F.-J.

Wortmann1
M. Stapels1
R. Elliott2
The Effect of Water on the Glass
L. Chandra2 Transition of Human Hair
1
DWI at RWTH Aachen
University, Pauwelsstrasse 8,
D-52074 Aachen, Germany
2
Unilever Research,
Port Sunlight, CH63 3JW
Wirral, UK
Received 19 August 2005;
revised 25 November 2005;
accepted 8 December 2005
Published online 15 December 2005 in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/bip.20429

Abstract: The glass transition of human hair and its dependence on water content were deter-
mined by means of differential scanning calorimetry (DSC). The relationship between the data is
suitably described by the Fox equation, yielding for human hair a glass transition temperature of
Tg ¼ 1448C, which is substantially lower than that for wool (1748C). This effect is attributed to a
higher fraction of hydrophobic proteins in the matrix of human hair, which acts as an internal plas-
ticizer. The applicability of the Fox equation for hair as well as for wool implies that water is homo-
geneously distributed in a-keratins, despite their complex morphological, semicrystalline structure.
To investigate this aspect, hair was rendered amorphous by thermal denaturation. For the amor-
phous hair neither the water content nor Tg were changed compared to the native state. These
results provide strong support for the theory of a quasi-homogeneous distribution of water within
a-keratins. # 2005 Wiley Periodicals, Inc. Biopolymers 81: 371–375, 2006
This article was originally published online as an accepted preprint. The ‘‘Published Online’’ date
corresponds to the preprint version. You can request a copy of the preprint by emailing the
Biopolymers editorial office at biopolymers@wiley.com

Keywords: human hair; differential scanning calorimetry; water content; glass transition; Fox
equation

INTRODUCTION is a decisive factor for an analogous phenomenon,

The glass transition of keratin fibers and particularly
its dependence on water content is of distinct practi-
cal relevance for textile as well as for cosmetic appli-
cations. For wool textiles the glass transition plays a
major role for the understanding of the (undesirable)
phenomenon of wrinkling1,2 while for human hair it
namely, the (desirable) stability of the water wave in
human hair.
Various approaches have been made to determine
the humidity dependence of the glass transition tem-
perature Tg in wool by mechanical tests1 as well as by
differential scanning calorimetry (DSC).3,4 Though it
may be argued that, in view of its relevance for me-

Correspondence to: F.-J.Wortmann, University of Manchester,

School of Materials, P. O. Box 88, Manchester MQ60 1QD, UK;
e-mail: f-j.wortmann@manchester.ac.uk
Biopolymers, Vol. 81, 371–375 (2006)
# 2005 Wiley Periodicals, Inc.

371
372 Wortmann et al.
chanical properties, Tg should best be measured by
mechanical means, DSC is a fast and readily applied
method offering the additional advantage that con-
stant conditions of water content for a sample are
more easily maintained.
The objective of the current investigation was to
determine the glass transition temperature for human
hair and its dependence on water content by DSC as
well as to find a suitable analytical description for the
data. Furthermore, the investigation was extended to
thermally denatured human hair in order to contribute
to our understanding of the distribution of the water
in the morphological structure of human hair.
EXPERIMENTAL
All experiments were conducted on commercial, medium-
brown, untreated European human hair (Fa. Kerling, Back-
nang, Germany) provided in the form of tresses. From these
tresses, fiber snippets (length 2–4 mm) were obtained. All
experiments were conducted with a DSC-7 instrument
(Perkin Elmer) using large-volume, pressure-resistant,
stainless steel capsules (Perkin Elmer).
It is well known that the physical history of a glassy ma-
terial, namely aging and annealing, has a major influence
on the measurement of the glass transition by DSC, espe-
cially by inducing an excess enthalpy peak. Therefore, a
procedure was chosen to control the history of the hair as
far as experimentally feasible by establishing stable condi-
tions of the water content for the samples, on the one hand,
while avoiding overly strong effects of aging due to long
storage on the other. To preserve this status, all mea-
surements were furthermore limited to conditions of less
than 20% water content to ensure that Tg was well above
the storage temperature under ambient room conditions
(20–258C).
Samples of about 100 mg of hair snippets were stored in
a desiccator first over P2O5 for 24 h and then for four days
at 208C over a suitable, concentrated salt solution, provid-
ing a constant relative humidity. For the DSC test, 5–10 mg
of material was rapidly transferred into a DSC capsule;
10
L of silicone oil was added and the capsule sealed. The
silicon oil was added to suppress as far as possible water
desorption from the sample during the measurement, which
is considered the major source of certain experimental
problems, as particularly observed by Kure et al.4
The DSC was calibrated, prior to each series of experi-
ments, using indium and palmitic acid, both of high purity.
For the measurements an empty container without the O-
ring rubber seal was used as reference. The optimum heat-
ing rate was found to be 58C/min, providing adequate signal
intensity while being on the safe side with respect to nonli-
nearities of the measurement, which are expected at higher
heating rates.
The water content of hair was determined on parallel
samples by DSC. Conditioned hair snippet samples were
weighed into gas-tight aluminum pans. Prior to the mea-
surement, the lid of the pan was perforated. The sample
was heated to 1358C (208C/min) and held at that tempera-
ture for 30 min, then cooled to room temperature under a
flush of nitrogen. In view of the discussion of the dry
weight of keratin,5 this procedure was chosen because the
completion of the desorption process of water was readily
observed in the DSC curve.
Weighing the sample before and after desorption yielded
its water content, which is the weight percentage of water
in the conditioned sample. This parameter is not to be con-
fused with the term regain, which relates to the dry weight
of the sample.
To enable the measurement of the glass transition for
totally amorphous hair material, the crystalline -helical
structures were irreversibly denatured.6 To achieve com-
plete denaturation, hair fiber snippets were submitted to a
DSC measurement in water in the temperature range 80–
1708C (58C/min),6 beyond the denaturation peak at around
1508C. Neither on cooling nor in a further DSC experiment
in water was any indication of a reformation of -helical
material found. After cooling, the capsules were opened;
the denatured hair snippets were removed, dried, and stored
under ambient conditions until used in DSC experiments
for measuring Tg in the same way as the native hair mate-
rial.
RESULTS AND DISCUSSION
Figure 1 shows a typical DSC curve for human hair
with a water content of 15%. The temperature range
of the transition is given by the onset and the offset
temperature, respectively, giving a span of about
158C. Following the considerations for the definition
of Tg, as reviewed by Peyser7 and as prescribed in
DIN-53765,8 the glass transition temperature is
defined as the midpoint between the lower and the
upper level of the DSC curve. This definition of Tg is
expected to correlate well with mechanical data.7
Due to the experimental procedure, the peaks for
excess enthalpy were generally small in the curves so
that this determination could be maintained for all ex-
perimental curves without introducing a relevant bias
and without the need to take recourse to integral
methods for the determination of Tg, as proposed by
Richardson and Savill.9
For the comparison of Tg measurements of keratin
fibers by DSC, in general it is important to note that
this standard practice for the determination of Tg is at
variance with the work by Phillips,3 Kure,4 and
Huson10 for wool and Ota et al.11 for human hair.
They determined the onset of the glass transition in
the DSC-curve as Tg, a practice covered by ASTM
D3418.12
Biopolymers DOI 10.1002/bip

