Clinical trial NCT02935894
Conditions: Stability & Variability of Lipid-derived Molecules in Sweat
Keywords: oxygenated lipids metabolic profiling sphingoid bases ceramides endocannabinoids non-invasive sampling
Sponsor: USDA, Western Human Nutrition Research Center
Start date: 2016-11-28; Completion date: 2017-06-08
Enrollment: 14 participants; Allocation: NA; Intervention model: SINGLE_GROUP

Brief summary:
The purpose of this study is to see what the differences are in sweat (amount and small molecule content) collected from different sites of the body and by different methods of sweat stimulation. Additionally, the investigators want to know whether the amount and small molecule content of the sweat is the same in an individual over time, and the same across individuals at a given time. Finally, the investigators want to know how consumption of over-the-counter anti-inflammatory drugs such as ibuprofen will affect the inflammatory mediator content of sweat and how that compares to blood. This information will help to better understand the composition and behavior of sweat and assess its potential utility as a routine clinical tool in skin research.

Detailed description:
Subjects will participate in 4 study day visits that will be scheduled about 1- week apart. During the first study visit, the investigators will compare sweat collected following pharmacological stimulation of sweating (using the drug pilocarpine) to sweat collected following physiological stimulation of sweating (using a stationary bicycle). During the second study visit, the investigators will collect sweat following stimulation of sweating by the drug pilocarpine from the inner part of the forearm (near the wrist) and also the upper surface of the thigh (near the knee). During the third study visit, the investigators will collect sweat following stimulation of sweating by the drug pilocarpine from the inner part of both forearms (near the wrist). During the fourth study visit, the investigators will collect a sweat sample following stimulation of sweating by the drug pilocarpine from the inner part of the forearm (near the wrist) and a blood sample (about one teaspoon) from the other arm. After blood and sweat collection, participants will consume 400 mg (two tablets) of ibuprofen. The investigators will then collect blood and sweat from 30 minutes, 2 hours and 4 hours after participants consume the ibuprofen.

Arms: arm 1: All subjects will participate in 4 study day visits in the same order.

1. compare sweat collected following pharmacological stimulation of sweating (using the drug pilocarpine) to sweat collected following physiological induction of sweating (exercise using a stationary bicycle).
2. compare sweat collected following stimulation of sweating by the drug pilocarpine from the inner part of the forearm (near the wrist) to upper surface of thigh (near the knee).
3. collect sweat following stimulation of sweating by the drug pilocarpine from the inner part of both forearms (near the wrist).
4. collect blood and sweat samples before and 30 minutes, 2 hours and 4 hours after consumption of 400 mg of ibuprofen.
Interventions: intervention 1: Ibuprofen intervention 2: Physiological induction of sweating intervention 3: Pilocarpine
Intervention descriptions: intervention 1: 400 mg ibuprofen given to inhibit cyclooxygenase metabolism intervention 2: 15 minutes exercise at 60-80% oxygen consumption to induce sweating intervention 3: Iontophoresis with a 1.5 milliampere (mA) current using a gel disc containing a 5% pilocarpine nitrate solution to induce sweating

Protocol documents:
SAP_002.pdf: 
Statistical Analysis Plan 
 
Official Title: Investigating the Stability, Variability and Mechanism of Incorporation of 
Lipid Mediators into Eccrine Sweat 
 
ClinicalTrials.gov ID: NCT02935894 
 
Unique Protocol ID: 929370 
 
Authors: Karan Agrawal and John W. Newman 
Institutions: University of California-Davis (K.A, J.W.N.); USDA-ARS-Western Human 
Nutrition Research Center (J.W.N.) 
 
Date: December 5, 2017 
 
 


Introduction 
The purpose of this study is to see what the differences are in sweat (amount and 
small molecule content) collected from different sites of the body and by different methods 
of sweat stimulation. Additionally, the investigators want to know whether the amount and 
small molecule content of the sweat is the same in an individual over time, and the same 
across individuals at a given time. Finally, the investigators want to know how 
consumption of over-the-counter anti-inflammatory drugs such as ibuprofen will affect the 
inflammatory mediator content of sweat and how that compares to blood. This information 
will help to better understand the composition and behavior of sweat and assess its 
potential utility as a routine clinical tool in skin research. 
Subjects will participate in 4 study day visits that will be scheduled about 1- week 
apart. During the first study visit, the investigators will compare sweat collected following 
pharmacological stimulation of sweating (using the drug pilocarpine) to sweat collected 
following physiological stimulation of sweating (using a stationary bicycle). During the 
second study visit, the investigators will collect sweat following stimulation of sweating by 
the drug pilocarpine from the inner part of the forearm (near the wrist) and also the upper 
surface of the thigh (near the knee). During the third study visit, the investigators will 
collect sweat following stimulation of sweating by the drug pilocarpine from the inner part 
of both forearms (near the wrist). During the fourth study visit, the investigators will 
collect a sweat sample following stimulation of sweating by the drug pilocarpine from the 
inner part of the forearm (near the wrist) and a blood sample (about one teaspoon) from 
the other arm. After blood and sweat collection, participants will consume 400 mg (two 
tablets) of ibuprofen. The investigators will then collect blood and sweat from 30 minutes, 
2 hours and 4 hours after participants consume the ibuprofen. 
 
Analysis Objectives and Sets 
Data from the first three study visits will be analyzed to assess the impact of 
different sweat stimulation techniques and collection sites on the sweat lipid mediator 
profile. Secondarily, data from pilocarpine-stimulated sweat collected from the volar 


forearm collected at each study visit will be evaluated to assess to compare the temporal 
stability and intra- and inter-individual variability pharmacologically stimulated sweat 
lipid mediators. Data from the fourth study visit will be analyzed to determine the impact 
of systemic ibuprofen administration on sweat lipid mediators as a means to evaluate the 
source of these metabolites. Differences in sweat lipid mediators at each collection time 
and correlations between sweat and plasma lipid mediator concentrations at baseline will 
be assessed to evaluate these outcomes. 
 
Data Analysis Workflow 
Visits 1–3 
Prior to statistical analyses, data will be curated such that analytes with >30% 
missing values across the data set are excluded. The remaining data will be screened for 
outliers using Huber’s maximum likelihood type estimates to determine the center and 
spread of each analyte [1]. Missing data will be imputed by least squares multivariate 
normal imputation [2], after which data will be transformed to normal using the 
generalized logarithm transform [3], and normality will be verified using the Shapiro-Wilk 
test [4]. All data pretreatment will be performed in JMP Pro 12 (SAS Institute, Inc., Cary, NC, 
USA). 
To prevent differential storage times, samples will be processed weekly along with a 
pooled sweat LRM. If batch effects are noted, data will be adjusted using the batch-to-
concentration correlation for each lipid mediator detected in the LRM, by calculating the 
batch-specific residuals and subtracting these from the measured data. Batch-adjusted data 
will be then transformed to normal as described above. All batch adjustments will be 
performed in Microsoft Excel 2010 (Redmond, WA, USA). 
Comparisons of mean lipid mediator concentrations in sweat stimulated by 
pilocarpine iontophoresis and exercise will be made by paired Student’s t-tests. 
Comparisons of collection sites (anterior distal thigh, lower back and volar forearm) will be 
made by one-way ANOVAs, while assessments of sweat lipid mediator temporal stability 
will be made by repeated measures ANOVAs. Subject identification will be included as a 


