Skin Research and Technology 1997; 3: 126-132
Printed in Denmark. All rights reserved
EEMCO guidance for the assessment of stratum comeurn
hydration: electrical methods
E. Berardescal
European Group for Efficacy Measurements on Cosmetics and Other Topical
Products (EEMC0)2
'Department of Dermatology, University of Pavia, IRCCS Policlinics S.Matteo, Pavia, Italy, 2Members of the EEMCO Group: P. Masson (Chairman),
France; L. Rodrigues (Secretary), Portugal; E. Berardesca, Italy; CL Gummer, UK; 1-L Liv@que,France; M . Loden, Sweden;G. Piirard, Belgium; 1/: Rogiers,
Belgium; D. Salter, UK; G. Sauermann, Germany; 1. Serup, Denmark; M . A . Trevethan, UK
Background/airns: The improvement of stratum corneum hy-
dration is one of the most important claims in the cosmetic in-
dustry.
Methods: Objective assessment of moisturization can be done
with devices based on electrical methods provided these instru-
ments are used in an appropriate manner.
Results: This paper deals with the biophysical basis behind
these techniques and describes the most important variables,
pitfalls and drawbacks related to measurements and current in-
strumentation. Individual-related and environment-related vari-
I N RECENT years, measurement devices have been
introduced into experimental dermatology and
cosmetology for quantitative evaluation of several
parameters related to skin function. Among these, de-
vices claiming to quantify skin hydration have be-
come very popular due to their relatively low cost and
ease of use. The use of these methods to assess skin
hydration has been documented in both normal and
diseased skin (1-4). However, it is also clear that many
factors may influence the measurements and can alter
the final interpretation of results (5-7). Furthermore,
reproducibility and variability between different in-
struments and methods can make it difficult to ad-
equately compare data obtained in different labora-
tories and experiments.
Biophysical Basis and Its Consequences
1. Physical mechanism
At least three different types of electrical charge car-
riers have been proposed to be involved in electrical
transport through keratinized tissue (1). Hence we
must consider:
a) Conduction by electrons and holes;
126
Cupyrrgkt 0 Munksgmrd 199i
Skin Research and Technology
lSSN 0909-752X
ables are also analyzed as well as study designs for predictive
or use tests.
Conclusions: Practical suggestions for standardization of meas-
urements are given.
Key words: hydration - stratum corneum - conductance - ca-
pacitance - skin bioengineering - impedance
0 Munksgaard, 1997
Accepted for publication 23 August 1996
b) Conduction exchange of protons all along the H-
bonded network of water molecules;
c) Conduction by ions larger than protons ("large
ions").
The evidence is that all of these are involved in elec-
trical conduction in skin, but to differing degrees (8,
9). Conduction by electrons and holes is likely to be
significant only in an abnormally dry state, and con-
duction by large ions only if the electrical field stim-
ulation is at frequencies below the MHz range. In vivo
human skin has a water content around 10% near the
stratum corneum surface, and in these circumstances
protonic conduction can be expected to dominate and,
therefore, is highly dependent on the water content.
It should be noted that conduction by all three types
of carrier will be governed by thermal activation, and
so will obey an exponential or Arhennius law of the
form o=o0 exp(-E/T) where the activation energy E
will vary with water content. This means that skin
conductance depends on both water content and skin
temperature.
The relationship between electrical conductance
and water content is not linear, but depends upon the
binding state of water molecules to the keratin chains,
which is itself described by the water sorption iso-

therm. It has been customary in the literature to define
three types of water according to their strength of
binding to the keratin: ”tightly bound water” for
water contents from 0 to around 7%, “bound water”
between about 7% and 35”0, and “free water ” above
35%).This division can be considered simplistic on the
basis of more detailed theory (8), but is generally
helpful. The consequence of variation in water bind-
ing strength is that there is no direct proportionality
between total water content and electrical conduc-
tance. Substances or treatments that interact with the
keratin-water network may, therefore, modify con-
ductance without, in fact, changing the water content
of the sample.
Because of the complex physics involved in this
situation, it is incorrect to refer to the electrical “ca-
pacitance” of the skin without specifying the stimulat-
ing frequency and other experimental conditions used
to make the estimate. What this literature generally
calls ”capacitance” is not actually electrical capaci-
tance in the usual scientific and engineering sense, be-
cause it is frequency dependent, unlike true electrical
capacitance.
