AC Griffonia Lysate Advanced Efficacy Data · 2020. 9. 10. · Griffonia Simplicifolia Seed Extract . CAS #: 999999-99-4 . EINECS #: 310-127-6. Name. of Study Results: Immediate Skin

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Code: 16634 INCI Name: Griffonia Simplicifolia Seed Extract CAS #: 999999-99-4 EINECS #: 310-127-6

Name of Study Results

Immediate Skin Lifting Benefits Assay

According to the results of the assessment, 66.7% of participants were able to perceive a significant tightening effect on the area around the eye where the experimental (5.0% AC Griffonia Lysate Advanced in base serum) was applied. Additionally, the experimental (5.0% AC Griffonia Lysate Advanced in base serum) delivered 36.2% more of a perceivable tightening effect than the base serum alone. The active in base serum demonstrated a statistically significant difference in perceived tightening, when comparing the percent change of experimental and base ratings.

ORAC Assay

AC Griffonia Lysate Advanced exhibited antioxidant activity comparable to 200μM Trolox®. The antioxidant capacity of AC Griffonia Lysate Advanced increased as the concentration increased, as a result we can assure that its ability to minimize oxidative stress is dose dependent.

Cellular Viability Assay

AC Griffonia Lysate Advanced exhibited positive results by increasing cell metabolism. The increase in fluorescent signal indicates an increase in cellular metabolism and viability post AC Griffonia Lysate Advanced treatment. For these reasons, we can assume AC Griffonia Lysate Advanced is suitable for cosmetic applications designed to increase cell viability and metabolism.

AC Griffonia Lysate Advanced Efficacy Data

file://Act-svcs-01/public/Marketing%20Materials/Formulations/Sprayable%20Lotion%20with%20NEC.docxfile://Act-svcs-01/public/Marketing%20Materials/Formulations/Sprayable%20Lotion%20with%20NEC.docx

Code Number: 16634

INCI Name: Griffonia Simplicifolia Seed ExtractINCI Status: ConformsCAS Number: 999999-99-4EINECS Number: 310-127-6

TRF#: 6306Lot Number(s): 72745

Suggested Use Levels: 1.0 - 10.0%Use Level for Assay: 5.0%

Sponsor: Active Concepts, LLC 107 Technology Drive Lincolnton, North Carolina 28092

Study Director: Maureen DanaherPrinciple Investigator: Kara Rivera

Suggested Applications: Anti-Aging, UV Protection, Tightening, Lifting

INTRODUCTIONAn in-vivo study was conducted to evaluate the immediately perceivable skin tightening benefits of AC Griffonia Lysate Advanced. Along with perceivable skin tightening, images were taken to visually assess the appearance of fine lines and wrinkles after a short term period. All participants were asked to evaluate the improvement in skin tightening compared to the control. Results indicate that this material is capable of significantly increasing perceivable skin tightening and minimizes the appearance of fine lines compared to the control.

MATERIALS AND METHODSThis study was conducted using 6 M/F subjects between the ages of 25-50. Half of the participant population was instructed to apply the 5.0% active loaded eye serum to their right under eye area and the other half to their left. The participants were then instructed to apply the base eye serum to their opposite under eye area. Each participant was supplied with ~2.0mg of product for each application.

Baseline photos were taken prior to application of the test materials. Photos were taken again three mintues following application of the test materials. Photographic assessments were performed using the VISIA Complexion Analysis System (Canfield Scientific., Fairfield, NJ, USA). The VISIA System, with a configurable head support, ensured consistent positioning of each subject’s head. The participants cleaned their skin with a gentle facial wipe (Daily Facial Towelettes – Paraben Free Formula by Kirkland Signature) before the image was obtained. Images were taken for each subject to quantify the feature counts for wrinkles.

Three minutes after application of test materials, participants were asked to rate the feeling of tightness on a scale of 1-10 for each under eye area, where 1 represents no perceived improvement. This data provides an assessment of perceivable skin tightening. The average ratings for the experimental and control were calculated, and the differences between time points were recorded and compared. For statistical analysis a two-sample t-test, assuming unequal variance, was performed to compare data. The significance threshold was set at 0.05.

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AC Griffonia Lysate Advanced

Benefits of AC Griffonia Lysate Advanced:

• Immediately Perceivable Results

• Neuro-Cosmetic

• Long Term Anti-Aging Benefits

• 5-HTP Marketing

in vivo Analysis - Immediate Skin Lifting Benefits

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Table 2. Participant assessment of tightness.

According to the results of the assessment, 66.7% of participants were able to perceive a significant tightening effect on the area around the eye where the experimental (5.0% AC Griffonia Lysate Advanced in base serum) was applied. Additionally, the experimental (5.0% AC Griffonia Lysate Advanced in base serum) delivered 36.2% more of a perceivable tightening effect than the base serum alone. The active in base serum demonstrated a statistically significant difference in perceived tightening, when comparing the percent change of experimental and base ratings. P-values were less than or equal to 5% indicating results of the study were not due to random chance.

Participant Assessment of Tightness

t-Test: Two-Sample Assuming Equal Variances

Variable 1 Variable 2Mean 6.166666667 4.333333Variance 0.805555556 5.222222Observations 7 7Pooled Variance 3.013888889Hypothesized Mean Difference 0df 12t Stat 1.975658322P(T

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RESULTS


AC Griffonia Lysate AdvancedTomorrow’s Vision... Today!®

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Figure 1. Participant #2 treated with 5.0% AC Griffonia Lysate Advanced in Base Serum displays a reduction (10.0%) in wrinkle feature counts from beginning of treatment (T=0) to T=3 minutes via VISIA Image Analysis. Images on the top are participant #2 with image enhancement, through VISIA, which provides higher visualization of feature changes. Images on the bottom are natural photos of participant #2.

