ACB Kale Protein Blend

Efficacy Data

Code: 20036
INCI Name: Hydrolyzed Kale Protein & Hydrolyzed Carrot Protein & Hydrolyzed Lemon Protein
CAS #: 100209-45-8 & 100209-45-8 & 100209-45-8
EINECS #: 309-353-8 & 309-353-8 & 309-353-8

Name of Study Results

The results of the assessment indicate that when

incorporated into a shampoo, ACB Kale Protein
Blend is capable of improving hair characteristic 44%
more than a control shampoo and 36% more than the
control conditioner. When used in both the shampoo
and conditioner at 2%, ACB Kale Protein Blend
improved shine, volume, dry and wet combability,
smoothness, hydration, softness and manageability by
37%, 59%, 54%, 50%, 32%, 35%, 55% and 38%
respectively, better than the control shampoo. These
results can be further supported by figures 1 through
10, where clearly the half of the subject's head treated
with 2.0% ACB Kale Protein Blend appears more
voluminous, shiny, soft and healthy.
When comparing hair characteristics of the baseline
Hair Testing assessments to the post style assessments, the
benefits of including 2% ACB Kale Protein Blend in
a shampoo and conditioner are even more apparent.
In relation to the baseline readings, the test-half of the
head improved all subjective parameters, improving
shine, volume, dry and wet combability, thickness,
smoothness, hydration, softness and manageability
by 41%, 47%, 75%, 87%, 14%, 47%, 30%, 30%, and
63% respectively, with an the test product improving
hair conditions on average by 48% when compared to
the untreated baseline.

For this reason, ACB Kale Protein Blend would be an

ideal product to use in cosmetic applications designed
for anti- aging hair care that yields voluminous, soft and
shiny hair aesthetics.

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Cellular Viability Assay
As shown in figure 1, ACB Kale Protein Blend exhibited
comparable results by increasing cell metabolism. An
increase in fluorescent signal indicates an increase in
cellular metabolism and viability. ACB Kale Protein
Blend does not appear to have negative effects on
cellular metabolism and can safely be used in cosmetic
materials

Both the innovative hydrolysis method

used in the production of ACB Kale
Protein Blend and the traditional method
of hydrolysis induced when
creating Wheat Hydrolysate yeild a protein hydrolysate
Hydration Assay capable of producing hair hydrating benefits at
virtually identical levels. This demonstrates a novel
approach in hydrolysis, such as the one used in the
manufacturing of ACB Kale Protein Blend provides
just as much moisture as the Wheat Hydrolysate with
proven hydration benefits.

As shown in figure 1, ACB Kale Protein Blend

exhibited similar strong antioxidant properties similar to
200μM concentration of Trolox®, our highest standard
used. The antioxidant capacity of ACB Kale Protein
Blend increased as the concentration increased, as a
result we can assure that its ability to minimize oxidative
stress is dose dependant. ACB Kale Protein Blend
ORAC Assay
was designed to provide hair and skin benefits such as
conditioning and moisturization and, in addition, act as
a film former. However with the present study we can
confirm that this unique ingredient is not only capable of
providing functional benefits but it is also capable of
providing potent antioxidant benefits when added to
cosmetic and personal skin and hair care applications.

As shown in Figure 1, results indicate continuous

improvements in the barrier of the skin throughout the
3 week test period. After one week, the solution
containing 2.0% ACB Kale Protein Blend decreased
TEWL 8% more effectively than the base lotion alone.
After three weeks, the solution containing 2.0% ACB
TEWL Kale Protein Blend demonstrated even more
effective barrier protection, decreasing TEWL 21%
better than the base lotion alone.
When compared to the untreated control, the solution
containing 2.0% ACB Kale Protein Blend decreased
transepidermal water loss by 19% after one week and
by 25% after three weeks.
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Typical Amino Acid Composition See attached

Immediate results showed and average increase in

hair diameter of 13.8% with an average increase of
13.4% four hours following the initial application. After
placing individual hair strands under a microscope,
Microscopy Imaging of the individual strands were
Volumizing Assay taken to visually demonstrate the increase in hair
diameter achieved when using ACB Kale Protein
Blend at 2.0% in a solution compared to the use of
Wheat Hydrolysate at 2.0% in a solution. ACB Kale
Protein Blend is able to effectively volumize the hair for
thicker and younger looking hair, a revolutionary step
for anti-aging hair care products.