Commercially available electrical methods for as-
sessing skin moisture are based on measuring impe-
dance or its reciprocal, conductance, as a function of
one or more different stimulating frequencies. Both
impedance and conductance are frequency-dependent
vector quantities, and the derivation from them of
standard physical parameters such as resistance, reac-
tance and capacitance is not always simple. Standard
textbooks of physics should be consulted. In the case
of heterogeneous and time-varying biological ma-
terials such as skin, the situation is significantly more
complicated, and a specialised review may be necess-
ary (10). It should be noted in particular that when
the equivalent electrical capacitance of skin is calcu-
lated, it is still a frequency-dependent property, unlike
the electrical capacitance of nonbiological dielectrics
described in the literature of engineering and physics.
For this reason the electrical ”capacitance” of skin will
be referred to here using inverted commas (thus: “ca-
pacitance”). It is not a true material property, but is
highly dependent on the method of measurement
used, and any estimates of this quantity should there-
fore also record the stimulating frequency or frequen-
cies at which they were obtained.
A simple view is that the impedance (or conduc-
tance) measures the opposition to (or ease of) the flow
of electric current through the skin, and depends
upon resistance and reactance. Capacitance is calcu-
lated from reactance. Skin conductance and ”capaci-
tance” are related, and generally both increase with
Electrical methods for assessment of SC hydration
moisturization, although the exact relationship is ex-
tremely complex and variable.
2. Some basic considerations
a) Dependence on properties of the skin s u f a c e
Because the electrodes in contact with the skin are
generally rigid, their areas of contact with the skin
may be dependent upon the pressure applied. Sub-
stances present between the electrodes and the skin
(hairs, oils, waxes) may affect the results and must be
taken into account. Conductance measurements may
also be influenced by changes in surface microto-
pography (11).
b) lntegratioii over depth
The distribution of the electric field through the dif-
ferent skin layers cannot be calculated adequately be-
cause of the complexity of the geometry and the com-
position of the materials involved. For example, the
field distribution partly depends on the complex per-
mittivity of the tissue, which is itself very dependent
on the distribution of polar materials (water, ions, di-
poles, etc.) with depth through the various skin
layers. It is possible to some extent to overcome this
theoretical difficulty by direct experimentation using
skin stripping. All non-invasive instruments give re-
sults that are to some extent an integration over
depth, with the precise dependence on depth varying
from instrument to instrument and also from skin site
to skin site. This source of variance can only be par-
tially overcome by calibration procedures involving
measurement of depth dependence in skin layer
models. It should therefore be noted that it is possible
for commercial instruments to give the same reading
from different water amounts if these amounts are
distributed according to different water depth pro-
files.
c) Water content and skiti temperatiire
It is possible for skin that is moist and cool to give
exactly the same electrical response at a single fre-
quency as dry and warm skin. To separate and specify
potentially confounding influences such as water con-
tent, temperature change and sweat gland activity, it
is necessary to use some form of electrical spectro-
scopic technique, that is, stimulation at three or more
different frequencies, or a time-domain approach fol-
lowed by Fourier transformation (5, 6). Therefore, it is
recommended to control skin and room temperature
to reduce the requirement for multi-frequency meas-
urements.
127
Berardesca et al.
Estimates of Hydration
A number of prototypes for measuring electrical
properties of skin have been manufactured in differ-
ent reserach laboratories. Some commercial devices
are also available. They work at differences frequen-
cies, using various sizes of probes that are applied
with distinct pressure on the skin. The influence of
these variables has been not thoroughly studied in the
literature. All of them give hydration values (ex-
pressed in arbitrary units) that are not linearly corre-
lated with the hydration state of the skin, and read-
ings are affected by the presence of salts and ions on
the skin (12).
Corneometer 820-825
This is described as being a "capacitance" measuring
device operating at low frequency (40-75 Hz), which
is sensitive to the relative dielectric constant (or per-
mittivity) of material placed in contact with the elec-
trode surface. Because increasing the water content of
the stratum corneum will in general increase its rela-
tive permittivity (although by a very complex and
variabie relationship), the device can therefore esti-
mate in about 20 ms the stratum corneum water con-
tent in arbitrary (relative) units (12). The probe exerts
a constant pressure on the skin surface of 3.5 N, and
covers an area of 49 mm2. It estimates water content
in the epidermis to an approximate depth ranging be-
tween 60 and 100 pm (5,13). The Corneometer has
been widely used to estimate the dryness of skin (2,
14-17). Results are strongly influenced by the qualities
of the skin surface, including surface roughness (11).
