Ref:NU-PI01-BC-AT01

EVALUATION OF THE EFFICACY OF AN ANTI-GRAY TREATMENT

FOR HAIR BY:

COLORIMETRIC IMAGE ANALYSIS
SELF-EVALUATION QUESTIONNAIRE

PRODUCT REFERENCE:
GREY AWAY LOTION
Batch E0112009



Test in vivo on subjets




Sponsor:
B2C
BP 617
F-59061 ROUBAIX Cedex 01, FRANCE




















SPINCONTROL
March 2001
238, rue Giraudeau
F-37000 TOURS, FRANCE

1. CONDITIONS OF REALIZATION

1.1. Site of study

Ste SPINCONTROL
238 rue Giraudeau - 37000 TOURS, FRANCE
Tel: +33 (0)2 47 37 62 79
Fax: +33 (0)2 47 37 58 28
e-mail: spincontrol@spincontrol.fr

1.2. Sponsor

B2C
BP 617
F-59061 ROUBAIX Cedex 01, FRANCE

1.3. Monitor

CATRYSSE S.
B2C
BP 617
F-59061 ROUBAIX Cedex 01, France
Tel.: +33 (0)3 20 26 27 48
Fax.: +33 (0)3 20 26 27 28


1.4. Investigator

BEAU P.
Director Ph.D.
SPINCONTROL

1.5. Scientific Director and Study Coordinator

PITTET J.C.
Ph.D.
SPINCONTROL
Tel: +33 (0)2 47 37 12 86
e-mail: pittet@spincontrol.fr
1.6. Technician

JACOB S.
Biometrology Technician
SPINCONTROL

1.7. Quality Assurance Manager

PERRIER C.
Quality Engineer
SPINCONTROL
Tel: +33 (0)2 47 37 09 49

2. SUMMARY OF STUDY

2.1. Objective

The aim of this test was to determine the efficacy of an anti-gray hair treatment:
Ref. Anti-Gray Lotion batch E0112009

This evaluation was done by:
Colorimetric analysis of hair images.
Self-evaluation questionnaire

The evaluation was carried out for a total period of 28 days of application.

2.2. Population

The study was conducted on 20 healthy adults. Equal numbers of men and women were recruited.
The subjects were required to strictly comply with the inclusion/exclusion criteria defined in section
4.1.1

For inclusion, the subjects had to have light to dark brown hair with about 50 to 80% gray hair,
evenly distributed if possible.
2.3. Timetable and determinations

The study was conducted from January to February 2002.

Timetable:
T0 T7 days T14 days T28
days


January 24 2002 January 31 2002 February 07 2002 February 21 2002
- Photogr
aphic acquisition of hair
- Telepho
ne interviews on product
acceptance
- Photogra
phic acquisition of hair
- Self-
evaluation
- Photogra
phic acquisition of hair
- Self-
evaluation
questionnaires questionnaires

Comments: - Carrying out the T28 days time point depended on the results obtained at T14 days.
- A telephone survey was conducted 7 days after the first application of
the product to verify color changes to the hair and subject satisfaction.

2.4. Choice of the study

- Open.
- Non-comparative study.
- Subjects were their own controls in relation to T0, i.e prior to treatment.




3. LEGAL ASPECTS

3.1. Qualification of personnel

The Investigator certifies that the Study Coordinator and all persons participating in this study were
qualified for the skills required for its realization.

3.2. Data storage

The Investigator stored the data on two supports, i.e. paper and computer.
Paper storage is in a special cabinet, locked and in proximity to a fire extinguisher.
Computer storage is on DAT tape stored for 10 years.
The Investigator will keep a copy of the protocol signed by the sponsor and himself, the case report
forms and all documents used to fill them out, study participation consent forms, original
documents of all types and all documents concerning the project for 10 years after the final report is
remitted. All these documents will remain available for inspection within a reasonable time period
for an authorized representative of the sponsor or by legal authorities.
The investigator will inform the sponsor when data destruction is planned after 10 years


3.3. Protocol amendment

The Sponsor is informed that any modification he introduces leading to a protocol amendment will
generate additional costs.

3.4. Insurance

The Sponsor covered this study with an insurance policy taken out with:

AXA No. 132.131.25.04

The Investigator took out a liability policy with:

AXA No. 79.191.004.

The investigator and the sponsor took out separate liability insurance policies to protect the
subjects.
Extract of modified law No. 88-1138 dated December 20, 1988, concerning the protection of
persons engaged in biomedical research (article L 209-7):
"For biomedical research without direct individual benefit, the Sponsor assumes even in the
absence of fault indemnity for the harmful effects of research on the person who is engaged and
that of his beneficiaries, without invoking a third party or the dropout of the person who initially
consented to participate in the research".