Version#2/06.07.18

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 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Tradename: ACB Kale Protein Blend

Code: 20036

CAS #:

Lot #: 150106C & 150106D

Test Request Form #: 787

Sponsor: Active Concepts, LLC – 107 Technology Drive, Lincolnton, North Carolina 28092
Study Director: Erica Segura
Principle Investigator: Meghan Darley

Abstract

The condition of the cuticle (the outer most layer of the hair) significantly affects both, manageability and volume
of our hair. Overtime as hair becomes damaged the cuticle often lifts as a result of a variety of influences including
environment and styling processes. This results in flat, dull hair that is difficult to manage. Improving the body of
the hair has been shown to instantly make it appear healthier and more youthful. Increasing combability not only
eases manageability, but also helps to minimize physical damage that perpetuates the loss of body and difficulty
in styling.

ACB Kale Protein Blend is a product designed to provide volume, hydration and conditioning properties to the
hair. However, this unique ingredient also enhances shine, dry and wet combability, manageability and the
smoothness of the hair. The purpose of this study was to confirm whether or not ACB Kale Protein Blend is
capable of providing these additional benefits.

A half head study was conducted to determine the comparison of using a shampoo incorporating ACB Kale
Protein Blend vs. a control shampoo, in addition to the comparison of using a conditioner incorporating ACB
Kale Protein Blend vs. a control conditioner. Each volunteer’s hair was photographed. The images of the half
head study were used in conjunction with a sensory assessment subjectively rating shine, volume, dry and wet
combability, thickness, smoothness, hydration, softness and manageability. This assessment was conducted
both before and after treatment. Based on the results obtained, ACB Kale Protein Blend is capable of enhancing
the volume and overall health of the hair perfect for use in anti-aging hair care product lines.

Materials and Methods

The study was conducted using five participants with a variety of hair types. Each subject had their baseline photo
taken prior to having their hair washed. The participant was also asked to complete a survey rating their hair prior
to treatment on a scale of 1 to 10, with 1 being the lowest and 10 being the highest, using the following parameters
shine, volume, dry and wet combability, thickness, smoothness, hydration, softness and manageability. Half of
the head was shampooed and conditioned with a base shampoo/ base conditioner + 2.0% ACB Kale Protein
Blend. The hair on the other half of the head was treated with the base shampoo and conditioner alone. After
the application and rinse of the test and positive control products, each participant’s hair was blown dry using a
round brush on both sides of the head.
Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
1
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 1 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Once the hair was completely dry, the participant was asked to again assess the volume, shine, dry combability,
thickness, smoothness, hydration, softness and manageability of both halves of her hair. Assessments were
made using a rubric from 1 to 10, with 1 being the lowest and 10 being the highest.

Results

Assessment of Assessment of Assessment of Assessment of

Baseline Half Head Half Head Half Head Half Head
Assessment of shampooed Shampooed Conditioned Conditioned with
Untreated Hair with Untreated with Kale with Untreated Kale Protein
Control Protein Blend Control Blend
Shine 5.00 4.00 7.00 7.00 10.00
Volume 4.00 4.00 7.00 3.00 8.00
Dry
Combability 2.00 2.00 6.00 6.00 10.00
Wet
Combability 1.00 2.00 5.00 6.00 10.00
Thickness 6.00 8.00 7.00 6.00 7.00
Smoothness 5.00 5.00 9.00 8.00 10.00
Hydration 7.00 6.00 10.00 7.00 10.00
Softness 7.00 3.00 10.00 6.00 10.00
Managability 3.00 4.00 7.00 6.00 9.00
Mean 4.44 4.22 7.56 6.11 9.33