The new version (825) allows continuous measure-
ments.
Nova DPM 900
This instrument uses measurements of phase angle
(10). Phase angle is one of the two (polar) coordinates
of impedance at each particular frequency of stimula-
tion. The device is described as using frequencies up
to 1MHz (18),and can have different diameter or cus-
tomized probes. Measurements may be sampled
"continuously", after a 5 s interval, or singly and im-
mediately (19). It measures the upper stratum corne-
um layers, like the Skicon 200. Readings are usually
higher than with the Corneometer, but in good corre-
lation (19). The Nova DPM has also been reported to
be less variable than the Corneometer, but less suit-
able for testing dry skin (19).
128
Skicon 200
This device employs measurements of conductance at
high frequency (3.5 MHz). The probe consists of two
concentric electrodes, of 2 and 4 mm diameter, separ-
ated by a suitable dielectric. The estimate is displayed
after 3 s. The Skicon measures a more superficial
range of depths than the Corneometer (2,14,16), and
may therefore seem more variable, even though it has
been reported to be more sensitive in measuring
highly hydrated skin and water sorption-desorption
processes (16). The readings are increased significant-
ly in the presence of glycerol and conductive materials
other than water, so very great care must be taken in
interpreting the meaning of the results.
Other instruments, such as the SCIM (surface char-
acterizing impedance monitor) can be used to assess
moisture according to the multifrequency model. The
SCIM instrument measures electrical impedance, both
magnitude and phase angle, at 31 different frequen-
cies. The probe has concentric electrodes and this is
used to select the range of depths over which the re-
sults are integrated (21, 22). Further data are needed
to evaluate and compare the usefulness of this instru-
ment in assessing skin moisturizers. The multifrequ-
ency approach has also been used to characterise skin
disease (8, 10, 21-23).
General Considerations Before Starting
a Study
Some practical advice, independent of the instrument
used, should be followed to perform accurate and re-
producible measurements. The probe should be
placed normal to the skin surface with slight pressure,
just sufficient to start the measurement process. Each
of the devices described above has a spring in the
probe to assure a reproducible pressure. A different
position of the probe, or too many hairs, can cause
unreliable readings (24). When necessary, shaving or
cutting the hairs is recommended before starting the
study (16). Several measurements of the same site can
cause occlusion, which results in an increase in the
values displayed. Therefore, it is recommended to
wait for at least 5 s before repeating a measurement
on the same site. It is advisable to measure at least
three times, once at each of three different but nearby
sites, in order to have more reproducible data.
Measuring failures may occur if the skin or probe sur-
face is wet, for instance by sweating, or if the surface
is contaminated by dirt particles, oil or grease.
Cleaning of the probe is recommended between meas-
urements.
It is also important to consider environmental in-

fluences. To obtain reproducible readings, it is desir-
able to perform measurements under appropriate am-
bient conditions (25), keeping the ambient tempera-
ture low enough to minimise sweat production (e.g.,
20°-22”C) and, ideally, relative humidity between 40%
and 60%. The weather conditions of recent days can
however be just as influential as the ambient con-
ditions to which the skin is exposed during the test
itself. It is also desirable to minimise air movement in
the vicinity of the test site. Volunteers should be relax-
ed and acclimatized to the measuring environment for
at least 20 min. The instrument itself should also be
acclimatized for at least this time. The test site should
be exposed to ambient air for at least 10 min before
taking measurements. The efficiency and limitations
of these various commercial devices for assessing skin
moisturization have been reported in the literature (2,
7, 14, 16, 20).
Variables
Variables related to the individual
Regional differences in stratum corneum water con-
tent have been reported (2, 7, 13, 25, 26); but when
interpreting such results, it must be remembered that
all these electrical devices give only relative, not abso-
lute, indications of water content. The actual relation-
ship between their outputs and true water content is
highly complex and also variable. Site-to-site differ-
ences in the same anatomical region (e.g., forearm) are
reported by some (3) but denied by others (14). High
hydration values appear to exist on the forehead and
palms; lower values are observed at the abdomen,
thigh and lower leg (27). The hydration of contra-
lateral pairs of sites is generally indistinguishable (7)
except on the dorsum of the hands. Hence compara-
tive left-right studies of the same selected skin sites
are recommended to avoid skin site differences. The
exact site can be located as YO of distance from the
wrist in order to provide left-right comparisons. The
influence of body hair should be noted (24), especially
if comparison is made between sites that are not
contralateral. No significant differences have been re-
ported between males and females when matched by
age and tested on the same skin sites (7). The effect of
age should also be considered (7, 28).
