34rd IFSCC Congress, Brazil, 14-17 October 2024

Automated In Vitro Analysis of UV Protection Performance of Powder

Sunscreen Products

Ribeiro, Claudio1; Baroni, Karin1; Yeung, Joey2; Yao, Ge2; Jin, Suxin3*; Chen, Ye2

1 Lubrizol Life Science, Lubrizol, San Paulo, Brazil; 2 Lubrizol Life Science, Lubrizol
Southeast Asia Pte Ltd, Singapore; 3 Lubrizol Life Science, Lubrizol Specialty Chemicals
Manufacturing (Shanghai) Co. Ltd., Shanghai, China.

* Chen, Ye, 44 Tanjong Penjuru, Singapore, 609032, +65-66636316, [email protected]

Abstract

The performance evaluation of sunscreen products is crucial to understand the

effectiveness of such products’ protection against solar radiation. Traditionally, the effectiveness

of sunscreens is assessed in vivo, which is expensive and has potential ethical issues. Hence,

in vitro methods have been developed to address the concerns. Unfortunately, the in vitro

methods acknowledged so far are not applicable to powder products, not to mention automation.

This work is about designing an effective automated in vitro method to evaluate the UV

protection performance of powder sunscreen products. In vivo claims were used as references

for method optimization to ensure correlation. SPF and UVA photoprotection were determined

by assessing UV transmittance through a thin film of powder sunscreen sample through

automated spreading on a roughened substrate, before and after exposure to a controlled dose

radiation from a defined solar exposure source.

The results showed that the SPF, UVAPF, and critical wavelength tested in vitro can be

well correlated to the in vivo claims. Proper sample treatment is necessary. The auto-application

can give consistent and better correlated SPF values compared with manual application.

Keywords: sunscreen; UV protection; powder product; in vitro; automation.

Introduction.
 34rd IFSCC Congress, Brazil, 14-17 October 2024

As a highly regulated product category, sunscreen products are subject to in vivo sun

protection factor (SPF) tests to assess their protection level against sun light and claimable SPF

to consumers. The current efficacy tests acknowledged in most countries are in vivo based, and

the new ISO 23675 (in vitro SPF testing) is still under development [1-3]. There is one in vitro

test method developed for UVA (ultraviolet A) claims, ISO 24443, which is applicable to most of

the product formats, however, excluding powder products [4].

On the other hand, with the increasing usage of sun care products, there are more

occasions that consumers prefer products of solid formats such as powder sunscreens, which

are easy to carry and use, with a dry and non-sticky feel.

Therefore, it’s of significance to design an in vitro method that can produce accurate and

repeatable results that are in vivo results correlated. With the help of automated spreading

machine, the human error can be well mitigated, while consistency well improved. This study

focuses on the method development based on data correlation with the in vivo claims.

Materials and Methods.

Materials: Liquid paraffin and glycerin were used as supply. Sandblasted (SB) PMMA plates

were used for in vitro analysis.

Market samples

I: Claim: SPF 20

Active ingredients: Titanium Dioxide [Nano], Ethylhexyl Methoxycinnamate

II: Claim: SPF 50+ PA ++++

Active ingredients: Ethylhexyl Methoxycinnamate, Bis-Ethylhexyloxyphenol Methoxyphenyl

Triazine, Titanium Dioxide, Ethylhexyl Triazone.

III: Claim SPF 32 PA +++

 34rd IFSCC Congress, Brazil, 14-17 October 2024

Active ingredients: Titanium Dioxide, Ethylhexyl Methoxycinnamate.

IV: Claim SPF 26 PA ++

Active ingredients: Ethylhexyl Methoxycinnamate, Titanium Dioxide, Zinc Oxide.

Sample treatment.

Three different approaches (A, B, and C) were conducted to examine if the sample treatment

process is practical and analysis results show good correlation with in vivo results.

A) Direct application with puff: Powder sunscreen sample (1.2 mg/cm2) is directly applied on a

SB PMMA plate using a puff. (Puff provided by the market sample.)

A-1: Sample spread on the PMMA plate manually.

A-2: Sample spread on the PMMA plate by automatic robot arm.

