International Journal of Cosmetic Science, 2009, 31, 119–129
Quartz plates for determining sun protection in vitro
and testing photostability of commercial sunscreens
J. Akrman*, L. Kubáč*, H. Bendová , D. Jı́rová  and K. Kejlová
*VUOS Research Institute of Organic Syntheses, Rybitvı́ 296, 53354 Pardubice 20 and  National Institute of Public
Health (NIPH), Šrobárova 48, 100 42 Prague 10, Czech Republic
Received 3 June 2008, Accepted 4 September 2008
Keywords: quartz testing plate, SPFin vitro, sunscreens, sun protection factor, UVAPFin
Synopsis
Testing plate made of optical quartz has been devel-
oped for the purpose of determination of sun protec-
tion factor (SPF)in vitro by the method of diffusion
transmission spectroscopy; the plates were coars-
ened by sanding and grinding to surface roughness
values (Ra) of 18 lm. The plate was coated with a
film of sunscreen by an application of 2 mg cm)2
as that used for determination of SPFin vivo by
the COLIPA method. The transmission values
measured were converted into the SPFin vitro
and the protection factor in ultraviolet A light,
UVAPFin vitro. The testing plate was tested with com-
mercial sunscreens. The found values of SPFin vitro
fit well with the values determined by means of the
COLIPA method in vivo. The plates coated with
sunscreen film were irradiated with light simulating
the sun radiation. The values of protection factors
obtained before and after irradiation were com-
pared, and the differences were used for estimation
of photostability of the UV filters included.
Résumé
Des plaques pour test en quartz optique ont été
développées dans le but de déterminer in vitro le
facteur de protection solaire par la méthode de
spectroscopie en diffusion transmission. Les pla-
ques ont été érodées par sablage et polissage pour
obtenir des surfaces de rugosité Ra de 18 lm. La
Correspondence: Jiřı́ Akrman, VUOS Research Institute of
Organic Syntheses, Rybitvı́ 296, 53354, Czech Republic.
Tel.: +420 466823137; fax: +420 466822971;
e-mail: jiri.akrman@vuos.com
ª 2009 The Authors. Journal compilation
ª 2009 Society of Cosmetic Scientists and the Société Française de Cosmétologie
doi: 10.1111/j.1468-2494.2008.00482.
 
