Application paper (2005) 1-5

EFFECT OF CENTRIFUGATION ON SAMPLE

HOMOGENEITY
H. Dihang1, P. Bru1, G. Meunier1, B. Pouligny2, P. Snabre2
Formulaction, 10 impasse Borde Basse, 31240 LUnion, France
2
Centre de Recherche Paul Pascal, avenue Albert Schweitzer, 33600 Pessac, France
1

Abstract
Centrifugation is a common method used in many industries to reduce the time for stability studies, as the acceleration field
leads to a quicker destabilization of the system. However, correlation with real shelf life of a colloidal system can prove to be
complex, as the stress applied to the sample is far from natural forces, which can be encountered in the normal life of a
product. This paper presents some data proving the detrimental effects of centrifugation on the destabilization process, as
important segregation between small and large particles is observed after centrifugation of the dispersion and not when the
sample is left at rest.
Keywords: stability, centrifugation, suspension, dispersion, shelf life, Turbiscan.

Introduction
Many industrial products available in the market
are in the form of emulsion or suspension. These
products are often complex due to the large number
of components in the formulation and their physical
stability is a critical parameter, which has to be
controlled thoroughly. Different techniques are
available to monitor the stability of such colloidal
systems and most of them offer the possibility to
accelerate the destabilization phenomena in order
to obtain results more quickly. Two methods are
commonly used to accelerate the destabilization
process: temperature and centrifugation. The
scientific community is regularly debating whether
these methods are realistic and can be correlated to
real shelf life of a product.
In this paper, experiments show evidences that
centrifugation
causes
segregation
among
polydispersed particles, which is not observed when
the sample is left still for the stability analysis. This
effect is highlighted using simple fluorescence
imaging experiments. The mechanism involved in
this process involves the jamming properties of the
percolation network, which can be applied to most
of the gel-like structures, corresponding to many
industrial products (cosmetic creams, toothpaste,
yogurt, cheese, paint, etc.).
Gel structures are between liquid and solid
states. The gel transition, or so-called sol-gel
transition involves a percolation threshold and
jamming transition, whereby a continuous path
between particles starts to form (as in Figure 1 for
weakly interacting PMMA particles). The jamming
transition can be caused either by an increase in
particle density (concentrated emulsion or
suspension) or physico-chemical interactions

between particles leading to a restriction of particle

motion in the dispersion.

Figure 1. Microscopy image of 10m 40m PMMA

beads in a viscous fluid.
The gelation network greatly influences the
rheology of the dispersion and leads to the
existence of a yield stress. The dispersion remains
static for low mechanical stress, but flows when the
stress exceeds the yield stress. This is the typical
example of toothpaste, which remains in the tube as
long as one does not press on it, the yield stress
corresponding to the stress necessary for the
toothpaste to flow out of the tube.

Experimental
1. Materials
The suspensions studied in this work are
composed of PMMA beads (density 1.19 g/cm3)
dispersed in a refractive index-matched fluid. The
fluid is a mixture of microscope oil immersion
(Sigma-Aldrich S150) and n-hexadecane (viscosity
25cP, density 0.88 g/cm3). The suspension is
transparent and therefore can be studied by optical