random effect in the one-way ANOVA models, as only two subjects produced sweat from all 
three collection sites. All univariate comparisons will be performed in JMP Pro 12 (SAS 
Institute, Inc.) and adjusted for multiple comparisons using the Benjamini-Hochberg 
procedure at q = 0.05 [5].  
Multivariate analyses will be conducted by partial least squares discriminant 
analysis (PLS-DA), using two different algorithms in the R statistical environment (Vienna, 
Austria). Models will be constructed using either the orthogonal scores algorithm with 
univariate scaling and leave-one-out cross-validation in imDEV version 1.42, a Microsoft 
Excel add-in [6], or using the pls and caret packages in R version 3.4.1 [7, 8]. In both cases 
sweat stimulation technique, collection site or study visit will be used as classifiers, and 
variables will be clustered by Spearman correlation coefficients using the Minkowski 
distance and Ward agglomeration. Models built using imDEV version 1.42 will be evaluated 
using the Q2 score, with Q2 ≥ 0.4 considered indicative of a predictive model [9]. Models 
built using the pls and caret packages in R version 3.4.1 will be evaluated using model 
accuracy, and the significance of model accuracy was evaluated using a chi-squared test. 
 
Visit 4 
Plasma and sweat data will be independently pretreated, and all data pretreatments 
will be performed in JMP Pro 13 (SAS Institute, Inc., Cary, NC, USA). Prior to statistical 
analyses, data will be curated such that analytes with >30% missing values across the data 
set will be excluded. The remaining data will be screened for outliers using Huber’s 
maximum likelihood type estimates to determine the center and spread of each analyte [1]. 
Missing data will be imputed by least squares multivariate normal imputation with 
covariance shrinkage [2], after which data will be transformed to normal using the 
procedures of Box and Cox [10], and normality was verified using the Shapiro-Wilk test [4].  
Linear mixed models evaluating the relationship between metabolite concentrations 
and time with a random intercept for subject, will be performed in R version 3.1.0 using the 
lme4 package [11] after verification of population homoscedasticity at each timepoint in 
plasma and sweat by Bartlett’s test using JMP Pro 13. Statistical significance of the fixed 


effect of time was determined by likelihood ratio tests of the full model against a null model 
(i.e., without the fixed effect of time, but with random intercept for subject). Results of 
likelihood ratio test will be considered significant if P < 0.05 and the false discovery rate 
(q) as estimated by the method of Storey and Tibshirani using the fdrtool package is < 0.2 
[12, 13]. Differences between metabolite concentrations at each timepoint will be assessed 
by Tukey’s post-hoc HSD test using the multcomp package [14].  
Comparisons of metabolite patterns in plasma and sweat will be performed in JMP 
Pro 13. Correlations between imputed but not transformed baseline (i.e. 0 h) lipid 
mediator concentrations in plasma and sweat will be estimated using Kendall rank 
correlation coefficients for analytes detected in both matrices. Additionally, relative 
abundances of all detected lipid mediators in each matrix will be estimated on both a per-
class (e.g. relative abundance of total alcohols detected regardless of fatty acid precursor) 
and by fatty acid precursor within a lipid mediator class (e.g. individual abundances of 
alcohol metabolite isomers of arachidonic acid), and will be compared using the Wilcoxon 
rank-sum exact test.  
 
References 
[1] 
P.J. Huber, E.M. Ronchetti, The Basic Types of Estimates, in: Robust Statistics, Second 
Edition, John Wiley & Sons, Inc., Hoboken, NJ, 2009, pp. 53-84. 
[2] 
SAS Institute Inc., Modeling Utilities: Exploring Data for Outliers, Missingness, and 
Strong Predictors, in: JMP(R) 12 Basic Analysis, SAS Institute Inc., Cary, NC, 2015, 
pp. 313-342. 
[3] 
B.P. Durbin, J.S. Hardin, D.M. Hawkins, D.M. Rocke, A variance-stabilizing 
transformation for gene-expression microarray data, Bioinformatics (Oxford, 
England) 18 Suppl 1 (2002) S105-10. doi: 10.1093/bioinformatics/18.suppl_1.S105. 
[4] 
S.S. Shapiro, M.B. Wilk, An analysis of variance test for normality (complete 
samples), Biometrika 52(3-4) (1965) 591-611. doi: 10.1093/biomet/52.3-4.591. 
[5] 
Y. Benjamini, Y. Hochberg, Controlling the false discovery rate: a practical and 
powerful approach to multiple testing, J R Stat Soc Series B Stat Methodol 57(1) 
(1995) 289-300. 


[6] 
D. Grapov, J.W. Newman, imDEV: a graphical user interface to R multivariate 
analysis tools in Microsoft Excel, Bioinformatics (Oxford) 28(17) (2012) 2288-90. 
doi: 10.1093/bioinformatics/bts439. 
[7] 
M. Kuhn, contributions from J. Wing, A. Williams, C. Keefer, A. Engelhardt, T. Cooper, 
Z. Mayer, B. Kenkel, the R Core Team, M. Benesty, R. Lescarbeau, A. Ziem, L. Scrucca, 
Y. Tang, C. Candan, T. Hunt, caret: Classification and regression training. R package 
version 6.0-76, 2017. https://cran.r-project.org/package=caret. 
[8] 
B.-H. Mevik, R. Wehrens, K.H. Liland, pls: Partial least squares and principal 
component 
regression. 
R 
package 
version 
2.6-0, 
2016. 
https://cran.r-
project.org/package=pls. 
[9] 
J.A. Westerhuis, H.C.J. Hoefsloot, S. Smit, D.J. Vis, A.K. Smilde, E.J.J. van Velzen, J.P.M. 
van Duijnhoven, F.A. van Dorsten, Assessment of PLSDA cross validation, 
Metabolomics 4(1) (2008) 81-89. doi:10.1007/s11306-007-0099-6. 
[10] 
G.E.P. Box, D.R. Cox, An analysis of transformations, J. R. Stat. Soc. Series B Methodol. 
26(2) (1964) 211-252. 
[11] 
D. Bates, M. Maechler, B. Bolker, S. Walker, contributions from R.H.B. Christensen, H. 
Singmann, B. Dai, G. Grothendieck, P. Green, lme4: Linear mixed-effects models using 
'Eigen' and S4. R package version 1.1-14, 2017. https://cran.r-project.org/web/ 
packages/lme4/lme4.pdf. 
[12] 
B. Klaus, K. Strimmer, fdrtool: Estimation of (local) false discovery rates and higher 
criticism. R package version 1.2.15, 2015. https://cran.r-project.org/web/ 
packages/fdrtool/fdrtool.pdf. 
[13] 
26. 
J.D. Storey, R. Tibshirani, Statistical significance for genomewide studies, 
Proc. Natl. Acad. Sci. 100 (2003) 9440-9445. doi: 10.1073/pnas.1530509100. 
[14] 
T. Hothorn, F. Bretz, P. Westfall, contributions from R.M. Heiberger, A. 
Schuetzenmeister, S. Scheibe, multcomp: Simultaneous inference in general 
parametric models. R package version 1.4-7, 2017. https://cran.r-project.org/web/ 
packages/multcomp/multcomp.pdf.  