Table 1. Average Results of Participant’s Assessment

Percent Difference -
Percent Difference - Percent Difference –
Comparison of Control
Comparison of Control Baseline Assessment
Conditioner and 2.0% ACB
Shampoo vs. 2.0% ACB Kale vs. Half-Head Kale
Kale Protein Blend +
Protein Blend + Shampoo Protein Blend
Conditioner
Shine 43% 30% 41%
Volume 43% 75% 47%
Dry
Combability 67% 40% 75%
Wet
Combability 60% 40% 87%
Thickness -14% 14% 14%
Smoothness 44% 20% 47%
Hydration 40% 30% 30%
Softness 70% 40% 30%
Managability 43% 33% 63%
Mean 44% 36% 48%
Table 2. Average Change Experienced for Participant’s Sensory Assessment

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
2
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 2 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Assessment of Hair Characteristics

12 Baseline Assessment of Untreated
Rating from 1 (worst) to 10 (best)

Hair
10
Assessment of Half Head shampooed
8 with Untreated Control

6 Assessment of Half Head Shampooed

with ACB Kale Protein Blend
4
Assessment of Half Head
2 Conditioned with Untreated Control

0 Assessment of Half Head

Conditioned with ACB Kale Protein
Blend

Graph 1. Rating of hair characteristics following sensory assessment

Half-Head Half-Head
Treated with Treated with
Positive Control 2.0% ACB Kale
Shampoo and Protein Blend
Conditioner + Base
Shampoo &
Conditioner

Figure 1. Full head Baseline, Untreated Hair Figure 2. Half-Head Treated

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
3
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 3 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Half-Head Half-Head
Treated with Treated with
Positive Control 2.0% ACB Kale
Shampoo and Protein Blend
Conditioner + Base Shampoo
& Conditioner

Figure 3. Full head Baseline, Untreated Hair Figure 4. Half-Head Treated

Half-Head Half-Head
Treated with Treated with
Positive Control 2.0% ACB Kale
Shampoo and Protein Blend
Conditioner + Base Shampoo
& Conditioner

Figure 5. Full head Baseline, Untreated Hair Figure 6. Half-Head Treated

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
4
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 4 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Half-Head Half-Head
Treated with Treated with
Positive Control 2.0% ACB Kale
Shampoo and Protein Blend
Conditioner + Base Shampoo
& Conditioner

Figure 7. Full head Baseline, Untreated Hair Figure 8. Half-Head Treated

Half-Head Half-Head
Treated with Treated with
Positive Control 2.0% ACB Kale
Shampoo and Protein Blend
Conditioner + Base Shampoo
& Conditioner

Figure 9. Full head Baseline, Untreated Hair Figure 10. Half-Head Treated

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
5
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 5 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Changes in Hair Characteristics from Baseline to Post Style

Shine
90%
80%
Managability 70% Volume
60%
50%
40%
30%
Softness 20% Dry Combability
10%
0%

Hydration Wet Combability

Smoothness Thickness

Graph 2. Hair Assessment results for sensory characteristics

Treated Half vs. Control Half After Application & Style

Shine
60%

Managability 50% Volume

40%

30%

20%
Softness 10% Dry Combability

0%

Hydration Wet Combability

Smoothness Thickness

Graph 3. Hair Assessment results for sensory characteristics observed between the positive control and test products

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
6
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 6 of 7 Version#1/02-27-2015/Form#76
 Active Concepts Salon
Half-Head Study
info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Discussion

The results of the assessment indicate that when incorporated into a shampoo, ACB Kale Protein Blend is
capable of improving hair characteristic 44% more than a control shampoo and 36% more than the control
conditioner. When used in both the shampoo and conditioner at 2%, ACB Kale Protein Blend improved shine,
volume, dry and wet combability, smoothness, hydration, softness and manageability by 37%, 59%, 54%, 50%,
32%, 35%, 55% and 38% respectively, better than the control shampoo. These results can be further supported
by figures 1 through 10, where clearly the half of the subject's head treated with 2.0% ACB Kale Protein Blend
appears more voluminous, shiny, soft and healthy.

