Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System
Considering the importance of the cutaneous tissue investigation and the need for the development of new protocols to non-invasively establish the safety and efficacy of dermocosmetics and topical products, we aimed at developing an HPLC-TBARS-EVSC (high performance liquid chromatography–thiobarbitu...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-04-01
|
| Series: | Scientia Pharmaceutica |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-0532/89/2/17 |
| _version_ | 1797536108447268864 |
|---|---|
| author | Rafael Sauce Claudinéia Aparecida Sales de Oliveira Pinto Carmen Ayala-Jara Zulita Adriana Prieto Maria Valéria Robles Velasco André Rolim Baby |
| author_facet | Rafael Sauce Claudinéia Aparecida Sales de Oliveira Pinto Carmen Ayala-Jara Zulita Adriana Prieto Maria Valéria Robles Velasco André Rolim Baby |
| author_sort | Rafael Sauce |
| collection | DOAJ |
| description | Considering the importance of the cutaneous tissue investigation and the need for the development of new protocols to non-invasively establish the safety and efficacy of dermocosmetics and topical products, we aimed at developing an HPLC-TBARS-EVSC (high performance liquid chromatography–thiobarbituric acid reactive species–ex vivo <i>stratum corneum</i>) assay for the lipid peroxidation measurement on subjects’ <i>stratum corneum</i> (SC) obtained by tape stripping; additionally, we applied the HPLC-TBARS-EVSC assay in an emulsified sunscreen system containing ethylhexyl triazone and bemotrizinol as UV filters. HPLC analysis was performed in isocratic mode with 35% methanol/65% phosphate buffer (pH 7.0) as the mobile phase. The diode detector was set at 532 nm to quantify the malondialdehyde (MDA)-TBA adduct. An ex vivo tape stripping method was applied in 10 volunteers in three pre-defined regions of the volar forearms: the control; the irradiated; and the site containing the sunscreen (2.0 mg·cm<sup>−2</sup>). Ten adhesive tapes per region were used for SC removal. An exclusive ex vivo protocol to measure SC lipid peroxidation was preliminarily developed with linearity and selectivity. The protocol suggested the use of an artificial irradiation dose (5506 KJ·m<sup>−2</sup>) to improve the assay response from the SC. The sunscreen system had a significative decrease in SC lipoperoxidative damage compared to the control. Our protocol can aid in the efficacy establishment of anti-UV and antioxidant agents, for instance, in studies that aim at elucidating the level of SC lipid peroxidation and even in carrying out baseline investigations characterizing different ethnicities and genders. |
| first_indexed | 2024-03-10T11:54:57Z |
| format | Article |
| id | doaj.art-58bc66b1203e4ea194ddae46e45ef0b8 |
| institution | Directory Open Access Journal |
| issn | 0036-8709 2218-0532 |
| language | English |
| last_indexed | 2024-03-10T11:54:57Z |
| publishDate | 2021-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Scientia Pharmaceutica |
| spelling | doaj.art-58bc66b1203e4ea194ddae46e45ef0b82023-11-21T17:22:11ZengMDPI AGScientia Pharmaceutica0036-87092218-05322021-04-018921710.3390/scipharm89020017Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen SystemRafael Sauce0Claudinéia Aparecida Sales de Oliveira Pinto1Carmen Ayala-Jara2Zulita Adriana Prieto3Maria Valéria Robles Velasco4André Rolim Baby5Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo 05508-900, BrazilDepartment of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo 05508-900, BrazilUniversidad Nacional de Trujillo Juan Pablo II Av. s/n, Trujillo 13011, PeruUniversidad Nacional de Trujillo Juan Pablo II Av. s/n, Trujillo 13011, PeruDepartment of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo 05508-900, BrazilDepartment of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo 05508-900, BrazilConsidering the importance of the cutaneous tissue investigation and the need for the development of new protocols to non-invasively establish the safety and efficacy of dermocosmetics and topical products, we aimed at developing an HPLC-TBARS-EVSC (high performance liquid chromatography–thiobarbituric acid reactive species–ex vivo <i>stratum corneum</i>) assay for the lipid peroxidation measurement on subjects’ <i>stratum corneum</i> (SC) obtained by tape stripping; additionally, we applied the HPLC-TBARS-EVSC assay in an emulsified sunscreen system containing ethylhexyl triazone and bemotrizinol as UV filters. HPLC analysis was performed in isocratic mode with 35% methanol/65% phosphate buffer (pH 7.0) as the mobile phase. The diode detector was set at 532 nm to quantify the malondialdehyde (MDA)-TBA adduct. An ex vivo tape stripping method was applied in 10 volunteers in three pre-defined regions of the volar forearms: the control; the irradiated; and the site containing the sunscreen (2.0 mg·cm<sup>−2</sup>). Ten adhesive tapes per region were used for SC removal. An exclusive ex vivo protocol to measure SC lipid peroxidation was preliminarily developed with linearity and selectivity. The protocol suggested the use of an artificial irradiation dose (5506 KJ·m<sup>−2</sup>) to improve the assay response from the SC. The sunscreen system had a significative decrease in SC lipoperoxidative damage compared to the control. Our protocol can aid in the efficacy establishment of anti-UV and antioxidant agents, for instance, in studies that aim at elucidating the level of SC lipid peroxidation and even in carrying out baseline investigations characterizing different ethnicities and genders.https://www.mdpi.com/2218-0532/89/2/17<i>stratum corneum</i> oxidative stressHPLCTBARStape stripping |
| spellingShingle | Rafael Sauce Claudinéia Aparecida Sales de Oliveira Pinto Carmen Ayala-Jara Zulita Adriana Prieto Maria Valéria Robles Velasco André Rolim Baby Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System Scientia Pharmaceutica <i>stratum corneum</i> oxidative stress HPLC TBARS tape stripping |
| title | Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System |
| title_full | Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System |
| title_fullStr | Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System |
| title_full_unstemmed | Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System |
| title_short | Preliminary Protocol Development of a HPLC-TBARS-EVSC (Ex Vivo <i>Stratum Corneum</i>) Assay for Skin Research: Application in a Sunscreen System |
| title_sort | preliminary protocol development of a hplc tbars evsc ex vivo i stratum corneum i assay for skin research application in a sunscreen system |
| topic | <i>stratum corneum</i> oxidative stress HPLC TBARS tape stripping |
| url | https://www.mdpi.com/2218-0532/89/2/17 |
| work_keys_str_mv | AT rafaelsauce preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem AT claudineiaaparecidasalesdeoliveirapinto preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem AT carmenayalajara preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem AT zulitaadrianaprieto preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem AT mariavaleriaroblesvelasco preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem AT andrerolimbaby preliminaryprotocoldevelopmentofahplctbarsevscexvivoistratumcorneumiassayforskinresearchapplicationinasunscreensystem |