Measurement of Agricultural and Dietary Glyphosate Exposure Among Pregnant Women
Phase: NA
Conditions: Exposure to Herbicides
Sponsor: Boise State University

 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
Study Protocol 
Measurement of Agricultural and Dietary Glyphosate 
Exposure among Pregnant Women 
NCT04155463 
June 30, 2022 

1 
 
TABLE OF CONTENTS 
 
1. INTRODUCTION ................................................................................ PAGE 4 
1.1 
Organization of this Document 
1.2 
General Information for Lab Members 
1.3 
Terminology 
1.4 
New Lab Member Training Requirements 
1.5 
Scope 
 
 
2. RECRUITMENT .................................................................................. PAGE 8 
2.1 
Summary of Methods 
2.2 
Materials* 
2.3 
Procedures 
 
 
3. ENROLLMENT & INITIALCONTACTLESS MEETING ............ PAGE 11 
3.1 
Summary of Methods 
3.2 
Materials* 
3.3 
Procedures 
 
 
4. WEEKLY URINE SAMPLE COLLECTION ................................. PAGE 15 
4.1 
Summary of Methods 
4.2 
Materials* 
4.3 
Procedures 

2 
 
5. WEEKLY URINE SAMPLE ANALYSIS& STORAGE ................. PAGE 18 
5.1 
Summary of Methods 
5.2 
Materials* 
5.3 
Procedures 
5.4 
Weekly Urine Sample Labeling System 
 
 
6. DIETARY INTERVENTION .............................................................. PAGE 21 
6.1 
Summary of Methods 
6.2 
Schedule 
6.3 
Materials* 
6.4 
Procedures 
 
 
7. DAILY URINE SAMPLE COLLECTION ........................................ PAGE 27 
7.1 
Summary of Methods 
7.2 
Materials* 
7.3 
Procedures 
 
 
8. DAILY URINE SAMPLE ANALYSIS& STORAGE… .................. PAGE 30 
8.1 
Summary of Methods 
8.2 
Materials* 
8.3 
Procedures 
8.4 
Daily Urine Sample Labeling System 

3 
 
9. URINE SAMPLE SHIPMENT & GLYPHOSATE ANALYSIS…. PAGE 35 
9.1 
Summary of Methods 
9.2 
Materials 
9.3 
Procedures 
 
 
*Materials Available Upon Request 

4 
 
1. 
INTRODUCTION 
1.1 Organization of this Document 
 
The standard operating procedures and associated materials for this studyare 
included in this compilation, outlining all of the fieldwork and laboratory 
activities to be carried out by the Curl Agricultural Health Lab as part of the 
study Measurement of Agricultural and Dietary Glyphosate Exposure 
among Pregnant Women. The documents are arranged in nine categories, 
each with the standard operating procedures for the following aspects of the 
study: 1) Introduction; 2) Recruitment; 3) Enrollment & Initial Contactless 
Meeting; 4) Weekly Urine Sample Collection; 5) Weekly Urine Sample 
Analysis & Storage; 6) Dietary Intervention; 7) Daily Urine Sample 
Collection; 8) Daily Urine Sample Analysis & Storage; and 9) Urine Sample 
Shipment & Glyphosate Analysis. 
1.2 General Information for Lab Members 
 
The procedures presented in these standard operating procedures are intended 
to ensure the collection of high-quality data, and lab members should follow 
them at all times. 
 
To ensure no COVID-19-specific risks, no more than two lab members will 
be in the lab at the same time. Such interaction with comply with all Boise 
State University policies, will only occur when required and will follow all 
standard COVID-19 safety guidelines, including face masks, social 
distancing, and frequent hand washing. Masks will be worn in the lab and in 
the Environmental Research Building (ERB) at all times, and social distancing 
protocols will be followed. Before returning to the Boise State University 
campus after scheduled breaks or campus closures, all lab members will 
follow university guidelines for COVID-19 testing and isolation, if required. 
All lab members will participate in a weekly or bi-weekly virtual meeting to 
remain up-to-date and aligned on tasks. There will be no face-to-face 
interactions between lab members and potential or enrolled study participants. 
 
All communication with participants and any potential participants will be 
done via their preferred method of communication (call or text) and in their 
preferred language. In most instances in this protocol it is assumed that text is 
the preferred method of communication for participants once enrolled in this 

5 
 
study. In addition to collecting and testing urine samples, as well as managing 
food ordering and delivery activities, lab members will also be responsible for 
keeping clear and thorough records of these activities. Accurate, detailed 
documentation is a vital part of the data collection process. The following 
guidelines will help lab members in completing the forms included in this 
document and logging activities: 
 
• 
Be compulsive in record keeping. It is better to record too much 
information than not enough. After the data are collected, these 
records will be the only way of knowing what occurred during 
sample collection and in the lab. 
 
• 
Always use pen (no pencils). 
 
• 
If a record on a form needs to be changed, draw a single line 
through the entry that needs to be changed, write in thecorrect 
information, and then write the date of the change and the 
initials of the lab member making the change. 
 
• 
If a mistake is made or there are deviations from protocol, 
indicate this on the associated form. This is very importantin 
helping the data analysts identify factors that may affect data 
quality. 
 
• 
Always use 24 hour (i.e. military) time. 
 
1.3 Terminology 
 
The following abbreviations are used throughout this document: 
 
IRB 
IRB stands for Institutional Review Board. This is a 
committee that protects the rights and welfare of human 
subjects recruited to participate in research conducted 
through Boise State University. 
 
ERB 
ERB stands for Environmental Research Building. This 
building is located on the Boise State University main 
campus, and houses the Curl Agricultural Health Lab. 

6 
 
CAHL 
CAHL stands for Curl Agricultural Health Lab. The 
CAHL is located in the ERB, room 3106, at Boise State 
University. 
 
WIC Clinic 
WIC stands for Women, Infant, and Children. Recruitment 
for this study will be completed in cooperation with WIC 
clinics in the Southwest, Central Southwest, Central, and 
South Central Health Districts of Idaho. This includes 
clinics in Caldwell and Nampa (Southwest); Boise, 
Meridian, and Garden City (Central); and Twin Falls, 
Shoshone, Gooding, Jerome, and Heyburn (South 
Central). 
 
SG 
SG stands for specific gravity. CAHL lab members will be 
testing the specific gravity of the participants’ urine 
samples. 
 
DI 
DI stands for deionized water. The refractometer used for 
measuring specific gravity will be cleaned using DI water. 
 
CDC 
CDC stands for the Centers for Disease Control and 
Prevention. This entity will be responsible for analyzing 
all urine samples collected in this study. At the conclusion 
of the urine sampling component of this study, all samples 
will be shipped to a laboratory at the CDC for glyphosate 
analysis. 
 