Variables related to the instrument
DPM and Skicon readings are usually higher than
those of the Corneometer (2, 14). Furthermore, it
should be remembered that, in general, different
samples of a same instrument model can give differ-
ent readings (29).In general, it appears that the DPM
Electrical methods for assessment of SC hydration
and Skicon are more useful in assessing relatively
high water content levels and desorption kinetics,
whereas the Corneometer is more sensitive when as-
sessing relatively dry skin (20).
Variables related to the environment
Temperature and relative humidity are known to in-
fluence stratum corneum water content (30,31). Under
some conditions a linear relationship has been found
between ”capacitance” and relative humidity (27).
Therefore, room humidity should be kept constant, or,
in cases of variation of relative humidity, this vari-
ation should be dealt with by the statistical analysis of
variance. Some normalisation procedures have been
proposed (27). Seasonal variations due to alterations
in temperature and relative humidity occur (31-33),
and day-to-day and even diurnal variations are also
reported (14).
Testing With Electrical Methods
Introduction
Experimental design of in vivo studies is a large and
complex subject, and for optimal results a relevantly
experienced professional statistician should be con-
sulted before the protocol is finalized. Here it is only
possible to make a few general remarks and recom-
mendations.
The number of volunteers participating in the study
must be sufficient to permit adequate statistical analy-
sis. Measurements should be taken with an awareness
of the possibility of cyclical dependence upon time of
day or time of year. The actual time of day as well as
the date should always be recorded for each measure-
ment.
Cleansing the skin should be taken into account and
TABLE 1. Study designs with drawbacks and advantages to evaluate
moisturizing properties of topical products
STUDY DESIGNS
Single application (4-6 hrs)
Measurements start after 30 min from application
Predicitve for product efficacy
Easy standardization of environment
Effects of nonadsorbed material (’wiping’)
Repeated application (3-4 weeks)
Difficult standardization of climatic variables
Improvement of hydration and/or skin condition expected
Difficult discrimination between products
More subjects may be needed
All studies
Use always a reference control (i.e., glycerol in water)
Standardize skin sites and environmental variables
Combine different techniques with visual and tactile evaluation
129
 Berardesca et al.
considered as a variable (7). If it is necessary, when
testing different products, to make comparisons with
other instruments or laboratories and reduce differ-
ences related merely to instrumentation, it is recom-
mended that a reference material should also be
tested (e.g., 10% glycerol in water).
Single application as predicitve test (Table 1 )
In single application studies the time course of hy-
dration is monitored for a few hours after the appli-
cation of a product. This makes this design less sensi-
tive to long-term changes in the environment, which
may in turn enable better discrimination between
products (34, 35). Measurements are taken for a base-
line at t=O and then at fixed intervals between 4 and
6 h after product application.
In this experimental design, the interactions be-
tween the product, the stratum corneum and the
probe should particularly be taken into account, and
the first measurement should be taken at least 30 min
after product application. Indeed, application of
emulsions results in immediately raised instrument
readings (presumably due to water present in the for-
mulation), and this effect tends to decrease after a 10
to 15 min “evaporation phase” (36, 37). Application
of occlusive moisturizers, such as pure petrolatum
without water, results in a slower increase of water
content due to the occlusive interaction between the
lipids and the stratum corneum (”lipidization phase”
(36, 38, 39). Indeed, the effect of nonabsorbed cream
components on ”capacitance” values can be demon-
strated (38-41), particularly when these components
are removed from the the skin surface (”wiping ef-
fect”). Application of various humectants increases
“capacitance” levels, but removal after 1 or 2 h in-
duces an immediate decrease in instrument readings,
indicating that ”capacitance” values do not only re-
flect the ”capacitance” of the skin itself but also that
of the product (42). Indeed, occlusive products allow
a build-up of water under the film but decrease the
magnitude of the responses, whereas humectants at-
tract water and increase “capacitance” levels. The
wiping effect should be taken into consideration be-
cause it alters the interpretation of the study. The
single application test could be predictive.
