Singlet Oxygen Quantification After Skin Exposure to Ultraviolet A (UVA) Light
Phase: NA
Conditions: Sun Damaged Skin
Sponsor: University of California, Davis

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
Page 1 of 9 
 
Revised: May 11, 2017 
 
 
1) 
Protocol title: Singlet oxygen quantification after skin exposure to 
ultraviolet A (UVA) light 
 
Protocol version date: May 11, 2017 
 
2) 
Objectives 
The objective of this study is quantification of singlet oxygen species in the 
skin after exposure to UVA light 
3) 
Background 
 
Ultraviolet (UV) radiation can lead to skin damage and cancer. UVA irradiation is 10 
to 100 times more abundant in natural sunlight than UVB, thus human skin is 
exposed to more UVA irradiation daily.(1-4) UVA irradiation is not completely 
filtered by clothing and it penetrates deeper into the dermis than UVB, potentially 
causing more damage.(3-5)  
 
It is believed that skin cancer, photo aging, and skin immunomodulation are 
mediated by the reactive oxygen species (ROS) that are generated in response to UV 
radiation.(6) Singlet oxygen (1O2) is one type of ROS.(6)  
 
The aim of this study is to quantify the level of singlet oxygen generated in the skin 
after exposure to UVA radiation. Previous studies have used low dose UVA1 
irradiation of 20 J/cm2, which mimics exposure to strong sunlight of approximately 
two hours.(4) This study will use UVA doses equivalent to or less than what humans 
are exposed to in daily life. Additionally, the aim is to quantify singlet oxygen 
produced in individuals of various skin types before and after application of 
sunscreens containing zinc oxide and avobenzone (SPF 30). To our knowledge, 
there is no method for quantifying singlet oxygen in human skin after exposure to 
UVA light. This is a novel method that may help us understand further the protective 
effects of various skin types, as well as sunscreens.  
 
4) 
Inclusion and Exclusion Criteria 
Inclusion criteria:  
• Subjects aged 18 years and older  
Exclusion criteria:  
• Those who are currently smoking or have smoked within the past 
3 years.  
• Aspirin use  
• Multivitamins and supplements that contain vitamin E  
• Adults unable to consent 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
• Individuals who are not yet adults (infants, children, teenagers) 
• Pregnant women 
• Prisoners 
5) 
Study Timelines 
The study will be conducted over the course of one year.  
The duration of each subject’s participation is approximately one week.  
 
6) 
Study Endpoints 
Primary study endpoints include: 
• To quantify singlet oxygen generated by the skin after exposure 
to three different UVA radiation doses (each equivalent to or less 
than UVA dose encountered in daily life) 
 
         Secondary study endpoints: 
• Difference in singlet oxygen generate between individuals of 
different skin types  
• Difference in singlet oxygen generation with two different 
sunscreens (zinc oxide and avobenzone, each SPF 30) vs control 
• Change in appearance of skin pigmentation 
 
 
 
 
 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
7) 
Procedures Involved 
 
Subjects meeting the inclusion criteria without any of the exclusion criteria 
will be enrolled into this study. Up to 17 subjects will be recruited into this 
study. 
 
Group A: N=7 Fitzpatrick skin type I-II 
Group B: N=5 Fitzpatrick skin type III-IV 
Group C: N=5 Fitzpatrick skin type V-VI 
 
All subjects will undergo the same procedures, as follows: 
 
 
Visit 1:  
• Informed consent  
• Review medical history and concomitant medications 
• Fitzpatrick skin type classification  
• Three spots measuring 2cm2 each will be marked on subject’s left 
dorsal forearm and right dorsal forearm 
•  Each spot will be assigned numbers 1, 2, or 3.  
• Each spot will be exposed to corresponding UVA dose. 
• UVA exposure at each site and singlet oxygen measurements will be 
recorded 
• Any one spot will be exposed to UVA for up to 5 minutes. The exposure 
at any one skin site will range from 0.33 J/cm2, 8.25 J/cm2, and 16.5 
J/cm2. The exposures used in this study are below those normally used 
in the literature.(4) 
• Subject will be compensated $10 (gift card) at completion of the visit  
 
 
Visit 2: May occur on the same day as Visit 1 up to 1 month after visit 1 
• Three spots measuring 2cm2 each will be marked on subject’s left 
dorsal forearm and right dorsal forearm 
•  Each spot will be assigned numbers 1, 2, or 3.  
o First spot: control (no sunscreen) 
o Second spot: 2mg/cm2 sunscreen with zinc oxide (SPF 30) 
will be applied 
o Third spot: 2mg/cm2 sunscreen with avobenzone (SPF 30) 
will be applied  
• Only one UVA dose will be used for all exposures. The dose will be 
chosen based on dose response during Visit 1 (either 8.25 J/cm2 or 
16.5 J/cm2) 
• Any one spot will be exposed for up to 5 minutes 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
• Subject will be compensated $15 (gift card) at completion of the 
visit  
• This completes subject’s involvement in the study 
 
