ac royal jelly extract efficacy data - active concepts,...

Tomorrow’s Vision… Today!®

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Code: 20478 INCI Name: Butylene Glycol & 10-Hydroxydecanoic Acid & Sebacic Acid & 1,10-Decanediol CAS #: 107-88-0 & 1679-53-4 & 111-20-6 & 112-47-0EINECS #: 203-529-7 & 216-848-1 & 203-845-5 & 203-975-2

Type of Study Results

High Resolution Ultrasound Skin Imaging

As evidenced in a 4-week efficacy study of AC Royal Jelly Extract on skin, skin density was improved by 6.16% after 1 week and by 17.69% after 4 weeks when compared to the untreated control. When compared to the base lotion AC Royal Jelly Extract improved skin density more than the base lotion alon. Results indicate that AC Royal Jelly Extract is capable of improving skin density when compared to the untreated control and the base lotion. AC Royal Jelly Extract has a positive effect on skin’s density when used at recommended use levels.

Moisturization Assay

As evidenced in a 4-week efficacy study of AC Royal Jelly Extract on the skin, moisture levels were improved by 24.08% after 24 hours and by 50.69% after 4 weeks when compared to the untreated control. When compared to the base cream AC Royal Jelly Extract improved moisturization by 27.15% after 4 weeks. The results indicate that AC Royal Jelly Extract is capable of increasing moisturization when compared to both the untreated control as well as the base lotion. The present study confirms that AC Royal Jelly Extract is capable of providing moisturizing and skin hydrating benefits when added to cosmetic applications.

ORAC Assay

As shown in figure 1, AC Royal Jelly Extract exhibited antioxidant activity comparable to 50μM Trolox®. The antioxidant capacity of AC Royal Jelly Extract increased as the concentration increased, as a result we can assure that its ability to minimize oxidative stress is dose dependent. With the present study we can confirm that this unique ingredient is capable of providing antioxidant benefits when added to cosmetic applications.

TEWL

When compared to the untreated control, the solution containing 2.0% AC Royal Jelly decreased transepidermal water loss by 2% after four weeks of application. AC Royal Jelly Extract is a functional material that can also decrease transepidermal water loss, thus providing great moisture retention.

AC Royal Jelly Extract Efficacy Data

Version#2/12-12-15/ Form#82

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High Resolution Ultrasound Skin Imaging Assay

i n f o @ a c t i v e c o n c e p t s l l c . c o m • + 1 ( 7 0 4 ) - 2 7 6 - 7 1 0 0 • F a x : + 1 ( 7 0 4 ) - 2 7 6 -7101

Version#1/08-12-15/Form#75

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.

Tradename: AC Royal Jelly Extract Code: 20478 CAS #: 107-88-0 & 1679-53-4 & 111-20-6 & 112-47-0 Test Request Form #: 920 Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC 28092

Study Director: Erica Segura

Principle Investigator: Meghan Darley

Test Performed: High Resolution Ultrasound Skin-Imaging Assay

Introduction

An in-vivo study was conducted over a period of four weeks to evaluate the effect on skin density AC Royal Jelly Extract. 10 M/F subjects between the ages of 23-45 participated in the study. Data gathered from the high resolution ultrasound imaging yielded results that indicate that this material is capable of significantly improving skin density compared to the control.

Materials

A. Equipment: DermaLab Skin Combo (Ultrasound Probe)

Methods High Resolution Ultrasound Skin imaging is based on measuring the acoustic response after an acoustic pulse is sent into the skin. The energy of the acoustic pulse is low and will not affect the skin in any way. When the acoustic pulse is emitted and hits different areas of the skin, part of the pulse will be reflected and part will be transmitted further into the skin. The reflected signal travels back and is picked up by the ultrasound transducer. After processing the signal, a cross-sectional image appears on the screen. This image represents an intensity, or amplitude, analysis of the signals. The intensity of the signals that are received refer to a color scale. Dark colors represent areas of the skin with low reflection. This means that there are no changes or very small changes in density between the structures in the skin. Bright colors represent areas with strong reflections, signifying substantial changes in density between structures. 10 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. The DermaLab ultrasound probe was used to determine the skin density of the subject’s volar forearms.

