-van der Waals forces acting across t.he
thin films and are discussed from this point of
view in anot.her paper (29). The specific
capacitances are nevertheless of some interest
in t.hemselves. Thus, for decane as the solvent,
the capacitances increase as the lipid chains
become more unsaturated. In this respect t.he
aqueous (Table II) and glycerol systems are
similar. For hexadecane, the situation is
qualit.atively similar [the capacitances for films
of monolinolein and l110nolinolenin in aqueous
0.1 M NaCI are 0.667 ± 0.006 and 0.803
± 0.009 ,uF/ cm2, respect.ively (33) J in that
great.er unsaturation again produces higher
capacit.ances but, in glycerol, the act.ual values
are considerably smaller.3 For example, in
glycerol the specific capacitance for monoolein
in hexadecane is only slight.ly greater than t.hat
hexadecane result is much larger. These effects
are apparently related t.o the adsorption and
conformation of the lipids in the various sys-
tems [see e.g. Refs. (15, 16, 33)J but a detailed
160 discussion is beyond the scope of this paper.
Other applications. In the Introduction,
reference \yas made to the occurrence of micro-
decane+
3 The capacitance of the monolinolenin + hexa-
decane films in glycerol could not be determined because
the contact angle was too large to be measured in the
present apparatus.
 Bulk phase inler- Observed ~lope Specific capacilance
facial lension of Eq. [23] from "Observed slope"
(dyne em-I) (V-') C/lF em-')

1Ionoolein + n-decane 4.14 ± 0.04 0.221 ± 0.005 0.366 ± 0.012

Monoolein + n-hexadecane 2.73 ± 0.04 0.334 ± 0.005 0.365 ± 0.012
Monoolinolein + n-decane 3.91 ± 0.04 0.285 ± 0.006 0.446 ± 0.016
Monolinolein +n-hexadecane 2.51 ± 0.04 0.526 ± 0.005 0.528 ± 0.013
Monolinolenin + n-decane 3.40 ± 0.04 0.407 ± 0.008 0.554 ± 0.017
MonolinoJenin + n-dodecane 2.44 ± 0.04 0.576 ± 0.017 0.563 ± 0.023

lenses of oil phase in black lipid films and the
problem \I'hich t.hese created in t.he determina-
t.ion of t.he true area of t.he black film, and hence
of the specific capacit.ance. This maHer has
been discussed by several aut.hors but., apart
from ,,'ait.ing for t.he lenses to coalesce ",it.h the
Plateau-Gibbs border (by ,,'hich time the
films have usually broken), no method of cir-
cumventing the experimental difficulties has so
far been found. The use of Eq. [23J is, how-
ever, such a method since no film area measure-
ment is involved in the estimation of the specific
capacitance. All that. is required is a reliable
value of t.he appropriate interfacial tension. At
least t\"O of t.he black films listed in Table II
(those cont.aining n-octane or 2: 2 :4-trimethyl-
pentane) did not embody visible microlenses.
If t.hese syst.ems may be taken to validate Eq.
[23J, then it is clear from the agreement be-
t.\\'een the· observed_ an(predict.ed slopes that
such microlenses as \"ere present in the films
containing higher alkanes could not have sig-
nificantly affected t.he estimat.es of their area.
One further application arises in connection
with the difficulties commonly encountered in
t.he determination of the interfacial tension in
systems containing surfactants. Thus, in the
Wilhelmy plate technique t.he contact angle
may not be exactly zero, while in the drop-
volume approach the rheology of the surface
films could introduce errors during the detach-
ment of the drops. It is therefore of interest to
note that the use of Eq. [23J, together \"ith
measurements of the specific capacitance of the
black films, yields a new and independent (if
some\vhat laborious!) method for estimating
interfacial tensions in certain types of system.
For the monoglyceride-alkane/electrolyte solu-

Q 08
-"-
d;'
0
' ) 06

<f!

~'" 0.'\

02

0 0 1.6

(E2-E~)'103 volt2

FIG, 8. Change in cos (j as a function of E2 for four phospholipid-decane black films' in 0,1 M NaC!,
., egg phosphatidyl choline; 0, 1,2-dioleyl phosphatidyl choline; ••• , brain phosphatidyl serine; 6,
bacterial phosphatidyl ethanolamine.
tions described in this paper it would seem that, 15. ANDREWS, D. M., !\IANEV, E. D., AND HAYDON.
given the validity of Eq. [23J, any error in the D. A., Special Disc. Faraday Soc. No.1, 46
(1970).
drop-volume technique must be less than ca.
16. FETTIPLACE, R., ANDREWS, D. M., AND HAYDON,
2%. D. A., 1. JIembrane Bioi. 5,277 (1971).
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18. PARSONS, R., ill "~Iodern Aspects of Electro-
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Cientificas throughout the course of this work.
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