ICF_001.pdf: 
Permission to Take Part in a Human Research Study
Page 1 of 9
Title of research study: Investigating the stability, variability and mechanism of incorporation 
of lipid mediators into eccrine sweat
Investigator: John W. Newman, Ph.D.
Why am I being invited to take part in a research study?
We invite you to take part in this research study because you are a generally healthy male, age 20-40 
years, who does not perform vigorous physical activity on a regular basis.
What should I know about a research study?
(Experimental Subject's Bill of Rights) 

Someone will explain this research study to you, including:
o
The nature and purpose of the research study.
o
The procedures to be followed.
o
Any drug or device to be used.
o
Any common or important discomforts and risks.
o
Any benefits you might expect.
o
Medical treatment, if any, that is available for complications.

Whether or not you take part is up to you.

You can choose without force, fraud, deceit, duress, coercion, or undue influence.

You can choose not to take part.

You can agree to take part now and later change your mind.

Whatever you decide it will not be held against you.

You can ask all the questions you want before you decide.

If you agree to take part, you will be given a signed and dated copy of this document.
Who can I talk to?
If you have questions, concerns, or complaints, or think the research has hurt you, talk to the research 
team: study coordinator: Karan Agrawal, M.Sc. at (530) 752-5217, principal investigator: John 
Newman, Ph.D. at (530) 752-1009
Document Revision Date: September 22, 2015
 
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This research has been reviewed and approved by an Institutional Review Board (“IRB”). 
Information to help you understand research is on-line at 
http://www.research.ucdavis.edu/policiescompliance/irb-admin/.You may talk to a IRB staff member at 
(916) 703-9151, hs-irbadmin@ucdavis.edu, or 2921 Stockton Blvd, Suite 1400, Room 1429, 
Sacramento, CA 95817 for any of the following:

Your questions, concerns, or complaints are not being answered by the research team.

You cannot reach the research team.

You want to talk to someone besides the research team.

You have questions about your rights as a research subject.

You want to get information or provide input about this research.
Why is this research being done?
The purpose of this study is to see what the differences are in sweat (amount and small molecule 
content) collected from different sites of the body and by different methods of sweat stimulation. 
Additionally, we want to know whether the amount and small molecule content of the sweat is the 
same in an individual over time, and the same across individuals at a given time. Finally, we want to 
know how consumption of over-the-counter anti-inflammatory drugs such as ibuprofen will affect the
inflammatory mediator content of sweat and how that compares to blood. This information will help 
us to better understand the composition and behavior of sweat and assess its potential utility as a 
routine clinical tool in skin research. 
How long will the research last?
We expect that you will be in this research study for approximately six months. You will be asked to 
attend four study visits at our research center. The first three visits will last between 1-2 hours. The 
fourth visit will last 5 hours. Each visit will be separated from the previous visit by at least one week.
How many people will be studied?
We expect about 17 people to enroll in this study.
Document Revision Date:September 22, 2015
 
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What happens if I say yes, I want to be in this research?
If you decide to volunteer, you will first be pre-screened by a member of our research team to see if you 
are eligible for enrollment. This screening visit will be conducted either in-person or on the telephone 
and will last about 20 minutes. During this pre-screen, the research team will fill out a questionnaire 
based on your responses to questions related to your health and supplement and medication use.
If you are eligible to be enrolled into the study you will be asked to participate in an approximately 
4-6 week long study that will be conducted at the USDA Western Human Nutrition Research Center 
located in 430 W Health Sciences Dr, Davis, CA 95616. You will be asked to participate in 4 study day 
visits that will be scheduled about 1- week apart. The first three study visits will last about 1-2 hours 
each and the fourth study visit will last about 5 hours. 
You will be asked to fast overnight (12 hours) before each study visit (no food or beverage, but you 
can drink as much water as you like) and avoid applying any cream or medication to your body for 24 
hours before your study visit. Addtionally, you will be asked to avoid consuming any non-steroidal anti-
inflammatory drugs such as aspirin, ibuprofen (Advil, Motrin, etc), acetaminophen (Tylenol) or 
naproxen (Aleve) for at least 48 hours before your study visit.
During each study visit, we will collect sweat using a device designed for sweat collection in infants.
For sweat collection:
1. Your skin will be cleaned with an alcohol wipe and wetted with water
2. Electrodes containing gel disks will be placed on two areas of your body and secured with straps 
or tape. The electrodes will be attached to a power source and a mild electric current, which is 
generally painless will be applied for 5 minutes.
3. One of the electrodes will be removed and the area cleaned with water while a marker will be 
used to mark the area where the other electrode is attached. The other electrode will then be 
removed and the area will be cleaned with water.
4. After the electrodes have been removed, a collection device consisting of a thin plastic tube 
coiled inside a disk will be attached to your skin (where the electrode previously was) using 
straps or tape and secured with an elastic band or bandage. You will be asked to sit for 30 
minutes while we collect sweat. The collection device will then be removed.  
During your first study visit, after obtaining informed consent, we will collect sweat from the inner 
part of one of your forearms (near the wrist) as described above. We will then ask you to ride a 
stationary bicycle for up to 15 minutes after which we will use a sweat collection device (step 4 above) 
on your other forearm to collect sweat for 30 minutes. While you are on the stationary bicycle, we will 
measure your heartrate and blood oxygen capacity using a non-invasive finger sensor and also measure 
your metabolic rate by asking you to breathing normally into a face mask fit snugly over your mouth and
nose.
During your second study visit, we will collect sweat from the inner part of one of your forearms 
(near the wrist) and also the upper surface of your thigh (near the knee). Both collections will occur 
almost simultaneously and will follow the procedure described above.
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During your third study visit, we will collect sweat from the inner part of both of your forearms 
(near the wrist) and your lower back. All collections will start at approximately 10 minute intervals and 
will follow the procedure described above.
Prior to your fourth study visit, you will undergo a video interview with the study physician, Dr Raja
Sivamani. During this interview, Dr Sivamani will review your medical history to make sure you can 
safely consume ibuprofen. Dr Sivamani will also go over the risks and side-effects associated with 
ibuprofen consumption during the interview.
During your fourth study visit, we will collect a sweat sample from the inner part of one of your 
forearms (near the wrist) using the procedure described above. We will also collect a blood sample 
(about one teaspoon) from your other arm. After we collect blood and sweat, you will be asked to 
consume 400 mg (two tablets) of ibuprofen. We will then collect blood and sweat from you 30 minutes, 
2 hours and 4 hours after you consume the ibuprofen. 
After four study visits, your participation in the study is complete and we will process your 
compensation. 
While you will remain fasted during each study visit, we will have pitchers of water available for 
you to consume at will. Addtionally, after each study visit, you will have access to the dining area where 
snacks such as juice, granola bars, oatmeal, dried fruit, coffee, tea and hot cocoa will be available for 
you to consume.
Blood and sweat collected from you will be used to measure metabolites derived from normal 
biological processes which will include inflammatory mediators. Addtionally, blood and sweat collected 
from you during visit 4 will also be used to measure ibuprofen levels at each collection time. 
What happens if I do not want to be in this research?
You may decide not to take part in the research and it will not be held against you.
What are my responsibilities if I take part in this research?
If you take part in this research, you will be responsible to:
1. NOT apply any cream or medication to your body 24 hours before you arrive for each study visit.
2. NOT consume any non-steroidal anti-inflammatory drugs such as aspirin, ibuprofen (Advil, Motrin, 
etc), acetaminophen (Tylenol) or naproxen (Aleve) for at least 48 hours before you arrive for each 
study visit.
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3. Come to four study sessions, after a 12 hour overnight fast, lasting between one to five hours.
4. Bring or wear shorts to the first two study sessions.
5. Inform the research team if you experience any discomfort at any time during the study.  
What happens if I say yes, but I change my mind later?
You can leave this research study at any time and it will not be held against you.
If you decide to leave the research, please contact the investigator so that the investigator can cancel 
your scheduled visit(s).
Data that has been collected during your time enrolled in the research study will be used for its 
intended purposes, when possible.
You will receive compensation for the portion of the study you do complete (see below). 
Is there any way being in this study could be bad for me?
There is a small risk of loss of balance, dizziness, or shortness of breath and discomfort associated 
with an increase in heart rate, increased body temperature, and sweating that could result from a 15 
min session on an exercise bicycle. Individuals with knee or hip pain, injuries, or weakness should 
not participate in this particular test. The session on the exercise bicycle is necessary to induce 
sweating by physiological methods, but may present additional risks to individuals in very poor 
cardiovascular health or with undiagnosed cardiovascular risk factors such as family history of heart
disease, hypertension, high cholesterol, diabetes, harmful use of alcohol, tobacco use, stress, physical 
inactivity, obesity or an unhealthy diet. Before you perform the test we will evaluate your risk level by 
asking you questions about your health and symptoms related to heart disease. It is very important 
that you answer these questions as truthfully as possible to avoid serious adverse events. If our staff 
determines that you are in a high risk category, we will not do the test, and exclude you from the 
study. Potential cardiovascular-related serious adverse events include stroke and heart attack, which 
could result in death.
You will be asked to fast for 12 hours prior to, and during each of your study visit days. You may 
experience a headache, feelings of hunger or low energy as a result of fasting or lack of caffeine.
There is a small risk for bruising as a result of blood sample collection. During the blood drawing 
procedure, there is the possibility that you might experience some momentary pain or stinging 
sensations or bruising, or very rarely an infection around where the needle was inserted. However, blood
will be taken only by experienced and licensed staff, and sterile methods will be used at all times.
The measurement of metabolic rate uses an air collection mask that fits snugly over the nose and 
mouth. This may create a feeling of claustrophobia in some individuals. Participants may request that the
mask be readjusted or removed at any time.
Document Revision Date:September 22, 2015
 