Subject compensation is outlined as follows: 
 
Visit 1: $10 
Visit 2: $15 
 
 
Protocol activity 
VISIT 1 
VISIT 2 
Informed Consent 
X 
 
Medical History 
X 
 
Review concomitant Medication 
X 
 
Mark 3 areas measuring 2cm2 each 
on right and left dorsal forearm 
X 
X 
Expose each of the 3 marked skin 
areas to one UVA dose (0.33 J/cm2, 
8.25 J/cm2, and 16.5 J/cm2)  
X 
 
Determine best UVA dose for 
subject based on dose response 
data  
X 
 
Weigh sunscreen 2mg/cm2 
 
X 
Apply zinc oxide containing 
sunscreen to one marked spot on 
each arm  
 
X  
Apply avobenzone containing 
sunscreen to one marked spot on 
each arm 
 
X 
Mark one spot as “control” (no 
sunscreen) 
 
X 
Expose each of the 3 marked skin 
areas to one UVA dose (as 
determined during visit 1) 
 
X 
 
 
8) 
Data and/or Specimen Management and Confidentiality 
All data will be coded for the safety of the study subjects. Research data will only be 
available to the listed research personnel. The research site will maintain 
appropriate medical and research records for this study in compliance with ICH E6, 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Section 4.9 and regulatory and institutional requirements for the protection of 
confidentiality of subjects.  
 
All of the study data will be coded and stored within a locked cabinet within a locked 
room with two levels of keyed entry required to gain access. Furthermore, data files 
will either be stored in password-protected files within a locked room that will 
require two levels of keyed entry for access.  
 
Precaution will be taken to maintain the privacy of the participants. These include 
the following: All subjects will be assigned a subject ID number after signing the 
consent form. This will be their only form of identification throughout the 
remaining of the study period. Each subject’s name, age, and medical record 
number will be recorded. The consent forms will be kept in a binder. The subjects 
will be entered into the data analysis sheets as subject ID numbers and a separate 
password-protected file will contain the key for these codes. All of the files will be 
saved on password-protected computers within locked rooms. All study related 
material will be kept in a locked cabinet within a locked room at UC Davis 
Department of Dermatology. Only the research team personnel will have access to 
study-related materials. Participant information will not be disclosed to third party 
individuals except for those authorized to oversee the research project. 
 
Data that are already collected will not be destroyed but will be stored in a codified 
format that will not allow linkage to identifying information.  
The singlet oxygen data are immediately stored after capture into a password-
protected computer that will be stored in a locked room within the Institute for 
Regenerative Cures (IRC) at UC Davis. All data stored in binders will be 
transported to the UC Davis Clinical Trials unit (CTU).  
 
Each of the subjects’ signed consents will be filed in a locked cabinet within a locked 
room. All of the subjects’ data will be entered into the data analysis sheets as codes 
and a separate password protected file will contain the key for these codes. All of 
the files will be saved on computers that will be within locked rooms. The research 
team will follow the UC Davis Institutional Policy for data security to ensure that all 
subjects and collected data are protected.  
 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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9) 
Data and/or Specimen Banking 
 
All patient information will be only accessible to authorized research personnel, and 
otherwise personalized information will be locked in a cabinet and password 
protected electronic drive at the UC Davis Department of Dermatology (IRC and 
CTU). After the study has been completed and analyzed, all patient information will 
be discarded. The code keys will be stored in a secure file in a password-protected 
computer in a locked room and this code key will be deleted upon completion of the 
analysis.  
 
10) Provisions to Monitor the Data to Ensure the Safety of Subjects 
 
The records and study materials will be maintained in a locked cabinet. 
Subjects will be asked about their health and any adverse events that may 
have occurred. Adverse effects will be related to the treating physician. Noted 
and serious adverse effects will be relayed to the IRB within 24 hours of the 
occurrence.   
 
11) Withdrawal of Subjects 
 
The subjects may withdraw consent at any time and they will be reassured 
that their clinical care will not be compromised if a subject were to withdraw. 
Subjects will be withdrawn from the study if they develop adverse side 
effects. The investigator can remove the subject from the study. Possible 
reasons for removal include: 
• Failure to follow protocol 
• Hypersensitivity to sunscreen 
 
12) Risks to subjects 
Subjects will be counseled that the physical risks from sunscreen application may 
include skin redness or irritation. The physical risks from UVA light may include 
mild redness or tanning of the spot exposed to UVA light. Subjects will be advised to 
inform the research team if they experience any side effects.  
Although provisions are put in place to protect identifiable data, we cannot 
guarantee absolute protection of the data collected from each subject. Accordingly, 
the subjects will be informed of this risk.  
 