K01 
K01 refers to the Mentored Research Scientist 
Development Award, the funding for this project. 
 
 
1.4 New Lab Member Training Requirements 
 
IRB Training Requirements 
 
In order to interact with human subjects, new lab members must complete 
the CITI training, the IRB Additional Personnel Form, and the IRB 
Modification Form. 

7 
 
Lab Training Requirements 
 
Lab Training 
 
All new lab members will receive general lab training prior to involvement in 
the study, although the manner in which this training occurs will depend on 
the current rules regarding COVID-19 safety. 
 
CAHL Shared Research Drive and K01 Database 
 
All electronic documentation will be stored in a protected Microsoft Access 
document entitled “K01 Database” located on a shared research drive 
operated through Boise State University. In order to access the shared research 
drive, all new lab members will email Ms. Spivak and ask her to request 
security access. 
 
CAHL Calendar and Google Sheets 
 
All scheduling will be electronically documented in the password protected 
CAHL Calendar, and all cryovial storage locations will be electronically 
documented in the password protected CAHL Google Sheets. 
 
1.5 Scope 
 
Glyphosate is the most commonly used herbicide in the world. This increase 
has been driven, in large part, by the introduction of glyphosate-resistant crops 
such as “Roundup Ready” corn and soybeans. In Idaho, nearly a quarter 
million acres of Roundup Ready sugarbeets are cultivated each year. 
Human exposure to glyphosate and its formulations may occur through 
dietary, agricultural and residential pathways. Despite its extensive use, 
frequent presence in foodstuffs and environmental media, and potential 
toxicity, current exposure levels in human populations are not well 
documented. However, several studies have suggested that in utero 
exposure to glyphosate and glyphosate formulations may have 
reproductive effects and adversely impact developing fetuses. This 
study therefore aims to quantify pathways of glyphosate exposure 
among pregnant women. 

8 
 
2. 
RECRUITMENT 
 
2.1 Summary of Methods 
 
This study will involve recruiting 40 pregnant women in their first trimester 
to measure residential and dietary glyphosate exposure throughout their 
pregnancies. This will be done in cooperation with local WIC clinics. 
 
2.2 
Materials 
 
WIC Clinic Collaboration 
 
• 
Introduction to Study and Recruitment Video for WIC Staff 
(https://youtu.be/pFH3MIELXNU) 
 
• 
Video for Potential Participants (English: 
https://youtu.be/sOG0lppKzUw or Spanish: 
https://youtu.be/p75mIj0hwrc) 
 
• 
Recruitment Flyer* 
 
• 
Index Card Flyer* 
 
• 
Slip for Potential Participant Contact Information* 
 
Contacting Potential Participants 
 
• 
Completed Slip for Potential Participant Contact Information 
 
• 
Recruitment Scripts* 
 
• 
Eligibility Questionnaire* 
 
• 
Cell phone with One Talk App 
• 
CAHL Calendar 
 
• 
K01 Database 

9 
 
2.3 Procedures 
WIC Clinic Collaboration 
WIC Clinic Collaboration 
 
The staff at the WIC clinics will watch the Introduction to Study and 
Recruitment Video for WIC Staff to provide them with general information 
about the study and the virtual study recruitment process. During 
appointments with clients, the staff will show the Video for Potential 
Participants to women who are thought to meet eligibility requirements based 
on available information, or this video may be emailed to these women from 
the WIC staff to watch at their convenience. This video will be provided to 
potential participants in their preferred language, either English or Spanish. 
This video will instruct them to inform their WIC staff if they are interested 
in participating. The WIC staff will then complete a Slip for Potential 
Participant Contact Information with the potential participant’s name and 
contact information in the participant’s preferred language, English or 
Spanish. The WIC clinic staff will fax the completed Slip for Potential 
Participant Contact Information to Dr. Curl or will call Dr. Curl to relay 
potential participant information. The Recruitment Flyer and Index Card Flyer 
can be used by the WIC staff to aid in recruitment either by emailing these 
documents to potential participants or printing these documents for use in the 
clinics should face-to-face appointments at the WIC clinics take place. This 
process will continue until recruitment is complete. 
 
Documentation 
 
Dr. Curl will record all information contained in each Slip for Potential 
Participant Contact Information in the K01 Database. Dr. Curl will also 
perform an initial eligibility screening of each potential participant’s address 
and due date. If a potential participant is eligible after the initial screening, a 
lab member will attempt to contact her. 
 
Contacting Potential Participants 
 
Contacting Potential Participants 
 
A lab member will contact the potential participant during her preferred 
contact time and in her preferred language via her preferred contact method 

10 
 
no more than two days after receiving the Slip for Potential Participant 
Contact Information. If the potential participant is contacted via text message 
(using the Text Message Recruitment Script), a phone call will be arranged 
for that time (if available) or for a later date to cover the Phone Recruitment 
Script and Eligibility Questionnaire. If contacted via phone call but the 
potential participant does not answer, a voicemail will be left in accordance 
with the Voicemail Recruitment Script if the potential participant has a 
voicemail option set-up. If contacted via phone call and the potential 
participant does answer, the lab member will cover the Phone Recruitment 
Script and Eligibility Questionnaire. However, another call can be arranged if 
the potential participant is not available to talk at that time. 
 
Determining Eligibility and Scheduling 
 
Once the potential participant is contacted via a phone call, the lab member 
will recite the Phone Recruitment Script. If the potential participant is 
interested in the study at this point, the lab member will administer the 
Eligibility Questionnaire, following the instructions on the form and asking 
all the necessary questions. This questionnaire will be completed on paper for 
proper study documentation purposes. If the participant is eligible, the lab 
member will then arrange a date and time to have an initial meeting. 
 
Documentation 
 
Scheduled initial contact meetings will be documented in the CAHL 
Calendar. All correspondence with any potential participant (regardless of 
interest in study or eligibility) will be recorded in the Potential Participant 
Contact Form in the K01 Database. This also includes any attempts to reach 
a potential participant by text or phone call, and if a voicemail is left. If the 
participant is uninterested or ineligible, the reason will also be recorded in 
that form and an “XX” will be placed in “Participant ID” field. If the 
participant is interested and eligible, all paper documents containing contact 
information, eligibility questions, and/or notes will be placed in the 
“Pending” file in the locked filing cabinet located in CAHL. If the 
participant is uninterested or ineligible, all paper documents containing 
contact information, eligibility questions, and/or notes will be placed in the 
“Uninterested/Ineligible” file located in the same locked filing cabinet. 