When comparing hair characteristics of the baseline assessments to the post style assessments, the benefits of
including 2% ACB Kale Protein Blend in a shampoo and conditioner are even more apparent. In relation to the
baseline readings, the test-half of the head improved all subjective parameters, improving shine, volume, dry and
wet combability, thickness, smoothness, hydration, softness and manageability by 41%, 47%, 75%, 87%, 14%,
47%, 30%, 30%, and 63% respectively, with an the test product improving hair conditions on average by 48%
when compared to the untreated baseline.

For this reason, ACB Kale Protein Blend would be an ideal product to use in cosmetic applications designed for
anti-aging hair care that yields voluminous, soft and shiny hair aesthetics.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
7
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 7 of 7 Version#1/02-27-2015/Form#76
 Cellular Viability Assay Analysis

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Tradename: ACB Kale Protein Blend

Code: 20036

CAS #: 100209-45-8 & 100209-45-8 & 100209-45-8

Test Request Form #: 1268

Lot Number: NC-150401

Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC 28092

Study Director: Erica Segura
Principle Investigator: Meghan Darley

Test Performed:
Cellular Viability Assay

Introduction

The cellular viability assay is useful for quantitatively measuring cell-mediated cytotoxicity, cell proliferation and
mitochondrial metabolic activity. Increased metabolism in a cell indicates ample cellular respiration and
adenosine triphosphate (ATP) production. ATP is the molecular energy of cells and is required in basic cell
function and signal transduction. A decrease is ATP levels indicates cytotoxicity and decreased cell function
while an increase in ATP levels indicates healthy cells.

The cellular viability assay was conducted to assess the ability of ACB Kale Protein Blend to increase cellular
metabolic activity in cultured dermal fibroblasts.

Assay Principle

The assay utilizes a nonfluorescent dye, resazurin, which is converted to a fluorescent dye, resorufin, in
response to chemical reduction of growth medium from cell growth and by respiring mitochondria. Healthy cells
that are in a proliferative state will be able to easily convert resazurin into resorufin without harming the cells.
This method is a more sensitive assay than other commonly used mitochondrial reductase dyes such as MTT.
An increase in the signal generated by resazurin-conversion is indicative of a proliferative cellular state.

This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied.
This information is offered solely for your investigation, verification, and consideration.

Page 1 of 3 Version#1/05-22-15/Form#64
 Cellular Viability Assay Analysis

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Materials

A. Kit: Cellular Senescence Assay Kit (Chemicon® International; KAA002)

B. Incubation Conditions: 37°C at 5% CO2 and 95% relative humidity (RH)
C. Equipment: Forma humidified incubator; ESCO biosafety laminar flow hood; Light
microscope; Pipettes
D. Cell Line: Normal Human Dermal Fibroblasts (NHDF) (Lonza; CC-2511)
E. Media/Buffers: Dulbecco's Modified Eagle Medium (DMEM); Penicillin-Streptomycin
(50U- 50mg/mL); Fetal Bovine Serum (FBS); Phosphate Buffered Saline (PBS)
F. Culture Plate: Falcon flat bottom 6-well tissue culture treated plates
G. Reagents: 5-bromo-4-chloro-indolyl-β-D-galactopyranoside (X-gal)
H. Other: Sterile disposable pipette tips; wash bottles; 15mL conical tubes, 1.5mL
microcentrifuge tubes

Methods

Human dermal fibroblasts were seeded into 96-well tissue culture plates and allowed to grow to confluency in
complete DMEM. A 10-fold serial dilution was performed resulting in ACB Kale Protein Blend concentrations
on 1%, 0.1%, and 0.01% in complete DMEM and incubated with fibroblasts for 24 hours.

Ten microliters of viability reagent was added to 90µL of cell culture media in culture wells.

This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied.
This information is offered solely for your investigation, verification, and consideration.

Page 2 of 3 Version#1/05-22-15/Form#64
 Cellular Viability Assay Analysis

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Results

The data obtained from this study met criteria for a valid assay and the controls performed as anticipated.

