Department of Transportation (DOT) and Department of Health and Human Services (DHHS) Urine Specimen Collection Training

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Authors: Garland E. Pendergraph, PhD, JD, MLS(ASCP)SM, HCLD/CC(ABB) and Ramona Holt, MT(ASCP)

Department of Transportation (DOT) regulated urine specimen collections are an important part of employment and workplace screenings. As a collector, you'll perform DOT urine specimen collections for drug and alcohol testing, return-to-duty testing, and more.

In order to perform DOT-regulated urine specimen collections, you'll need to complete qualification training. MediaLab's online course in Department of Transportation (DOT) Regulated Urine Specimen Collection satisfies the requirements for qualification training as outlined in Federal Regulations, 49 CFR Part 40.

Important: This course requires you to find a qualified observer to observe five required mock collections. This course is normally used by hospitals, laboratories, and physician offices that already have a collector on staff who can be an observer. If you are a private individual, this course may not be appropriate for you. MediaLab & LabCE do not provide observers or maintain a list of observers for you to contact.

What does this course include?

This course includes three components:
  1. Online reading material covering DOT rules and procedures (about 1 - 2 hours to complete)
  2. A short exam covering the reading material (about 10 - 15 minutes to complete)
  3. A checklist to record completion of five mock collections (time varies)

How to become qualified to perform DOT urine collections

There are two steps to becoming qualified for DOT urine collection:
  1. Complete a qualification training course, like this online course from MediaLab.
  2. Complete five mock collections under various scenarios in the presence of a qualified observer.
MediaLab's online DOT urine collection training course includes an integrated checklist feature to document completion of these required mock collections, but MediaLab does not provide observers for the mock collections. You will need to locate a qualified collector locally who can attest that you have completed the mock collections.

Who can observe the mock collections for DOT qualification training?

A qualified collector must perform the observations. A qualified collector must either (a) regularly perform DOT-regulated urine specimen collections, (b) regularly observe collections for DOT training requirement, or (c) have completed a "train the trainer" course. Observations must be conducted in person by a qualified collector and cannot be performed online or remotely.

How do I find an observer?

This course is normally used by hospitals, laboratories, and physician offices that already have a collector on staff who can be an observer. If you are a private individual without access to an observer, this course may not be appropriate for you.

How does an observer report completion of the mock collections to MediaLab?

An observer will complete your online checklist and provide his / her contact information. The completed checklist, including the observer's contact information, becomes part of your certification documentation. MediaLab does not validate or audit completed observations. An individual or company that wants to validate your credentials may need to contact your observer.

Am I DOT-certified after completing this course?

Once you have completed the qualification training from MediaLab and documented that you have completed the five required mock collections, then you are qualified to perform DOT urine specimen collections. You will receive a certificate from MediaLab attesting that you have completed qualification training, and you will be able to print out documentation that you have completed the mock collections. The DOT does not issue certificates, licenses, or collector numbers, so you will not receive any materials from the DOT itself.

How long is the certification good for?

After completing the qualification training and five mock collections, you must complete refreshing training within five years. You are responsible for keeping up with changes to DOT regulations and making sure that you comply with any changes.

Is this course acceptable for refresher training under DOT Rule 49 CFR Part 40 Section 40.33 Subpart C?

Yes. You must complete both the qualification training (reading, video, exam) and complete the five mock collections in order to satisfy the requirements for refresher training.

Continuing Education Credits


  • Describe the three types of urine specimen collections.
  • Identify the different types of custody and control forms and explain how to complete them properly.
  • Describe the correct sequence of a proper drug screen collection.
  • Identify and explain how to handle unusual collection situations.