FIGURE 1 Typical DSC curve of human hair with 15%
water content (heating rate 58C/min). The graphical defini-
tion of the glass transition temperature is given.
An initial observation of the systematic change of
the glass transition of the keratin fiber wool with
water content was made by Wortmann et al.1 based
on measurements of torsional recovery of wool fibers,
corroborated by data from a variety of other tests. It
was shown that the data for water content between 9
and 20% were well described by the Fox equation13:
1=Tg ¼ w1 =Tg1 þ w2 =Tg2 ;
where w is the weight fraction and the subscripts 1
and 2 refer to dry wool and pure water, respectively.
Fox13 expected his equation to apply to systems
that are compatible and not too strongly polar. Equa-
tion (1) implies that the depression of Tg with solvent
concentration is fully explained by the Tg differences
of the constituents of the system, regardless of spe-
cific interactions. Features such as crystallinity or
crosslinking only play a role insofar as they affect dil-
uent distribution.14,15
For keratins, the size of the step in the DSC curve,
which is the change of heat capacity DCp at the glass
transition, is experimentally quite variable (0.03–
0.2 J g
1 K
1) and shows a mean of only about
0.1 J g
1 K
1 over the humidity range considered.
This is quite small compared to DCp  0.4 J g
1 K
1
found for other proteins.16 For this reason, the DSC
values for Tg for hair are somewhat more variable
than those obtained for the mechanical measurements
for wool.1 The size and variability of the change in
heat capacity at Tg furthermore constrained the use of
other multiparameter approaches, such as the Couch-
man–Karasz17 or the Gordon–Taylor18 equation.
Biopolymers DOI 10.1002/bip
Effect of Water on Glass Transition of Hair 373
To stabilize the fit of the Fox equation to the data
and for ease of comparison with wool, the Tg value
for water 125 K (
1488C) is taken from Ref. 1, yield-
ing for the dry hair Tg1 ¼ 417 6 3 K (6 standard
error), equivalent to 1448C.19 The solid line in Figure 2
shows the fit through the DSC results.
The value for the glass transition temperature of
amorphous solid water has been subject of consider-
able debate, as reviewed by Kalichevsky et al.20
There appears to be a general consensus, however,
that the Tg of water falls into the region between 130
and 145 K at normal DSC heating rates. Considering
the wide range for the extrapolation, the applied value
for the Tg of water is in acceptable agreement with
the literature data.
The glass transition temperature of hair (1448C) is
308C smaller than for wool (1748C). As a conse-
quence, the Fox equation curve for Tg lies systemati-
cally lower compared to wool as shown in Figure 2.
Looking at the shift between the curves in terms of
water content yields a value of 2–3%. Human hair thus
‘‘softens’’ at lower water content and temperatures
than wool. This is a rather unexpected result, because
human hair is generally considered more hydrophobic
than wool, due to a higher fraction of hydrophobic
high-glycine-tyrosine (HGT) proteins.21,22
The applicability of the Fox equation for the
human hair/water as well as for the wool/water sys-
tem,1 without the need to consider, e.g., crystallinity,
implies that water, despite the complex morphologi-
cal, semicrystalline structure of keratin fibers, acts as
if it is homogeneously distributed throughout the fiber
structure.
FIGURE 2 Glass transition temperature Tg vs. water
content for native (*) and amorphous (^) hair samples.
The curves relate to the fit of the Fox -equation for native
hair (solid line) and sheep’s wool (broken line, Ref. 1),
respectively. The lines are limited on the x-axis by the
anticipated maximum water content of the materials.
374 Wortmann et al.
In this context it is important to note that the
effects of water on a somewhat analogous polymer,
namely nylon 6, do not follow the Fox equation.23 In
nylon 6 as well as 66, water sorption drops with the
degree of crystallinity.24 The same principle seems to
apply for a variety of partly -helical proteins, such
as lactoglobulin, zein, and others.25
This does not apply for ovalbumin, silk fibroin,
and wool, for which Mellon et al.26 showed that even
severe treatments, that lead to denaturation of the
higher protein structures, have very little effect on
water sorption.
