The DOT 5-panel drug test is defined by the Department of Transportation in 49 CFR Part 40 — the federal regulation that governs all workplace drug and alcohol testing for safety-sensitive transportation employees. The test is performed on a urine specimen at an HHS-certified laboratory, and the results are reviewed by a Medical Review Officer (MRO) before being reported to the employer.
The 5 Drug Panels
The following five drug classes are required on every DOT-mandated urine drug test, regardless of which DOT agency (FMCSA, USCG, FTA, FAA, PHMSA) oversees your program.
Marijuana
THC
What it detects
Delta-9-tetrahydrocannabinol (THC)
Analyte(s) tested
THC-COOH (11-nor-delta-9-THC-9-carboxylic acid)
Initial cutoff
50 ng/mL
Confirmatory cutoff
15 ng/mL
Positive even if marijuana is legal in your state. DOT testing is federal — state law does not apply.
Cocaine
COC
What it detects
Cocaine and its metabolite benzoylecgonine (BE)
Analyte(s) tested
Benzoylecgonine
Initial cutoff
150 ng/mL
Confirmatory cutoff
100 ng/mL
Cocaine metabolites remain detectable in urine for 2–4 days after use in most individuals.
Opioids
OPI
What it detects
Six opioid analytes across three sub-groups
Analyte(s) tested
Codeine · Morphine · 6-AM · Hydrocodone · Hydromorphone · Oxycodone · Oxymorphone
Initial cutoff
Varies by analyte (see table below)
Confirmatory cutoff
Varies by analyte (see table below)
The opioid panel was significantly expanded in 2017 to include semi-synthetic opioids like oxycodone and hydrocodone — the most-abused prescription drugs in the country.
Amphetamines
AMP
What it detects
Amphetamine, Methamphetamine, MDMA, and MDA
Analyte(s) tested
Amphetamine · Methamphetamine · MDMA · MDA (methylenedioxyamphetamine)
Initial cutoff
500 ng/mL (all analytes)
Confirmatory cutoff
250 ng/mL (all analytes)
A legitimate Adderall prescription can be reported as a negative result by the MRO if the prescription is valid — but the employee must disclose it to the MRO, not to their employer.
Phencyclidine
PCP
What it detects
Phencyclidine (PCP)
Analyte(s) tested
Phencyclidine (parent compound)
Initial cutoff
25 ng/mL
Confirmatory cutoff
25 ng/mL
PCP has no accepted medical use and is a Schedule II controlled substance. It is detectable in urine for up to 8 days in occasional users.
Opioid Panel — Analyte-by-Analyte Cutoffs
Because the opioid panel was expanded in 2017, it has multiple sub-analytes with different cutoff concentrations. These are the exact values from 49 CFR §40.87.
| Analyte | Initial Cutoff | Confirmatory Cutoff |
|---|---|---|
| Codeine | 2,000 ng/mL | 2,000 ng/mL |
| Morphine | 2,000 ng/mL | 2,000 ng/mL |
| 6-Acetylmorphine (6-AM / heroin marker) | 10 ng/mL | 10 ng/mL |
| Hydrocodone | 300 ng/mL | 100 ng/mL |
| Hydromorphone | 300 ng/mL | 100 ng/mL |
| Oxycodone | 300 ng/mL | 100 ng/mL |
| Oxymorphone | 300 ng/mL | 100 ng/mL |
Source: 49 CFR Part 40, §40.87 — U.S. Department of Transportation
How the Test Works: From Collection to Result
DOT testing follows a strict chain-of-custody process defined in Part 40. Here's each stage:
Urine specimen collected
A collector at a DOT-approved collection site supervises the process and seals the specimen following 49 CFR Part 40 chain-of-custody procedures.
Lab immunoassay screen
A federally certified HHS lab (SAMHSA-certified) runs an initial immunoassay screen. Any result at or above the initial cutoff is sent for confirmation.
GC/MS or LC/MS/MS confirmation
A different technique — gas chromatography/mass spectrometry or liquid chromatography tandem mass spectrometry — confirms the specific drug and quantity against the confirmatory cutoff.
Medical Review Officer (MRO) review
A licensed physician trained in substance abuse reviews the confirmed result, contacts the donor to check for a legitimate medical explanation, and issues the final verified result.
Result reported to employer
The MRO reports the final result — Negative, Positive, Refusal, Cancelled, or Test Not Performed — to the employer or C/TPA. Only the final verified result is shared.
Key rules employers and employees must know
- All DOT-mandated tests must use a federally certified (HHS/SAMHSA) laboratory — no exceptions.
- The specimen is split into two: a primary specimen (A bottle) and a split specimen (B bottle). Employees may request the B bottle be tested at a second certified lab. APCA does not cover the cost of a split specimen retest — this cost is passed on to the employer or employee.
- An MRO reviews every confirmed positive before the result is transmitted to the employer. The employer never receives raw lab data.
- State marijuana laws, medical marijuana cards, and recreational use in legal states have no bearing on a DOT test result.
- Dilute specimens (creatinine 2–20 mg/dL, specific gravity 1.0010–1.0030) may be reported as Dilute Negative or Dilute Positive and may require a recollection.
Frequently Asked Questions
Does a valid prescription protect you on a DOT test?
It depends. A valid prescription for a Schedule II or III controlled substance may allow the MRO to verify the result as negative — but only if the medication is prescribed to you by a licensed provider and you disclose it to the MRO during the review call. Marijuana prescriptions (including medical marijuana cards) currently never change the outcome — DOT testing is federal, and cannabis remains a Schedule I substance federally. Note: the DEA has proposed reclassifying marijuana to Schedule III, which could affect this rule in the future, but no change has taken effect as of publication.
Is a 10-panel test the same as the DOT 5-panel?
No. The DOT-mandated test for safety-sensitive transportation workers is the 5-panel test defined in 49 CFR Part 40. A 10-panel test is a non-DOT workplace test that adds barbiturates, benzodiazepines, methadone, propoxyphene, and methaqualone to the standard five. Only the 5-panel test (performed under 49 CFR Part 40 procedures) satisfies DOT regulatory requirements.
Can an employer add more drugs to the DOT test?
No. Employers subject to DOT testing must use only the 5-panel test and the procedures in 49 CFR Part 40 for their DOT-mandated tests. They may run a separate, parallel non-DOT test with additional panels — but the DOT result and the non-DOT result are two separate processes with different rules.
What happens if an employee refuses to test?
A refusal to test is treated exactly the same as a verified positive under DOT regulations. The employee must be immediately removed from safety-sensitive duties and referred to a Substance Abuse Professional (SAP). Refusals include not showing up, leaving the collection site, providing an insufficient specimen without medical explanation, and tampering.
When does the DOT 5-panel become a 6-panel?
DOT has proposed adding fentanyl as a sixth analyte to the mandatory panel. The proposed rule was published and comment periods have closed, but as of publication the final rule had not yet taken effect. When it does, all DOT-mandated tests will be required to screen for fentanyl as well.
Need a DOT-compliant testing program?
APCA manages 5-panel DOT testing programs for USCG, FMCSA, and non-DOT employers. Setup takes less than 24 hours.
Regulatory Sources
- 49 CFR Part 40 — Procedures for Transportation Workplace Drug and Alcohol Testing Programs
- 49 CFR §40.87 — What are the cutoff concentrations for initial and confirmatory drug tests?
- DOT Office of Drug and Alcohol Policy and Compliance (ODAPC)
- SAMHSA Mandatory Guidelines for Federal Workplace Drug Testing Programs