11 
 
3. ENROLLMENT & INITIAL CONTACLESS MEETING 
 
3.1 
Summary of Methods 
 
After a woman has been determined eligible for the study and has indicated a 
willingness to participate, a lab member will schedule and attend an initial 
contactless meeting that will include the Informed Consent process, 
Demographics Questionnaire, Pesticide Exposure Questionnaire, collection of 
the first weekly urine sample, and an explanation of the process for the 
remaining weekly urine sample collections. A Urine Chain of Custody Form 
has been developed to track the movement of all urine samples from collection 
to final storage in the lab. Lab members will initial each step of sample 
transport on the log, and note any observations, deviations from sample 
handling protocols, and/or problems with each sample. Once back at CAHL 
the lab member will analyze and store the first weekly urine sample. 
3.2 
Materials 
 
• 
Two Informed Consent Forms* 
 
• 
Video for Informed Consent (English: 
https://youtu.be/cI-wEKWKVso or Spanish: 
https://www.youtube.com/watch?v=sL3WqtcT7Zk) 
• 
Demographics Questionnaire* 
 
• 
Pesticide Exposure Questionnaire* 
 
• 
Video for Urine Collection Instructions (English: 
https://youtu.be/Gf-xqsx3mLI or Spanish: 
https://www.youtube.com/watch?v=WQ9MjRADA_o) 
 
• 
Cell phone with One Talk App 
 
• 
Supplies for first and second weekly urine sample collections: 
 
a) Specimen cups with barcoded label for the first and 
second week’s collections (week numbers 1 and 2, 
respectively) in separate sealable plastic bags 

12 
 
 
b) Extra specimen cup 
 
c) Two pairs of nitrile gloves 
 
d) Two coolers 
 
e) Two ice packs 
 
f) Two Sharpies and a pen 
 
g) Urine Chain of Custody Form* 
 
h) $10 Gift Card 
 
i) Urine Collection Instruction Form* 
 
j) Urine Collection Reminder Poster* 
 
• 
CAHL Calendar 
 
• 
K01 Database 
 
3.3 
Procedures 
 
Supplies Preparation 
 
A lab member will compile and set out the materials listed above. Hard copies 
of all necessary papers will be placed in a folder labeled with the potential 
participant’s ID number. The lab member will take all supplies and this folder 
to the initial meeting. 
Scheduling 
 
All scheduling, including any scheduling changes, will be recorded in the 
CAHL Calendar and in the K01 Database. The lab member will call or text 
(based on the potential participant’s preference) the potential participant one 
day prior to the scheduled meeting as a reminder. 

13 
 
Initial Contactless Meeting 
 
A lab member will conduct the initial contactless meeting with the potential 
participant in her preferred language. For any exchange of materials, the 
participant should be inside her home with the door shut while the lab member 
drops off materials at her door while wearing a mask. Once the lab member is 
back inside the car, the participant may come out and pick up the materials. 
Only one lab member should be present for this meeting so that two people 
are not traveling in the car together. This lab member will be fluent in the 
potential participant’s preferred language. At this meeting, the lab member 
will arrive and drop of the two copies of the Informed Consent Forms (in the 
preferred language) at the potential participant’s door that have already been 
signed and dated by the lab member. The lab member will then return to the 
car and call the potential participant to introduce themselves, instruct her to 
pick up the Informed Consent Forms, and tell her that these forms will be 
explained via video. The lab member will then text the link for the Video for 
Informed Consent to the potential participant in her preferred language, either 
English or Spanish. After the potential participant watches the video, the lab 
member will review the document and answer questions (if necessary) over 
the phone. If the potential participant agrees to participate in the study, the lab 
member will then instruct the participant to sign and date both copies of the 
Informed Consent Form and keep one copy for herself and place one copy 
back outside for the lab member to collect and keep. The lab member will put 
this copy in the car to bring back to the lab. At this time, this woman is 
considered an official study participant, and will be assigned a participant ID 
number. 
 
The lab member will then conduct the Demographics Questionnaire and 
Pesticide Exposure Questionnaire in the participant’s preferred language over 
the phone, following the instructions provided at the top of these documents. 
After completion of both questionnaires, the lab member will inform the 
participant that she can now provide the first weekly urine sample and explain 
that the urine sample collection process will be explained via video. The lab 
member will then text the link for the Video for Urine Collection Instructions 
to the participant in her preferred language, English or Spanish. While the 
participant is watching this video, the lab member will drop off at the 
participant’s door the following supplies: 1) specimen cup with barcoded label 
for the first week’s collection (week number 1) in a sealed plastic bag; 2) a 
pair of nitrile gloves; and 3) a Sharpie. The video will instruct theparticipant 

14 
 
to call the lab member to discuss any questions or concerns, including if there 
is an accident and an extra specimen cup is needed. 
 
After the participant provides the first urine sample, she will place the 
specimen cup outside her door in the sealed plastic bag. The lab member will 
then collect the sample, ensure the collection date and time have been 
provided and that the cup is labeled with the Participant ID number, and place 
it in the lab cooler with the lab ice pack to bring back to the lab. The lab 
member will also complete the appropriate sections of the Urine Chain of 
Custody Form. The lab member will then leave a $10 gift card and the 
following supplies for the second week’s collection (week number 2) at the 
participant’s door: 1) specimen cup with barcoded label for the second week’s 
collection in a sealed plastic bag; 2) a cooler for the participant to keep; 3) an 
ice pack for the participant     to      keep; 4) extra specimen cup to be used  
in case of an accident and only when barcoded cup is not available; and 5) the 
Urine Collection Instruction Form and Urine Collection Reminder Poster 
(both in the preferred language). Once back in the car, the lab member will 
call the participant and instruct her to pick up the supplies and will review the 
process for subsequent weekly urine sample collections as described in 
Section 4 of this document. The participant can use the Urine Collection 
Instruction Form as a reminder on how to complete the collection process, and 
the Urine Collection Reminder Poster as a reminder to collect samples 
weekly. The lab member will schedule subsequent weekly urine sample 
collections over the phone. 
Documentation upon Returning to the Lab 
 
After the initial contactless meeting, the lab member will return to the CAHL. 
There, the lab member will place the first weekly urine sample in the 
refrigerator and complete the appropriate sections of the Urine Chain of 
Custody Form. This sample will remain in the refrigerator for no more than 
24 hours until analyzed. The cooler will be returned to the storage shelf and 
the ice pack will be placed back in the freezer. 
 
The lab member will then document participant information in the K01 
database and paper copies of materials will be placed in the locked filing 
cabinet. At this time the lab member should analyze and store the weekly urine 
sample as described in Section 5 of this document. 