ACB Kale Protein Blend exhibited significant effects on cellular metabolism compared to the control.

Cellular metabolism results are expressed as a percentage of the control.

122
105
Viability (% of Control)

89

Figure 1: Cellular Metabolism of ACB Kale Protein Blend-treated fibroblasts

expressed in terms of percent of control.

Discussion

As shown in figure 1, ACB Kale Protein Blend exhibited comparable results by increasing cell metabolism. An
increase in fluorescent signal indicates an increase in cellular metabolism and viability. ACB Kale Protein Blend
does not appear to have negative effects on cellular metabolism and can safely be used in cosmetic materials.

This information is presented in good faith but is not warranted as to accuracy of results. Also, freedom from patent infringement is not implied.
This information is offered solely for your investigation, verification, and consideration.

Page 3 of 3 Version#1/05-22-15/Form#64
 Tomorrow’s Vision… Today!®

ACB Kale Protein Blend Hydrolyzed Protein

Hydration Comparison Assay

Code: 20036
INCI Name: Hydrolyzed Kale Protein & Hydrolyzed
Carrot Protein & Hydrolyzed Lemon Protein
Suggested Use Levels: 1.0 - 10.0%
TRF#: 787

Abstract Results

Hydrolyzed proteins, such as Oat, Soy and Wheat have Increase in Hair Hydration
been used in hair care as a traditional means to hydrate 18
the hair and provide strengthening properties. Until 16
Percent (%) Increase

recently, hyrolysis was induced using acid, water, or 14

12
fermentation. Active Concepts has impelemented an
10
innovative hydrolysis approach to the newest and most
8
bio-available vegetable protein on the market, ACB Kale
6
Protein Blend. This microorganism prompted hydrolysis 4
creates the by product, lactic acid, as a secretion which 2
provides comparable hydrating benefits to the leading 0
hydrolyzed proteins on the market. Untreated Control 2.0% ACB Kale Protein 2.0% Wheat
Blend Hydrolysate

Materials and Methods
The hair samples used in this study were tested using
identical intervals and percentages of two protein
hydrolysates, ACB Kale Protein Blend and Wheat
Hydrolysate. The materials used in the procedure
to determine the diameter of each strand were an
untreated control hair sample, the control hair sample
(2.0% Wheat Hydrolysate in an Aqueous Solution), and
the sample treated with the test material (2.0% ACB
Kale Protein Blend in an Aqueous Solution). Using
a 9003 DPM Nova Impedance Meter, hydration levels
of each strand of hair were measured. Both the 2.0%
Wheat Hydrolysate Aqueous Solution and the 2.0%
ACB Kale Protein Blend Aqueous Solution were
shown to increase moisture levels by comparable
amounts in the respective hair strands.
Figure 1.Percent increase in hair hydration using ACB Kale
Protein Blend and Wheat Hydrolysate
Discussion
Both the innovative hydrolysis method used in
the production of ACB Kale Protein Blend and
the traditional method of hydrolysis induced when
creating Wheat Hydrolysate yeild a protein hydrolysate
capable of producing hair hydrating benefits at
virtually identical levels. This demonstrates a novel
approach in hydrolysis, such as the one used in the
manufacturing of ACB Kale Protein Blend provides
just as much moisture as the Wheat Hydrolysate with
proven hydration benefits.

Page 1 of 1
Version 1/ 05.22.15/ Form 2

info@activeconceptsllc.com • +1 (704)-276-7100 • Fax: +1 (704)-276-7101

 Oxygen Radical Absorbance Capacity (ORAC) Assay

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Tradename: ACB Kale Protein Blend

Code: 20036

CAS #: 100209-45-8 & 100209-45-8 & 100209-45-8

Test Request Form #: 1267

Lot Number: NC-150401

Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC 28092

Study Director: Erica Segura
Principle Investigator: Meghan Darley

Test Performed:
Oxygen Radical Absorbance Capacity (ORAC) Assay

Introduction

Reactive oxygen species (ROS) are generated by normal cellular processes, environmental stresses, and UV
irradiation. ROS are dangerous to cellular structures and functional molecules (i.e DNA, proteins, lipids) as
they act as strong oxidizing agents or free radicals. The oxygen radical absorbance capacity (ORAC) assay
is a standard method used to assess antioxidant capacity of physiological fluids, foods, beverages, and natural
products. The assay quantitatively measures a sample’s ability to quench free radicals that have the potential
to react with and damage cellular components.