Repeated application (Table 1 )
This is a more realistic test. Repeated application
studies are used to evaluate the improvement of skin
condition after repeated long-term use (generally a
few weeks) of the products to be tested. At least 8-12
h should elapse between the last application of the
product and the final measurements. Several repeated
130
application studies report improvement of skin con-
dition after 4 weeks of treatment (4345). Long-term
studies have the possibility that varying environmen-
tal influences could affect volunteers during treatment
(e.g., changes of climate or season during the experi-
ment), and this must be taken into account. Therefore,
more subjects may be needed to distinguish statisti-
cally between products of similar moisturizing poten-
tial.
Statistics
The statistical analysis of data obtained from a study
of electrical impedance or conductance in vivo should
be straightforward, provided the recommendations
outlined above have been followed. Again, the com-
ments here are intended merely as an introduction.
An experienced statistical practitioner, or at the very
least a textbook, should always be consulted for as-
sistance in interpreting the results.
A power test should be performed before starting a
study in order to enroll the adequate number of sub-
jects. When distribution of the data is not normal, it
may be desirable to apply data transformations to
achieve normality or to present the data with median
and confidence intervals. For normally distributed
data, analysis of variance (ANOVA) for repeated
measures can be used to compare different times and
(for short-term studies) the kinetics of moisturization.
Alternatively, non-parametric tests, such as that of
Friedman, may be used. For normal data, product
comparisons at the same time interval can be made
by an unpaired Student’s t test for two samples, or
ANOVA for three or more samples. Multiple sub-
group comparisons may be necessary to detect further
differences between products.
In long-term studies, initial values can be compared
to final values by a paired Student’s t test for normal
data, or a Wilcoxon signed rank test for non-
parametric data.
Many statistical packages running on personal com-
puters provide these capabilities.
Conclusions
Water plays an important role in the physical prop-
erties of the stratum corneum, and reliable quantifi-
cation of water in the corneum and its interaction with
topically applied products is essential for understand-
ing skin physiology and developing efficient skin care
formulations. Conductance can readily differ by four
orders of magnitude between dry and hydrated
samples (42), and thus offers an approach to assessing

stratum corneum water, even though the exact re-
lationship between instrument readings and actual
stratum corneum water content is complex and vari-
able, preventing accurate calibration and opening the
possibility of misinterpretation.
These and many similar considerations mean that
electrical approaches to assessing skin hydration can
give only relative indications, not absolute measures,
and even then only if great care is taken in the design,
conduct and interpretation of the test. False positive
and false negative results may arise if these ap-
proaches are used simplistically, but much valuable
information is available if they are designed and used
with due attention to their biophysical basis.
For these reasons we recommend that electrical ap-
proaches be used with the care that is appropriate to
the quality of information required from them; in par-
ticular, in standard studies, it is acceptable to use only
a single stimulating frequency, as do most commer-
cially available instruments, provided that it is poss-
ible to ensure an effective standardization of skin tem-
perature changes, sweat gland activity, skin sites,
water depth profile changes, etc. A statistically-justi-
fied number of volunteers should be employed and
the potential confounding variables should be iden-
tified and controlled whenever possible. The use of a
suitable control is essential because absolute cali-
bration that relates instrumental readings to actual
water content in vivo is not possible for present com-
mercial electrical devices. The only calibration pro-
cedure that may be useful is a relative calibration in
which instrumental measurements are taken at differ-
ent times on some assumedly constant reference ma-
terial. Research studies require an increased level of
sophistication, and therefore a multi-technique ap-
proach is required.
New methods, more experimental and expensive,
are emerging; they could improve upon the perform-
ance of electrical approaches (6), but for the forseeable
future electrical techniques will remain the more basic
and widespread methodology, and will remain useful
provided that their limitations are clearly understood
and experimental conditions are designed to over-
come the associated difficulties as much as possible.
Finally, it should be remembered that no single type
of instrumental test of skin is sufficient; other clinical
and instrumental assessments should always be used
to guard against errors of interpretation.
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Address:
Enzo Berardesca
Dept. uf Dernzatolugy
University of Pavia
lRCCS Policlinico S. Matteu
27100 Pavia
ltaly
E-nzail:berarde@ipv36.unipv.it