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There may be some risk with ibuprofen intake. In certain individuals ibuprofen has been known to 
cause an upset stomach including nausea, bloating and gas; diarrhea; constipation; headaches; dizziness; 
nervousness; itching skin or a rash; blurred vision or ringing ears.
During sweat collection, we will use a device that delivers pilocarpine to your skin using a mild 
electric current. The pilocarpine gel may cause some skin irritation such as redness or contact dermatitis.
There is a very small risk of a burn from the device. The use of the device will also require pressing on 
your skin. If you bruise easily, you may develop a bruise at the location of the measurements although 
this would be unusual. However, this device is used frequently on infants for sweat collection. 
There may also be risks to your privacy. The research team will limit the collection of personally 
identifying information on the study visit days and your forms will be identified only by a randomly 
generated participant identification code. The code, linked to your personal information, will be stored in
a locked file in a locked and secure location. Your date of birth will only be used to calculate age and 
none of the data collected beyond the initial screening interview will be identified by your name. 
However, just like with other personal information kept by your health care providers, banks, and others,
even these safeguards cannot guarantee absolute protection of the data. Although rare, there are reported 
cases of breaches that have resulted in discrimination in insurance or employment.
For more information about risks and side effects, contact the research study team.
Will being in this study help me in any way?
We cannot promise any benefits to you or others from your taking part in this research. The outcome of 
the research may lead to the development of better ways of analyzing the skin features of the human 
body.
What happens to the information collected for the research?
Efforts will be made to limit use or disclosure of your personal information (see description above), 
including research study and medical records, to people who have a need to review this information. We 
cannot promise complete confidentiality. Organizations that may inspect and copy your information 
include the IRB and other University of California representatives responsible for the management or 
oversight of this study. The Unites States Department of Agriculture (USDA), who sponsors this study, 
may have access to your participation records.
During your participation in this research, data will be collected about you. The de-identified data 
and any specimens, such as blood or tissue that are taken from you for this study, they will become the 
property of the University of California or USDA. The specimens may be used in this research, may be 
used in other research, and may be shared with other organizations. The specimens could lead to 
discoveries or inventions that may be of value to the University of California or to other organizations. 
Under state law you do not have any right to money or other compensation stemming from products that
may be developed from the specimens. 
Document Revision Date:September 22, 2015
 
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If you agree that biological specimen(s) collected from you may be shared or used in other 
research, please initial here: ____________
Otherwise, your specimen will be destroyed at the end of this study.
Federal law provides additional protections of your medical records and related health information. 
These are described in the UC Davis Health System Notice of Privacy Practices 
(http://www.ucdmc.ucdavis.edu/compliance/pdf/notice.pdf) and in an attached document..
Can I be removed from the research without my OK?
The person in charge of the research study or the sponsor can remove you from the research study 
without your approval. Possible reasons for removal include health risks, an inability to complete 
critical study components, lack of cooperation with study personnel, etc. We will tell you about any 
new information that may affect your health, welfare, or choice to stay in the research.
What else do I need to know?
There is no charge for you to participate in this study. Neither you nor your insurance carrier will be 
charged for your taking part in the research. All costs associated with the study will be paid by the 
sponsor/department.
It is important that you promptly tell the person in charge of the research if you believe that you have
been injured because of taking part in this study. If you are 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 your 
insurance company just like 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, you may
call the IRB Administration at (916) 703-9151 or email at HS-IRBAdmin@ucdavis.edu.
If you agree to take part in this research study, we will compensate you by check for up to $75, if 
you complete all aspects of the study, for your time and effort. Should you choose to withdraw from the 
study or are unable to complete all procedures, your compensation will be pro-rated to reflect your time 
and effort contributed. The rate of pay offered is scheduled as follows: 
Visit
Hours
Compensation
Study Visit 1
2
$10.00
Study Visit 2
1
$10.00
Study Visit 3
1
$10.00
Document Revision Date:September 22, 2015
 
APPROVED by the Institutional Review
Board at the University of California, Davis
Protocol
APPROVED
929370
February 13, 2017


Permission to Take Part in a Human Research Study
Page 8 of 9
Study Visit 4
5
$45.00
Total
Approximately 9
$75.00
You may be asked for your social security number for payment purposes. It will not be used for any 
other purpose without your permission
The results of this study, including specimens collected, may have commercial value to the sponsors,
UC Davis, and/or the researchers. You will have no legal or financial interest in any commercial 
development resulting from the research or from the information or materials collected.
Are there other research opportunities?
If you are interested in being contacted for future research, please provide your phone number and/or 
email. This is completely optional.
            (initials) Yes, I am willing to be contacted for future research opportunities. My phone number 
and/or email is:          
 
            
 
            
 
            
 
            
 