Oxygen Radical Absorbance Capacity (ORAC) assay was conducted to assess the antioxidant capacity of ACB
Kale Protein Blend.

Assay Principle

This assay is based upon the effect of peroxyl radicals generated from the thermal decomposition of 2, 2’-
azobis-2-methyl-propanimidamide dihydrochloride (AAPH) on the signal intensity from the fluorescent probe,
fluorescein, in the presence of an oxygen radical absorbing substance. The degree of change is indicative of
the amount of radical damage and the presence of antioxidants results in an inhibition in the free radical
damage to the fluorescein. The antioxidant protection of the sample can be calculated by comparing it to a
set of known standards. Trolox®, a water soluble vitamin E analog, with known antioxidant capabilities is used
in this ORAC assay as the standard for measuring the antioxidant capacity of unknown substances. ORAC
values, expressed in µM of Trolox® equivalents (TE), are calculated using the area under the curves (AUC)
of the test product, Trolox®, and the control materials. Trolox equivalency is used as the benchmark for
antioxidant capacity of mixtures since it is difficult to measure individual components.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
1
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 1 of 3 Version#1/05-22-15/Form#56
 Oxygen Radical Absorbance Capacity (ORAC) Assay

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Materials

A. Equipment: Synergy H1 Microplate reader (BioTek Instuments, Winooski, VT); Gen5

software (BioTek Instuments, Winooski, VT); Pipettes
B. Buffers: 75mM Potassium Phosphate (pH 7.4); Deionized H2O
C. Reagents: 2,2′-Azobis(2-methylpropionamidine) dihydrochloride (AAPH) (153mM);
6-Hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox®);
Fluorescein Sodium Salt (4nM)
D. Preparation: Pre-heat (37°C) Synergy H1 Microplate reader; Prepare Trolox®
standards, sample dilutions, fluorescein solution, and AAPH.
E. Microtitre Plates: Corning 96 Well Black Side/Clear Bottom Microplates

Methods

Solutions of ACB Kale Protein Blend and Trolox® (positive control) were prepared in 75mM potassium
phosphate buffer. Materials were prepared at three different concentrations/dilutions. Trolox® was used as a
reference for antioxidant capacity and prepared at a concentrations ranging from 12.5µM to 200µM in 75mM
potassium phosphate buffer.

For the ORAC assay, 25µL of test material and Trolox® were combined with 150µL of fluorescein in 75mM
potassium phosphate buffer and incubated in the Synergy HT Microplate reader at 37˚C for 30 minutes. At the
end of the incubation period, 25µL of AAPH were pipetted into each well. Fluorescent measurements were then
taken every 2 minutes for approximately 2 hours at an excitation wavelength of 485nm and an emissions
wavelength of 520nm.

The AUC and Net AUC values of the standards and samples were determined using Gen5 2.0 Data Reduction
Software using the below equations:

𝑅2 𝑅3 𝑅4 𝑅𝑛
𝐴𝑈𝐶 = 0.5 + + + +⋯+ → 𝑊ℎ𝑒𝑟𝑒 𝑅 𝑖𝑠 𝑓𝑙𝑢𝑜𝑟𝑒𝑠𝑐𝑒𝑛𝑐𝑒 𝑟𝑒𝑎𝑑𝑖𝑛𝑔
𝑅1 𝑅1 𝑅1 𝑅1

𝑁𝑒𝑡 𝐴𝑈𝐶 = 𝐴𝑈𝐶𝑠𝑎𝑚𝑝𝑙𝑒 − 𝐴𝑈𝐶𝑏𝑙𝑎𝑛𝑘

The standard curve was obtained by plotting the Net AUC of different Trolox® concentrations against their
concentration. ORAC values of samples were then calculated automatically using the Gen5 software to
interpolate the sample’s Net AUC values against the Trolox® standard curve. ORAC measurements for the test
material were expressed in micro moles Trolox® equivalents per mL (µMTE), where 1 ORAC unit is equal to 1
µMTE.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
2
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 2 of 3 Version#1/05-22-15/Form#56
 Oxygen Radical Absorbance Capacity (ORAC) Assay