vitro
plaque a été revêtue avec un film d’écran solaire à
raison de 2 mg cm)2, comme il est coutume de le
faire pour déterminer un facteur de protection
solaire SPFin vivo par la méthode du COLIPA. Les
valeurs de transmission mesurées ont été conver-
ties en facteur de protection solaire SPFin vitro et en
facteur de protection dans l’UVA, UVAPFin vitro.
La plaque a été testée avec des écrans solaires
commerciaux. Les valeurs de SPFin vitro trouvées
correspondent bien aux valeurs déterminées par la
méthode in vivo du COLIPA. Les plaques revêtues
avec les films d’écran solaire ont été irradiées avec
une lumière simulant les radiations solaires. Les
valeurs des facteurs de protection obtenues avant
et après irradiation ont été comparées et les diffé-
rences ont été utilisées pour estimer la photostabi-
lité des filtres UV inclus dans les formules.
Introduction
The method of determination of sun protection fac-
tor (SPF) of sunscreens ‘in vivo’ [1] in a group of
volunteers is based on irradiation of skin by fil-
tered light from a xenon discharge lamp for a per-
iod of time necessary for inducing reddening of the
skin (the minimum erythema dose – MED). The
SPF value of a product is defined as the ratio of
the MED on product protected skin to the MED on
unprotected skin in the same subject.
The application of spectrophotometric mea-
surements of transmission to the determination of
SPFin vitro is based on the fact that the SPF is a rela-
tive number. For example, if a layer of sunscreen
transmits one-third of incident ultraviolet radiation,
then it is assumed that the sunscreen has SPF = 3.
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Quartz plates for determining sun protection factors
The SPFin vitro is calculated from the transmission
measurements using the following expression [2]:
R
400nm
S  E  
290nm
SPF ¼
R
400nm
S  E  T  
290nm
where Sk is spectral radiation of sunlight at partic-
ular conditions, e.g. South Europe in the middle of
summer at noon.
Ek is relative effectiveness of ultraviolet radiation
(relative harmfulness) at the wavelength k for for-
mation of delayed erythema on human skin.
Kk is the wavelength step.
Numerical values of Sk and Ek are known [2].
Hence, weighted values of transmission are used
with regard to the harmfulness and intensity pro-
portion of radiation of a particular wavelength.
In analogy, newly used is the measurement of
transmission in the wavelength range of 320–
400 nm for determination of protection factor
in vitro in UVA region, i.e. the protection factor
UVAPF [3].
The UVAPF protection factor in vitro is calcu-
lated from the transmission values by means of
the following equation:
R 400nm
320nm P  I  
UVAPF ¼ R 400nm
320nm P  I  T  
where Pk = persistent pigment darkening action
spectrum, relative number (Pk = 1 for 320 nm).
Ik = spectral radiation obtained from UV source
(320–400 nm), W m)2 nm)1.
Tk = transmission of sunscreen sample at the
wavelength k.
Kk = wavelength step.
The spectrophotometric measurement is
demanding with regard to the precision of appara-
tus, for e.g. a sunscreen with a protection factor of
20 transmits only 5% of incident radiation. It is
necessary to use a spectrophotometer with integra-
tion attachment (Ulbricht sphere), which detects
not only direct radiation but also the scattered one.
Strictly speaking, the development of method for
determination of SPFin vitro consists of finding a
suitable substrate for spreading the tested sun-
screen. Another parameter that can be chosen is
the amount of the sunscreen applied. The in vivo
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120
J. Akrman et al.
COLIPA method uses 2 mg sunscreen per 1 cm2
skin. Human skin has a jagged topography, and
the substrate for measuring SPF should also have
jagged surface with projections and depressions.
The substrate recommended in literature is the
ð1Þ
porous surgical band Transpore 3M with the sun-
screen dose of 2 mg cm)2 [2]. Porous substrates
are covered with a film of sunscreens with frac-
tions of different optical thickness instead of a film
with homogeneous thickness. SPF calculated from
transmissions is lower in the case of porous sub-
strates [4]. Other recommended substrates are
polymethylmethacrylate plates or quartz glass
plates coarsened with carborundum powder to a
roughness of 3–7 lm with applied sunscreen dose
of 0.8 mg cm)2 [5]. Very high values of SPF are
obtained when using amount rate of sunscreens of
2 mg cm)2. Also recommended are polymethyl-
methacrylate plates matted by blasting with glass
ballotini. A roughened plate with a homogeneous
layer of glycerol was used as a reference substrate
[3, 5, 6].
This paper deals with development of substrate
for spectrophotometric determination of SPFin vitro
of sunscreens. Its first part describes modification
of surface of quartz plate by sanding and grinding,
which enables application of sunscreens at an
amount of 2 mg cm)2. The prepared quartz plate
was tested with 20 commercial sunscreens. The
plates coated with the tested sunscreen were irra-
ð2Þ diated with artificial sunlight, whereupon changes
were determined in effectiveness of protection pro-
vided by the sunscreens, which were because of
photodegradation.
Materials and methods
Preparation of quartz testing plates
Experiments on treatment of surface was per-
formed on optical quartz plates with dimensions
of 10 cm · 10 cm · 2 mm. Grinding of quartz
plate with carborundum powder was performed
by spreading the powder on the plate surface and
moistening, whereupon the grinding itself was
carried out by means of circling movement of an
applied glass plate. The grinding of quartz plate
with coarse carborundum powder is not a suit-
able technique. The coarse-grained carborundum
powder is brittle and at the same time, quartz is
very hard, and the grinding necessitates applica-
tion of a certain pressure. The uniformity of
ª 2009 The Authors. Journal compilation
International Journal of Cosmetic Science, 31, 119–129
Quartz plates for determining sun protection factors
ground surface is not perfect. Inspection of plate
by magnifying glass reveals locally less roughened
places.
In cooperation with Development Optical Work-
shop of Czech Academy of Sciences in Turnov, we
chose the method of blasting of grinding agent by
means of a stream of compressed air. From among
the tested grinding agents – corundum powder,
glass-making sand and two types of steel detritus –
the optimum result was obtained with the glass-
making sand. Blasting of quartz sand produces
holes of 40–60 lm diameters in the surface. The
inner surface of the holes is not smooth: micro-
scopy reveals radial scratches of 3 lm diameter.
However, at some spots also larger holes are
formed with smooth inner surface, with a diameter
up to 300 lm. Steel detritus produced on a surface
that is highly irregularly devastated. Plate blasted
with corundum powder was uniform, but slightly
roughened with valleys of 3–7 lm.
The sand-blasting operation was performed with
a blasting apparatus TB90 made by Toušeňská
Ltd. (Turnov, Czech Republic): the nozzle distance
was 25 cm, its diameter was 12 mm and the air
pressure was 400 kPa. The grain size of sorted
glass-making sand was 0.3–1.0 mm. As quartz is
very hard, there was a risk of breaking the quartz
plate. Therefore, the quartz plate to be ground was
affixed on a glass plate and protected on sides with
glued glass plates.
Besides grinding we also used the technique of
etching with hydrofluoric acid. The hydrofluoric
acid of 20% or 40% concentration applied to
sand-blasted plate gradually smoothened back its
surface. Therefore, we used an etching mixture
composed of ammonium fluoride in 90% sulphuric
acid, which is used in glass industry for refining
sand-blasted glass surfaces. This etching acid pref-
erentially dissolves sharp edges. Microscopic exami-
nation of the surface showed that the etching with
acid smoothened sharp projections as well as
scratches on the surface of depressions. The surface
of sand-blasted plate exhibits sharp projections.
These sharp peaks were carefully removed by
means of emery paper. This after-grinding trans-
formed the sharp peaks into small facets, which
were not smooth, but having 3 lm scratches.
Plates were tested with one commercial sun
cream. Testing cream was selected for its
SPFin vivo = 18 as well as for the content of UV fil-
ters commonly used in sunscreens: 6% ethylhexyl
methoxycinnamate (EHMC), 1% butyl methoxy-
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International Journal of Cosmetic Science, 31, 119–129
J. Akrman et al.
dibenzoylmethane (BMDBM) and 2% titanium
dioxide (data from producer). Four plates were
prepared for testing: 1 – sand-blasted and finely
