STEPS SPF ISO 24444 Protocol Testing

The ISO 24444 SPF test is now Note that is some countries,
alternative methods are accepted.
adopted in nearly 60 countries
Product Application
Supportable Claims
A very accurately measured and
• Sunscreen Efficacy controlled amount (2 mg/sq cm)
• SPF Number of product is applied to a marked out
area of skin. The product is evenly
Applicable Countries spread, using a standardised
technique. The product is then
Harmonisation of this test allows us
1.Product Application allowed to dry for 15 to 30 minutes.
to certify for the counties below...

Water Resistance
Australia
For most countries, 40 minutes or 80
New Zealand minutes. Up to 4 hrs is permitted in
Australia.
European Union 28 countries Solar Simulation
UK A Solar simulator, which has been
designed and calibrated to imitate
India the spectrum of sunlight, is used to
apply small incremental doses of
2. Water Resistance Challenge Japan light to the protected area. An
unprotected area and an area with a
Taiwan Standard Sunscreen applied, are
also exposed. Overnight, a mild
Korea erythema (slight sunburn) develops
at test sites where the SPF has been
MERCOSUR 6 countries exceeded.
Canada
References
ASEAN 10 countries
[Monographed Methods}
South Africa ISO 24444 : 2010
3. Solar Simulator Mexico

Chile

Russia
Eurofins Dermatest Pty Ltd
Israel 20 to 22 King St
Rockdale NSW Australia
India ph 61 2 9556 2601
Russia [email protected]

Japan www.dermatest.com.au
4. Reading of Results
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 PROTOCOL DETAILS

SPF TEST ACCORDING TO THE ISO 24444 PROTOCOL

1. Objective
The test panel will be convened to evaluate the effectiveness of a test material as a
sunscreen product by determining the Sun Protection Factor (SPF) on human skin as
described in the document: International Standard ISO 24444 - Cosmetics - Sun
Protection Test methods -in-vivo determination of the sun protection factor (SPF).

2. Test Material Handling

The record of the sample is entered into a log identifying the lot number, sample
description, batch number, sponsor, date received and tests requested. Samples are
retained for a period of two years beyond final report generation.

3. Ethical Principles for Conduct of Study

3.1 The study will be conducted in accordance with the principles as described in the
document WMA Declaration of Helsinki – for Medical Research Involving Human
Subjects.

4. Standard for Inclusion of a Panelist in a Study

4.1 Individuals over the age of consent.
4.2 Individuals free of any dermatological or systemic disorder which would interfere
with the results, at the discretion of the investigator.
4.3 Individuals who have completed a preliminary medical history evaluation.
4.4 Individuals who have read, understood and signed an informed consent document
relating to the specific study to which they are subscribing.
4.5 Individuals with no known abnormal response to sunlight.

5. Standard for Exclusion of a Panelist from a Study

5.1. Individuals taking medication which in the opinion of the investigator would
mask or interfere with the results.
5.2. Individuals with chronic skin allergies.
5.3. Individuals with suntan or sunburn.
5.4. Individuals with abnormal reaction to the sun.
5.5. Pregnant or lactating females.
5.6 Subjects accustomed to using sun beds.
5.7 Subjects who have participated in an SPF study within the last two months.

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6. Informed Consent and Medical History Forms
An informed consent will be obtained from each volunteer prior to initiating the study
describing reasons for the study, possible adverse effects, associated risks and
potential benefits of the treatment and their limits of liability. Panelists sign and date
the informed consent document to indicate their authorisation to proceed and
acknowledge their understanding of the contents. Each subject is assigned a
permanent identification number and completed an extensive medical history form.
These forms along with the signed consent forms, are available for inspection only on
the premises of Dermatest Pty Ltd and during normal office hours.

7. Panel Composition
a minimum of ten healthy volunteers, between the age of consent and 70 years will be
recruited for this study. The panel consisted of fair skin individuals with Fitzpatrick
skin types I, II or III.

8. Solar Simulation
The light source employed is a small beam 150 watt Xenon Arc Solar Simulator
(Solar Light Co., Philadelphia, Pennsylvania, Model 16S or Model 601) having a
continuous emission spectrum in the UV range from 290 to 400 nm and compliant
with the spectral performance requirements of the Annex B of the ISO protocol.
Xenon arc is selected on the basis of its black body radiation temperature of 6000oK
which produces continuous UV spectra (all wavelengths) substantially equivalent to
that of natural sunlight. This device is equipped with a dichroic mirror (which reflects
all radiation below 400nm) and works in conjunction with a 1 mm thick Schott
WG320 filter (which absorbs >99% of radiation below 290nm) to produce simulation
of the solar UVA©UVB spectrum. A 1 mm thick UG 11 filter (black lens) is added
to remove reflected (infra-red, greater than 700 nm) heat and remaining visible
radiation. UV radiation is monitored continuously during exposure using a Sunburn
UV Meter/Dose Controller System (Solar Light Co) . Measurements are taken at a
position within 8 mm from the surface of the skin. The field of irradiation is >0.5 cm
in diameter, with at least 1 cm between each adjacent site. Realignment of the Light
Sources and calibration of the sunburn meters are conducted by independent
certification facilities and adjustment to light source power supply only by the
Director. The calibration data for each solar simulator will appear on the test report in
the following tabular format.