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Following initial measurements, all subjects were asked to apply 2 mg of each test material on their volar forearms. Measurements were taken 24 hours after application of test materials and then weekly for 4 weeks. The test material consisted of 2.0% AC Royal Jelly Extract 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.

Results AC Royal Jelly Extract showed improvements in skin density at a 2.0% concentration. Please note each value is an average of three consecutive readings per test site. Individual Raw Data:

t = 0 1 week 2 weeks 3 weeks 4 weeks

Subject 1-Test 55 53 40 49 49

Base Lotion 57 63 56 58 59

Untreated Control 64 68 56 63 64

Subject 2-Test 30 61 53 57 61

Base Lotion 46 39 42 46 36


Subject 3-Test 60 43 37 49 48

Base Lotion 62 54 53 50 50


Subject 4-Test 72 73 71 78 78

Base Lotion 69 62 59 68 70


Subject 6-Test 59 74 55 71 77

Base Lotion 64 69 47 70 65


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T = 0 1 week 2 weeks 3 weeks 4 weeks

Subject 7-Test 66 44 44 50 52

Base Lotion 48 54 41 52 50


Subject 8-Test 75 88 72 86 87

Base Lotion 67 74 45 50 55


Subject 9-Test 43 49 52 57 65

Base Lotion 49 52 43 55 53


Subject 10-Test 72 62 71 72 75

Base Lotion 69 62 57 60 65


Raw Data:

t=0 1 week 2 week 3 week 4 week

Experimental (2.0% AC Royal Jelly Extract in Base Lotion)

60.1 60.3 55.77 64.6 65.77

Base Lotion 58.1 56.8 49.55 56.6 55.88

Untreated Control 77.9 82.5 71.22 79.6 74.11

Base Lotion vs. Untreated Control

-25.42% -31.15% -30.42% -28.89% -24.59%

Experimental (2.0% AC Royal Jelly Extract in Base Lotion) vs. Base Lotion

-22.85% -26.91% -21.68% -18.84% -11.24%

Experimental (2.0% AC Royal Jelly Extract in Base Lotion) vs. Untreated

3.44% 6.16% 12.56% 14.13% 17.69%

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Figure 1: Percent difference in skin density recordings

between test materials

Discussion As evidenced in a 4-week efficacy study of AC Royal Jelly Extract on skin, skin density was improved by 6.16% after 1 week and by 17.69% after 4 weeks when compared to the untreated control. When compared to the base lotion AC Royal Jelly Extract improved skin density more than the base lotion alon. Results indicate that AC Royal Jelly Extract is capable of improving skin density when compared to the untreated control and the base lotion. AC Royal Jelly Extract has a positive effect on skin’s density when used at recommended use levels.

-40.00%

-30.00%

-20.00%

-10.00%

0.00%

10.00%

20.00%

30.00%

T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks

Pe

rce

nt(

%)

Ch

ange

Comparative Analysis of Skin Density

Base Lotion vs.Untreated Control

2% Royal Jelly Extractvs. Base Lotion

2% Royal Jelly Extractvs. Untreated Control

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Moisturization/Hydration Assay




Tradename: AC Royal Jelly Extract

Code: 20478 CAS #: 107-88-0 & 1679-53-4 & 111-20-6 & 112-47-0 Test Request Form #: 920 Sponsor: Active Concepts, LLC; 107 Technology Drive Lincolnton, NC 28092



Test Performed: Moisturization/Hydration Assay

Introduction

An in-vivo study was conducted over a period of three weeks to evaluate the moisturization benefits of AC Royal Jelly Extract. 10 M/F subjects between the ages of 23-45 participated in the study. Results indicate that this material is capable of significantly increasing moisturization compared to the control. The moisturization assay was conducted to assess the moisturizing ability of AC Royal Jelly Extract.