Course Outline

  • Introduction
      • Intent of This Program
      • Mock Collections
      • Requirements for Retraining
  • Collections
      • Types of Collections
      • Standard Collection
      • Monitored Collection
      • Monitored Collection, continued
      • Directly Observed Collection
      • Prosthetic Devices
      • Directly Observed Collection, continued
      • Directly Observed Collection, continued
      • In BOTH the monitored and observed collections, the collector MUST be the same gender as the donor.
  • Custody and Control Forms
      • Federal Drug Testing Custody and Control Form (CCF)
      • Electronic Federal Drug Testing Custody and Control Form (eCCF)
      • Non-Federally Regulated Custody and Control Form
      • Custody and Control Form
      • Exceptions
      • When a Federally Regulated Custody and Control form is not available, in all situations it is perfec
  • Requirements for the Collection Site
      • Collector Responsibilities
      • Prerequisites for Proper Collection
      • Collection Site Security Requirements
      • Other Collection Site Requirements
      • Collection Supplies
  • Complete and Accurate Documentation
      • Completing the Federal Drug Testing Custody and Control Form (CCF)
      • Completing Step 1 of the Federal Drug Testing CCF
  • Donor Identification
      • Acceptable Forms of Identification
      • Self-Employed Donor Without Proper Identification
      • Unacceptable Forms of Identification
      • Positive Identification Must Be Obtained Before Collection
  • Steps for Typical Urine Collection
      • Steps for Typical Urine Collection
      • Donor Preparation
      • Hand Washing
      • Collection Kit
      • Donor Provides the Specimen
      • Donor Gives Specimen to Collector
      • Specimen Integrity
      • Completing Step 2 of the Federal Drug Testing CCF: Recording Urine Specimen Temperature
      • Pouring the Collected Specimen into the Specimen Bottle(s)
      • Collector Applies Tamper-Evident Seals
      • Collector Dates Seals; Donor Initials Seals
      • Completing Step 5 (copy 2) of the Federal Drug Testing CCF: Donor Certification Statement
      • Completing Step 4 of the Federal Drug Testing CCF: Chain of Custody
      • Collector Packages Specimen
      • Distribution of Custody and Control Form (CCF)
      • If there are several donors waiting to have a drug screen and two or more restrooms are available at
  • Special Collection Situations
      • When Problems Occur
      • Indwelling Catheter and External Urine Bag
      • Medications
      • Refusal to Test
      • Refusal to Test, continued
      • Situations Not Considered as Refusal
      • Donor Refuses to Complete Paperwork
      • Shy Bladder Procedures
      • Shy Bladder Procedures, continued
      • Shy Bladder Procedures, continued
      • Observed Collection
      • Observed Collection Procedure
      • Observed Collection Procedure, continued
      • Observed Collection Scenario #1
      • Observed Collection Scenario #2
      • Observed Collection Scenario #3
      • Observed Collection Scenario #4
      • Observed Collection Scenario #5
      • Broken Security Seal
  • Fatal Flaws
      • Accidental Spills
      • Fatal Flaws and Correctable Flaws
  • Some Final Thoughts
      • Responsibilities and requirements for collectors
      • Differences between Department of Transportation (DOT) and Department of Health and Human Services (HHS) Mandatory Guidelines for Federal Drug Testing...
      • References

Additional Information

Level of Instruction: Basic.
Intended Audience: Personnel who will be conducting DOT urine drug screen collections.
Author Information:
Garland E. Pendergraph, PhD, JD, MLS(ASCP)SM, HCLD/CC(ABB) is a retired clinical laboratory director but presently serves part-time as the director for three laboratories. He is currently licensed as a laboratory director in the State of Georgia. Dr. Pendergraph received his MSPH from the University of Kentucky in Lexington, his PhD in medical parasitology/entomology and mycology from the University of North Carolina in Chapel Hill and his law degree with a concentration in health care law from Concord Law School, Purdue University. He also did a Fellowship in Tropical Medicine at Louisiana State University School of Medicine and received a Certificate in Health Care Risk Management from Florida State University. He is the author of a textbook in phlebotomy, a number of scientific articles, plus internet training programs. He has over 25 years experience in urine and saliva drug screen collections and breath alcohol testing.
Ramona Holt, MT(ASCP) is Supervisor for Patient Service Centers, Quest Diagnostics in the Albany-Valdosta, Georgia area. Previously, she was employed as a laboratory manager by Doctors Laboratory, Inc. and Solstas Lab Partners. She received her baccalaureate degree in medical technology from the Medical College of Georgia, in Augusta, Georgia. She has 23 years experience in urine drug screen collections.
Course Description: The course follows the "Urine Specimen Collection Guidelines" for the U.S. Department of Transportation Workplace Drug Testing Programs (49 CFR Part 40). 

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A urine filled sample cup
Donor initials the tamper-evident seal
White CCF and samples are sealed in a bag
Prosthetic device
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