DOT Urine Specimen Collection Procedures


Chain of custody” is the method of tracing each urine specimen to maintain control from initial collection to final disposition. To prevent mistakes when collecting urine samples, it is vital to follow these steps.


  1. The collector verifies the identity of the employee by checking his/her photo identification;
  2. The collector instructs the employee to remove any outer garments (e.g., jacket, coat, hat) and leave their personal belongings such as purses or briefcases in a locked box;
  3. The collector instructs the employee to empty their pockets and display the contents;
  4. The employee is given the option to keep their wallet after being inspected;
  5. The collector explains the basic collection procedure/shows donor instructions on the back of the CCF;
  6. The collector completes Step 1 of the CCF (parts of this may be pre-printed);
  7. The collector instructs the donor to wash and dry their hands;
  8. The collector unwraps a sealed collection container in front of the donor;
  9. The donor is provided with a single-use plastic container from the collection kit;
  10. The donor remains in the presence of the collector until entering the privacy enclosure to provide the specimen;
  11. The collector ensures that privacy for urination is secure and a bluing agent is in the toilet.
  12. The donor then provides a specimen and brings the specimen to the collector—
  13. The collector observes that the specimen quantity is at least 45mL and checks the split specimen box in Step 2;
  14. The collector determines the temperature reading and checks off the temperature box in Step 2 within 4 minutes of the collection taking place;
  15. The collector pours at least 30 mL of urine into the primary specimen bottle and at least 15 mL in the second specimen bottle;
  16. The collector securely places tamper-evident bottle seals over the bottle caps/lids;
  17. The collector dates specimen seal bottles only after they are affixed to the specimen bottles;
  18. The donor initials specimen seal bottles only after they are affixed to the specimen bottles;
  19. The collector folds away Copy 1 of the CCF to reveal Copy 2 (the MRO Copy), with Copies 3, 4, and 5 underneath as well) and instructs the donor to read and complete Step 5 of the CCF
  20. The collector reviews Step 5 for completion and accuracy;
  21. Collector returns to Copy 1 and completes Step 4, (prints and signs his/her name, notes the time and date of the collection and the delivery system transporting the specimen to the lab);
  22. The collector puts the specimen bottles and Copy 1 of CCF in a leak-proof plastic bag and seals the bag;
  23. The collector distributes Copy 5 to the employee and retains the collector copy (Copy 3);
  24. The collector faxes or otherwise transmits Copy 2 to the MRO and Copy 4 to the DER;
  25. The collector puts the sealed specimen bag in the secure designated area to be picked up by a courier.

Follow the steps in order, no shortcuts.

This means that the DOT compliant order of the collection is Step 1, then the actual collection of the specimen, followed by Steps 2, 3, 5, and finally Step 4 (making the required order 1, 2, 3, 5, THEN 4).

Collector training is required before performing DOT urine specimen collections. View our training courses to see current specimen collection training available online.

DOT Urine Specimen Collection Procedures - Joe Reilly & Associates Inc

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