Materials A. Equipment: DermaLab Skin Combo (Hydration/ Moisture Pin Probe)

Methods The moisture module provides information about the skin’s hydration by measuring the conducting properties of the upper skin layers when subjected to an alternating voltage. The method is referred to as a conductance measurement and the output is presented in the unit of uSiemens (uS). A moisture pin probe is the tool used to gather hydration values. 10 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 Corneometer was used to measure the moisture levels on the subject’s volar forearms. The Corneometer is an instrument that measures the amount of water within the skin. The presence of moisture in the skin improves conductance therefore results in higher readings than dry skin. Therefore the higher the levels of moisture, the higher the readings from the Corneometer will be. Baseline moisturization readings were taken on day one of the study.

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Following initial measurements, all subjects were asked to apply 2 mg of each test material on their volar forearms. Measurements were taken immediately after application of test materials and then weekly for 4 weeks. The test material consisted of 2% AC Royal Jelly Extract 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.

Results AC Royal Jelly Extract showed very high moisturizing capabilities at a 2.0% concentration. Please note each value is an average of three consecutive readings per test site.

Individual Raw Data:

T=0 T=24 1 week 2 weeks 3 weeks 4 weeks

Subject 1-Test

Experimental 25 61 63 65 68 70

Base Lotion 45 72 47 53 64 59

Untreated Control 43 67 35 49 44 51

Subject 2-Test

Experimental 43 75 72 97 99 108

Base Lotion 67 67 73 70 86 86


Subject 3-Test


Base Lotion 18 35 41 44 54 59


Subject 4-Test


Base Lotion 43 72 85 82 86 85


Subject 5-Test

Experimental 57 77 85 123 149 148

Base Lotion 55 91 89 95 115 116


Subject 6-Test

Experimental 35 61 65 69 98 101

Base Lotion 39 51 53 75 61 72

Untreated 53 53 63 61 59 55

Subject 7-Test

Experimental 37 72 78 84 105 121

Base Lotion 39 65 72 67 73 70


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T=0 T=24 1 week 2 weeks 3 weeks 4 weeks

Subject 8-Test


Base Lotion 29 72 69 74 79 82


Subject 9-Test


Base Lotion 28 43 45 57 66 66


Subject 10-Test


Base Lotion 26 45 66 55 57 60


# of Subjects 10 10 10 10 10

Results of Group:

t=0 t=24 1 week 2 weeks 3 weeks 4 weeks Experimental (2.0% AC Royal Jelly Extract in Base Lotion)

37 62 67 88 90 107

Base Lottion 39 61 64 75 74 84


Base Lotion vs. Untreated -12.58% 24.08% 25% 32.02% 40.61% 50.69%

Experimental (2.0% AC Royal Jelly Extract in Base Lotion) vs. Untreated

-16.12%

25.10% 31.25% 55.79% 70.58% 91.61%

Experimental (2.0% AC Royal Jelly Extract in Base Lotion) vs. Base Lotion

-4.11% .815% 5% 18% 21.32% 27.15%

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Figure 1: Moisturization Results.

Figure 2: Comparative Moisturization Results.

0

20

40

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80

100

120

T=0 T=24 T=1 Week T=2 Weeks T=3 Weeks T=4 Weeks

Mo

istu

riza

tio

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µSi

em

en

s)

Average Moisturization

Untreated Control

Base Lotion

Experimental (2% RoyalJelly Extract in baselotion)

-40

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20

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60

80

100

T = 0 T = 24 Hours T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks

Pe

rce

nt

(%)

Dif

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Comparative Moisturization

Base Lotion vs.Untreated Control

2% Royal Jelly Extractin Base Lotion vs.Untreated Control

2% Royal Jelly Extractin Base Lotion vs. BaseLotion

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Discussion As evidenced in a 4-week efficacy study of AC Royal Jelly Extract on the skin, moisture levels were improved by 24.08% after 24 hours and by 50.69% after 4 weeks when compared to the untreated control. When compared to the base cream AC Royal Jelly Extract improved moisturization by 27.15% after 4 weeks. The results indicate that AC Royal Jelly Extract is capable of increasing moisturization when compared to both the untreated control as well as the base lotion. The present study confirms that AC Royal Jelly Extract is capable of providing moisturizing and skin hydrating benefits when added to cosmetic applications.