after-ground, 2 – sand-blasted, 3 – sand-blasted
and etched, 4 – smooth plate.
An amount of cream applied to plates was
searching at which SPF measured gives value 18,
in accordance with SPF in vivo. The smooth quartz
plate gives very steep dependence of amounts vs.
protection factor. The sand-blasted plate provides
the SPF values that are still very high although
there is a considerable shift when compared with
the smooth plate (curves 2 and 4 in Fig. 1). The
fine after-grinding of sand-blasted plate with emery
paper leads to further lowering of the measured
SPF values of the sunscreen tested. On the other
hand, the etching of sand-blasted plate leads to an
increase of SPF (curves 2 and 3 in Fig. 1).
Following procedure was adopted to monitor the
stepwise decrease of measured SPF because of
grinding with emery paper. The plate was weighed
and spectrophotometric baseline was determined
for film of glycerol, 2 mg cm)2. Plate was cleaned
in ultrasonic water bath and protection factor for
quantity of cream, 2 mg cm)2, was determined.
After cleaning the cream, the plate was carefully
ground with emery paper. Then the plate was
cleaned again from quartz powder and a loss of
the weight because of grinding was determined.
Glycerol was applied to the plate and new baseline
was set up. The plate was cleaned and an amount
Figure 1 Dependence of amount rate of sunscreen milk
on protection factor in vitro, mean value from 16
measurement spots and standard deviation. Testing plate:
1 – sand-blasted and finely after-ground, 2 – sand-
blasted, 3 – sand-blasted and etched, 4 – smooth plate.
SPF, sun protection factor.
121
Quartz plates for determining sun protection factors
Figure 2 Step-wise grinding of plate with emery paper
followed by decrease in protection factor and applied
amount of cream, 2 mg cm)2. SPF, sun protection factor.
)2
of 2 mg cm cream was spread on the plate. SPF
was measured and found to be lower than before
grinding.
These procedures were repeated: gradual grind-
ing resulted in stepwise decreases in the SPF value
measured. The changes in SPF values measured
depending on the way of processing are shown in
Fig. 2. Grinding of 73 mg of quartz led to decrease
in measured SPF from a value of 39 to 26. Fig-
ures 3 and 4 presents a micrograph of the plate
that was sanded and ground with emery paper
(darker areas are valleys). This roughened quartz
plate was prepared for testing commercial sun-
screens.
Determination of surface roughness (Ra) was
conducted on plate using portable tester (Surftest
SJ-201; Mitutoyo Corp., Kawasaki, Japan). The
average surface roughness, Ra = 17.8 ± 1.2, peak
Figure 3 Micrograph of surface of quartz plate, magnifi-
cation 160·.
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122
J. Akrman et al.
Figure 4 Micrograph of surface of quartz plate, magnifi-
cation 64·.
to valley height surface roughness, Ry = 104.2 ±
20.5 and root mean square roughness, Rq =
22.9 ± 2.4, were assessed at six random spots in
accordance with ISO standard 4287.
Spectrophotomeric method of measurement
The measurement of transmission was carried out
with a spectrophotometer Varian Cary 50 Sola-
screen (Varian Inc., Mulgrave, Australia) equipped
with extension Varian Fabric Protection in the
range of wavelength from 290 to 400 nm in steps
of 5 nm. The spectrophotometer extension is desig-
ned for measuring the SPF of textiles and sun
glasses [7, 8]. The spectrophotometer has a hori-
zontal table on which the plate with sunscreen is
placed. The plate can easily be moved in horizontal
directions, which allows transmission measure-
ments at various spots of the plate. With one
10 cm · 10 cm plate, 16–25 measurements on dif-
ferent spots can be made. The table is springy and
presses the plate at the beginning of the integration
sphere, which is located in an arm above the table.
Application of sunscreen cream to testing plate
The procedure of application, spreading and drying
the cream film has a large effect upon the accu-
racy of the SPF measurement. It is necessary to
use a 10 cm · 10 cm plate. With a smaller plate,
e.g. 4 cm · 4 cm, it is difficult during the spread-
ing to achieve uniform film thickness in the middle
and at the edges of the plate. The plate is placed
on lab scales, and the cream is spotted from a
glass rod uniformly over 25 dots on the plate (the
pattern formed by 5 · 5 rows of dots). The
ª 2009 The Authors. Journal compilation
International Journal of Cosmetic Science, 31, 119–129
Quartz plates for determining sun protection factors
procedure must be quick as water readily evapo-
rates from the cream and the weight of plate with
cream decreases. The cream is spread with a circu-
lar finger motion using a slight pressure detected
by the scales as a value of about 15 g. Then the
cream is spread by transversal movements using a
stronger pressure in a stream of warm air (40–
45C) from hair drier. The spreading is continued
for about 20–50 s, until it is visually evident that
the emulsion has broken. After drying, the plate is
weighed again to determine the approximate value
of dry matter in the cream.
In our experience, about 10 mg cream remains
on the finger, the amount being the same with
adopting the rubber finger cot. Therefore, for
achieving, e.g. the cream amounts of 2 and
1.2 mg cm)2 on the 10 cm · 10 cm plate, the
necessary amounts weighed are 210 and 130 mg,
respectively. Alternatively, the weighed amount of
cream can also be spread with a finger that was
saturated with the cream by rubbing the index fin-
ger against thumb.
The baseline for photometric measurement of
transmission is adjusted by means of a plate with
glycerol film of 2.0 mg cm)2. Before reuse, the
plates are cleaned with brush and warm water
containing a detergent, whereupon it is necessary
to remove the residues of UV filters (particularly
titanium dioxide) in an ultrasonic bath containing
a detergent (50C for a period of 15–40 min).
Finally, the plate is rinsed with water and ethanol
from washing bottle and dried.
Procedure of measurement of SPFin vitro using
quartz plate
The given procedure was developed for all the
measurements performed on the testing quartz
plates.
1 Transmission measurement of pure plate at 16
spots. The purity check of the surface serves the
purpose of detection of potential residues of tita-
nium dioxide or other UV filters from previous
measurement. The transmission of pure plate at
various spots is 72–75% for the radiation of
290 nm wavelength (referred to the plate with
glycerol; the curve is compared with that stored
in the PC memory from previous measurement).
2 Adjustment of baseline (the transmission of
100%) using a plate with glycerol film of
2 mg cm)2 at a selected spot on the plate and
adjustment of baseline with blocked transmission
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International Journal of Cosmetic Science, 31, 119–129
J. Akrman et al.
of light through the sample (the transmission of
0%) using the black plate delivered with the
instrument. Testing of uniformity of surface of
the plate at 16 additional spots. The transmis-
sion at the wavelength of 290 nm is 98–100%.
It is useful to label spots with different transmis-
sion on the plate.
3 Measurement of plate with film of tested sun-
screen. Figure 5 presents the diagram from the
measurement of sun milk 15 (Table I) displayed
by means of the software Cary WinUV version
3.00 supplied with the spectrophotometer
Varian. The transmission measurement was
carried out at 16 different spots on the plate.
The mean value of the determined factor
was SPFin vitro = 16.4 (SD of 0.8). The value
in vivo of this sunscreen was also determined:
SPFin vivo = 15.0. The UV filters present in this
sun milk are photostable; the protection factor
does not decrease on irradiation.
The data of values wavelength vs. transmission
are transferred into an Excel spreadsheet specially
arranged for calculation of the values of SPF,
UVAPF, SPF/UVAPF ratio, UVA/UVB ratio and
critical wavelength. The calculated values are dis-
played directly, except for the value of critical
wavelength, which is read manually from the dis-
played graph of absorbance area integral vs. wave-
length for 90% of area integral.
Besides the protection factor UVAPFin vitro, the
protection in UVA light is expressed also by the
critical wavelength and by the UVA/UVB ratio [9].
Figure 5 Graph from measurement of sunscreen 15,
check of surface, plate purity and transmission measure-
ments. 1 – pure plate against plate with glycerol film.
2 – plate with glycerol film. 3 – plate with sunscreen film
against plate with glycerol film.
123
Quartz plates for determining sun protection factors J. Akrman et al.