            
 
            
 
            
 
            
 
            .
Your signature documents your permission to take part in this research.
Signature of subject
Date
Printed name of subject
Signature of person obtaining consent
Date
Printed name of person obtaining consent
Document Revision Date:September 22, 2015
 
APPROVED by the Institutional Review
Board at the University of California, Davis
Protocol
APPROVED
929370
February 13, 2017


Permission to Take Part in a Human Research Study
Page 9 of 9
Document Revision Date:September 22, 2015
 
APPROVED by the Institutional Review
Board at the University of California, Davis
Protocol
APPROVED
929370
February 13, 2017

Prot_000.pdf: 
PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 1 of 12 
  
1) Protocol Title Investigating the stability, variability and mechanism of incorporation of 
lipid mediators into eccrine sweat 
 
2) Author(s) of Protocol John W. Newman, Ph.D. -USDA Western Human Nutrition 
Research Center and Department of Nutrition, University of California Davis  
 
 
 UC Davis Researcher 
 Researcher from other institution 
 Private Sponsor 
 Cooperative Group 
 Other: __________ 
 
3) IRB Review History 
N/A 
 
4) Objectives   
Validate the utility of sweat as a non-invasive sample for the pharmacological investigation of 
skin biology 
 
Specific Aim 1: Develop and Report Validated Protocols for the Collection and Quantification of 
Lipid Mediator Profiles in Sweat by evaluating: 
1) Comparison of pharmacological and physiological methods of sweat stimulation (i.e. 
comparing the lipid mediator profile of sweat collected following stimulation by pilocarpine 
nitrate to sweat collected following stimulation by light exercise) 
2) Site-specificity of sweat lipid mediator composition (i.e. comparing the lipid mediator profile 
of sweat collected from the volar forearm to sweat collected from the anterior distal thigh 
and/or lower back) 
3) Inter- and intra-individual variability of the sweat mediator lipidome 
 
Specific Aim 2: Evaluating the impact of systemic cyclooxygenase inhibition on the plasma and 
sweat mediator lipidome   
 
5) Background 
Blood and urine represent common human biofluids that have been extensively studied in the 
context of pharmacokinetic and metabolomic analyses. However, the collection of these 
biofluids is either physically or culturally invasive and therefore subject compliance is 
sometimes difficult to achieve. The use of non-invasive matrices such as hair, oral fluid, sweat 
and tears has improved subject compliance in pharmacokinetic studies, and recently there has 
been interest in conducting metabolomic analyses using sweat for the same reason. While sweat 
has been previously used to diagnose cystic fibrosis in neonates and infants and to detect illicit 
drugs in forensic settings, it has rarely been used in metabolomic analyses due to a lack of 
uniform sweat collection protocols that provide a reproducible and measureable volume of 
sweat. With advances in technology, the Macroduct® sweat collection system was developed 
and validated by Wescor, Inc. for the reproducible collection of neonatal sweat for the diagnosis 
of cystic fibrosis. More recently, the Macroduct® system was used in a proof-of-principle study 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 2 of 12 
  
to conduct an untargeted metabolomic analysis of human sweat, and has since been used to 
develop a screening tool for lung cancer based on the sweat metabolite profile. 
 
As can be seen from the cited studies above, sweat has predominantly been used to study 
pulmonary diseases. However, given its proximity to the skin, there is also the potential for sweat 
to be an informative non-invasive matrix in cutaneous research. Current cutaneous research 
methods are dependent on tissue biopsies, which are again invasive methods, and therefore not 
suitable for repeated temporal sampling. To the best of our knowledge, only a single study exists 
that examines the composition of sweat in the context of cutaneous disease, and this study 
demonstrated that subjects with atopic dermatitis, psoriasis or hyperhidrosis exhibited increased 
prostaglandin E2 (PGE2) levels relative to healthy control.  
 
PGE2 is a member of a class of compounds known as bioactive lipid mediators, which regulate 
inflammatory and immune responses. These mediators, which include an array of oxygenated 
lipids (“oxylipins”), endocannabinoids and ceramides, are generally produced locally via a 
variety of biosynthetic pathways in response to extracellular stimuli and function similarly to 
local hormones or autacoids. Additionally, ceramides play an important structural role in the 
epidermal barrier. Lipid mediators have been studied previously in cutaneous research, but have 
depended on a limited number of analytical targets and/or focused on a single class of analytes, 
preventing the identification of mediator pathway cross-talk. Our laboratory has developed 
methods to quantify over 125 bioactive lipid targets, most not previously studied in the context 
of cutaneous biochemistry. Understanding the actions and interactions of these lipid mediators in 
the skin could improve treatment modalities, thus improving skin care and reducing the burden 
of skin conditions on subjects with cutaneous diseases. 
 
Recently, we conducted a study which demonstrated differences in the sweat lipid mediator 
profiles of subjects with and without atopic dermatitis using the Macroduct® sweat collection 
system on the volar bilateral forearm of human subjects (IRB #605131). However, little is known 
about the source of lipid mediators in sweat, the effects of different sweat stimulation techniques 
on lipid mediator composition as well as the site-specificity of the sweat mediator lipidome. 
Collecting such baseline data is critical for the evaluation of routine mediator lipidomic analyses 
of sweat as a non-invasive tool to support studies investigating cutaneous inflammatory 
responses, dietary manipulation, and skin diseases lacking biomarkers and therapeutic targets. 
 
 
6) Inclusion and Exclusion Criteria 
Inclusion criteria 
• 20-40 y 
• Male 
• Weight > 110 pounds 
 
Exclusion criteria 
• Diagnosed active chronic diseases for which the individual is currently taking daily 
medication, including but not limited to: 
o Diabetes mellitus 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 3 of 12 
  
o Cardiovascular disease 
o Cancer 
o Gastrointestinal disorders  
o Kidney disease 
o Liver disease 
o Bleeding disorders 
o Asthma 
o Autoimmune disorders 
o Hypertension 
o Osteoporosis  
• Recent minor surgery (within 4 wk) or major surgery (within 16 wk) 
• Recent antibiotic therapy (within 4 wk) 
• Recent hospitalization (within 4 wk) 
• Use of over-the-counter or prescription medications at the time of the study that directly 
affect endpoints of interest (e.g. hyperlipidemia, glycemic control, steroids, statins, anti-
inflammatory agents, and weight loss aids) 
• Adults who are not able to consent 
• Under current medical supervision 
• Ibuprofen intolerance or allergy 
• Those with a bleeding disorder 
• Current enrollee in a clinical research study. 
• Individuals with blood clotting or platelet defect disorders 
• Individuals with orthopedic limitations or cardiovascular risk that preclude participation in 
the physiological stimulation of sweat by light exercise portion of the study 
• Individuals who are trained athletes or that regularly perform physical activity defined as 
“vigorous” by the Centers for Disease Control and Prevention 
 
 
7) Number of Subjects 
The target sample size is 10 participants that complete the study visits. In addition, we aim to 
enroll additional participants not to exceed 20% of the calculated sample size to account for 
participants contributing incomplete or invalid data for various reasons including study 
withdrawal, compromised sample conditions, etc. Thus a total of 12 people will be enrolled. 
However due to subject withdrawal and non-compliance, we plan to now enroll 5 additional 
subjects, bringing the total number of enrolled subjects to 17. 
 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 4 of 12 
  
8) Recruitment Methods 
Participants will be recruited from Davis, Woodland, and the greater Sacramento area using the 
following methods:  
• WHNRC study subject referrals 
• Personal contact 
 
WHNRC Human Studies office maintains a list of people who have participated in previous 
studies, and who wish to be contacted about new studies. In addition, Dr. Sivamani has 
conducted previous studies from which there is a list of interested persons. The contact will be in 
the form of an email providing a study description. It will be up to the individual to respond is 
they wish to participate. 
 