3.5. Declaration to the CNIL (National Data Protection Commission)

In application of the "Data and Personal Freedom" law dated January 6, 1978, the database required
for carrying out all SPINCONTROL studies has been declared to the National Commission on Data
and Personal Freedom.



3.6. Subject anonymity

The subjects were identified for the sponsor by a 5-character alphanumeric code and by a number.

3.7. Consent for participation in the study

A form was remitted to each subject in order to define:
- the aim of the study, its methodology and duration,
- any expected cosmetic benefits, restrictions related to the study and predictable risks,
including interruption of the study before its termination,
- the non-inclusion period, the amount of indemnity if applicable, the possibility for the
subject to verify the accuracy of the information in their file and its subsequent destruction
This information enabled the subject to sign a form for free, informed and express consent with full
knowledge.

3.8. Confidentiality

All information, data and results of the study are confidential.
All persons having access to this information, data and results have been informed that they are
confidential.
All information on the health status of subjects and the results of clinical examinations conducted
before the study for their recruitment, selection and admission are subjected to the rules of medical
secrecy compliant with article 226-13 and the following of the Penal Code and the Medical Ethics
Code (decree No. 95-1000 of December 6, 1995): in no case can they be communicated to the
Sponsor with their identity.

"Investigators and all persons cooperating in trials are bound by professional confidentiality
especially concerning the nature of the products studied, trials, persons included and the results
obtained" (extract of article R 5120 of application decree No. 90-872 dated September 27, 1990 of
modified law No. 88-1138 dated December 20, 1988, concerning the protection of persons engaged
in biomedical research: "Huriet" law).

3.9. Quality Assurance

The entire study brief (protocol, results, report and all associated documents) has been controlled by
Quality Assurance in compliance with regulatory stipulations and existing procedures. The
procedures for the complete audit of this study are described in the "Procedure for Procedures" No.
PRE-ASQ-1.
The investigator is ready to receive and cooperate with any audit designated by the sponsor to verify
that the study was conducted in compliance with the protocol and existing procedures.

3.10. Regulations

This study without direct therapeutic finality for the subject was conducted in compliance with the
most recent recommendations of the World Health Association (1964 Helsinki Declaration
amended at Somerset West, Republic of South Africa, 1996).
This study was not within the scope of French law No. 88-1138 dated December 20, 1988 modified
by laws No. 90-86 dated January 23, 1990 and No. 94-630 of July 25, 1994, as indicated in the
"Guide to legislative and regulatory texts" (BOMS (Health Ministry Official Bulletin) No. 91/13
bis) concerning the protection of persons engaged in biomedical research. Thus, no information was
communicated to the national file of persons engaged in biomedical research without direct
individual benefit and the opinion of the Institutional Review Board (Ethics Committee) was not
sought, but the spirit of decree no. 90-872 dated September 27, 1990 on the application of this law
was heeded.

3.11. Practical considerations

Prior agreement between the Investigator and the Sponsor of the present study will be required for
any publication, regardless of which party so desires it.

4. METHODOLOGY

4.1. Subject selection

The SPINCONTROL panel was composed of subjects having previously responded to a
computerized questionnaire concerning their medical history, allergies, health and cosmetic care
practices, and including various administrative information. A licensed esthetician was responsible
for questionnaires.

The procedure for recruitment, selection, and admission of willing subjects, after free, informed and
express consent for study participation, was prepared to guarantee clear and precise information,
enabling them to determine the aim and consequences of their consent.
In particular, this procedure included:
- A preliminary interview where they were informed of the objective of the study protocol,
indemnity procedures where applicable, and the expected cosmetic benefits, study-related
limitations and predictable risks.
- Reading an information sheet covering the essential parts of the study and the signature of a
free, informed and express consent form by the subject: this enables the subjects to decide
upon their inclusion based on the conditions proposed.
- This consent enabled subjects to be covered by the liability insurance taken out by the
sponsor and investigator once the subject was definitively admitted to the study by the
coordinator.

Subjects recruited were selected under the responsibility of the investigator and study coordinator
on the basis of the inclusion/non-inclusion criteria and proscriptions/restrictions defined in the
protocol (copy in appendix).

There were 20 subjects selected for this study.
In order to compensate for possible withdrawals of one or several subjects, two additional subjects
were recruited. The results are presented for all subjects present at the last examination time in the
study (T28 days).

4.1.1. Review of principal inclusion criteria

Concerning the study:
Male or female subjects;
Caucasian;
30 to 80 years of age (no limit);
Having dark hair (light to dark brown, black);
With gray hair evenly distributed over the entire scalp with at least about 50% gray hair;
Not having applied hair coloring for at least 8 months;
Accepting to wash their hair the day before measurements with the shampoo provided;
Having healthy skin (absence of psoriasis, eczema, erythema, edema, scars, wounds, lesions), on
the scalp in particular;
Not pregnant nor breast feeding for at least 3 months;
Not under a hormone treatment for a pathology, e.g. a thyroid problem.