Table I Characterization of sunscreens

Sample Declared Dry matter

No. SPF value content [%] Active ingredients INCI names

1 40 42 Titanium dioxide, methylene bis-benzotriazolyl tetramethylbutylphenol, octocrylene, ethylhexyl

triazone, bis-ethylhexyloxyphenol methoxyphenyl triazine
2 25 45 Titanium dioxide, ethylhexyl triazone, ethylhexyloxyphenol methoxyphenyl triazine
3 16 38 Ethylhexyl methoxycinnamate, titanium dioxide, diethylamino hydroxybenzoylhexyl benzoate
4 8 33 Ethylhexyl methoxycinnamate, diethylamino hydroxybenzoylhexyl benzoate
5 8 100 Ethylhexyl methoxycinnamate, butyl methoxydibenzoylmethane
6 12 32 Octocrylene, titanium dioxide, bis-ethylhexyloxyphenol methoxyphenyl triazine,
butyl methoxydibenzoylmethane
7 12 100 Ethylhexyl methoxycinnamate, butyl methoxydibenzoylmethane, titanium dioxide, zinc oxide
8 12 26 Ethylhexyl methoxycinnamate, titanium dioxide, methylene
bis-benzotriazolyl tetramethylbutylphenol
9 8 23 Ethylhexyl methoxycinnamate, benzophenone-3
10 12 40 Ethylhexyl methoxycinnamate, butyl methoxydibenzoylmethane
11 14 12 Ethylhexyl methoxycinnamate, ethylhexyl dimethyl paraminobenzoic acid,
butyl methoxydibenzoylmethane, methylbenzylidene camphor
12 16 38 Ethylhexyl methoxycinnamate, ethylhexyl triazone, butyl methoxydibenzoylmethane
13 18 30 Ethylhexyl methoxycinnamate, titanium dioxide, butyl methoxydibenzoylmethane
14 20 32 Ethylhexyl methoxycinnamate, methylene bis-benzotriazolyl tetramethylbutylphenol,
titanium dioxide
15 15 50 Octocrylene, titanium dioxide
16 15 40 Titanium dioxide, zinc oxide
17 25 38 Ethylhexyl methoxycinnamate, ethylhexyl triazone, titanium dioxide,
butyl methoxydibenzoylmethane
18 15 43 Ethylhexyl methoxycinnamate, methylbenzylidene camphor, butyl methoxydibenzoylmethane
19 40 72 Titanium dioxide
20 40 70 Titanium dioxide

SPF, sun protection factor.

