Detecting drug use involving benzodiazepines such as alprazolam, chlordiazepoxide, clonazepam, diazepam, midazolam, oxazepam, temazepam, clobazam, flunitrazepam, flurazepam, lorazepam, prazepam, triazolam, and zolpidem
Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)
7-Amino Flunitrazepam (Metabolite of Flunitrazepam) (Rohypnol)
7-Aminoclonazepam
Alprazolam (Xanax)
Ativan (Lorazepam)
Benzodiazepines
Chlordiazepoxide (Librium)
Clonazepam
Dalmane (Flurazepam)
Diazepam (Valium)
Flunitrazepam (Rohypnol)
Flurazepam (Dalmane)
Halcion (Triazolam)
Hydroxy-Ethyl Flurazepam (Metabolite of Flurazepam)
Librium (Chlordiazepoxide)
Lorazepam (Ativan)
Oxazepam (Serax)
Restoril (Temazepam)
Rohypnol (Flunitrazepam)
Serax (Oxazepam)
Temazepam (Restoril)
Triazolam (Halcion)
Valium (Diazepam)
Xanax (Alprazolam)
Urine
For situations where chain of custody is required, a Chain-of-Custody Kit (T282) is available. For chain-of-custody testing, order BNZX / Benzodiazepines Confirmation, Chain of Custody, Random, Urine.
Additional drug panels and specific requests are available. Call 800-533-1710 or 507-266-5700.
If urine creatinine is required or adulteration of the sample is suspected, also order ADULT / Adulterants Survey, Random, Urine in addition to this test.
Supplies: Sarstedt Aliquot Tube, 5 mL (T914)
Collection Container/Tube: Plastic urine container
Submission Container/Tube: Plastic, 5-mL tube
Specimen Volume: 1 mL
Collection Instructions:
1. Collect a random urine specimen.
2. No preservative
Additional Information:
1. No specimen substitutions.
2. STAT requests are not accepted for this test.
3. Submitting less than 1 mL will compromise our ability to perform all necessary testing.
If not ordering electronically, complete, print, and send a Therapeutics Test Request (T831) with the specimen.
1 mL
Gross hemolysis | Reject |
Icterus | Reject |
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Urine | Refrigerated (preferred) | 7 days | |
Frozen | 14 days |
Detecting drug use involving benzodiazepines such as alprazolam, chlordiazepoxide, clonazepam, diazepam, midazolam, oxazepam, temazepam, clobazam, flunitrazepam, flurazepam, lorazepam, prazepam, triazolam, and zolpidem
Benzodiazepines are any of a group of compounds having a common molecular structure and acting similarly as depressants of the central nervous system. As a class of drugs, benzodiazepines are among the drugs most frequently prescribed in the western hemisphere because of their efficacy, safety, low addiction potential, minimal side effects, and high public demand for sedative and anxiolytic agents.