Application paper (2005) 1-5

methods. A first system is composed of a mixture of

180m 200m (volume fraction b=0.25) and
10m 40m PMMA beads (volume fraction
s=0.25) dispersed in the viscous fluid. A second
system consists in polydispersed small 10m
40m PMMA beads (volume fraction =0.25 or
=0.5), also immersed in the same fluid (Figure 1).
The suspension is put in a glass cell (11 mm
internal diameter, 8 cm sample height), which can
later be centrifuged. Particles are heavier than the
fluid and settle in the cell.
Small particles from the same batch (10m
40m PMMA beads) are made fluorescent by
impregnation of an organic dye. Tracers are dyed
with Rhodamine B and excited with the 514 nm line
of an Argon ion laser.
The samples were centrifuged using a sorvall
machine (Dupont instruments, average distance
between sample and axis of rotation 15cm).
Samples were tested under three average
accelerations : 10g, 150g and 1000g.
2. Fluorescence imaging
The samples were analyzed by fluorescence
imaging. A laser beam shaped as a sheet
illuminates a vertical plane across the glass cell.
The fluorescence of PMMA labelled beads is
observed through a filter to view the vertical
repartition of small tracers (volume fraction 0.005).
Quantitative concentration fields of species can be
obtained from maps of fluorescence intensities.
3. Turbiscan technology
The heart of the optical scanning analyser,
Turbiscan, is a detection head, which moves up
and down along a flat-bottom cylindrical glass cell
(Figure 2). The detection head is composed of a
pulsed near infrared light source ( =880 nm) and
two synchronous detectors. The transmission
detector (at 180) receives the light, which goes
through the sample, while the backscattering
detector (at 45) receives the light scattered
backward by the sample. The detection head scans
the entire height of the sample, acquiring
transmission and backscattering data every 40 m.
The Turbiscan LAb can be thermo-regulated from 4
to 60C and linked to a fully automated ageing
station (Turbiscan ags) for long-term stability
analyses.

Results
1. Settling at rest of PMMA suspensions
When the suspension is left to settle at rest,
the following fluorescence images are obtained
(Figure 3).
t=0

t=3 hours

t=10 hours

t=24 hours

Figure 3. Fluorescence of small labelled particles

during the settling at rest of bidispersed PMMA
beads (b=0.25 and s=0.25).
The

fluorescence

field

shows

no

significant

segregation of the particles during the settling

process. After three hours, a weak increase in the
small particle concentration becomes visible at the
fluid / suspension interface. The small particle
concentration profiles (Figure 4) indeed show a
narrow peak at the top of the sample.

t=0
t = 3h
t = 24h

z (cm)
Figure 4. Small particle concentration profiles along
the vertical direction during the settling of a
bidispersed PMMA suspension. Z=0 denotes the
vertical position of the air/fluid interface.
Looking at the second system consisting of
small polydispersed PMMA beads, at 25% and 50%
volume fraction, similar results are obtained (Figure
5), showing uniform fluorescence field during
particle settling.

Figure 2. Principle of Turbiscan measurement

The Turbiscan makes scans at various preprogrammed times and overlays the profiles on one
graph in order to show the destabilisation.

Application paper (2005) 1-5

=25%

t=0

t=3 days

=50%

t=0

t=3 days

Figure 5. Fluorescence images of small labelled

particles during the settling, at rest, of small
polydispersed PMMA beads.

of the sample is highly concentrated in small

particles (s=0.5-0.55). For higher accelerations
(150g or 1000g), a third phase appears at the very
top of the sample, showing strong fluorescence.
This phenomenon is due to the polydispersity of
small PMMA beads (10m - 40m), since the small
sized fraction segregates to the top.
Similar results are obtained with suspensions of
small polydispersed PMMA beads, showing a higher
concentration of the smallest particles at the top.
We only show the concentration profiles (Figure 8
and 9).

s
G=0
G= 150g

2. Settling under centrifugation

On the other hand, fluorescence images
obtained after centrifugation of the bidispersed
sample (Figure 6) show spectacular segregation
effects even for relatively small acceleration (10g).

z (cm)

G=
G=10g
G=150g
G=1000g
Figure 6. Fluorescence images of small labelled
particles during the settling under centrifugation of
bidispersed PMMA beads (b=0.25 and s=0.25).

Figure 8. Small particles concentration profiles

along the vertical direction during the settling of 25%
polydispersed PMMA beads (10m-40m) under
centrifugation.

s
G=0
G= 150g

Centrifugation leads to vertical segregation of

particles and formation of a highly fluorescent top
layer of small PMMA beads. Segregation occurs as
a result of small particle transport in the fluid
backflow and is particularly visible when looking at
the small particle volume fraction profiles (Figure 7).