9) Compensation to the Subjects 
Participants will be compensated a reasonable hourly wage for their time contributions to the 
study.  Participants will be compensated $10 for Visit 1, $10 for Visit 2, $10 for Visit 3, and $45 
for Visit 4. Total possible compensation will be $75 per participant and will be pro-rated based 
on time involved. The compensation will be provided in the form of a check written to the 
volunteer. 
 
10) 
Study Timelines 
The study will be conducted over a one-year period. Each subject’s participation will consist of 
four visits. The first three visits will last approximately 1 hour each, the fourth visit will last 
approximately 4 hours. The duration of a single subject’s participation in the study is not 
expected to exceed six months. 
 
Additionally, subjects expressing an interest in the study will be screened in-person or by 
telephone and eligibility for enrollment in the study will be determined by verbal completion of 
the eligibility screening questionnaire with a trained study staff member. If a subject is 
determined to be eligible for participation, they will be given further information about the study 
procedures and consent protocol and emailed a copy of the consent document. Time will be 
allocated at the first study visit to complete the consent document and answer any questions 
related to the study.  
 
11) 
Study Endpoints 
 
• Primary endpoints: 
o Changes in sweat oxidized lipid, endocannabinoid and ceramide concentrations 
following exercise-induced sweating relative to sweat collection following pilocarpine 
iontophoresis 
o Changes in sweat oxidized lipid, endocannabinoid and ceramide concentrations due to 
sweat collection from the anterior distal thigh and lower back compared to sweat 
collected from the volar forearm 
o Changes in sweat oxidized lipid, endocannabinoid and ceramide concentrations 
following consumption of ibuprofen relative to baseline 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 5 of 12 
  
o Changes in plasma oxidized lipid, endocannabinoid and ceramide concentrations 
following consumption of ibuprofen relative to baseline 
 
• Secondary endpoints:  
o Changes in plasma and sweat ibuprofen concentrations following oral administration 
o Relative abundance of sweat metabolites derived from primary metabolism covering 
carbohydrates and sugar phosphates, amino acids, hydroxyl acids, free fatty acids, 
purines, pyrimidines, aromatics, exposome-derived chemicals 
o Relative abundance of sweat metabolites derived from complex lipids covering 
ceramides, sphingomyelins, cholesteryl esters, oxysterols, lyso- and phospholipids, 
mono-, di- and triacylglycerols, galactosyl- and glucuronyllipids 
 
12) 
Procedures Involved 
 
Prior to enrollment in study 
All volunteers will be subjected to either an in-person interview at the WHNRC or to a telephone 
interview during which their eligibility and willingness to participate in the study will be 
determined. Twelve subjects eligible and willing to participate will be enrolled in the study and 
scheduled to begin participation at least seven days after enrollment.  
 
 
Study Design and Visits 
Subjects will participate in four study visits, separated by at least one week. On each of the study 
days, participants will arrive in the morning after an overnight 12 hour fast (ad lib water 
consumption will be allowed and encouraged). Collection procedures will be performed at 
approximately the same time of the day to avoid circadian effects. Subjects will also be asked to 
not apply any cream or medication to their body 24 hours before each study visit and refrain 
from use of non-steroidal anti-inflammatory drugs such as ibuprofen for at least 48 hours prior to 
each study visit. 
 
• Study Visit 1: After obtaining written informed consent, sweat will be collected using the 
Macroduct® sweat collector from the volar forearm following stimulation of sweating by 
pilocarpine iontophoresis as part of the Webster Sweat Induction System. Following sweat 
collection, subjects will be given a 15-minute break after which they will undergo up to 15-
minutes of light exercise on a stationary bicycle to stimulate sweat by physiological methods 
(see below). Following the light exercise, sweat will be collected from the opposite volar 
forearm using the Macroduct® sweat collector. The study visit is complete after the second 
sweat collection. 
 
• Study Visit 2: Sweat will be collected using the Macroduct® sweat collector from the volar 
forearm and anterior distal thigh following stimulation of sweat by pilocarpine iontophoresis 
as part of the Webster Sweat Induction System. Collections will occur simultaneously at both 
sites. The study visit is complete after both sweat collections. 
 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 6 of 12 
  
• Study Visit 3:  Sweat will be collected using the Macroduct® sweat collector from both volar 
forearms and the lower back following stimulation of sweat by pilocarpine iontophoresis as 
part of the Webster Sweat Induction System. Collections will begin at approximately 10 
minute intervals at all three sites. The study visit is complete after all three sweat collections.  
 
• Study Visit 4: A baseline sweat sample will be collected using the Macroduct® sweat 
collector from the volar forearm following stimulation of sweating by pilocarpine 
iontophoresis as part of the Webster Sweat Induction System. A blood sample (~ 6 mL) will 
be collected in EDTA-treated Vacutainer® tubes following venipuncture of the opposite 
antecubital vein. Following sweat and blood collection, subjects will consume 400 mg of 
ibuprofen orally. Blood and sweat collections will occur at 30 minutes, 2 hours and 4 hours 
post-ibuprofen consumption. The study visit is complete after the blood and sweat collections 
at 4 hours post-ibuprofen consumption. Total blood drawn per participant over the course of 
the study will be ~ 24 mL. Prior to Study Visit 4, participants will undergo a video interview 
with the study physician (and co-PI of the study) Dr Raja Sivamani, during which time Dr 
Sivamani will obtain a medical history from the subjects and review the risks and side effects 
of ibuprofen with the subjects.  
 
Participants will remain fasted during study visits (ad lib water consumption will be allowed and 
encouraged). At the end of the study visit, subjects will be allowed access to the WHNRC dining 
room where snacks such as juice, granola bars, oatmeal, dried fruit, coffee, tea and hot cocoa will 
be made available for their consumption. 
 
All blood and sweat samples collected will be measured for metabolites derived from normal 
biological processes which will include inflammatory mediators. Additionally, blood and sweat 
collected during visit 4 will also be used to measure ibuprofen levels at each collection time. 
 
Description of the “light exercise” 
Sweat stimulation by light exercise, such as riding a stationary bicycle with resistance, has been 
used in several recent studies to collect sweat for ionic analysis. In the proposed study, subjects 
will ride a cycle ergometer (Monark ergometer) with resistance for 10-15 minutes in a 
temperature and humidity monitored room. Resistance (in kg*m/min) will be set so as to elevate 
heart rate to that evinced during moderate (60-80% VO2max) steady state aerobic exercise and 
pedal cadence will be set using a metronome at 70 rpm. Heart rate will be continuously 
monitored by sensors during the exercise. Additionally, subjects will be fitted with air collection 
masks connected to a metabolic cart in order to continuously monitor oxygen consumption, 
respiratory quotient and substrate use in order to ensure achievement of steady state aerobic 
exercise. All procedures are non-invasive and should cause minimal subject discomfort. 
 