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Results

ACB Kale Protein Blend exhibited very potent antioxidant activity at a 0.5% concentration.

199 206

100
83

13

Figure 1: Antioxidant capacities

Discussion

As shown in figure 1, ACB Kale Protein Blend exhibited similar strong antioxidant properties similar to 200µM
concentration of Trolox®, our highest standard used. The antioxidant capacity of ACB Kale Protein Blend
increased as the concentration increased, as a result we can assure that its ability to minimize oxidative stress
is dose dependant.

ACB Kale Protein Blend was designed to provide hair and skin benefits such as conditioning and moisturization
and, in addition, act as a film former. However with the present study we can confirm that this unique ingredient
is not only capable of providing functional benefits but it is also capable of providing potent antioxidant benefits
when added to cosmetic and personal skin and hair care applications.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
3
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 3 of 3 Version#1/05-22-15/Form#56
 Transepidermal Water Loss Study

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Tradename: ACB Kale Protein Blend

Code: 20036

CAS #: 100209-45-8 & 100209-45-8 & 100209-45-8

Test Request Form #: 1269

Lot Number: NC-150401

Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC 28092

Study Director: Erica Segura
Principle Investigator: Meghan Darley

Test Performed: Transepidermal Water Loss Study

Introduction
An in-vivo study was conducted over a period of three weeks to evaluate the ability of ACB Kale Protein
Blend to enhance barrier function through reduction in Transepidermal Water Loss (TEWL). Results indicate
that this material is capable of efficiently reducing TEWL which allows moisture retention.

Materials

A. Equipment: DermaLab Skin Combo

Methods

Ten volunteers M/F between the ages of 23 and 45 and who were known to be free of any skin pathologies
participated in this study. A Dermalab Combo was used to measure TEWL on the subject’s volar forearms.
The instrument consists of a probe that is based upon the vapor gradient with an open chamber. This open
chamber design maintains the free natural evaporation from the skin without interfering with the
environment over the measurement area. This ensures unbiased and accurate readings. Operation of the
water loss module is fully menu drive, allowing for pre-setting and standard deviation or measurement time.
Baseline TEWL readings were taken on day one of the study.

Following initial measurements, all subjects were asked to apply 5milligrams of each test material on their
volar forearms. Measurements were taken immediately after application of the test materials and then
weekly for three weeks. The test material consisted of 2% ACB Kale Protein Blend in a base lotion.

For added perspective, measurements of an untreated test site and a site treated with a base lotion
(Cetaphil Moisturizing for All Skin Types) were recorded.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
1
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 1 of 2 Version#1/05-22-15
 Transepidermal Water Loss Study

info@activeconceptsllc.com • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Results

Figure 1: Improvements in barrier function following application of the test

materials after a period of 3 weeks.

Discussion

As shown in Figure 1, results indicate continuous improvements in the barrier of the skin throughout the 3
week test period. After one week, the solution containing 2.0% ACB Kale Protein Blend decreased TEWL
8% more effectively than the base lotion alone. After three weeks, the solution containing 2.0% ACB Kale
Protein Blend demonstrated even more effective barrier protection, decreasing TEWL 21% better than the
base lotion alone.

When compared to the untreated control, the solution containing 2.0% ACB Kale Protein Blend decreased
transepidermal water loss by 19% after one week and by 25% after three weeks.

