1

A new natural and sustainable eco-friendly peeling agent for

cosmetic use that brings performance while respecting skin

barrier

Jinhzu XU1, Isabelle TERRISSE

3,Simon TROUILLE

1,, Brice BONNET

1, Christèle

RIBAUD1, Yégor DOMANOV

1, Anissa AZOUAOUI

1, Christian TRAN

1, Caroline

FRAISSE3, Catherine BREDOUX

3, Corinne FERRARIS

3, Hèléne MERCAT

1, Nisrine

ZAARAOUI1, Jean- Jacques SCHOONJANS

2, Sébastien DUPRAT de PAULE

2, Anaide

AGHA2, Arthur CROIZIER

2, Julie SONGEUR

3, Isabelle CASTIEL

1, Christian

ORESAJO4, Catherine MARION

3

1L’Oreal Research & Innovation, 1 Av Eugène Schueller, BP 22 93601 Aulnay sous Bois

France

2CHIMEX, 16 rue Maurice Berteaux 95 500 Le Thillay , France

3L’Oreal Research & Innovation, 188 rue Paul Hochart BP 553, 94152 Chevilly- Larue,

France

4L’Oreal Research & Innovation, 30 Terminal Ave, Clark, 07066 New Jersey, USA

Key words: Skin physiology/structure; Skin barrier; Cell culture; Exfoliation;

Chenopodium quinoa Willd;

SUMMARY

Chenopodium quinoa Willd, commonly known as Quinoa, is a well‐known major and

common food ingredient in the Andean communities. It is also an alternative food source in

other regions of the world, with regard to its higher nutritional value than that of most

common cereals. The outer husk of the grain, which is removed before consumption and

discarded, is particularly rich in saponins.

A Quinoa Husk Extract (QHE) was initially identified as a new skin friendly desquamating

agent, usable in cosmetic products, as a complement to dermatological peeling. Such an

approach illustrates our aims at searching new and effective cosmetic ingredients of natural

origin with accessible cost and minimal environmental impact(s).

In vitro studies with 1.5% Quinoa Husk Extract showed exfoliating effect (dihydroxyacetone

test and chromametry) versus placebo with good tolerance, preservation of both the skin

barrier integrity (TEWL) and hydration (corneometry).

These results have been confirmed in a double-blinded clinical study versus lipohydroxy acid

(C8-LHA) whose the exfoliating and rejuvenating properties are well documented. Beneficial

effects on the skin quality have been proven on skin texture with a statistically significant

improvement of skin grain fineness (clinical scoring) at all times vs T0. Statistically

significant decrease of the cutaneous microrelief (measured roughness parameters) and

asymmetry of the diffusion revealing a smoother skin (GonioluxTM acquisitions) were also

evidenced after 12 weeks. Furthermore, a statistically significant improvement of pore

visibility, complexion radiance and pigmentary homogeneity (clinical scoring) was also

demonstrated at all times vs T0. As the comparative agent has well known exfoliating

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properties good results have been shown also on most of the recorded parameters in this

group. But some items as the pores visibility and improvement of complexion (decrease of

dull complexion) are statistically in favor of this new agent QHE. Quinoa Husk Extract could

be appropriate to complete peeling procedures used by dermatologists and to reinforce long

term benefits.

INTRODUCTION

Quinoa (Chenopodium quinoa Willd.) is a well-known staple food of the Andean zone,

principally used in the same manner as wheat and rice [1]. Its nutrition facts and functional

potential have been reviewed [2], revealing a wide spectrum of activities. Most of the quinoa

phytochemicals are reported to be saponins and phenolic acids [3]. Quinoa extracts are

reported to have many beneficial effects, including the anti-inflammatory properties of its

saponins [4], anti-oxidant activity [5], inhibition of skin collagenase [6] and matrix

metalloprotease [7].

The outer husk of the quinoa seeds, particularly rich in saponins (20-30%), is removed prior

to consumption in order to eliminate its bitter taste. It is considered as a by-product with no

(or very limited) commercial value [8].