B) With spatula: Powder sunscreen samples is taken out with a spatula and applied on a SB

PMMA plate.

B-1: 0.0300±0.0005 g Sample spread on the PMMA plate manually.

B-2: 0.0300±0.0005 g Sample spread on the PMMA plate by automatic robot arm.

B-3: 0.0300±0.0005 g of sample spread on the PMMA plate by automatic robot arm.

C) Mixing with liquid paraffin: Powder sunscreen samples is taken out with a specular and mix

well with liquid paraffin (1:1, 1:2, and 2:1) and applied on a SB PMMA plate.

C-1: 0.0300±0.0005 (0.015 g) of solution sample is applied on the PMMA plate and spread by

automatic robot arm.

C-2: 0.0600±0.0005 (0.030 g) of solution sample is applied on the PMMA plate and spread by

automatic robot arm.

 34rd IFSCC Congress, Brazil, 14-17 October 2024

Test Procedure for Method A and B:

1. Used an air duster to blow the flint on the PMMA plate away.

2. Measured 0.03 g/0.06 g of sample on SB plate using respective methods using analytical

balance.

3. Used manual or automatic robot arm application to apply 0.03 g/0.06 g of sample on the

SB plate.

4. Measured the weight change after application using analytical balance.

5. Waited for 30 minutes before conducting pre-irradiation UV transmittance analysis.

6. Placed plate in solar simulator for stated exposure time in the software.

7. Conducted post-irradiation UV transmittance analysis after taking out plate.

Test procedure for Method C:

1. Measured powdered sunscreen with liquid paraffin in a 20 ml vial with different dilution

levels.

2. Mixed the solution together with a magnetic stirrer at 500 rpm until homogenous.

3. Used air duster to blow the flint on the PMMA plate away.

4. Measured 0.03 g/0.06 g of solution on SB plate using micropipette using analytical

balance.

5. Used automatic robot arm to apply 0.03 g/0.06 g of sample on the PMMA plate.

6. Measured the weight change after application using analytical balance.

7. Waited for 30 minutes before conducting pre-irradiation UV transmittance analysis.

8. Placed plate in solar simulator for stated exposure time in the software.

9. Conducted post-Irritation UV transmittance analysis after taking out the plate.

Results.
 34rd IFSCC Congress, Brazil, 14-17 October 2024

Market sample-I was used to test the effect of sample treatment. When the untreated

sample was applied directly on the PMMA plate with a puff, it was difficult to ensure an even

distribution of the sample on the plate (Figure 1-a). Moreover, powders are light and fluffy so

they can easily come off when spread under force. Using a spatula can better control the amount

put on the plate compared with the puff method (Figure 1-b). However, the same issue due to

the fluffy nature of the product occurred during the spreading phase. Both puff and spatula

methods gave a weight loss about or greater than 50% (Table II). When mixed with liquid paraffin,

a fine liquid sample can be obtained and applied easily on the PMMA plates. The auto spreading

can be utilized in a similar manner of standard liquid sunscreen samples (Figure 1-c).

Figure 1. Samples weighed on the PMMA plates and after spreading. (a) Using provided puff to

apply samples on the plates and after spreading (A-1manual and A-2 auto). (b) Using spatula to

take out sample and add on to the plates and after spreading (B-1 manual and B-2, 3 auto). (c)

Using liquid paraffin to dilute the powder and after auto spreading.
 34rd IFSCC Congress, Brazil, 14-17 October 2024

The in vitro SPF and UVAPF results (in Table II) based on the transmittance show that

the manual application gave lower SPF and UVAPF compared with auto application for puff and

spatula methods. All SPF values obtained by puff and spatula methods are below claimed SPF

value of 20. Coefficient of variation (COV) of SPF and UVAPF of auto application is greater than

manual application for puff and spatula methods. Whereas, mixing the powder with liquid paraffin

gave much smaller weight loss, narrower COVs for both SPF and UVAPF. The SPF reading of

C-1 is 20.57, as claimed SPF of the product.

Table II. Test Results of Different Application Methods.