These values are calculated from the area below
the absorbance curve (after converting transmis-
sion into absorbance: A = )log T). The UVA/UVB
ratio is the ratio of the areas under absorbance
curve in the UVA range (320–400 nm) and the
UVB range (290–320 nm), each area divided by
the range of wavelengths involved. The critical
wavelength means the wavelength up to which
the sunscreen can be considered effective. A wide-
spectrum sunscreen is the one having the critical
wavelength of 370 nm.
Measurement of photostability of sunscreens
It is known that some UV filters, such as BMDBM
or EHMC, present in sunscreens undergo photode-
gradation within several minutes in sunlight [10,
11]. The spectrophotometric method of determina-
tion of SPFin vitro finds the initial SPF. Quartz plates
enable measurements of protection factor of sun-
screen after irradiation in a suitable apparatus.
Hence, measurement of SPF can be used for test-
ing photostability of sunscreens.
The testing plates with sunscreen were irradiated
in an apparatus Q-Sun Xenon Test Chamber (Q-Sun
Laboratories, Cleveland, OH, USA). The instrument
is equipped with a xenon discharge lamp, and the
emitted radiation passes through a daylight filter.
Intensity of radiation was measured by built-in radi-
ometer at 340 nm. Supplied calibration radiometer
RC-20 also measures radiation at wavelength of
340 nm. In standard operation Q-Sun simulates
daylight in summer around noon at intensity of
0.68 W m)2 nm (340 nm). The working area is
cooled by means of ventilators. The instrument does
not allow water cooling of sample tray, so appara-
tus does not allow working at temperatures close to
human skin and does not meet requirements of UV
source as defined in COLIPA Guideline 2007 [3]. To
decrease thermal exposition of samples, the mea-
surement was conducted at intensity of radiation of
0.51 W m)2 nm (340 nm). The temperature of

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124 International Journal of Cosmetic Science, 31, 119–129
Quartz plates for determining sun protection factors J. Akrman et al.

working area was 53C and the temperature of
black panel was 70C. The quartz plate was placed
on a glass plate of 2 mm thickness to prevent poten-
tial irradiation of the sample from below by the UVB
light reflected from the sample tray. Time of irradia-
tion was fixed for all samples at 3 h and 45 min
(dose around of 15 MED).
Results
Measurement of sun protection factor in vitro of
20 commercial sunscreens
Tables I–V present the data of evaluation of sunsc-
reens that were commercially available in the
Czech Republic in 2006. Table I presents composi-
tion of its UV filters and the SPF value given on
the package. Sunscreens were tested in vivo in a
group of volunteers using the COLIPA method.
The SPF for the product was calculated as the
arithmetic mean of all individual SPF values
obtained from all subjects in the test. Each sample
was tested in a group of 14 volunteers (age 18–
60, gender female : male = 3 : 1, phototype II–IV).
The data in Table I are supplemented by the
determined values of dry matter content in the
sunscreens. The dry matter content values ranged
from 100% (for oil 5), 70% for the heavy creams
19 and 20, 45% for medium creams and 23–33%
for light creams. Calculation showed that a film of
5–20 lm thickness is formed after application
of the product on an even surface on an amount
of 2 mg cm)2 cream applied and evaporation of
the water was present.
The SPF values measured with roughened
quartz testing plate prepared are presented in
Table I. SPF was measured with three applied
amounts, namely, 1.1, 1.3 and 2.0 mg cm)2.
Transmission measurements were performed on
one plate at 16 spots. The SPF values declared
by the manufacturers are confronted with the
SPFin vivo values obtained by the standard COLIPA
method (Table II). The found values of SPFin vivo
agree well with the values declared by the manu-
facturers on the package, except for sunscreen 18,
for which both the value of SPFin vivo obtained by
the COLIPA method and value of SPFin vitro was
concordantly much lower than the value declared
by the manufacturer.
The eight sunscreens were overestimated in
SPFin vitro measurements under applied amount of
2.0 mg cm)2 compared with four overestimated
and six underestimated products for amount of
1.3 mg cm)2. In the case of application of 1.1 mg

Table II Declared SPF, SPFin vivo

and spectrophotometric measure- Sample Declared SPFin vivo SPFin vitro SPFin vitro SPFin vitro
ment of SPFin vitro on quartz testing No. SPF value COLIPA 2.0 mg cm)2 1.3 mg cm)2 1.1 mg cm)2
plate using various amount of sun-
screen 1 40 43.4 ± 6.8 39.4 ± 2.4 38.1 ± 2.6 35.1 ± 2.2
2 25 25.4 ± 4.8 31.2 ± 1.6 28.3 ± 1.5 22.3 ± 1.2
3 16 15.2 ± 1.6 16.4 ± 1.1 14.8 ± 0.9 11.7 ± 1.0
4 8 7.0 ± 1.1 6.1 ± 0.8 5.9 ± 0.4 5.3 ± 0.4
5 8 5.6 ± 1.7 10.4 ± 0.4 9.1 ± 0.3 7.2 ± 0.3
6 12 13.4 ± 1.7 14.5 ± 0.9 13.0 ± 0.6 11.0 ± 0.5
7 12 10.0 ± 1.7 17.4 ± 1.1 13.8 ± 1.1 9.8 ± 1.0
8 12 9.5 ± 1.5 10.5 ± 1.3 10.1 ± 1.3 12.7 ± 1.4
9 8 8.0 ± 1.1 13.5 ± 0.6 11.3 ± 0.7 8.1 ± 0.6
10 12 10.7 ± 1.7 8.4 ± 0.3 7.1 ± 0.2 6.0 ± 0.3
11 14 13.3 ± 1.7 16.5 ± 1.6 12.1 ± 1.3 9.8 ± 1.1
12 16 14.5 ± 1.6 14.3 ± 1.1 11.7 ± 0.9 9.8 ± 1.2
13 18 17.0 ± 1.0 22.4 ± 1.3 17.0 ± 0.8 16.6 ± 1.1
14 20 20.5 ± 3.4 23.5 ± 2.3 16.4 ± 1.6 15.1 ± 1.3
15 15 15.0 ± 2.0 16.4 ± 0.8 14.2 ± 1.1 9.8 ± 0.9
16 15 13.7 ± 1.6 14.3 ± 0.4 11.5 ± 0.4 9.7 ± 0.3
17 25 23.0 ± 2.0 38.1 ± 3.2 32.3 ± 3.1 29.6 ± 3.2
18 15 8.4 ± 1.1 7.1 ± 0.3 4.8 ± 0.3 3.3 ± 0.3
19 40 40.5 ± 7.1 39.1 ± 2.4 33.2 ± 2.3 26.6 ± 2.3
20 40 37.9 ± 6.8 42.1 ± 2.3 46.6 ± 3.8 35.7 ± 3.4