15 
 
4. WEEKLY URINE SAMPLE COLLECTION 
 
4.1 Summary of Methods 
 
As with the initial urine sample, all subsequent weekly urine samples will be 
collected, analyzed, and stored by lab members in a completely contactless 
manner. Weekly urine samples will be numbered sequentially. Each week, 
each study participant will provide a first morning void sample on a pre- 
scheduled day, and will place the sample in her cooler with her ice pack 
outside her home. A lab member will then collect that week’s sample, provide 
the participant with a specimen cup for the following week, and bring that 
week’s sample back to CAHL in a lab cooler with a lab ice pack. Once back 
at CAHL, the lab member will analyze and store the weekly urine sample. 
 
4.2 
Materials 
 
• 
Specimen cup with barcoded label 
 
• 
Extra specimen cup 
 
• 
Nitrile gloves 
 
• 
Cooler 
 
• 
Ice pack 
 
• 
Urine Chain of Custody Form for the correct participant ID 
 
• 
$10 Gift Card 
 
• 
One Sharpie and pen 
 
• 
Cell phone with One Talk App 
 
• 
CAHL Calendar 
 
• 
K01 Database 

16 
 
4.3 
Procedures 
 
Supplies Preparation 
 
A lab member will compile and set out the materials listed above. The Urine 
Chain of Custody Form will be taken from the folder in the locked filing 
cabinet labeled with the participant’s ID number. The lab member will take 
all supplies to the urine collection. 
Scheduling 
 
For weekly urine sample collections, participants will be informed at the 
initial contactless meeting of a pre-established date and time for weekly 
sample pick ups, which will be decided a priori by lab staff based on CAHL 
staff availability to travel to certain locations on certain days and times. 
Scheduling changes will be considered should the pre-set day and time for 
weekly sample collection not work for a study participant. All scheduling will 
be recorded in the CAHL Calendar. The lab member will call or text the 
participant one day prior to the scheduled pick up as a reminder. 
 
Weekly Urine Sample Collections 
 
Each week on the pre-scheduled day, participants will be instructed to provide 
a first morning void in the specimen cup with the barcoded label for that week, 
which was provided to them at the prior week’s collection. The participants 
will be instructed to use their previously provided Sharpie to record the date 
and time of collection directly on the specimen cup. The sample should be 
provided the morning of the collection day (first morning void), and then 
placed in the sealable plastic bag and into the previously provided cooler with 
the ice pack. This cooler will then be placed outside the participant’s home. A 
lab member will then travel to each participant’s home to collect the urine 
sample. If the lab member arrives at a participant’s home and finds that the 
weekly urine sample has not been provided, the lab member will make 
reasonable attempts to contact the participant and collect the urine sample. 
 
At collection, the lab member will ensure that the cup is labeled with the 
correct Participant ID number and correct week number. The cup will then be 
placed in the lab cooler with the lab ice pack to be taken back to CAHL. The 
lab member will leave in the participant’s cooler: 1) specimen cup with 
barcoded label for the following week’s collection in sealable plastic bag; 2) 

17 
 
extra specimen cup to be used in case of an accident and only when barcoded 
cup is not available; and 3) a $10 gift card for the sample just provided. At 
collection time, the lab member will complete the appropriate sections of the 
Urine Chain of Custody Form. The lab member will ensure that the participant 
filled in the date and time of collection on the cup label, and if this did not 
occur this will be noted in the Urine Chain of Custody Form, along with any 
other problems or deviations from protocol. 
 
After each sample collection, a text message will be sent to each participant 
in her preferred language. 
 
If a participant does not provide a urine sample for a given week, the lab 
member will document the missed week on the Urine Chain of Custody Form. 
This will be considered a missed week and that week number will not be 
reused the following week, and instead the next sequential week number will 
be used. 
 
Documentation upon Returning to the Lab 
 
After collecting the sample, the lab member will return back to CAHL. There, 
the lab member will place the weekly urine sample in the refrigerator and 
complete the appropriate sections of the Urine Chain of Custody Form. This 
sample will remain in the refrigerator for no more than 24 hours until analyzed 
for color, clarity and specific gravity, and stored at -80˚C. The lab member 
will then document all information in the K01 Database. The lab member will 
file the Urine Chain of Custody Form in the folder labeled with that 
participant’s ID number in the locked filing cabinet. At this time, the lab 
member should analyze and store the weekly urine sample as described in 
Section 5 of this document. 
Final Weekly Urine Sample Collection 
When each participant is approximately 4 weeks away from her due date, a 
text message will be sent to the participant in her preferred language to 
remind her to let us know once she gives birth. Once the participant informs 
us that her baby has been born, no more weekly urine samples will be 
provided or collected. Each participant will receive a $50 gift card for 
completion of the study. 

18 
 
Gift Cards 
 
All gift card purchases and documentation will follow Boise State 
University rules and regulations. 
 
 
5. WEEKLY URINE SAMPLE ANALYSIS & STORAGE 
 
5.1 Summary of Methods 
 
Weekly urine samples will be collected from participants and returned to 
CAHL in a lab cooler with a lab ice pack. Samples will be stored for no 
more than 24 hours under refrigeration at CAHL until analyzed and stored. 
During analysis, lab members will determine urine color, clarity, and 
specific gravity. Urine sample storage will occur immediately after urine 
sample analysis. Each sample will be aliquoted into four 4mL weekly 
cryovials for storage. These will be stored in a weekly cryobox in the CAHL 
-80˚C freezer. At the conclusion of an individual’s participation in the study 
(ideally when they give birth), one of the four 4 mL weekly cryovials from 
each week will be sent to the CDC for glyphosate analysis. The remaining 
three cryovials are duplicate samples that will be stored in thebiorepository. 
 
5.2 
Materials 
 
Weekly Urine Sample Analysis 
 
• 
Absorbent pads 
 
• 
Kimwipes 
 
• 
Cup containing ethyl alcohol 
 
• 
Squirt bottle containing DI water 
 
• 
Nitrile gloves 

19 
 
• 
Pen 
 
• 
Urine Sample Characteristics Log* 
 
• 
Color and Clarity Guide* 
 
• 
Transfer pipettes 
 
• 
Atago Urine Specific Gravity Refractometer, PAL 10-S 
 
• 
K01 Database 
 
Weekly Urine Sample Storage 
 
• 
Absorbent pads 
 
• 
Nitrile gloves 
 
• 
Eppendorf Plus Research Pipette and tip 
 
• 
Four weekly cryovials with barcoded labels 
 
• 
Weekly Cryobox 
• 
Empty Cryobox at the lab station 
 
• 
Google Drive Cryobox Document 
 
 
5.3 
Procedures 
 
Weekly Urine Sample Analysis 
 
Supplies Preparation 
 
Prior to starting urine sample analysis, the lab member will set up a lab station 
by placing the following supplies on an absorbent pad: 1) Kimwipes; 2) cup 
containing ethyl alcohol; 3) squirt bottle containing DI water; 4) Atago Urine 

20 
 
Specific Gravity Refractometer, PAL 10-S; 5) transfer pipettes; and 6) the 
appropriate urine specimen cup from the refrigerator. The lab member will 
also have out the Color and Clarity Guide and a Urine Sample Characteristics 
Log. A pair of nitrile gloves will be worn when handling urine samples. 
 