Based on the reported beneficial properties of quinoa seeds and in particular the high saponin

content of its husks, a quinoa husk extract (QHE) was prepared. Its potential to provide a safe,

gentle and soft cosmetic exfoliation, complementary to traditional peeling was evaluated.

As this effect is of the outmost interest for women with coarse skin, large and too visible

pores, dull and heterogeneous complexion associated with pigmentary disorders, clinical

studies were then performed to validate its exfoliating activity in vivo and evaluate the effect

of a formulation of QHE on women having this skin issues.

MATERIAL AND METHODS:

In vitro studies

Desquamating effect:

Activation of proteolytic enzyme: QHE was incubated for 2 hours with Plantar SC powder.

Desmoglein 1 released by endogen proteases was quantified using Elisa assay.

QHE was applied topically on skin explant at 5% to 10% for 48H. Histological analyses were

performed to quantify corneocyte cohesion. Expression of Epidermal differentiation markers

was evaluated using immunostaining on skin sections.

The acute skin irritancy potential was assessed using the human reconstructed epidermis

(EpiskinSM model) and the acute eye irritant potential was assessed using the corneal opacity

and permeability method (BCOP) with 1% and 10% dilution of the test substance.

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In vivo studies

Proof of Concept Desquamating clinical study (Study 1)

A monocenter double-blind, randomized and comparative proof of concept study was carried

out on 35 women aged from 21 to 40 years old, with Fitzpatrick skin types II to III.

In the study design, each subject was its own control: each subject received 1.5% QHE and

0.3% LHA in comparison with placebo, 10% glycolic acid (positive control) and an untreated

area according to the randomization list.

5 areas of 6.25 cm² (2.5*2.5 cm) were defined on the anterior face of the upper arms and

forearms (10 areas in total). 5 of the areas were located at the right upper arm and forearm and

5 of the areas each at the left upper arm and forearm. The investigational products and the

untreated area were randomized on the 5 areas of the upper arms and on the 5 areas of the

forearms. (See Figure 1)

The study products were applied once a day, except week-ends and days off, on each area at

the investigational center, , during one month.

Figure 1: Treatment randomization,

one of the possible assignments

Efficacy assessment consisted of clinical scoring and chromametry after DHA application,

measurement of trans-epidermal water loss and corneometry.

The epidermis turn-over was assessed by the dihydroxy-acetone test: the investigator scored

the skin color after DHA application every day during the first week and every two days

between D8 and D29, except week-ends. Chromametry measurements were also done in order

to follow the kinetic of skin discoloration.

The Skin barrier function was controlled by following the measurement of the TEWL and

the skin hydration by corneometry. The measurements were done every day during the first

week and every two days between D8 and D29, except week-ends.

QHE

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Real conditions Skin Quality clinical study (Study 2)

A double-blind randomized 2-parallel group study was performed to evaluate efficacy and

safety of a formulationwith this new cosmetic ingredient. One hundred and ten Caucasian

women, aged 35 to 50 with Fitzpatrick skin types I to IV, were recruited after having given

their written consent. All women had a coarse skin with a dull skin tone (with a clinical

scoring of at least 4) and inhomogeneous tone (Dermascore pigmentary scale of at least 3),

very visible pores (Dermascore scale of at least 4), rough skin (clinical scoring of at least 4).

Fifty percent of these people have declared a “sensitive” skin.

The included volunteers applied according to randomization either the studied formulation

(containing QHE 1.5% in a simple cosmetic basis) or the comparative formulation in the same

cosmetic basis (see “tested products”). The product was appliqued twice a day on the whole

face and the neck for 3 months.

Efficacy assessment contained clinical evaluation, analysis of images provided by a

Goniolux® device, Silflo TM

imprints and self-assessment.

Clinical evaluations were performed in a blind manner by the same dermatologist throughout

the whole duration of the study at T0, Tim, T1M, T2M & T3M (For criterion, areas, principle

and evaluation time points,

For “heterogeous complexion or pigmentary heterogeneity” and pore visibility, an

improvement corresponds to a reduction of values.