SPF, sun protection factor.

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Quartz plates for determining sun protection factors J. Akrman et al.

Table III Correlation of SPFin vivo and SPFin vitro, slope of Some creams containing titanium dioxide were
linear regression and coefficient of determination R2 inhomogeneous, which is valid for samples 19 and
20. White lumps of titanium dioxide were visible
Applied 2.0 1.3 1.1 during spreading of cream on plate. White lumps
amount mg cm)2 mg cm)2 mg cm)2 were preferentially rubbed in the valleys on
surface of the plate. This phenomenon of inhomo-
Slope 1.080 0.987 0.836 geneity of creams with content of titanium dioxide
R2 0.963 0.959 0.956 probably influenced measured SPF. However, a
repeated measurement gives the same values of
SPF, sun protection factor. SPF.

cm)2, three products were overestimated and 12

creams were underestimated.
Correlation between SPFin vivo and SPFin vitro sig-
nifies that measured values are overestimated for
applied amount of 2.0 mg cm)2 and underesti-
mated for amounts of 1.3 and 1.1 mg cm)2
(Table III).
Sample 17 exhibited a very high value of mea-
sured SPF of 38 at dose of 2.0 mg cm)2 compared
to in vivo determination of SPF of 20. Some creams
showed unexpectedly relationship between applied
amount and measured SPF. Sunscreens 1 and 2
produced oily films with high refractive index after
evaporation of water. Probably reflection and a
backward scattering of light play a role with spec-
trophotometric measurement of transmission.
Photostability of commercial sunscreens
The photostability tests of commercial sunscreens
were carried out on the same plate by determining
the initial and final protection factors (the latter
after a 3.45-hour exposition). The changes in UVA
protection are expressed by the UVA/UVB ratio
and critical wavelength as well as protection factor
UVAPFin vitro (Tables IV and V).
The exposition of plates with sunscreens in the
Q-Sun apparatus resulted in a dramatic decrease
in effectiveness in vitro. This particularly applies to
preparations 5, 7, 11, 13 and 17, which are based
on a mixture of UVB filter, EHMC, and UVA filter,
BMDBM. These creams are characterized by over-
estimation of initial SPFin vitro when compared with

Table IV Protection factors in vitro

Sample SPF SPF UVAPF UVAPF at the beginning and after exposi-
No. initial exposed initial exposed tion to artificial sun radiation,
applied amount 2 mg cm)2 and
1 39.4 ± 2.4 35.8 ± 2.6 11.8 ± 1.0 12.0 ± 0.9 mean value of 16 measure-
2 31.2 ± 1.6 29.3 ± 2.1 11.4 ± 0.4 11.2 ± 0.7 ments ± SD
3 16.4 ± 1.1 14.2 ± 1.0 4.2 ± 0.3 5.0 ± 0.4
4 6.1 ± 0.8 5.2 ± 0.3 4.1 ± 0.2 3.2 ± 0.2
5 10.4 ± 0.4 6.4 ± 0.2 2.8 ± 0.2 1.3 ± 0.1
6 14.5 ± 0.9 9.8 ± 0.8 7.3 ± 0.3 5.0 ± 0.4
7 17.4 ± 1.1 9.3 ± 0.8 9.2 ± 0.6 2.2 ± 0.2
8 10.5 ± 1.3 6.4 ± 0.9 2.0 ± 0.3 2.1 ± 0.2
9 13.5 ± 0.6 10.7 ± 0.7 2.0 ± 0.2 1.8 ± 0.3
10 8.4 ± 0.3 6.2 ± 0.3 5,6 ± 0.3 2.1 ± 0.4
11 16.5 ± 1.6 4.5 ± 0.9 2.0 ± 0.1 1.3 ± 0.1
12 14.3 ± 1.1 9.8 ± 0.8 5.6 ± 0.3 1.1 ± 0.1
13 22.4 ± 1.3 8.2 ± 1.0 7.1 ± 0.4 1.9 ± 0.3
14 23.5 ± 2.3 15.4 ± 1.8 3.1 ± 0.2 2.8 ± 0.3
15 16.4 ± 1.0 16.3 ± 0.8 4.8 ± 0.3 4.4 ± 0.5
16 14.3 ± 0.4 12.2 ± 0.4 4.1 ± 0.4 3.0 ± 0.3
17 38.1 ± 3.2 20.3 ± 2.1 4.6 ± 0.3 2.3 ± 0.4
18 7.1 ± 0.3 6.9 ± 0.3 5.2 ± 0.4 1.4 ± 0.5
19 39.1 ± 2.4 36.7 ± 2.0 7.2 ± 0.5 7.4 ± 0.4
20 42.1 ± 2.3 46.8 ± 2.4 6.8 ± 0.3 7.0 ± 0.5

UVAPF, UVA protection factor; UVBPF, UVB protection factor; SPF, sun protection factor.