Information contained in this technical literature is believed to be accurate and is offered in good faith for the benefit of the customer. The company, however, cannot assume any liability or risk
2
involved in the use of its chemical products since the conditions of use are beyond our control. Statements concerning the possible use of our products are not intended as recommendations to
use our products in the infringement of any patent. We make no warranty of any kind, expressed or implied, other than that the material conforms to the applicable standard specification.

Page 2 of 2 Version#1/05-22-15
 Tomorrow’s Vision… Today!®

ACB Kale Protein Blend Hair Volumizing Assay

Hydrolyzed Protein Comparison

Code: 20036
INCI Name: Hydrolyzed Kale Protein & Hydrolyzed
Carrot Protein & Hydrolyzed Lemon Protein
Suggested Use Levels: 1.0 - 10.0%
TRF#: 787

Abstract Results

Hydrolyzed proteins, such as Oat, Soy and Wheat have Effects on Hair Diameter
been used in hair care as a traditional means to hydrate 16
13.8 13.4
the hair and provide strengthening properties. Until Percent (%) Increase
14
recently, hydrolysis was induced using acid, water, or 12
fermentation. Active Concepts has implemented an 10

innovative hydrolysis approach to the newest and most 8

bio-available vegetable protein on the market, ACB Kale 6

Protein Blend. This microorganism prompted hydrolysis 4

2.2
2
creates the by product, lactic acid, as a secretion which 0.49
0
provides volumizing and anti-aging benefits measured.
Immediately Following Application 4 Hours After Application

ACB Kale Protein Blend Wheat Hydrolysate

Materials and Methods Figure 1.Percent increase in hair diameter over time after
treatment with ACB Kale Protein Blend and Wheat
The hair samples used in this study were tested using Hydrolysate

identical intervals and percentages of two protein

hydrolysates, ACB Kale Protein Blend and Wheat Microscopy Imaging of Individual Hair Strands
Hydrolysate. The materials used in the procedure
to determine the diameter of each strand were an
untreated control hair sample, the control hair sample
(treated with 2.0% Wheat Hydrolysate Solution), and
the sample treated with the test material (2.0% ACB
Kale Protein Blend). Each hair was imaged and
measured before a solution was applied. The hairs
were then removed from the slide and either placed in
the 2.0% solution of the Wheat Hydrolysate or the 2.0%
solution of ACB Kale Protein Blend. Each hair was
removed, measured and imaged then placed aside. Figure 2. Individual strand Figure 3. Individual strand
immediately following immediately following
After four hours, each hair was reimaged and measured treatment with Wheat treatment with ACB Kale
to demonstrate the sustained volume potential of each Hydrolysate (note beading) Protein Blend
respective hydrolyzed protein.
Page 1 of 2
Version 1/ 05.22.15/ Form 2

info@activeconceptsllc.com • +1 (704)-276-7100 • Fax: +1 (704)-276-7101

 Tomorrow’s Vision… Today!®

ACB Kale Protein Blend Hair Volumizing Assay

Hydrolyzed Protein Comparison

Code: 20036
INCI Name: Hydrolyzed Kale Protein & Hydrolyzed
Carrot Protein & Hydrolyzed Lemon Protein
Suggested Use Levels: 1.0 - 10.0%
TRF#: 787

Results (Cont.)

Figure 4. Individual strand Figure 5. Individual strand

four hours after treatment with four hours after treatment with
Wheat Hydrolysate ACB Kale Protein Blend

Discussion

Immediate results showed and average increase in

hair diameter of 13.8% with an average increase of
13.4% four hours following the initial application. After
placing individual hair strands under a microscope,
Microscopy Imaging of the individual strands were
taken to visually demonstrate the increase in hair
diameter achieved when using ACB Kale Protein
Blend at 2.0% in a solution compared to the use of
Wheat Hydrolysate at 2.0% in a solution. ACB Kale
Protein Blend is able to effectively volumize the hair
for thicker and younger looking hair, a revolutionary
step for anti-aging hair care products.

Page 2 of 2
Version 1/05.22.15/ Form 2

info@activeconceptsllc.com • +1 (704)-276-7100 • Fax: +1 (704)-276-7101