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9. Determination of Water Resistance (Where Conducted)
This does not form part of ISO 24444, but is a requirement of AS/NZS 2604 (2012)
for those products making a water resistance claim. This test is employed to
determine the substantivity of a test product and its ability to resist water immersion.
The procedure described in Appendix B of AS/NZS 2604 is followed. The immersion
schedule is listed as follows:
9.1 Subject immersed in the Spa pool for 20 min with 4 min of air agitation.
9.2 5 minutes rest period out of the water. This sequence is repeated for the required
time period. Immersion is achieved indoors in a circulating whirlpool tub through
1.5 cm diameter ports. The water is maintained at an average temperature of 33oC+/-
2oC. The pH of the water is maintained between 6.8 and 7.2.
The test area is air dried prior to exposure from the solar simulator using 25%
increments. The exact series of exposures given is determined by the control MED
and the expected SPF of the product.

Water Resistance Category Description for AS/NZS 2604 (2012)

10. Determination of the Sun Protection Factor

One test site area of 40 sq.cm serves to determine each subject's Minimal Erythema
Dose (MEDu). This is executed by exposing the back to a series of 5 timed

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incremental UV exposures at 112% or 125% intervals. The individual subject's
MEDu is the shortest time of exposure that produces perceptible unambiguous
redness at 16 to 24 hours post irradiation. The test area is described as the
infrascapular area of the back to the right and left of the midline. The application area
is 40 sq.cm. The product is delivered to the test site through plastic volumetric
syringes. The material is evenly applied to a rectangular area for a final covering of
2.0 mg/sqcm +/- 2.5%.
The product is deposited in a series of evenly distributed spots and then spread evenly
with a fingercot. Product application, UV exposures and measurements of responses
will be conducted in stable environmental conditions with the room temperature
maintained between 18oC and 26oC.
15 to 30 minutes after application, a series of 5 UV light exposures in 112.5%
increments calculated from previously determined MED's bracketing the expected
SPF will be administered from the solar simulator to subsites, each with an area of not
less that 1 cm sq and spaced at 1 cm separation within the treated area. On the actual
day of testing another series of exposures is administered to an adjacent untreated site
of unprotected skin to re-determine the MED. An adjacent test site is then selected to
perform a static determination on the test substance.
A reference sunscreen, as described in Annex c of the ISO 24444 Standard, is also
applied to each of the test subjects, utilising an application exposure procedure which
is the same as that utilised for the test product.
Following UV exposure to all test sites, the product is gently removed using moist
soft tissue together with ethanol if needed.

11. Evaluation of Response

The volunteers are instructed to return to the testing facility sixteen to twenty four
hours post exposure, for evaluation of delayed erythemic response. The smallest
exposure or the least amount of energy required to produce unambiguous redness
(MEDp) in the treated site is recorded. The SPF is then calculated by the equation:
MEDp ( Protected Skin ) / MEDu (Unprotected Skin) = SPFi (calculated to one
decimal place).
12. Calculations and Statistics
The SPFi values from an initial panel of the first 10 test subjects are sequentially
evaluated in order to determine a provisional mean Sun Protection Factor (SPFn1).
The statistical criteria described in the test method were then applied to determine a
confidence interval and statistical variance. Where necessary, additional subjects will
be tested according to the protocol if the first 10 results are not found to be within the
specified range.
13. Rejection Criteria
Panelist's results will be rejected and the panelist replaced if:
13.1. The responses on the treated test site are randomly absent or out of sequence.
This is an indication that the products are not spread uniformly.
13.2. An MED could not be obtained due to elicited response at all
exposure sites.

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13.3. The exposure series failed to elicit an MED response on either the
untreated or the applied skin areas.
The test is then considered a technical failure and the subject's data is discounted.
14. Individual Panelist Results
These will be set out in the attached report in ISO 24444 spreadsheet format.
15. Observations
No adverse effects or unexpected reactions of any kind will be recorded.
16. Archiving: All original samples, raw data sheets, technicians notebooks,
correspondence files and copies of final reports and remaining specimens are
maintained on premises of Dermatest Pty Ltd in limited access storage files. A
duplicate disk copy of final reports is archived separately off site.
17. Colour Discrimination Test
All technical employees of Dermatest Pty Ltd who are involved in scoring of exposed
skin spots are required to take and pass a visual colour discrimination examination
using the Farnsworth-Munsell 100 Hue Test.

18. Certification for E.U. Compliance

Where requested, an additional certification, indicating compliance with the E.U.
requirements for sunscreens, can be provided.

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19.Certification for AS/NZS 2604 (2012)
Where requested, an additional certification,
indicating compliance with AS/NZS
2604 (2102) requirements for SPF and
Water Resistance requirements for sunscreens,
can be provided.

References
1. Dermatest SOP – 010 Procedure for Conducting an SPF Study.
2. International Standard ISO 24444 (2010) - Cosmetics - Sun Protection Test
methods -in-vivo determination of the sun protection factor (SPF) Factor (SPF).
3. Australian New Zealand Standard AS/NZS 2604 (2012) – Sunscreen Products –
Evaluation and classification.
4. Commission Recommendation of 22 September 2006 on the efficacy of sunscreen
products and the claims made relating thereto. Official Journal of the European
Union L 265/39
5. WMA Declaration of Helsinki – for Medical Research Involving Human Subjects.

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