Measurement of Color and Clarity 
 
The color and clarity of each urine sample will be noted through comparison 
with the Color and Clarity Guide and recorded on the Urine Sample 
Characteristic Log. 
 
Measurement of Specific Gravity 
 
SG will be measured following the manufacturer’s instructions that 
accompany the refractometer. 
Documentation 
The lab member will complete the Urine Sample Characteristics Log, 
Information from the Urine Sample Characteristics Log will be recorded in 
the K01 Database. The paper copy of the Urine Sample Characteristics Log 
will then be filed in the folder labeled with the participant’s ID in the locked 
filing cabinet. 
 
Weekly Urine Sample Storage 
 
Supplies Preparation 
 
Urine sample storage should occur immediately after analysis. Prior to 
beginning storage procedures, the lab member will set the following supplies 
out on a lab station on an absorbent pad (this can be the same absorbent pad 
used during analysis): 1) Eppendorf Plus Research Pipette (set at 4.0 mL) 
and tip; 2) four 4 mL weekly cryovials with the appropriate barcoded labels; 
3) one cryobox at the lab station; and 4) the appropriate urine specimen cup. 
A pair of nitrile gloves will be worn when handling urine samples. 
 
Weekly Urine Sample Storage 
 
Four mL of the urine sample will be pipetted into four separate 4 mL weekly 
cryovials using the Eppendorf Plus Research pipette and tip. Once all four of 

21 
 
the weekly cryovials have been filled, the lab member will remove the 
weekly cryobox from the -80˚C freezer. This weekly cryobox will be placed 
at the lab station, and the newly filled weekly cryovials will then be 
transferred from the cryobox at the lab station and into the freezer weekly 
cryobox. The weekly cryobox in the freezer will only be removed from the 
freezer briefly for daily cryovial transfer, and it will be immediately returned 
to the freezer as quickly as possible. 
 
Documentation 
 
The lab member will record the specific cryovial label and cell number 
where each of the four weekly cryovials were placed in the freezer weekly 
cryobox. This will be recorded in the CAHL shared Google Drive Cryobox 
Document. 
 
6. DIETARY INTERVENTION 
 
6.1 Summary of Methods 
 
The dietary intervention will take place over the course of 17 days, from June 
14 to June 30. Participants will receive one week of conventional groceries 
and one week of organic groceries, the order of which will be randomly 
assigned. Each week, each study participant will shop for up to $150 worth of 
groceries using an online account at a local grocery store, and grocery orders 
will be delivered to participants. Participants will track their food intake each 
week using a Food Log. During the dietary intervention, participants will not 
provide weekly urine samples and will instead provide 14 daily urine samples. 
 
6.2 Schedule 
 
The dietary intervention will be divided into two weeks referred to as “Week 
One” and “Week Two.” Key dates during the dietary intervention, as well as 
key dates prior to starting the dietary intervention, are: 

22 
 
May 24-28: Dietary Intervention Reminder Flyers left in participant’s 
coolers 
 
June 3-June 9: Personalized Week One Grocery Flyer left in 
participant’s coolers 
 
June 7-11: Week One Food Logs left in participant’s coolers 
 
June 13: Week One grocery orders placed 
 
 
June 14: Week One grocery orders delivered, participants begin to eat 
Week One groceries 
 
June 15: Washout day 
 
June 15-21: Participants complete Week One Food Log 
June 16-22: Week One daily urine sample collections 
June 17-June 18: Personalized Week Two Grocery Flyer left in 
participant’s coolers 
 
June 21: Week Two grocery orders placed, Week Two Food Logs left 
in participant’s coolers 
 
June 22: Week Two grocery orders delivered, participants stop eating 
Week One groceries and begin to eat Week Two groceries 
 
June 23: Washout day 
 
June 22-29: Participants complete Week Two Food Log 
June 24-30: Week Two daily urine sample collections 
June 24: Week One Food Logs collected 
June 30: Week Two Food Logs collected 

23 
 
6.3 Materials 
 
• 
Shared Research Drive 
 
• 
Cell phone with One Talk App 
 
• 
Dietary Intervention Reminder Flyer* 
• 
Supplies for Week One: 
 
a) Personalized Week One Grocery Flyer* 
 
b) Week One Food Log* 
 
• 
Supplies for Week Two: 
 
a) Personalized Week Two Grocery Flyer* 
 
b) Week Two Food Log* 
 
6.4 Procedures 
 
Supplies Preparation 
 
Prior to beginning the dietary intervention, a lab member will create a new 
email account through Gmail for each participant. The lab member will then 
use this email to create an online account for each participant at the local 
grocery store. 
 
Grocery Ordering and Delivery 
 
During weekly urine sample collections from May 24-May 28, a lab member 
will leave the Dietary Intervention Reminder Flyer in the participants’ coolers 
in their preferred language. 
 
Participants will be provided their own local grocery store account username 
and password by a lab member. Participants will receive a personalized flyer 
(described in more detail below) with instructions regarding how to log into 

24 
 
their account and shop for up to $150 worth of groceries (either conventional 
or organic, as randomly assigned) and select their preferred grocery delivery 
time. 
 
Participants will also be instructed to call or text a lab member via the One 
Talk App once grocery shopping is complete. The lab member will then 
review the order to ensure that all items are available and correspond to the 
week in which the participant has been randomized (i.e., conventional or 
organic), and that no prohibited items have been added (e.g., no alcohol, non- 
food items, items from the “Personal Care & Health” department, cigarettes, 
or lottery tickets). The lab member will contact the participant if there are 
questions or concerns about the order. After the lab member has reviewed the 
order, he/she will contact the participant to confirm her delivery address and 
preferred delivery time. The participant or a member of her household must 
be home at the time of grocery delivery. The grocery order will be placed by 
the lab member. 
 
Participants will also be given a brief food log for each week, in which they 
will be asked to note any food or drinks they consume from food outside of 
their grocery order, including any meals from restaurants. Participants will 
also be asked to complete a Likert scale question each day with the following 
options: “Everything I ate was fromthe study,” “Most of what I ate was from 
the study,” “About half what I ate was from the study,” “A little bit of what I 
ate was from the study,” or “None of what I ate was from the study.” 
 