To evaluate skin “micro-relief”, SilfloTM

imprints were performed on the cheek (right or left

according to randomization) at basal, 1 and 3 months and analysed by QuantilineTM

software

GonioluxTM

provides images which can be analysed and gives measurement of the light

intensities re-emitted by the skin in all directions of space. The studied parameters are

“specular light quantity” (which gives information on the pure shininess of the skin), “global

diffused light quantity” (information on the “satiny” aspect of the skin), “global luminous

assessment” (total quantity of light re-emitted by the skin) and “asymmetry of diffusion”

(anisotropy of all reflections for 2 directions right and left) These images were performed on

the right or left cheek, opposite to the one chosen by the dermatologist for the clinical

evaluation at T0, T1M and T3M.

Self-Assessment was performed with a questionnaire on skin texture, complexion,

homogeneity, pores relief and the overall skin’s appearance. These criteria were assessed Tim

and T1M, T2M and T3M. Answers to the questions were categorized as « agree », «

somewhat agree », « neither agree nor disagree », « somewhat disagree», and « disagree ».

Percentage of positive responses (addition of « agree », & « somewhat agree » categories)

was compared between groups.

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Tested Products

For Study1: the studied product is a basic formulation containing the Quinoa Husk Extract

(QHE) at 1.5% (concentration chosen according to the results of in vitro studies).

The comparative product contains a derivative of hydroxy acids (LHA at 0.3%). The positive

control contains 10% of glycolic acid. All the investigational products are simple gel

formulations.

For Study 2: the product is a formulation containing the Quinoa Husk Extract (QHE) at 1.5%

(concentration chosen according to the results of in vitro studies in an O/W fluid emulsion.

The comparative product contains a derivative of hydroxy acids which are well known for

their exfoliating and rejuvenating properties. But the lipophilic nature of this C8-lipohydroxy

acid and its relatively slow penetration in the skin afford its efficacy at low concentrations.

In this study 0.3% was chosen in order to be efficient and to ensure a good tolerance for the

whole duration of the study.

RESULTS

Process development of quinoa husk extract

Quinoa husk represents about 10% w/w of the grain and is still considered as a by‐product of

no (or very limited) commercial value.

An industrial process was set up for developing this promising desquamating agent. The

sourcing of the husks was secured to ensure their compliance, reproducibility and

sustainability. The solvent used was optimized as 50/50 ethanol (food grade)/water integrating

the ethanol re‐use. The waste volume was minimized to reduce the environmental impact of

the process and an effective extraction yield was achieved through optimizing

temperature/duration of the extraction, solid/liquid separation, fine particle elimination and

solvent evaporation. Sterilization was further processed to guarantee an imperative

microbiological quality of the final extract.

From design to manufacturing, our eco‐responsible processes integrate a large range of

voluntary approaches including improvement in productivity, waste reduction, minimal use of

energy and water, and sustainable sourcing. All these steps represent a unique reference

system for assessing the ecological footprint of manufacturing processes and their

performances from an environmental point of view [9], [10].

Figure 2: Chimex eco-footprint

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The “EcoFootprint” is a very valuable tool for the assessment, improvement and valorization

of our processes and provides a technical and social overview of the benefit for customers.

This tool has been designed to be pedagogical and to point out main issues regarding

environmental impact. It can be used at each step of the process and can compare the

continuous improvement. The “EcoFootprint” focuses on two sets of criteria: Manufacturing

Footprint and Eco‐Design Footprint.

Figure 3: Composition of QHE

In vitro studies

On isolated plantar SC, 0.1% Quinoa Husk Extract treatment leads to a significant release of

desmoglein 1 suggesting that desquamating enzymes were activated.

On human skin explant, topical application of both 5% and 10% QHE solution revealed a

significant decrease of corneocyte cohesion as compared to the vehicle after 48H of treatment.

Analysis of terminal epidermal differentiation markers showed that the differentiation process

was maintained and reinforced as shown by an increased expression of filaggrin, a terminal

differentiation marker. Furthermore, analysis of the intercellular space lipids using DSC test

on SC showed no modification of their organization.

