Split specimen procedures
A split specimen is a mandatory step in any DOT collection under 49 CFR Part 40. It is the result of dividing one sample into two testing specimens for the lab. This back-up sample is used in the event the primary specimen is unsuitable for testing or the test result comes back as a positive, substituted, or adulterated test result and the driver wishes to contest the results.
Collection process
A collector automatically knows that a DOT collection requires a split specimen. The collector communicates this to the lab by indicating on Step 2 of the Federal Drug Testing Custody and Control Form (CCF) that it is a split specimen collection.
For urine specimens:
- The collector, not the driver, must pour at least 30 mL of urine from the collection container into a specimen bottle, to be used for the primary specimen. The collector then pours at least 15 mL of urine from the collection container into the second specimen bottle to be used as the split specimen.
- Both bottles are secured with lids/caps by the collector, and tamper-evident bottle seals are placed over the bottle caps/lids and down the sides of the bottle. The collector then writes the date on the tamper-evident bottle seals. The driver initials the tamper-evident bottle seals for the purpose of certifying that the bottles contain the specimen that the driver provided. Any excess urine must be discarded, unless used in conjunction with a DOT physical examination. Other than the DOT medical exam, no further testing must be done on the excess urine.
Both specimen bottles are placed in a sealed plastic bag in a shipping container (e.g., courier box) and sent to the lab.
For oral-fluid specimens, the collector follows the manufacturer’s instructions to package the split specimen collections:
- A volume of at least 1 mL of undiluted (neat) oral fluid is collected for the specimen designated as “Bottle A,” and a volume of at least 1 mL of undiluted (neat) oral fluid is collected for the specimen designated as “Bottle B,” or an otherwise sufficient amount of oral fluid is collected to permit an HHS-certified laboratory to analyze the specimen(s).
- In the presence of the employee, the collector places a tamper-evident seal from the CCF over the cap of each specimen container, taking care not to obstruct the expiration date on the collection containers. The collector must record the date of the collection on the tamper-evident seals, after they are affixed to the specimen containers.
- The collector instructs the employee to initial the tamper-evident seals on each specimen container. If the employee declines to do so, the collector must note this in the “Remarks” line of the CCF (Step 2) and complete the collection process.
Both specimen bottles are placed in a sealed plastic bag in a shipping container (e.g., courier box) and sent to the lab.
Protocols at the lab
Incoming specimens are processed by the lab using the protocols outlined in 40.83.
The lab must inspect each specimen and CCF for the following “fatal flaws:”
- There is no CCF;
- In cases where a specimen has been collected, there is no specimen submitted with the CCF;
- There is no printed collector’s name and no collector’s signature;
- Two separate collections are performed using one CCF;
- The specimen ID numbers on the specimen bottle and the CCF do not match;
- The specimen bottle seal is broken or shows evidence of tampering, unless a split specimen can be redesignated;
- There is an insufficient amount of specimen in the primary bottle for analysis, unless the specimens can be redesignated;
- For an oral-fluid collection:
- The collector used an expired device at the time of collection.
- If the collector failed to enter the expiration date in Step 4 of the CCF and the laboratory is unable to determine the expiration date by inspecting Bottles A and B.
When the lab finds a specimen meeting the fatal flaw criteria, it must document the findings and stop the testing process. The result is reported in accordance with 40.97(a)(3).
If the primary specimen cannot be tested because of leakage or a broken seal, the split specimen may be redesignated as the primary specimen to be tested. This is only a possibility if the lab believes a sufficient amount of specimen exists in the split specimen to conduct all the appropriate primary laboratory testing.
If the split specimen was inadvertently labeled as the primary specimen, the laboratory may redesignate the bottles as they should be. In the event the lab accidentally opens the split specimen instead of the primary specimen and the primary specimen’s seal remains in tact, the lab may continue processing the split specimen as the primary providing there is enough of a sample to conduct all the appropriate primary laboratory testing. The original primary specimen is then redesignated as the split specimen.
In any of the above cases, the lab would redesignate the two samples by crossing out A (primary) and marking B (split specimen) and vice versa. The laboratory employee will initial and date the changes on the bottles. The laboratory employee must also make a notation on Copy 1 of the CCF, Step 5a, and on any internal chain of custody documents.
If the split specimen is found to be unavailable for testing or appears to be insufficient, the lab must still process the primary specimen without providing the Medical Review Officer (MRO) details regarding the unavailable split specimen. In the event the MRO instructs the lab to forward the split specimen in these circumstances, the lab would then provide the MRO with the details for the unavailability. How the MRO handles a request for a split specimen analysis in this situation is covered in the MRO results subheading below.