G=0
G= 10g
G=150g
G= 1000g

z (cm)
Figure 9. Small particles concentration profiles
along the vertical direction during the settling of 50%
polydispersed PMMA beads (10m-40m) under
centrifugation.

z (cm)
Figure 7. Small particles concentration profiles
along the vertical direction during the settling of a
bidispersed suspension under centrifugation.
The blue curve, corresponding to 10g acceleration
show two regions : in the bottom part of the tube,
the smallest particles partly occupy the free space
between large particles (s=0.2) while the top part

3. Turbiscan analysis
The bidispersed PMMA beads suspension has
also been studied with the Turbiscan LAb at 30C
for 2 days. The following profiles are obtained in
transmission (Figure 10).

Application paper (2005) 1-5

Fluid/sediment
interface layer
Clarification

Centre part of
the top layer
sediment

Figure 10. Turbiscan analysis of small labelled

particles during the settling at rest of bidispersed
PMMA beads (b=0.25 and s=0.25).
It shows typical sedimentation, with an increase of
the transmission at the top, characteristic of a
clarification, and a decrease of the transmission in
the rest of the sample, due to an increase of
concentration. The vertical shape of the profiles,
indicates that no segregation is visible. If
segregation was taking place the clarification phase
would display distinct peaks for the different
population of particles.
The kinetics of sedimentation can be plotted
(Figure 11) in order to quantify the extend of
particles migration.

Figure 11. Kinetics of clarification of small labelled

particles during the settling at rest of bidispersed
PMMA beads (b=0.25 and s=0.25).
It shows that the sedimentation is finished after 10
hours. The slope of the curve indicates the velocity
of migration (1.86mm/h).

Discussion
The segregation phenomenon, which is
observed under centrifugation is corroborated by
microscopy analysis of samples from different layers
of the sediment (Figure 12).

Bottom part of the

sediment

Figure 12. Microscopy images of PMMA samples

from different layers of the sediment after
centrifugation of the bidispersed suspension (150g).
It highlights the issue of using centrifugation to
accelerate destabilisation phenomena. Even with
small acceleration (10g) the particles undergo
important segregation, with the smallest particles
going to the top and the largest ones remaining at
the bottom. At rest, no segregation occurs during
particle settling as observed with fluorescence and
Turbiscan analyses.
In order to understand the influence of
centrifugation upon the mechanical stability of the
particle network, rheological experiments have been
performed for bidispersed suspensions. These
experiments indicate a yield stress behaviour in
relation with weak attractive interactions between
small PMMA particles. Microscopy images of small
PMMA beads indeed show the formation of
reversible flocs (Figure 1). Above the percolation
threshold of small particles, the yield stress of the
bidispersed suspension increases from 0.1 Pa for
s 0.15, up to a few Pa for high volume fraction s
=
0.45.
For
suspensions
settling
under
centrifugation, the mechanical input is enough to
break the percolation network and allow segregation
effects. Even with small acceleration, the
mechanical stress exerted upon the particle network
exceeds the yield stress of the suspension, which
contributes in reducing the life time of the
percolation network.
The segregation phenomenon is not observed
when the sample is left to settle naturally because of
the jamming state of the percolation network. The
slight segregation, which can be observed at rest
results from a bleeding process, well known in the
concrete industry. Local and transient failures of the
percolation network result in local fluid upflows
conveying the smallest particles to the top of the
suspension.

Conclusion
This study proves the issue of using
centrifugation to accelerate the destabilization
process of emulsions or suspensions. Excess

Application paper (2005) 1-5

mechanical stress exerted upon the sample can

weaken the percolation network and destroy the
jamming state of the suspension microstructure,
then
leading
to
segregation
phenomena.
Segregation effects are less likely observed when
the sample settles at rest, which is the normal life of
most of the industrial products available on the
market. This paper only focuses on the influence of
centrifugation upon segregation, but other effects
can also be observed, such as coalescence of
emulsion droplets when using this method of
destabilization acceleration.
Using the Turbiscan technology, the physical
stability of the sample is analyzed in real condition
(no mechanical stress)and results can be correlated
to the normal shelf life of the studied formulations.