Puff Spatula Mixture

Method
A-1 A-2 B-1 B-2 B-3 C-1 C-2
Weight
53.95 48.14 51.50 50.66 74.34 8.91 22.35
change (%)
SPF 6.02 12.11 4.64 7.86 9.61 20.57 32.57
SPF COV
4.48 40.34 6.64 20.54 5.16 13.19 13.99
(%)
UVAPF 4.88 4.94 4.29 10.18 7.79 3.39 3.48
UVAPF COV
0.81 24.19 2.11 23.55 5.03 3.62 2.32
(%)
λ Critical (nm) 385.00 383.50 383.90 383.90 383.90 376.20 376.50

Therefore, mixing powder product with liquid paraffin was selected for sample treatment

before in vitro analysis. The dilution levels of 1:2 and 2:1 was also tested. With 2:1 dilution, the

powder samples cannot be mixed well with good spreadability, whereas 1:2 sample was too

liquid and showed phase separation overnight. Hence, 1:1 dilution ratio was selected as the best

ratio for the test.

Four market samples were then analyzed with in vitro SPF method, 4 SB plates for each

sample (0.0300±0.005 g liquid paraffin diluted sample per plate). The results in Table III show a
 34rd IFSCC Congress, Brazil, 14-17 October 2024

good correlation between tested SPF and UVAPF values and the claims of the commercial

products.

Table III. Test Results of Market Samples.

Market sample I II III IV

In-Vitro SPF 19.18 48.59 30.21 49.23
In-Vitro SPF STD 1.41 7.70 4.17 6.66
In-Vitro SPF COV 7.35% 15.84% 13.79% 13.53%
SPF claimed 20 50+ 32 26
UVAPF 3.97 5.33 9.25 7.67
UVAPF 95% CI 15.82% 3.32% 5.43% 5.60%
UVAPF claimed NA PA ++++ PA +++ PA ++
UVA balance 16% 9% 28% 14%
Critical wavelength (nm) 378.80 367.80 384.80 379.70
UVA/UVB ratio 0.458 0.379 0.641 0.502

Discussion.

The commonly used UV filters in powder sunscreens are Titanium Dioxide and

Ethylhexyl Methoxycinnamate. Although not many UV filters are used in powder sunscreens, a

variety of SPF and UVAPF claims can be achieved, covering low, moderate, and high protection.

Dilution with liquid paraffin can greatly improve the weighing process of the powder sample and

enhance the consistency of sample application (fulfilled by automated sample spreading).

Moreover, liquid paraffin won’t affect the reading of UV transmittance.

The total amount of sample applied on the PMMA plate is half the amount of liquid sample

for in vitro method, which is sensible because the amount of powder applied on skin is not the

same as liquid sample either.

Automated spreading procedure has proved its advantage over manual spreading in

terms of repeatability, reproducibility, and reliability [5-7]. With puff and spatula method, the dry

samples stuck on the roughened surface of the PMMA plate and made it difficult to spread. When

spread manually, operators applied different forces to spread the sample over the plate. However,

with constant pressure, force, and route, robot arm may not be able to adjust the force nor route
 34rd IFSCC Congress, Brazil, 14-17 October 2024

to intentionally spread the sample evenly. Hence, the COV values of automated spreading are

higher than manual ones. On the other hand, when the sample can be evenly spread on the

PMMA plate with constant force and fixed route, robot arm displayed narrower COV compared

with manual spreading.

In vitro SPF and UVAPF values can be highly correlated with the in vivo claims when the

samples were well distributed and spread on the PMMA plates. Thus, in vitro method can be

applied to evaluate the UV protection performance of powder samples. With this method, the

screening of sunscreen prototypes and assessment of formulation variable become feasible.

Conclusion.

The UV protection performance of powder sunscreen products can be evaluated by

automated in vitro method. Dilution of powder sunscreen sample with liquid paraffin is necessary.

The subsequent measurement of SPF, UVAPF, and critical wavelength can follow the in vitro

method for standard liquid samples. The test method developed can be used as a reference for

evaluating the effectiveness of powder sunscreen products, extending the use of non-invasive

in vitro method and greatly facilitate the product development of powder sunscreens.

Acknowledgments.

The authors thank Lubrizol Life Science for support for this work. The authors gratefully

acknowledge the assistance of Noelle Kung on the experimental work.

Conflict of Interest Statement.

NONE.

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