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126 International Journal of Cosmetic Science, 31, 119–129
Quartz plates for determining sun protection factors J. Akrman et al.

Table V Ratio of UVA/UVB and critical wavelength at

the beginning and after exposition to artificial sun radia-
tion

Critical Critical
wavelength wavelength
Sample UVA/UVB UVA/UVB initial exposed
No. initial exposed [nm] [nm]

1 0.57 0.61 377 378

2 0.60 0.59 375 374
3 0.50 0.54 369 371
4 0.63 0.70 372 375
5 0.36 0.18 366 333
6 0.61 0.60 374 373
Figure 6 Transmission of sunscreen 14 on quartz plate.
7 0.61 0.26 375 342
1 – before irradiation SPFin vitro = 23, critical wave-
8 0.32 0.38 366 370
9 0.33 0.34 350 350 length = 365 nm, ratio UVA/UVB = 0.35, 2 – after irra-
10 0.73 0.37 380 367 diation SPFin vitro = 15, critical wavelength = 372 nm,
11 0.14 0.18 342 360 ratio UVA/UVB = 0.40. SPF, sun protection factor.
12 0.59 0.20 377 337
13 0.53 0.35 377 363
decrease from the value of 23 to the value of 15.
14 0.35 0.40 365 372
15 0.48 0.48 372 372 The transmission in the wavelength range of 345–
16 0.41 0.41 367 368 380 nm covered by MBBT and titanium dioxide
17 0.35 0.24 370 362 did not change. The radiation in the wavelength
18 0.68 0.21 377 334 range of 380–400 nm caused a minimum skin
19 0.43 0.44 367 369
reaction; therefore, the protection factor UVAPF
20 0.43 0.46 367 367
remained unchanged at the value of 3.
Similar characteristics are exhibited by sun-
the determined SPFin vivo. Filters EHMC and screen 4, which contained simple mixture of insta-
BMDBM are photolabile. It is well known that the ble UVB filter EHMC and stable UVA filter,
photostability of BMDBM is further decreased in its diethylamino hydroxybenzoylhexyl benzoate. Prep-
mixtures with EHMC. The decomposition of aration 11, sunscreen foam, had the form of tur-
BMDBM had the consequence that the sunscreen bid oil, and its irradiation on the plate resulted in
loses it protection ability in the UVA region. This its separation into two phases.
was manifested by lowering of critical wavelength, The heavy creams 19 and 20 contained high
e.g. from the value of 370 nm to the value of amounts of titanium dioxide as the UV filter. They
330 nm. Likewise, decreased are the protection are two batches of the same preparation. The
factor UVAPF as well as the UVA/UVB ratios (sun- micronized inorganic filter, titanium dioxide, is
screens 5, 12 and 18). photostable. Nevertheless, the exposition changed
The decomposition of some formulations gave their protection factors: the SPF of cream 19 was
products that decreased transmission in the UVA decreased whereas that of cream 20 was
region. This mechanism was clearly visible in the increased. We have found out that these fluctua-
case of sunscreen 14 on the diffusion transmission tions in SPF are because of evaporation of water
spectra of the cream on plate before and after irra- residues from the emulsion film during irradiation.
diation (Fig. 6). The same SPF values were obtained after placing
Ratio of UVA/UVB had increased from value the plates in an drying oven at the temperature of
0.35 to 0.40 after irradiation. Sunscreen 14 con- 53C for a period of 3 h. The mass loss because of
tains, besides the photolabile EHMC, a photostable drying was 5% for both the creams. Figure 7 pre-
broad-band filter, methylene bis-benzotriazolyl sents a comparison of spectra of the creams dis-
tetramethylbutylphenol (MBBT) and a smaller solved in ethanol. The spectrum of cream 20 had
amount of titanium dioxide. The EHMC was in a higher absorbance and maximum at the wave-
part decomposed by irradiation. The transmission length of 308 nm and a shoulder was present at
increase in the UVB region resulted in an SPF 370 nm. The spectrum of cream 19 had a lower