Week One 
 
A lab member will leave a Personalized Week One Grocery Flyer in the 
participant’s cooler in her preferred language during weekly urine sample 
collections from June 3-June 9. Each Week One Grocery Flyer will be 
personalized for each participant. This flyer will clearly indicate: 1) the 
participant’s username and password for her online local grocery store 
account, 2) whether she should shop for conventional or organic groceries for 
this week, 3) how to complete her grocery shopping; 4) how to select her 
preferred grocery store location and delivery time frame, 5) the deadline for 
completing her grocery shopping, and 6) instructions to text or call lab 
members on the One Talk phone number once her grocery shopping is 
complete. 

25 
 
On the day before the participant is scheduled to complete her Week One 
grocery shopping, the lab member will send a text to the participant in her 
preferred language reminding her to complete her order by 5:00 PM the 
following day, if she has not done so yet. 
 
Grocery orders for Week One will be delivered to participants on June 14. 
Participants will begin to eat their Week One groceries on June 14. 
 
The Week One Food Log will be left in each participant’s cooler in her 
preferred language by a lab member during urine sample collections on June 
7-11. Participants will complete the Week One Food Log from June 15-June 
21. Participants will be instructed to leave the completed Week One Food Log 
in their coolers along with their daily urine samples on June 24, and a lab 
member will collect it on June 24. 
 
On June 14, after groceries have been delivered for Week One, the lab 
member will contact the participant in her preferred language to ensure the 
groceries were received, to remind her to complete the Week One Food Log, 
and to remind her that daily urine sample collection will begin on June 16. 
 
Week Two 
 
A lab member will leave the Personalized Week Two Grocery Flyer in the 
participant’s cooler in her preferred language during daily urine sample 
collection on June 17 or June 18. This flyer will be similar to the Personalized 
Week One Grocery Flyer, but with appropriate changes made for Week Two. 
 
On the day before the participant is scheduled to complete her Week Two 
grocery shopping, the lab member will send a text to the participant in her 
preferred language reminding her to complete her order by 5:00 PM the 
following day, if she has not done so yet. 
 
Grocery orders for Week Two will be delivered to participants on June 22. 
Participants will begin to eat their Week Two groceries on June 22. 
 
On the day before the participant is scheduled to receive her Week Two 
groceries, the lab member will send a text to the participant in her preferred 
language reminding her to collect her urine sample as a first morning void on 
the morning of June 22, prior to consuming any Week Two groceries. 

26 
 
The Week Two Food Log will be left in each participant’s cooler in her 
preferred language by a lab member during urine sample collections on June 
21. Participants will complete the Week Two Food Log from June 22-29. 
Participants will be instructed to leave the completed Week Two Food Log in 
their coolers along with their daily urine samples on June 30, and a lab 
member will collect it on June 30. 
 
On June 22, after the groceries have been delivered for Week Two, the lab 
member will contact the participant in her preferred language to ensure the 
groceries were received, to remind her to stop eating Week One groceries and 
begin eating Week Two groceries, and to remind her that there will be no daily 
urine sample collection the following day, June 23, and daily urine sample 
collection will resume on June 24. 
 
On June 23, the lab member will contact the participant in her preferred 
language to remind her that daily urine sample collection will resume the next 
day on June 24, to remind her to place her completed Week One Food Log in 
her cooler along with her daily urine sample, and to remind her to complete 
the Week Two Food Log. 
 
On June 24, in addition to collecting the daily urine sample, the lab member 
will pick up the completed Week One Food Log. 
 
On June 29, the lab member will contact the participant in her preferred 
language to remind her to place her completed Week Two Food Log in her 
cooler along with her final daily urine sample the following morning, and to 
remind her that the study will resume weekly urine sample collections the 
following week. 
 
On June 30, in addition to collecting the final daily urine sample and picking 
up the completed Week Two Food Log, the lab member will leave the 
materials to resume weekly urine sampling the following week. The necessary 
weekly urine sample materials should be determined individually for each 
participant based on the week number last recorded prior to the dietary 
intervention. 

27 
 
7. DAILY URINE SAMPLE COLLECTION 
 
7.1 Summary of Methods 
 
Daily urine samples will be provided during the dietary intervention during 
Week One, from June 16-22, and during Week Two, from June 24-30. 
Participants will provide a total of 14 daily urine samples. Daily urine samples 
will be numbered sequentially. Each day, each study participant will provide 
a first morning void sample and will place the sample in her cooler with her 
ice pack outside her home. A lab member will then collect that day’s sample, 
provide the participant with a specimen cup for the following day, and bring 
that week’s sample back to CAHL in a lab cooler with a lab ice pack. Once 
back at CAHL, the lab member will analyze and store the daily urine sample. 
 
Daily urine sample collections will follow the same protocol as described in 
Section 4 of this document, with the following major deviations: 
 
• 
Lab members will collect and deliver daily urine samplecups, 
rather than weekly urine sample cups 
 
• 
Lab members will complete a Daily Urine Chain of Custody 
Form, rather than the Urine Chain of Custody Form. 
 
• 
For non-local collections two days of daily urine samples will 
be collected during a two-day trip. On the first day a lab 
member will drive to that area, collect the daily urine samples 
for that day, transport the daily urine samples in a lab cooler 
with a lab ice pack to a hotel, and store the daily urine samples 
in the study refrigerator at the hotel. The lab member will spend 
the night at the hotel, and then on the second day the lab 
member will collect the daily urine samples for that day and 
bring the urine samples for both days back to CAHL in a lab 
cooler with a lab ice pack 
 
• 
Participants will receive one $50 gift card at the end of the 
dietary intervention period if they provided all 14 daily urine 
samples, rather than receiving a $10 gift card for each sample 
collected 

28 
 
7.2 Materials 
 
• 
Specimen cup with barcoded label in a sealable plastic bag (day 
one) 
 
• 
Specimen cup with barcoded label for the following day’s 
collection in a sealable plastic bag 
 
• 
Extra specimen cup 
 
• 
Nitrile gloves 
 
• 
Cooler 
 
• 
Ice pack 
 
• 
Daily Urine Chain of Custody Form for the correctparticipant 
ID* 
 
• 
One sharpie and pen 
 
• 
Cell phone with One Talk App 
 
• 
CAHL Calendar 
 
• 
K01 Database 
 
• 
Study refrigerator 
 
• 
Hotel confirmation number 
 
• 
$50 gift card (for the last day only) 

29 
 
7.3 Procedures 
 
Supplies Preparation 
 
The specimen cup with the barcoded label for the first day’s collection (day 
one) in a sealable plastic bag will be left in each participant’s cooler during 
June 7-11, the last weekly urine sample collection before starting the dietary 
intervention. 
 