Not presenting skin hypersensitivity to cosmetic products, to hair colorants in particular;
Not having atopic skin;
Pledging not to undergo UV exposure in an institute throughout the study;
Pledging not to tint or bleach the hair throughout the study;
Pledging not to have a haircut throughout the study.

4.1.2. Review of principal proscriptions and restrictions

No other cosmetic product for coloring, bleaching or treating hair was to be applied throughout the
study.
No shampoo other than that furnished 48 hours before the examination days.
In no case were the subjects to have a haircut throughout the study.


4.1.3. Acceptance of the use of images (Appendix 2)
Each subject was asked to sign an agreement for the commercial use of the photos taken and for
his/her possible participation for testimonial on a television show.

4.2. Description of the product

The product to test was provided by the sponsor. The product was packaged in a 200 ml bottle with
pump dispenser.
The sponsor was responsible for the manufacture, packaging, identification, determination of purity,
composition, safety and other characteristics of the product tested.
The sponsor agreed to provide an attestation of absence of toxicity of the product tested.

Ref.: ANTI-GRAY LOTION batch E0112009

The sponsor has provided to the coordinator with complementary quantities of products required for
the study.

The sponsor provided for the study:
The number of products necessary for the subjects participating in the study.
A sufficient quantity of products for the additional subjects in the study.
One product per reference and per batch for the Spincontrol sample collection.


4.2.1. Application of products, frequency, procedures

The product had to be applied 2 to 3 times a week, i.e. every 3 days on average.
Application was by carried out by the subjects.
The procedure involved first washing the hair with the shampoo provided, drying, and then
application of the product to all the hair. After application, gentle friction distributed the product
uniformly, although avoiding direct contact with the scalp.

4.3. Choice of the study

This was an "open" study, i.e. both the investigator and the subjects knew what product was
being applied.
The study was non-comparative
The subjects were their own controls in comparison to the time before treatment, T0.

4.4. Evaluation techniques


4.4.1. Measurement of hair color

The aim of the study was to quantify the change in hair color in a region previously defined and
identical at all examination times for the same subject.

The technique involved acquiring a photographic profile of the entire head of hair in
standard "daylight" illumination conditions. After this, a 40 x 50 mm region of interest, i.e.
about 20 cm, was defined on the image. Image analysis of this acquisition involved
extracting the images mean of gray levels as well as the proportion of white pixels after
thresholding (S=120 on a gray scale from 0 to 255). This proportion is directly related to the
"number of white hairs in the region examined. A comparison of this proportion at the
different time points was used to determine darkening of the hair and the disappearance of
gray hair resulting from the treatment.


T0 T14 days T28 days

Fig 1: Position of the region for measuring the gray scale and the number of pixels higher than the threshold (S=120).

4.4.2. Self-evaluation questionnaire

A self-evaluation was conducted both at T14 days and T28 days on the items defined in the
protocol.
These questionnaires were prepared and processed using ScanBac software (3SI). They were read
with a Fujitsu M3096GX scanner and the same software. The data were then analyzed and presented
using Excel (Microsoft).

4.5. Examination schedule

The effect of the treatment was determined after 14 and 28 days. A telephone survey was conducted
on day 7 to determine recoloration progress and subject satisfaction.

Dates:
T0: January 24, 2002
T7 days: January 31, 2002
T14 days: February 07, 2002
T28 days: February 21, 2002

4.6. Presentation of the results

Two types of presentation were adopted:
- overall: corresponding to the mean values of the entire panel and
expressed in tables and graphs.

- per subject: tabular summary of all values acquired during the
examinations.

4.6.1. Overall results with the entire panel

This presentation provides a rapid and overall view of changes in the parameters studied.
These overall results are presented for all the subjects present at the last measurement (T28 days).

4.6.2. Individual results

All raw measurements of the mean gray level and of the number pixels above and below the
threshold are presented for each subject.

4.6.3. Statistics

I nstrumental techniques

The significance of the results was determined using Student's t test for paired data (provided
product conditions of application are verified). The test was applied to raw data and to the changes
of parameters over time.

The acceptance threshold was set at 5%.

Self-evaluation

The frequency tables expressing all the qualitative variables will be established by tallying the total
test population and calculating the frequency of their various possible responses (given in
percentages). For each question, the results are presented as tables (population and frequencies).
The statistical difference of frequencies (%) was determined with a chi-square () test.
For questions involving agreement or disagreement, 1 type of distribution is presented:

"Agree" distribution
- the 4 responses were clustered in two non-symmetrical classes

Agree entirely AGREE
Agree
Do not agree entirely

Disagree
DISAGREE

The latter presentation expresses all the "agrees," regardless of the degree of acceptance.