Notification of due process
A Medical Review Officer (MRO) will contact a driver in the event of a positive, adulterated, or substituted result, informing the driver of the right to test the split specimen to prove an error in testing. The driver has 72 hours after being told of this right to request the split specimen.
The MRO will make all attempts to contact the driver so that the option is made available. The MRO will make three reasonable efforts within 24 hours (documenting them).
If the driver cannot be contacted, the MRO will contact the designated employer representative (DER) to get in touch with the driver on the MRO’s behalf. The DER is not told the reason why the MRO needs to speak with the driver personally. If the DER’s documented attempts (three reasonable efforts in another 24-hour period) do not result in a contact, the MRO reports the results as positive.
If the driver does not contact the MRO within 72 hours of being told the verified result, the primary specimen’s test result remains official. There MRO may review an explanation why the driver did not get back to the MRO within the 72 hours. This could include a serious illness, injury, or inability to contact the MRO, lack of actual notice of a verified positive test, or other circumstances unavoidably preventing the driver from contacting the MRO. If the MRO feels the explanation is legitimate, the MRO will direct the analysis of the split specimen be performed. If the there is no legitimate explanation why the driver did not contact the MRO within the 72 hours, then a split specimen does not have to be tested.
If the driver requests an analysis of the split specimen within 72 hours of having been informed of a verified positive result, the MRO directs the laboratory, in writing, to ship the split specimen to another DHHS-certified laboratory for analysis. The specimen must be sent regardless of payment for the testing. The employer may recoup the cost, but cannot refuse the employee of the right to test the split specimen based on payment. Even if the employer never gets reimbursed, the test must be done.
The reporting of the verified positive result is not delayed pending the split-specimen analysis. The employer would still be forced to remove the driver from the safety-sensitive function in accordance with 382.501 once notified of the verified positive on the primary specimen.
Testing of the split specimen
The laboratory that originally received both the primary and split specimens must forward the following items to the second laboratory:
- The split specimen in its original specimen bottle, with the seal intact;
- A copy of the MRO’s written request; and
- A copy of Copy 1 of the CCF, which identifies the drug(s)/ metabolite(s) or the validity criteria to be tested for.
The lab must not send to the second laboratory any information about the identity of the employee. Inadvertent disclosure does not, however, cause a fatal flaw. The regulations do not prescribe who gets to decide which HHS-certified laboratory is used to test the split specimen. That decision is left to the parties involved.
Based on the primary specimen result, the split specimen is processed in the following way:
- Drug(s) and drug metabolite(s) — The lab testing the split specimen must test for the drug(s)/drug metabolite(s) detected in the primary specimen. This split specimen, however, is not tested within the cutoff concentration in 40.85 and 40.91, as applicable. If the test does not reconfirm the presence of the drug(s)/drug metabolite(s) that were reported in the primary specimen, the lab must conduct validity tests. This will aid in determining if there is a reason why the retest failed to confirm the presence of the drug(s) found originally in the primary specimen. The lab is required to follow the validity testing procedures set forth in 40.87 or 40.93 . If the test or the validity testing fails to confirm the primary specimen results, the lab has the option of sending out the specimen or an aliquot of it to yet another HHS-certified laboratory for another reconfirmation test.
- Adulteration — Laboratories testing split specimens for an adulterant must use the same criteria as the primary specimen in accordance with 40.89 or 40.93, as applicable. The result of the primary specimen is reconfirmed if the split specimen meets these criteria. If the test fails to reconfirm the adulterant result reported in the primary specimen, the lab may send the specimen or an aliquot of it for testing at another HHS-certified laboratory that has the capability to conduct another reconfirmation test.
- Substitution — Laboratories testing split specimens for substitution must use the same criteria as the primary specimen in accordance with 40.88. The result of the primary specimen is reconfirmed if the split specimen meets these criteria.
Lab results
A laboratory responsible for testing the split specimen must report split specimen test results by checking the “Reconfirmed” box or the “Failed to Reconfirm” box (Step 5(b)) on Copy 1 of the CCF and by providing clarifying remarks using current HHS Mandatory Guidelines requirements. The laboratory-certifying scientist signs and dates the CCF.
The lab must report the split specimen results directly to the MRO at the MRO’s place of business. The results will not be reported through a DER or another service agent. The lab must fax, courier, mail, or electronically transmit a legible image or copy of the fully-completed Copy 1 of the CCF, which has been signed by the certifying scientist. It must transmit the laboratory result to the MRO immediately, preferably on the same day or next business day as the result is signed and released.
MRO results
The MRO will receive split specimen results that fall into five categories. The MRO must take the following action, as appropriate, when a laboratory reports split specimen results.