Sweat collection following stimulation of sweating by pilocarpine iontophoresis using the 
Webster Sweat Induction System 
The Macroduct® sweat collection system and the Webster Sweat Inducer are FDA-approved 
medical devices routinely used for sweat collection in neonates and infants in order to diagnose 
cystic fibrosis. The protocols for these devices are well established and freely available at 
http://wescor.com/translations/Translations/M2551-7A-EN.pdf. In brief, the collection site will 
be cleansed with a 70% solution of isopropanol followed by distilled water, and sweating will be 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 7 of 12 
  
stimulated using the Webster Sweat Inducer. This procedure consists of attaching positive and 
negative electrodes, each containing a manufacturer-supplied pilocarpine gel disk, to the 
collection site and applying a 1.5 mA current across the electrodes for five minutes. The area 
under the positive electrode will be marked using a felt-tip marker and electrodes will then be 
removed and the collection site will be cleansed with distilled water. The Macroduct® sweat 
collector will then be attached to marked site and secured using a tourniquet or bandage to 
maximize cutaneous contact. The collection device will remain in place for up to 30 minutes 
after which it will be removed and the collected sweat will be exuded into a collection vial and 
stored until analysis.  
  
 
13) 
Data and Specimen Banking 
All banked specimens will be stored in -20 oC or -80 oC freezers.  Participants will be given the 
option to provide consent for future, undetermined use of their specimens by initialing the 
designated section on a signed consent document and a verbal agreement of ‘yes’ during the 
discussion of consent for participation at the enrollment visit.  Any archived blood or sweat 
samples of participants having provided the described consent may be used at a later date for 
purposes not specified by this study.  All identifiable subject information will be removed except 
for assigned subject ID codes. If the subject does not agree to future use of specimens, all 
remaining specimens will be destroyed.   
 
14) 
Data Management and Confidentiality 
Identifiable information linked with the participant identification code will be stored separately 
from any data collected during the study and will only be made accessible to the study Principal 
Investigators and the Coordinator.  Data will be collected on hard copy forms and computer-
based questionnaires using only the participant identification code.  Completed forms will be 
kept in locked files at the WHNRC in rooms that are also locked.  Electronic files will be kept in 
folders on the WHNRC fileserver, which is password-protected and will be limited to access for 
key study personnel only.  The WHNRC fileserver is located behind a hardware firewall and is 
backed up off-site weekly, at a minimum. In the case that data needs to be stored on a laptop 
computer, the laptop will be encrypted.  Data entered manually will be double entered and 
compared to identify data entry errors.   
 
15) 
Provisions to Monitor the Data to Ensure the Safety of Subjects 
This study is an observational study and involves an intervention with an FDA-approved over-
the-counter non-steroidal anti-inflammatory drug. The Co-Investigator for the study, Dr. Raja 
Sivamani, will provide oversight for this part of the research and review the study participants’ 
medical history prior to prescribing the ibuprofen to each subject.  Nevertheless, it does present 
several challenges and various methods of data collection that participants will be subject to.  In 
order to ensure the safety of participants during administration of challenges or specimen 
collection, all aspects of the study will be carried out by trained study personnel. The other 
collections involve non-invasive minimal risk activities.   Participants will spend the study visit 
day in private rooms in the Metabolic Research Unit of the WHNRC containing hospital beds for 
rest and privacy during data and specimen collection.  A unit monitor will remain in the unit (but 
outside of the participant room) as a contact person for participants should any reactions, 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 8 of 12 
  
concerns, questions, or emergencies arise throughout the visit.  Participants will never be left 
alone in the Metabolic Research Unit. 
 
The exercise-induced sweat stimulation (exercise bicycle) will be administered by a qualified 
physiologist and staff who, prior to the test, will assess the subject’s risk level using the risk 
factors established by the American College of Sports Medicine: a) family history of premature 
heart disease, cigarette smoking, hypertension, dyslipidemia, impaired fasting glucose, obesity, 
and sedentary lifestyle. Those subjects falling in the low and moderate risk categories will be 
further evaluated by the physiologist who will verbally administer the Physical Activity 
Readiness Questionnaire prior to the test.  If the subject responds ‘yes’ to any of the questions, 
they will not perform the test. For those cleared for testing, heart rate, pulse, and blood pressure 
will be measured prior to administration of the challenge. During the test, participants will be 
asked to rate their level of perceived exertion, and if level reaches a level of ‘extremely hard’ or 
if the participant loses balance, becomes dizzy, or other adverse reaction occurs during the test, it 
will be stopped immediately.    
 
Venipuncture blood draws will be conducted by licensed, trained and experienced phlebotomists. 
If excessive bruising or discomfort occurs, no further venipunctures will be conducted. 
 
Sweat-stimulation by pilocarpine iontophoresis may cause minor discomfort to subjects in the 
form of slight stinging or prickling at the site of administration. Subjects will be asked 
periodically during iontophoresis if the discomfort is unbearable. If the subject responds ‘yes’, 
the iontophoresis procedure will be stopped immediately. 
 
If at any time a participant refuses to continue participation or refuses to participate in a 
particular aspect of the study, they will be excluded from all or part of the remaining procedures 
according to their wishes.  
 
16) 
Withdrawal of Subjects 
During the consent process, participants will be informed that they can withdraw from the study 
at any time without any risk of repercussion or penalty.  Compensation for participant time in the 
study will be pro-rated to reflect the withdrawal time.  Specimens previously collected during the 
study will be used as specified.   
 
17) 
Risks to Subjects 
There is a small risk of loss of balance, dizziness, or shortness of breath that could result from 
the 30 min light exercise to stimulate sweat production (exercise bicycle).  The use of an exercise 
bicycle is expected to result in increased heart rate and sweating but may present additional risks 
to individuals in very poor cardiovascular health or with undiagnosed cardiovascular risk factors 
such as family history of heart disease, hypertension, high cholesterol, type 2 diabetes, harmful 
use of alcohol, tobacco use, stress, physical inactivity, obesity and unhealthy diet.  All of these 
risk factors will be evaluated prior to having participants engage in the test. Potential 
cardiovascular-related serious adverse events include stroke and myocardial infarction, which 
could result in death. 
 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 9 of 12 
  
There is a small risk of bruising as a result of blood sample collection.  A licensed, trained 
phlebotomist will collect blood samples to minimize bruising.  
 
The measurement of metabolic rate uses an air collection mask that fits snugly over the nose and 
mouth. This may create a feeling of claustrophobia in some individuals. Participants may request 
that the mask be readjusted or removed at any time. 
 
Sweat collection procedures pose minimal risks to the subjects as they are all noninvasive. The 
Webster Sweat Inducer uses electricity for iontophoresis based induction of sweating. Physical 
risks of the sweat collection may include: 
• The use of the device will require pressing on the skin. If a subject bruises easily, they may 
develop a bruise at the location of the measurements although this would be unusual. 
• Iontophoretic induction of sweating using the Webster Sweat Inducer may cause contact 
dermatitis that may be adequately managed with topical steroids. It is also possible, although 
unlikely, that the iontophoretic procedure may cause burns to the subject. The device 
manufacturer has estimated that the risk of burns to be 1 in 50,000 iontophoretic procedures. 
It should be noted that both the Macroduct device and Webster Sweat Inducer have been 
cleared for sweat collection by the FDA and is frequently used on infants and neonates with 
rare adverse effects reported. 
• Iontophoresis may also cause minor discomfort to subjects in the form of slight stinging or 
prickling at the site of administration. However, the procedure has been approved by the 
FDA and any discomfort should cease immediately after iontophoresis is completed (~ 5 
min).  
 