T = 0 T = 3 Minutes

Wrinkles (63) Wrinkles (57)

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RESULTS



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RESULTS

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Figures 1 through 3 illustrate the changes in wrinkle characteristics on participants from baseline to three minutes following application of the base serum containing 5.0% AC Griffonia Lysate Advanced. The VISIA feature counts demonstrate a reduction in the overall appearance of wrinkles. According to the results of the panelist images and ratings assessment, it can be concluded that AC Griffonia Lysate Advanced can be used to visibly reduce the appearance of fine lines and wrinkles while providing a noticeable tightening effect on skin. For this reason, this exceptional ingredient is ideal to add to cosmetic and personal care applications where immediate lifting benefits are desired.

It should be noted that the 2020 Global Health Crisis has limited the panelist size and age demographic for this study. As the average participant age for this study is 26 years, AC Griffonia Lysate Advanced would be ideal for the ‘Prejuvenation’ trend. Treating the signs of aging before they set in has become an increasingly popular approach to dealing with wrinkles and other issues associated with getting older. ‘Prejuvenation’ encompasses both the preventative and restorative approach to skin care that Millennials and Generation Z are adopting. Instead of trying to turn back the effects of time, an increasing number of young adults are focused on an effective skin care regimen earlier in order to stop time in its tracks. The immediate tightening effect of AC Griffonia Lysate Advanced can help to maintain the health and aesthetics of youthful skin while also enhancing it with the active’s antioxidant capabilities and collagen production benefits.

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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 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 recom-mendations 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. Freedom from patent infringement is not implied. All information is for investigative purposes only.

Active Concepts LLC, AsiaKaohsiung, Taiwan

www.activeconceptsllc.comTel + 886 73599900

[email protected]

Active Concepts, LLCLincolnton, NC. USA

www. activeconceptsllc.comOffice: +1 (704) 276 7100

[email protected]

Active Concepts S.r.l. Milano ITALY

www.activeconcepts.it Tel +39 02 90360719

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[email protected]

DISCUSSION



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Oxygen Radical Absorbance Capacity (ORAC) Assay

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 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.

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[email protected] • Phone: +1-704-276-7100 • Fax: +1-704-276-7101

Tradename: AC Griffonia Lysate Advanced

Code: 16634

CAS #: 999999-99-4

Test Request Form #: 395

Lot #: 27203

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)

Introduction

Reactive oxygen species (ROS) are generated by normal cellular processes, environmental stresses, and UV irradiation. ROS are detrimental 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 AC Griffonia Lysate Advanced.

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.

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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 AC Griffonia Lysate Advanced 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:

𝐴𝑈𝐶 = 0.5 + 𝑅2

𝑅1+

𝑅3

𝑅1+

𝑅4

𝑅1+ ⋯ +

𝑅𝑛

𝑅1→ 𝑊ℎ𝑒𝑟𝑒 𝑅 𝑖𝑠 𝑓𝑙𝑢𝑜𝑟𝑒𝑠𝑐𝑒𝑛𝑐𝑒 𝑟𝑒𝑎𝑑𝑖𝑛𝑔

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 (µMTE), where 1 ORAC unit is equal to 1 µMTE.

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Results AC Griffonia Lysate Advanced began exhibiting antioxidant activity at a 0.05% concentration.

Figure 1: Antioxidant capacities

Discussion As shown in figure 1, AC Griffonia Lysate Advanced exhibited antioxidant activity comparable to 50µM Trolox®. The antioxidant capacity of AC Griffonia Lysate Advanced increased as the concentration increased, as a result we can assure that its ability to minimize oxidative stress is dose dependent. AC Griffonia Lysate Advanced was designed to provide anti-aging, tightening, and lifting properties. 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 applications.

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Cellular Viability Assay Analysis

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.


Tradename: AC Griffonia Lysate Advanced

Code: 16634

CAS #: 999999-99-4

Test Request Form #: 368

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 AC Griffonia Lysate Advanced 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.





Materials

A. Kit: PrestoBlue™ Cell Viability Reagent (Invitrogen, A13261)

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 96-well tissue culture treated plates

G. Reagents: PrestoBlue™ reagent (10X)

H. Other: Sterile disposable pipette tips

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 AC Griffonia Lysate Advanced

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.





Results

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

AC Griffonia Lysate Advanced at all concentrations is able to increase cellular metabolism compared to the

control.

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

Figure 1: Cellular Metabolism of AC Griffonia Lysate

Advanced-treated fibroblasts expressed in terms of percent of

control.

Discussion

As shown in figure 1, AC Griffonia Lysate Advanced exhibited positive results by increasing cell metabolism.

The increase in fluorescent signal indicates an increase in cellular metabolism and viability post AC Griffonia

Lysate Advanced treatment. For these reasons, we can assume AC Griffonia Lysate Advanced is suitable

for cosmetic applications designed to increase cell viability and metabolism.

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16634 AC Griffonia Lysate Advanced

Documents

AC Griffonia Lysate Advanced Efficacy Data · 2020. 9. 10. · Griffonia Simplicifolia Seed Extract . CAS #: 999999-99-4 . EINECS #: 310-127-6. Name. of Study Results: Immediate Skin