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Oxygen Radical Absorbance Capacity (ORAC) Assay




Tradename: AC Royal Jelly Extract Code: 20478 CAS #: 107-88-0 & 1679-53-4 & 111-20-6 & 112-47-0 Test Request Form #: 419 Lot #: 24407 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 Royal Jelly Extract. 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 Royal Jelly Extract 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:

𝐴𝐴𝐴𝐴𝐴𝐴 = 0.5 + 𝑅𝑅2𝑅𝑅1

+𝑅𝑅3𝑅𝑅1

+𝑅𝑅4𝑅𝑅1

+ ⋯+𝑅𝑅𝑅𝑅𝑅𝑅1

→ 𝑊𝑊ℎ𝑒𝑒𝑒𝑒𝑒𝑒 𝑅𝑅 𝑖𝑖𝑖𝑖 𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑓𝑒𝑒𝑒𝑒𝑖𝑖𝑓𝑓𝑒𝑒𝑅𝑅𝑓𝑓𝑒𝑒 𝑒𝑒𝑒𝑒𝑟𝑟𝑟𝑟𝑖𝑖𝑅𝑅𝑟𝑟

𝑁𝑁𝑒𝑒𝑁𝑁 𝐴𝐴𝐴𝐴𝐴𝐴 = 𝐴𝐴𝐴𝐴𝐴𝐴𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠𝑠 − 𝐴𝐴𝐴𝐴𝐴𝐴𝑏𝑏𝑠𝑠𝑠𝑠𝑏𝑏𝑏𝑏 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 Royal Jelly Extract began exhibiting antioxidant activity at a 2.5% concentration.

Figure 1: Antioxidant capacities

Discussion As shown in figure 1, AC Royal Jelly Extract exhibited antioxidant activity comparable to 50µM Trolox®. The antioxidant capacity of AC Royal Jelly Extract increased as the concentration increased, as a result we can assure that its ability to minimize oxidative stress is dose dependent. With the present study we can confirm that this unique ingredient is capable of providing antioxidant benefits when added to cosmetic applications.

56

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37

13

10

10

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30

40

50

60

50µM Trolox 6.25µM Trolox 10% 20478AC Royal Jelly

Extract

5% 20478AC Royal Jelly

Extract

2.5% 20478AC Royal Jelly

Extract

Antio

xida

nt C

apac

ity (µ

MTE

)

20478 AC Royal Jelly Extract ORAC

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Transepidermal Water Loss Study


Version#1/08-12-15


Tradename: AC Royal Jelly Extract Code: 20478 CAS #: 107-88-0 & 1679-53-4 & 111-20-6 & 112-47-0 Test Request Form #: 920

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



Test Performed: Transepidermal Water Loss Study

Introduction An in-vivo study was conducted over a period of three weeks to evaluate the ability of AC Royal Jelly Extract 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% AC Royal Jelly Extract 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.

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Transepidermal Water Loss Study


Version#1/08-12-15


Results

Figure 1: Improvements in barrier function following application of the test materials after a period of 4 weeks.

Discussion As shown in Figure 1, results indicate continuous improvements in the barrier of the skin throughout the 4-week

test period. After one week, the solution containing 2.0% AC Royal Jelly Extract decreased TEWL 11% more

effectively than the base lotion alone. After three weeks, the solution containing 2.0% AC Royal Jelly extract

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% AC Royal Jelly decreased transepidermal

water loss by 2% after four weeks of application. AC Royal Jelly Extract is a functional material that can also

decrease transepidermal water loss, thus providing great moisture retention.

0.00%

5.00%

10.00%

15.00%

20.00%

25.00%

30.00%

35.00%

T = 0 T = 1 Week T = 2 Weeks T = 3 Weeks T = 4 Weeks

Per

cen

t (%

) D

ecre

ase

Comparative Transepidermal Water Loss

2% Royal JellyExtract vs.Untreated Control

2% Royal JellyExtract vs. BaseLotion

ac royal jelly extract efficacy data - active concepts,...

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