As the amount of ibuprofen provided (400 mg) is within the recommended over-the-counter 
dosage for this product, we do not anticipate any risks associated with administration of this 
drug. However, ibuprofen intake has been known to produce an upset stomach including nausea, 
bloating and gas; diarrhea; constipation; headaches; dizziness; nervousness; itching skin or a 
rash; blurred vision or ringing ears in certain individuals. Prior to prescription of the ibuprofen, 
Dr Raja Sivamani will review the participants’ medical history and inform them of of the risks 
and potential side effects associated with ibuprofen intake. 
 
We will do our best to make sure that the personal information in the subjects’ study record will 
be 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. 
 
18) 
Potential Benefits to Subjects 
There are no direct benefits to the participants. Results from this study will benefit the scientific 
and medical communities and provide information that may be used to develop non-invasive 
sampling methods to support future cutaneous research studies. 
 
19) 
Vulnerable Populations 
UC-Davis Students, Employees and USDA Employees 
 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 10 of 12 
  
20) 
Multi-Site Research 
N/A 
 
21) 
Community-Based Participatory Research 
N/A 
 
22) 
Sharing of Results with Subjects 
No results will be shared with participants. 
 
23) 
Setting 
All in-person contact visits will occur at the USDA Western Human Nutrition Research Center 
on the UC Davis campus (430 West Health Sciences Dr., Davis, CA  95616).  
 
The facilities at the Western Human Nutrition Research Center were designed specifically to 
support human trials and include a clinical wing with private bedrooms for all-day study visits, a 
physiology laboratory with instrumentation for anthropometric measurements, body composition 
testing, and a reception area for screening potential participants.  In addition, the second floor is 
laboratory space for processing, analyzing and storing biological specimens.   
 
 
24) 
Resources Available 
Dr. John W. Newman is a senior scientist at the USDA Western Human Nutrition Research 
Center and holds an adjunct appointment in the Department of Nutrition at UC Davis. Dr. 
Newman will serve as PI for this project and is considered a leading expert in the field of 
metabolomics/lipomic analyses.  
 
Dr Raja K. Sivamani is an assistant professor of clinical dermatology at the UC Davis School of 
Medicine and a board certified dermatologist. His expertise centers on general dermatology 
including medical, surgical, and cosmetic services. Dr Sivamani will serve as co-PI for this 
project and oversee use of the Macroduct sweat collection and Webster sweat induction devices 
as well as oversee administration of ibuprofen to study volunteers. 
   
The human studies coordinator at the WHNRC, Dr. Ellen Bonnel, will be assisting the 
investigators with all phases of this study. Dr. Bonnel has held this position since 1999. In 
addition, to provide additional support for this study the WHNRC has a full complement of 
personnel with hands-on experience with phlebotomy expertise, and laboratory stress training.  
All necessary research equipment for this study is available at the WHNRC.  
 
Justin Waller, M.S., is the physiologist and physiology support laboratory supervisor at the 
WHNRC since February 2016. Mr. Waller has previous clinical and exercise physiology 
research experience with a variety of populations, including endurance athletes, post-bariatric 
surgery patients and breast cancer patients. His areas of expertise are in cardiovascular, 
endocrine and metabolic responses to aerobic exercise and he has previous experience validating 
commercial heart rate sensors and wearables.   


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 11 of 12 
  
 
25) 
Prior Approvals 
N/A 
 
26) 
Provisions to Protect the Privacy Interests of Subjects 
Each subject will be assigned a unique numerical identification code which will be linked to all 
data files. Data is managed through the Western Human Nutrition Research Center’s Laboratory 
Information Management System, using the identification codes only.  Only the investigators, 
the sponsors, and the WHNRC analytical laboratory director will have access to this system.  
Personal identifiers such as name, date of birth, social security number, driver’s license number, 
address, phone number, email address etc. will not be linked to the data files.  While the study is 
active, this personal information will be stored in a locked file cabinet in the Western Human 
Nutrition Research Center’s Human Studies Office, which is a secure office, along with signed 
consent forms. Only those individuals who are listed as study personnel responsible for 
consenting subjects will have access to these files. All records containing personal information 
will be destroyed upon completion of the study. 
 
27) 
Compensation for Research-Related Injury 
If subjects are injured as a result of participating in this study, USDA and the University of 
California will provide necessary medical treatment. The USDA and the University of California 
do not normally provide any other form of compensation for injury. 
 
28) 
Economic Burden to Subjects 
Transportation to the USDA WHNRC in Davis and related costs will be the responsibility of 
participants.  Complimentary on-site parking will be provided.  Compensation for participation 
sufficient to outweigh transportation costs. 
 
29) 
Consent Process 
Eligibility for participation in the study will be determined upon participant completion of a 
telephone interview with study personnel.  Eligible volunteers will be invited to visit the USDA 
WHNRC for an in-person discussion of the consent document with authorized study personnel.  
Volunteers will be allowed time for personal review of the document and given the opportunity 
for a question and answer session, as needed.  If the volunteer wishes to proceed with 
participation in the study, the consent form must be signed and dated by both the volunteer and 
the authorized study personnel. 
 
30) 
Process to Document Consent in Writing 
SOP: Written Documentation of Consent (HRP-091) procedures will be followed.   The consent 
form must be signed and dated by both the volunteer participant and authorized study personnel. 
A copy of signed consent forms will be provided to the subject upon request and the originals 
will be kept in a locked cabinet at the WHNRC human studies office. 
 
31) Drugs and Devices 


PROTOCOL TITLE: Investigating the stability, variability and mechanism of 
incorporation of lipid mediators into eccrine sweat 
 
Page 12 of 12 
  
Subjects will be administered 400 mg ibuprofen orally at study visit 4. Ibuprofen is an FDA-
approved over-the-counter non-steroidal anti-inflammatory drug. Although the drug is not being 
used for an anti-inflammatory effect, it is being utilized in FDA approved dosing. Ibuprofen will 
be sourced from the UC Davis Health System’s Investigational Drug Pharmacy following 
prescription to the study subjects by Dr Sivamani and will be stored in a locked medication 
container at room temperature at the WHNRC prior to administration to the study subjects. The 
ibuprofen will be packaged by the Investigational Drug Pharmacy into individual doses labelled 
with subject ID numbers, and will be transfered to the subjects by the study coordinator, Mr. 
Karan Agrawal under instructions from Dr Sivamani. 
 
Sweat collections will be conducted using the Macroduct® sweat collector, an FDA-approved 
medical device routinely used for sweat collection in neonates and infants. The device will be 
used according to manufacturer instructions and for its designated purpose. 
 
Sweat stimulation will be achieved by pilocarpine iontophoresis using the Webster Sweat 
Inducer, FDA-approved medical device routinely used for sweat collection in neonates and 
infants. The device will be used according to manufacturer instructions and for its designated 
purpose.