As early as 1959, Feughelman27 proposed a two-
phase filament/matrix model for the interpretation of
the mechanical properties of keratins. The axially ori-
ented, mechanically effective filaments have been
identified as the -helical material (30 vol %) in the
intermediate filaments (IF). The matrix comprises the
remainder of the morphological components, includ-
ing the nonhelical fraction of the IFs, the intermediate
filament associated proteins (IFAPs), which form the
classical27 matrix, as well as other morphological
components, such as cuticle, cell membrane complex,
nuclear remnants, etc. (70 vol %).28 It is well estab-
lished29,30 that water has a strong plasticizing effect
on the matrix phase, while the modulus of the fila-
ments remains unaffected by the uptake of water. In
this view there appears to be a conflict with the
assumption of effectively homogeneously distributed
water, as derived from the applicability of the Fox
equation.
Following previous investigations,1,3,10 Kure et al.4
again determined by DSC the glass transition tempera-
ture of wool as a function of regain. Applying the
implication of the two-phase model, they introduced
the hypothesis that water is only located in the matrix
phase of the fiber, while the filaments take up no water.
Consequently, they calculated the water content that
was relevant for the analysis of their Tg data for wool
on the basis of the matrix fraction only, thus introduc-
ing a bias compared to the usual values. The same bias
was subsequently introduced by Pierlot31 for the analy-
sis of the water sorption of wool. Such a bias would
obviously have rather far-reaching consequences
beyond representing just a calculational issue, affecting
the description and interpretation of a wide variety of
water-dependent properties of keratins such as mechan-
ical, thermal, sorption, etc.
To further investigate the question of the water
distribution in human hair, Tg measurements were
conducted on thermally denatured, amorphous hair. If
the crystalline phase (30%) is transferred into an
amorphous state, it is expected to absorb water simi-
larly as the matrix and thus increase the water content
at a given relative humidity. This would be expected
to lead to significant changes of the humidity depend-
ence of the glass transition.
However, the determination of water content for
native and amorphous hair, conducted in the context
of the Tg determination, showed no significant differ-
ences between the two materials. This experimental
result is in conflict with the hypothesis put forward
by Kure et al.4 and Pierlot,31 but in agreement with
the investigations by Mellon et al.26 The results for
Tg for amorphous hair are summarized in Figure 2.
The data fit well into the data set for the native hair,
showing that hair crystallinity has no effect on water
sorption and on Tg.
CONCLUSIONS
Hair takes up systematically less water than wool at
equal relative humidity and should thus be generally
less susceptible to the effects of water. In contrast to
this expectation, it is observed that the glass transition
for hair is systematically about 308C lower for hair
compared to wool. For both materials, the change of
Tg with water content is well described by the Fox
equation.
The decreased glass transition of human hair com-
pared to wool is attributed to a difference in composi-
tion of the matrix proteins. These contain a higher frac-
tion of hydrophobic HGT proteins in the case of hair,
which may induce a lower amount of hydrogen bond-
ing in the matrix that stabilizes the glassy state. These
proteins are thus acting as ‘‘internal’’ plasticizer.
That the partly -helical, native, and the denatured
amorphous hair shows the same dependence of Tg
with water content leads to the conclusion that water
is in fact quasi-homogeneously distributed in native
hair, where the adsorption to the surface of the -hel-
ical filaments is similar to the amount absorbed into
the matrix. Some insight into the interaction between
-helical proteins and water has been given by Manas
et al.32 on the basis of IR analysis. The special per-
formance of -keratin is attributed to the filamentous
arrangement of the -helical material in the IFs, lead-
ing to a large specific surface with good accessibility
for water.
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Reviewing Editor: Laurence Nafie