Additionally, in comparison with classical surfactants, Chenopodium quinoa husk extract

showed a better dermal profile even at high concentration (10%).

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Figure 4: Release of desmoglein 1 fragments

(n=3 samples 2 independent experiments)

Figure 5 Means and medians of scorage of stratum corneum

cohesion treated with QHE solutions (n=9)

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Figure 6: Histological slides of treated excised skin samples of one skin donor

Figure 7: Thermogrammes of intercellular lipids of isolated human sc treated by qhe

solutions (n=2)

Vehicle Control QHE 0.5%

QHE 5%

QHE 10%

Reference salicylic derivativ

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Figure 8: Episkin™ validated skin irritation test based on MIT reduction assay (n=3)

In vivo studies

In terms of efficacy results, 1.5% QHE showed a moderate exfoliating effect

(dihydroxyacetone test and chromametry) vs placebo, with good tolerance and maintenance of

skin barrier integrity (TEWL) and hydration (corneometry).

Additionally to this proof of concept/efficacy as desquamating agent, topical 1.5% QHE was

further investigated in a clinical study focused on skin quality in an optimized formulation. 49

women aged from 35 to 50 years, with I to IV phototypes and a coarse skin, a dull and

heterogeneous complexion and enlarged pores, have applied the product on the face and neck,

twice a day during 12 weeks. Clinical & instrumental evaluations were performed before

application (T0) and after 4, 8 and 12 weeks.

The efficacy of QHE was demonstrated on skin texture with a statistically significant

improvement of skin grain fineness (clinical scorage) at all times vs T0. Statistically

significant decrease of the cutaneous microrelief (measured roughness parameters) and

asymmetry of the diffusion revealing a smoother skin (GonioluxTM acquisitions) were also

evidenced after 12 weeks.

Furthermore, a statistically significant improvement of pore visibility, complexion radiance

and pigmentary homogeneity (clinical scorage) was also demonstrated at all times vs T0.

0

20

40

60

80

100

120

Neg. Cont. SolventCont.

Pos. Cont. AQHE 1% AQHE 10% QHE 100%

% v

iab

ility

(M

TT)

Irritant

Non irritant

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Figure 9: Clinical scoring of skin color after DHA application: 1.5% QHE has a moderate

exfoliating effect in the DHA test vs placebo and versus 0.3% LHA.

Figure 10: Skin color – L* parameter

Figure 11: Clinical scoring

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CONCLUSION

Quinoa Husk Extract (QHE) belongs to a new generation of natural active ingredient chosen

to fulfill the necessity of sparing resources in a full respect of local environment. Its mild

exfoliating properties have been demonstrated in vitro versus placebo with a good tolerance

without drying the skin while maintaining skin barrier integrity. These results have been

confirmed in vivo double-blinded clinical study versus a lipohydroxy acid (C8-LHA) whose

exfoliating and rejuvenating properties are well documented. Beneficial effects on the skin

quality have been proven on skin texture, pore visibility, complexion radiance and pigmentary

homogeneity.

References

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2. Vega-Galvez, A., Miranda, A., Vergara, J., Puente, L., and Martinez, E.A. Nutrition facts and

functional potential of quinoa (Chenopodium quinoa willd.), an ancient Andean grain: a review. J. Sci.

Food Agric. 90. 2541-2547 (2010).

3. G’omez-Caravaca, A.M., Segura-Carretero, A., Fernandez-Gutierrez, A. and Caboni, M.F.

Simultaneous determination of phenolic compounds and saponins in quinoa (Chenopodium quinoa

willd.) by a liquid chromatography diode array detection-electrospray ionization-time-of-flight mass

spectrometry methodology. J. Agric. Food Chem. 59. 10815-10825 (2011).

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Food Sci. 79. 1018-1023 (2014).

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quinoa (Chenopodium quinoa willd.). Int. J. Mol. Sci. 15. 19307-19318 (2014).

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