ª 2009 The Authors. Journal compilation

ª 2009 Society of Cosmetic Scientists and the Société Française de Cosmétologie
International Journal of Cosmetic Science, 31, 119–129 127
Quartz plates for determining sun protection factors
Figure 7 Spectra of creams 19 and 20 dissolved in
ethanol (100 mg L)1 ethanol, 40C, 1-cm cell). 1 – cream
20, 2 – cream 19.
absorbance on the whole, and the absorbance
maximum and shoulder at higher wavelengths,
namely 312 nm and 376 nm. These phenomena
are because of light scattering on the microparti-
cles of titanium dioxide. An indicator of particle
size is the absorbance ratio E308nm/540nm = 3.14
for cream 20 and ratio E312nm/540nm = 2.22 for
cream 19, where from it can be deduced that the
content of titanium dioxide in both the creams
was approximately the same, but the titanium
dioxide particles in cream 19 are larger. Cream 19
exhibited a lower protection effect when compared
with cream 20, especially after evaporation of the
water residues. Creams 1 and 2 contained mix-
tures of modern and stable UV filters. These two
creams guarantee high protection against UVA
radiation – the factor UVAPFin vitro is 11–12.
With the creams whose irradiation only destroys
the UVB filter EHMC, the protection factor SPF
decreased but the factor UVAPF did not change
very much. On the other hand, some creams based
on the UVA protection imparted by MBDBM com-
pletely lost their UVA protection ability after irradi-
ation (preparations 5, 7, 11, 12, 18 in Tables IV
and V). The photostability of BMDBM could be
increased by addition of a large amount of the
UVB filter, Octocrylen; however, this combination
was not present in the 20 preparations tested
probably because of patent barriers.
The only cream wherein the effectiveness
remained unchanged after irradiation was cream
15 which contained a mixture of octocrylen and
titanium dioxide. Product 6 contained octocrylene,
titanium dioxide, BMDBM and BEMT (bis-ethyl-
hexyloxyphenol methoxyphenyl triazine), thus this
ª 2009 Society of Cosmetic Scientists and the Société Française de Cosmétologie
128
J. Akrman et al.
Figure 8 Correlation of SPFin vitro vs. SPFin vivo. 1 – before
irradiation, y = 1.080x, R2 = 0.963; 2 – after irradiation
y = 0.916x, R2 = 0.947. SPF, sun protection factor.
formulation would be expected as photostable.
However, both in vitro SPF and UVAPF were
decreased after irradiation. Besides the strong con-
ditions of irradiation, the explanation of decompo-
sition of UV filters could be found in improper
choice of titanium dioxide as it is known that
some brands of micronized titanium dioxide exhi-
bit photocatalytic action.
Evaluated set of 20 sunscreens includes a num-
ber of photolabile formulations. Correlation of SPFin
vivo vs. SPFin vitro turned down after irradiation.
Slope of linear regression decreased from a value of
1.080 to 0.916, and coefficient of determination
R2 decreased from 0.963 to 0.947 (Fig. 8). Data
after irradiation are underestimated; this implies
that radiation dose was too high. Experiment on
the irradiation revealed that an initial overestima-
tion of SPF in vitro on products 5, 7, 11, 17 and
13 was because of its photounstability.
Discussion
The most significant finding of this paper is the
fact that the surface of testing plate should be
roughened to roughness value of about 18 lm. In
addition, the surface of plate must be treated in
more complex way to resemble human skin in its
topography with valleys and facets. After such
treatment, the measurement of protection factor
can be performed with amount of 2 mg cm)2 of
sunscreen preparation that is used in the COLIPA
method in vivo. We adopted sand-blasting of quartz
plate and grinding with emery paper. Of course,
the preparation of testing plate can be further
developed. We used only sand for the roughness
ª 2009 The Authors. Journal compilation
International Journal of Cosmetic Science, 31, 119–129
Quartz plates for determining sun protection factors
quality. The measurement of SPFin vitro on a sand-
blasted quartz plate can clearly distinguish
between sunscreen preparations with low level of
protection, i.e. SPF = 6–10, those with medium
level of protection (SPF = 15) and the creams with
high level of protection, i.e. SPF = 25 and higher.
EC Recommendation 2006/647/EC on the effi-
cacy of sunscreen products stated that the ratio of
the protection factor measured in the persistent
pigment darkening test is at least 1/3 of the factor
measured by the SPF testing method. Recom-
mended minimum critical wavelength is 370 nm.
Efficacy of products should be indicated on label
by category ‘low protection’, ‘medium protection’,
‘high protection’ or ‘very high protection’ [12].
From set of 20 tested sunscreens, only seven
products meet requirements after irradiation. Sun-
screens 1 should be labelled ‘high protection SPF
30’ and sunscreen 2 should be labelled ‘medium
protection SPF 25’. Two sunscreens, samples 3
and 6, should be labelled ‘low protection SPF 10.
Sunscreens 7, 8 and 10 should be labelled ‘low
protection SPF 6’.
The method of measurement of SPFin vitro repre-
sents a powerful tool for designers of sunscreens.
The method is quick and minimizes the necessity of
performing in vivo tests on volunteers. At the same
time, the in vitro method also quickly determines
the sun protection in UVA region. After determin-
ing the initial SPFin vitro, it is possible to irradiate
the testing plate with the sunscreen film by means
of a suitable light source and thus determine the
photostability of the given cosmetic composition
under real conditions of its application. The
method can be used for studies of mutual interac-
tions in mixtures of UV filters. In our tests, the irra-
diation was performed under such conditions that
simulate the application of cream in direct sunlight
for a period of 3.45 h. The conditions of irradiation
chosen seem too strong and consequently there is
an underestimation of the SPFin vitro compared with
the SPFin vivo values. The irradiation of testing plate
can induce, besides the photodegradation of UV fil-
ters, also other physical changes that affect the
SPF values. We examined the effect of evaporation
of water residues.
Acknowledgements
The project was supported by Ministry of Industry
of the Czech Republic – grant project no.: FT-TA3/
050.
ª 2009 The Authors. Journal compilation
ª 2009 Society of Cosmetic Scientists and the Société Française de Cosmétologie
International Journal of Cosmetic Science, 31, 119–129
J. Akrman et al.
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