For daily urine sample collections, a lab member will compile and set out the 
materials listed above. The Daily Urine Chain of Custody Form will be taken 
from the folder in the locked filing cabinet labeled with the participant’s ID 
number. The lab member will take all supplies to the urine collection. 
Daily Urine Sample Collections 
 
Each day, participants will be instructed to provide a first morning void in the 
specimen cup with the barcoded label for that day, which was provided to 
them at the prior day’s collection. The participants will be instructed to use 
their previously provided Sharpie to record the date and time of collection 
directly on the specimen cup. The sample should be provided the morning of 
the collection day (first morning void), and then placed in the sealable plastic 
bag and into the previously provided cooler with the ice pack. This cooler will 
then be placed outside the participant’s home. 
 
Two lab members will collect local daily urine samples as described in Section 
4 of this document. The two lab members will travel separately and divide the 
number of collections. 
 
One lab member will collect non-local daily urine samples as described in 
section 4 of this document, with the exception that non-local daily urine 
sample collections will occur daily during a two-day trip. For each two-day 
trip, a lab member will drive on the first day (this will occur on June 16, 18, 
20, 24, 26, and 28), collect the daily urine samples for that day, transport the 
daily urine samples in a lab cooler with a lab ice pack to the hotel, and store 
the samples in the study refrigerator at the hotel. The lab member will spend 
the night at the hotel, and then on the second day the lab member will collect 
the daily urine samples for that day (June 17, 19, 21, 25, 27, and 29). The lab 
member will then bring the urine samples for both days back to CAHL in a 

30 
 
lab cooler with a lab ice pack. Daily urine sample collected from the non-local 
area on June 22 and June 30 will be completed by one lab member in one day. 
 
The lab member will text the participant one day prior to each daily pick up 
as a reminder. After each sample collection, a text message will be sent to 
each participant. 
 
If a participant does not provide a urine sample for a given day, the lab 
member will document the missed day on the Daily Urine Chain of Custody 
Form. This will be considered a missed day and that day number will not be 
reused the following day; the next sequential day number will be used. 
 
Documentation upon Returning to the Lab 
 
After collecting samples, the lab member will return back to CAHL either 1) 
on the same day for local urine sample collections, or 2) on the second day of 
each two-day trip for non-local urine sample collections. There, the lab 
member will place the daily urine samples in the refrigerator and complete the 
appropriate sections of the Daily Urine Chain of Custody Form. The samples 
will remain in the refrigerator for no more than 24 hours until analyzed. 
Samples collected on the first day of each two-day trip must be prioritized for 
processing on the same day they return to the lab. 
 
Gift Cards 
 
A $50 gift card will be given to participants who provide all 14 daily urine 
samples. 
 
8. DAILY URINE SAMPLE ANALYSIS & STORAGE 
 
8.1 Summary of Methods 
 
Daily urine samples will be collected from participants and returned to CAHL 
in a lab cooler with a lab ice pack. Each day, one or more lab member(s) will 
be responsible for analyzing and storing all daily urine samples arriving in the 
lab that day. During analysis, the lab member will determine urine color, 
clarity, and specific gravity. Urine sample storage will occur immediately 
after urine sample analysis. Each sample will be aliquoted into two 4mL daily 

31 
 
cryovials for storage. These will be stored in the daily cryobox in the CAHL 
-80˚C freezer. In addition, 0.6 mL of each urine sample will be aliquoted into 
two 4 mL composite cryovials created for each participant for each week. 
These will be stored in the composite cryobox in the CAHL -80˚C freezer. At 
the end of the dietary intervention, each participant will have two 4 mL 
composite cryovials that contains 0.6 mL of each daily sample for each week, 
representing a weekly composite sample. At the conclusion of an individual’s 
participation in the study (ideally when she gives birth), one of the two 4 mL 
composite cryovials from each week will be sent to the CDC for glyphosate 
analysis (up to 2 total composite cryovials per participant: one for week one 
and one for week two). The remaining composite cryovials, as well as the two 
daily 4 mL cryovials, will be stored in the biorepository. 
 
8. 2 Materials 
Daily Urine Sample Analysis 
 
• 
Absorbent pads 
 
• 
Kimwipes 
 
• 
Cup containing ethyl alcohol 
 
• 
Squirt bottle containing DI water 
 
• 
Nitrile gloves 
 
• 
Pen 
 
• 
Urine Sample Characteristics Log* 
 
• 
Color and Clarity Guide* 
 
• 
Transfer pipettes 
 
• 
Atago Urine Specific Gravity Refractometer, PAL 10-S 
 
• 
K01 Database 

32 
 
Daily Urine Sample Storage 
 
• 
Absorbent pads 
 
• 
Nitrile gloves 
 
• 
Eppendorf Plus Research Pipette and tip 
• 
Four cryovials with barcoded labels (two daily cryovialsand 
two composite cryovials) 
 
• 
Daily Cryobox and Composite Cryobox 
 
• 
Google Drive Cryobox Document 
 
8. 3 Procedures 
 
Daily Urine Sample Analysis 
Lab members will analyze the daily urine samples as described in Section 
5.3 of this document. 
 
Daily Urine Sample Storage 
 
Supplies Preparation 
 
Urine sample storage should occur immediately after analysis. Prior to 
beginning storage procedures, the lab member will set the following supplies 
out on a lab station on an absorbent pad (this can be the same absorbent pad 
used during analysis): 1) Eppendorf Plus Research Pipette (set at 4.0 mL and 
then changed to 0.6 mL) and tip; 2) two 4 mL daily cryovials with the 
appropriate barcoded labels and two 4 mL composite cryovials with the 
appropriate barcoded labels; 3) one cryobox at the lab station; and 4) the 
appropriate urine specimen cup. A pair of nitrile gloves will be worn when 
handling urine samples. 

33 
 
Daily Urine Sample Storage (Daily Cryovials) 
 
Four mL of the urine sample will be pipetted into two separate 4 mL daily 
cryovials using the Eppendorf Plus Research pipette and tip. Once both of the 
daily cryovials have been filled, the lab member will remove the daily cryobox 
from the -80˚C freezer. This daily cryobox will be placed at the lab station, 
and the newly filled daily cryovials will then be transferred from the cryobox 
at the lab station and into the freezer daily cryobox. The daily cryobox in the 
freezer will only be removed from the freezer briefly for daily cryovial 
transfer, and it will be immediately returned to the freezer as quickly as 
possible. 
 
Daily Urine Sample Storage (Composite Cryovials) 
 
For the first daily urine sample collection of each week, 0.6 mL of the urine 
sample will be pipetted into two separate 4 mL composite cryovials using the 
Eppendorf Plus Research pipette and tip. Once both of the composite 
cryovials have been filled, the lab member will transfer the composite 
cryovials from the cryobox at the lab station into the freezer composite 
cryobox. The composite cryobox will be immediately placed in the freezer as 
quickly as possible. 
 