Category 1
The laboratory reconfirmed one or more of the primary specimen results. The MRO must report to DER and the employee the result(s) that was/were reconfirmed.
- In the case of a reconfirmed positive test(s) for drug(s) or drug metabolite(s), the positive is the final result.
- In the case of a reconfirmed adulterated or substituted result, the refusal to test is the final result.
- In the case of a combination positive and refusal to test results, the final result is both positive and refusal to test.
Category 2
The laboratory failed to reconfirm all of the primary specimen results because, as appropriate:
- Drug(s)/drug metabolite(s) were not detected;
- Adulteration criteria were not met; and/or
- Substitution criteria were not met.
The MRO must report to the DER and the employee that the test must be canceled. The MRO must inform the DOT of the failure to reconfirm using the format in Appendix F to Part 40.
In a case where the split failed to reconfirm because the substitution criteria were not met and the split specimen creatinine concentration was equal to or greater than 2mg/dL but less than or equal to 5mg/dL, the MRO must, in addition to informing the DOT of the failure to reconfirm, direct the DER to ensure the immediate collection of another specimen from the employee under direct observation, with no notice given to the employee of this collection requirement until immediately before the collection.
In a case where the split failed to reconfirm and the primary specimen’s result was also invalid, the MRO will direct the DER to ensure the immediate collection of another specimen from the employee under direct observation, with no notice given to the employee of this collection requirement until immediately before the collection.
Category 3
The laboratory failed to reconfirm all of the primary specimen results, and also reported that the split specimen was invalid, adulterated, and/or substituted.
In the case where the laboratory failed to reconfirm all of the primary specimen results and the split was reported as invalid, the MRO must:
- Report to the DER and the employee that the test must be canceled and the reason for the cancellation.
- Direct the DER to ensure the immediate collection of another specimen from the employee under direct observation, with no notice given to the employee of this collection requirement until immediately before the collection.
- Inform the DOT of the failure to reconfirm using the format in Appendix F to Part 40.
In the case where the laboratory failed to reconfirm any of the primary specimen results, and the split was reported as adulterated and/or substituted, the MRO must:
- Contact the employee and inform the employee that the laboratory has determined that the split specimen is adulterated and/or substituted, as appropriate.
- Follow the procedures of 40.145 to determine if there is a legitimate medical explanation for the laboratory finding of adulteration and/or substitution, as appropriate.
If the MRO determines that there is a legitimate medical explanation for the adulterated and/or substituted test result, the MRO reports to the DER and the employee that the test must be canceled and informs ODAPC of the failure to reconfirm using the format in Appendix F to Part 40.
If the MRO determines that there is not a legitimate medical explanation for the adulterated and/or substituted test result, the MRO must take the following steps:
- Report the test to the DER and the employee as a verified refusal to test. Inform the employee that the employee has 72 hours to request a test of the primary specimen to determine if the adulterant found in the split specimen is also present in the primary specimen and/or to determine if the primary specimen meets appropriate substitution criteria.
- Except when the request is for a test of the primary specimen and is being made to the laboratory that tested the primary specimen, follow the procedures of 40.153, 40.171, 40.173, 40.179, 40.181, and 40.185, as appropriate.
The laboratory that tests the primary specimen to reconfirm the presence of the adulterant found in the split specimen and/or to determine that the primary specimen meets appropriate substitution criteria, it reports the result to the MRO on a photocopy (faxed, mailed, scanned, couriered) of Copy 1 of the CCF.
If the test of the primary specimen reconfirms the adulteration and/or substitution finding of the split specimen, the MRO must report the result as a refusal to test.
If the test of the primary specimen fails to reconfirm the adulteration and/or substitution finding of the split specimen, the MRO must cancel the test.
Category 4
The laboratory failed to reconfirm one or more — but not all of the primary specimen results — and also reported that the split specimen was invalid, adulterated, and/or substituted. The MRO, in the case where the laboratory reconfirmed one or more of the primary specimen result(s), must follow procedures in paragraph 40.187(a) (Category A) and:
- Report that the split was also reported as being invalid, adulterated, and/or substituted (as appropriate).
- Inform the DER to take action only on the reconfirmed result(s).
Category 5
The split specimen was not available for testing or there was no laboratory available to test the specimen. The MRO must:
- Report to the DER and the employee that the test must be canceled and the reason for the cancellation;
- Direct the DER to ensure the immediate recollection of another specimen from the employee under direct observation, with no notice given to the employee of this collection requirement until immediately before the collection; and
- Notify the DOT of the failure to reconfirm using the format in Appendix F to Part 40.
MRO Recordkeeping
For all split specimen results, the MRO must: