SUN CARE 33
TABLE 3: SPF 30 SUNSCREEN CONTAINING 7% BACILLUS LYSATE Phase Ingredients / INCI
A Octocrylene (PARSOL® 340) Homosalate (PARSOL® HMS)
Ethylhexyl Salycilate (PARSOL® EHS) Avobenzone (PARSOL® 1789) Caprylic Capric Triglycerides
Glyceryl Stearate (and) PEG-100 stearate
Cetearyl alcohol (and) Ceteareth 20 (Procol L CS-20-D) Cetyl alcohol
C15-19 Alkane (Emogreen L-19) B Deionized water
Dimethicone (XIAMETER® PMX-200 Silicone Fluid 10 cSt) EDTA (Protacide Na3 EDTA)
Phenoxyethanol (and) Ethylhexylglycerin (Euxyl PE 9010) Carbomer (Carbopol® 940 Polymer) Glycerin
Polysorbate 20 Propanediol (Zemea) Polyglyceryl-4 Caprate (Tegosoft PC 41) C
Sodium Hyaluronate (and) water (Actique™ Hyal 1%) Hydrolyzed Jojoba Esters (and) Water (Floraesters K-20W) Triethanolamine 99% Tocopherol clear
D Bacillus Lysate 11. Jedwards
1 1 1
2 2 2 2 3 4
2 5 6 7 2 6 8 9 6
Supplier % w/w 1
TABLE 4: SPF 30 SUNSCREEN CONTAINING 10% BACILLUS LYSATE Phase Ingredients / INCI
5.000 5.000 5.000 3.000 5.000 3.000 3.000 2.500 4.500 1.000
44.100 0.100 1.000 0.200 3.000 1.500 2.000 0.500 1.500 1.500
10 0.500 11
0.100 7.00
Procedure: Heat phase A to 85°C while stirring. Heat phase B to 80°C and add to phase A while stirring and homogenizing the emulsion. Cool down the emulsion to 40°C and add phase C. Add phase D and homogenized.
Suppliers: 1. DSM 2. Protameen 3. Seppic 4. Dow Chemicals 5. Shulke 6. Ess. Ingredients 7. Pride 8. Evonik 9. Jarchem 10. Jeen Internat.
of the Bacillus Lysate was determined using the Pierce Coomassie Plus (Bradford) Assay Kit (Thermo Scientific, Cat# 23236) following the manufacturers provided instructions. The Pierce Bovine Serum Albumin Standard Kit
(Thermo Scientific, Cat# 23209) was used as the protein sample to generate the standard curve.
UV absorption profile The UV absorption profile was obtained using a Molecular Devices SpectraMax Plus 384 microtiter plate reader equipped with SoftMax Pro analytical software (Molecular Devices, Version 5.46). Briefly, varying dilutions of the Bacillus Lysate
(150 uL) were transferred to a NuncTM 96-well UV flat bottom microtiter plate (ThermoFisher Ref#8404) and scanned between 190 nm – 700 nm. Plots of the UV absorption profile were generated using GraphPad Prism Version 9.5.1 (GraphPad Software, La Jolla, CA).
5 4 3 2 1
0 200 300 400 500 Wavelength
Figure 2: UV absorbing profile of Bacillus Lysate between 200nm-700nm, compared to oxybenzone
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A Octocrylene (PARSOL® 340) Homosalate (PARSOL® HMS)
Ethylhexyl Salycilate (PARSOL® EHS) Avobenzone (PARSOL® 1789) Caprylic Capric Triglycerides
Glyceryl Stearate (and) PEG-100 stearate
Cetearyl alcohol (and) Ceteareth 20 (Procol L CS-20-D) Cetyl alcohol
C15-19 Alkane (Emogreen L-19) B Deionized water
Dimethicone (XIAMETER® PMX-200 Silicone Fluid 10 cSt) EDTA (Protacide Na3 EDTA)
Phenoxyethanol (and) Ethylhexylglycerin (Euxyl PE 9010) Carbomer (Carbopol® 940 Polymer) Glycerin
Polysorbate 20 Propanediol (Zemea) Polyglyceryl-4 Caprate (Tegosoft PC 41) C
Sodium Hyaluronate (and) water (Actique™ Hyal 1%) Hydrolyzed Jojoba Esters (and) Water (Floraesters K-20W) Triethanolamine 99% Tocopherol clear
D Bacillus Lysate 11. Jedwards
1 1 1
2 2 2 2 3 4
2 5 6 7 2 6 8 9 6
Supplier % w/w 1
5.000 5.000 5.000 3.000 5.000 3.000 3.000 2.500 4.500 1.000 41.100 0.100 1.000 0.200 3.000 1.500 2.000 0.500 1.500 1.500
10 0.500 11
0.100 10.00
Procedure: Heat phase A to 85°C while stirring. Heat phase B to 80°C and add to phase A while stirring and homogenizing the emulsion. Cool down the emulsion to 40°C and add phase C. Add phase D and homogenized.
Suppliers: 1. DSM 2. Protameen 3. Seppic 4. Dow Chemicals 5. Shulke 6. Ess. Ingredients 7. Pride 8. Evonik 9. Jarchem 10. Jeen Internat.
Formulation and testing of Bacillus Lysate-formulated chemical UV filter sunscreen To determine the SPF-boosting capability of the Bacillus Lysate, various percentages (3%, 7%, and 10%) were added to a SPF 30 sunscreen formulation. The sunscreen formulation and production process can be found in Tables 1-4. In vivo SPF determination for each
formulation was carried out following the International Standard - ISO 24444:2019 protocol. The intrascapular area of the back to the right and left of the midline were used as test sites for SPF validation. Each test site was cleaned using a dry cotton pad and a 30 cm2
area was delineated using a
gentian violet surgical marker. Care was taken to ensure that each site had uniform pigmentation, skin tone, and texture, absence of warts, moles, nevi, scars, blemishes, and dermal lesions. The complete protocol is described by the
Bacillus Lystate Bacillus Lystate (50%) Bacillus Lystate (10%) Bacillus Lystate (1%) Oxybenzone (0.1 mg/mL)
5 4 3 2 1
600 700 0 270 280 290 300 Wavelength (nm)
Figure 3: UV absorbing profile of Bacillus Lysate for UVB wavelength, compared to oxybenzone
October 2023 PERSONAL CARE
Internal Standard ISO 24444:2019 (Second edition 2019-12) - Cosmetic - Sun Protection Test Methods - In vivo Determination of Sun Protection Factor (SPF). One unprotected site was used to determine
a subject’s Minimal Erythemal Dose (MED), which was noted as the lowest erythemal effective radiant exposure that produced the first perceptible unambiguous erythema with defined borders appearing over >50% of the exposed site 16 to 24 hours after UV exposure. A minimum of five progressive UV light doses were administered within each test site. Test material and SPF standards were
swirled with a glass rod prior to use and evenly applied using a spatula to the rectangular test site areas using a minimum of 30 cm2 concentration of 2.0 ± 0.05 mg/cm2
. The test mate-rial and reference sunscreen
test sites were randomly distributed on the back of each subject to reduce error arising
Bacillus Lystate Bacillus Lystate (50%) Bacillus Lystate (10%) Bacillus Lystate (1%) Oxybenzone (0.1 mg/mL)
for a final
310
320
Absorbance
Absorbance
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