Each day when the next daily urine sample is collected from a participant, 
another 0.6 mL from that sample will be aliquoted into the two separate 4 mL 
composite cryovials, on top of the previously frozen sample. This process will 
be repeated until the composite cryovials contains 0.6 mL from each of the 
seven days, representing a weekly composite cryovial. The composite 
cryobox in the freezer will only be removed from the freezer briefly for 
aliquoting, and it will be immediately returned to the freezer as quickly as 
possible. 
 
Documentation 
 
The lab member will record the specific cryovial label and cell number 
where each of the two daily cryovials were placed in the freezer daily 
cryobox. This will be recorded in the CAHL shared Google Drive Cryobox 
Document. The lab member will also record the specific cryovial label and 
cell number where each of the two composite cryovials were placed in the 

34 
 
freezer composite cryobox. This will also be recorded in the CAHL shared 
Google Drive Cryobox Document. 
 
9. URINE SAMPLE SHIPMENT & GLYPHOSATE ANALYSIS 
 
9.1 Summary of Methods 
 
Once all weekly and daily urine samples have been collected, analyzed, and 
stored for all 40 participants, two lab members will complete a quality 
assurance (QA) check of each cryovial to ensure that it is labeled with the 
correct barcoded label and that it is placed in the correct cryobox and correct 
cell, and that this information correlates with the information in the K01 
Database, as well as with the Google Drive Cryobox Documents. The QA will 
be done thoroughly, but quickly, to prevent the thawing of the cryovials. 
Following the QA, the first of the four cryovials from every other week and 
the first composite cryovials from each week in the dietary intervention, along 
with a number of quality control vials and randomly selected cryovials, will 
be sent to the CDC for glyphosate analysis. These samples will be packaged 
by three lab members and then sent on dry ice to the CDC via Overnight Air 
Priority Shipping with tracking. 
 
9.2 Materials 
 
Quality Assurance 
 
• 
Google Drive Cryobox Document 
 
• 
K01 Database 
 
• 
Nitrile gloves 
 
• 
Frozen cryoboxes 
 
Sample Shipment 
 
• 
Twelve cryovial carrier boxes (labeled as box 1-12) 

35 
 
• 
Twenty-four small absorbent pads 
 
• 
Twenty-four rubber bands 
 
• 
Twelve biohazard bags 
 
• 
Shipping cooler 
 
• 
Shipping box 
 
• 
Shipping stickers 
 
• 
Shipping tape 
 
• 
Nitrile gloves 
 
• 
All weekly, composite, and daily cryoboxes in the -80˚C freezer 
 
• 
Four absorbent pads 
 
• 
Five ice packs 
 
• 
Five pounds of dry ice 
 
9.3 Procedures 
 
Quality Assurance 
 
Weekly Urine Samples Electronic Documentation Check 
 
The electronic database documents containing information on the stored 
weekly urine sample collections and cryovials will be exported from their 
original source. All changes to any of these documents will be done on the 
exported copies and will be tracked. No changes will be made the original 
data sources. 
 
The lab member will cross-check the exported forms with the exported aliquot 
boxes document. The lab member will highlight any information on the 

36 
 
exported aliquot boxes document that does not match or that seems 
implausible. The lab member will briefly describe the issue in the margins of 
the exported aliquot boxes document. The lab member will refer to the Urine 
Chain of Custody Form as needed. 
 
Weekly Urine Sample Cryovials Check 
 
After completion of the Electronic Documentation Check, two lab members 
will cross-check the now highlighted exported aliquot boxes document with 
the frozen cryovials in order to 1) confirm that each cryovial is in the correct 
cell; 2) confirm that each cryovial is labeled correctly; and 3) clarify any typos 
or resolve other issues in the exported aliquot boxes document. 
 
Documentation and Correction 
 
After all the weekly cryoboxes have been checked and returned to the -80˚C 
freezer, the lab members will review all the highlighting and notes in the 
exported aliquot boxes document. Typos will be corrected as appropriate. 
 
Quality Assurance For Daily and Composite Urine Samples 
 
QA for the daily and composite urine samples will follow the same procedures 
as QA for the weekly urine samples as described above. 
 
Sample Shipment 
 
Sample Selection 
 
Dr. Curl will select 600 cryovials to send to the CDC. This selection will 
include the first of the four cryovials from every other weekly urine sample 
collected from each participant throughout the study. In addition, based on the 
weekly urine samples being sent to the CDC, Dr. Curl will randomly select 
participant IDs and week numbers to serve as weekly urine sample quality 
controls. For these urine samples the second of the four cryovials will also be 
sent to the CDC. Dr. Curl will also send the first of the two composite 
cryovials from each week of the dietary intervention from each participant to 
the CDC, as well as all 14 daily cryovials from one participant fromeach day 
of the dietary intervention to serve as composite sample qualitycontrol. 

37 
 
Supplies Preparation 
 
Prior to the shipment of urine samples to the CDC, three lab members will set 
up a shipping station in the lab with the following supplies: 1) twelve cryovial 
carrier boxes; 2) twenty-four small absorbent pads; 3) twenty-four rubber 
bands; 4) twelve biohazard bags; 5) shipping cooler; 6) shipping box; 7) 
shipping tape; 8) shipping stickers; and 9) four absorbent pads. The lab 
members will also have five ice packs in the freezer and five pounds of dry 
ice in the -80˚C freezer, and will have the Aliquot Boxes Spreadsheet 
available electronically. 
 
Packaging and Shipping 
 
Packaging will begin with weekly cryovials, then composite cryovials, and 
then daily cryovials. Cryoboxes will be pulled from the -80˚C freezer one at 
a time and the selected cryovials from that cryobox will be transferred to a 
cryovial carrier box. 
 
The lab members will place the five ice packs from the freezer at the bottom 
of the shipping cooler and place two absorbent pads on top. The lab members 
will then remove all twelve cryovial carrier boxes in biohazard bags fromthe 
-80˚C freezer and place them into the shipping cooler along with five pounds 
of dry ice. Finally, two more absorbent pads will be placed on top and the lid 
to the shipping cooler will be taped closed with one single strip of shipping 
tape running horizontally across the lid. The shipping cooler will be placed 
into the shipping box and the shipping box will be taped closed with one single 
strip of shipping tape running horizontally across the box. No more tape 
should be used on the shipping cooler or shipping box as the dry ice in the 
shipment must not be completely sealed off from oxygen. 
 
Appropriate shipping stickers will be placed on the shipping box, and the 
box will be transported for Overnight Air Priority Shipping with tracking 
with early morning arrival. The lab members will email the CDC to provide 
the tracking information, and will also request that the CDC confirm receipt 
of the samples and that all samples arrived frozen once the shipment arrives.