13) Potential Benefits to Subjects 
There are no direct benefits to the subjects.  

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
This study will help us add to the general scientific knowledge that may 
benefit individuals in the future.   
 
14) Sharing of Results with Subjects 
Any peer reviewed manuscripts that result from the study will be made 
available to the subjects on request.  
 
15) Provisions to Protect the Privacy Interests of Subjects 
 
Steps will be taken to protect subjects’ privacy interests. Subjects will only interact 
with approved study personnel who have successfully completed human subjects 
training. This includes those involved in the study visits, consent, and in any study 
related procedures. We will do our best to make sure that the personal information 
in the subject’s record is kept private. However, no study can guarantee 100% 
protection of private information despite all of our measures to protect each 
subject’s privacy. Subjects will be made aware of this during the consent process.  
 
 
16) Compensation for Research-Related Injury 
 
If a subject is injured as a result of being in this study, the University of California 
will provide necessary medical treatment. Depending on the circumstances, the 
costs of the treatment may be covered by University or the study sponsor or may be 
billed to the subjects’ insurance company just like any other medical costs. The 
University and the study sponsor do not normally provide any other form of 
compensation for injury. For more information about compensation, call the IRB 
administration at (916) 703-9151 or email at IRBAdmin@ucdmc.ucdavis.edu. 
 
17) Economic Burden to Subjects 
 
Standard of care and other routine costs will be billed to the patient or the patient’s 
insurance carrier, Medicare, or Medi-Cal where appropriate. Only the costs of 
research and experimental procedures will be paid by the sponsor/department. 
Subjects may incur a cost in relation to travel needed to get to the UC Davis 
Dermatology Department for each visit.  
 
18) Drugs or Devices 
☒ I confirm that all investigational drugs will be received by the IDS 
pharmacy, which will store, handle, and administer those drugs so that 
they will be used only on subjects and be used only by authorized 
investigators.  

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
 
☒ I confirm that all investigational devices will be labeled in accordance with 
FDA regulations and stored and dispensed in such a manner that they will 
be used only on subjects and be used only by authorized investigators. 
 
 
Device used in the study:  
SINGLET OXYGEN MONITOR FOR UVA-SKIN STUDIES 
 
The UVA-produced Singlet Oxygen monitor detects the weak, near infrared 
luminescence from the a-X system in molecular oxygen. It uses an ultra-sensitive 
photomultiplier tube (PMT) coupled to a photon counting system to monitor this weak 
emission produced by the UVA source that is also part of the system. This non-imaging, 
point sensor is an ultra-sensitive portable and automated real-time detection system for 
singlet O2. The current system includes (1) a fiber –coupled CW UVA source module, (2) 
PMT detector module, and (3) data acquisition with photon counter board. 
 
A block diagram of the major components of the singlet oxygen sensor is shown in 
Figure 5. 
 
Figure 1. Block diagram of the singlet oxygen monitor. 

Protocol Title: Singlet oxygen quantification after skin 
exposure to ultraviolet A (UVA) light  
 
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Revised: May 11, 2017 
 
 
Figure 2. Fiber coupled UVA light source and light box. Subjects insert their arms 
through the right side of the light box. The equipment rack including the PC screen is 
shown in the background. 
 
 
 
References: 
 
 
 
1. 
Tewari A, Grage MM, Harrison GI, Sarkany R, Young AR. UVA1 is skin deep: 
molecular and clinical implications. Photochem Photobiol Sci. 2013;12(1):95-103. 
2. 
Tuchinda C, Srivannaboon S, Lim HW. Photoprotection by window glass, 
automobile glass, and sunglasses. J Am Acad Dermatol. 2006;54(5):845-54. 
3. 
Menter JM, Hatch KL. Clothing as solar radiation protection. Curr Probl 
Dermatol. 2003;31:50-63. 
4. 
Wang F, Smith NR, Tran BA, Kang S, Voorhees JJ, Fisher GJ. Dermal damage 
promoted by repeated low-level UV-A1 exposure despite tanning response in human 
skin. JAMA Dermatol. 2014;150(4):401-6. 
5. 
Sklar LR, Almutawa F, Lim HW, Hamzavi I. Effects of ultraviolet radiation, 
visible light, and infrared radiation on erythema and pigmentation: a review. Photochem 
Photobiol Sci. 2013;12(1):54-64. 
6. 
Hanson KM, Clegg RM. Observation and quantification of ultraviolet-induced 
reactive oxygen species in ex vivo human skin. Photochem Photobiol. 2002;76(1):57-63.