Negative
Cutoff concentrations by liquid chromatography tandem mass spectroscopy:
Alprazolam: 10 ng/mL
Alpha-hydroxyalprazolam: 10 ng/mL
Chlordiazepoxide: 10 ng/mL
Clonazepam: 10 ng/mL
7-Aminoclonazepam: 10 ng/mL
Diazepam: 10 ng/mL
Nordiazepam: 10 ng/mL
Midazolam: 10 ng/mL
Alpha-hydroxy midazolam: 10 ng/mL
Oxazepam: 10 ng/mL
Temazepam: 10 ng/mL
Clobazam: 10 ng/mL
N-Desmethylclobazam: 10 ng/mL
Flunitrazepam: 10 ng/mL
7-Aminoflunitrazepam: 10 ng/mL
Flurazepam: 10 ng/mL
2-Hydroxy ethyl flurazepam: 10 ng/mL
Lorazepam: 10 ng/mL
Prazepam: 10 ng/mL
Triazolam: 10 ng/mL
Alpha-hydroxy triazolam: 10 ng/mL
Zolpidem: 10 ng/mL
Zolpidem phenyl-4-carboxylic acid: 10 ng/mL
Benzodiazepines are extensively metabolized, and the parent compounds are not detected in urine. This test screens for (and confirms) the presence of:
-Alprazolam
-Alpha-hydroxyalprazolam (metabolite of alprazolam)
-Chlordiazepoxide
-Clonazepam
-7-Aminoclonazepam (metabolite of clonazepam)
-Diazepam (separate prescribable drug and metabolite of medzazepam)
-Nordiazepam (metabolite of clorazepate, halazepam, prazepam, diazepam and medazepam)
-Midazolam
-Alpha-hydroxy midazolam (metabolite of midazolam)
-Oxazepam (separate prescribable drug and metabolite of clorazepate, halazepam, prazepam, medazepam, temazepam, and diazepam)
-Temazepam (separate prescribable drug and metabolite of medazepam and diazepam)
-Clobazam
-N-Desmethylclobazam (metabolite of clobazam)
-Flunitrazepam
-7-Aminoflunitrazepam (metabolite of flunitrazepam)
-Flurazepam
-2-Hydroxy ethyl flurazepam (metabolite of flurazepam)
-Lorazepam
-Prazepam
-Triazolam
-Alpha-hydroxy triazolam (metabolite of triazolam)
-Zolpidem
-Zolpidem phenyl-4-carboxylic acid (metabolite of zolpidem)
The clearance half-life of long-acting benzodiazepines is more than 24 hours. It takes 5 to 7 half-lives to clear 98% of a drug dose. Therefore, the presence of a long-acting benzodiazepine greater than the limit of quantification indicates exposure within a 5 to 20-day interval preceding specimen collection. Following a dose of diazepam, the drug and its metabolites appear in the urine within 30 minutes. Peak urine output is reached between 1 and 8 hours.
For additional information, including metabolism, clearance (half-life), and approximate detection times, see Drugs of Abuse Testing Guide.
No significant cautionary statements
1. Gudin JA, Mogali S, Jones JD, Comer SD. Risks, management, and monitoring of combination of opioid, benzodiazepines, and/or alcohol use. Postgrad Med. 2013;125(4):115-130. doi:10.3810/pgm.2013.07.2684
2. Dowell D, Ragan KR, Jones CM, Baldwin GT, Chou R. CDC Clinical Practice Guideline for Prescribing Opioids for Pain-United States. MMWR Recomm Rep. 2022;71(No. RR-3):1-95
3. Baselt RC. Disposition of Toxic Drugs and Chemical in Man. 12th ed. Biomedical Publications; 2020
4. Langman LJ, Bechtel LK, Holstege CP. Clinical toxicology. In: Rifai N, Chiu RWK, Young I, Burnham CAD,Wittwer CT, eds. Tietz Textbook of Laboratory Medicine. 7th ed. Elsevier; 2023:chap 43
Benzodiazepines are extensively metabolized by the liver and subsequently exist in the urine primarily as conjugated esters (-glucuronides). The conjugated metabolites are cleaved during a mild hydrolysis utilizing the enzyme glucuronidase. Stable isotope forms of the compounds are added as internal standards to account for extraction losses. An aliquot of the hydrolyzed sample is diluted, and the analytes are separated by liquid chromatography tandem mass spectroscopy and analyzed by multiple reaction monitoring.(Unpublished Mayo method)
Monday through Friday
This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.
80347
80339
80368
G0480 (if appropriate)
Test Id | Test Order Name | Order LOINC Value |
---|---|---|
BNZU | Benzodiazepines Confirmation, U | 90890-5 |
Result Id | Test Result Name |
Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
|
---|---|---|
608256 | Alprazolam by LC-MS/MS | 59615-5 |
608257 | Alpha-Hydroxyalprazolam by LC-MS/MS | 16348-5 |
608258 | Chlordiazepoxide by LC-MS/MS | 20522-9 |
608259 | Clonazepam by LC-MS/MS | 16229-7 |
608260 | 7-aminoclonazepam by LC-MS/MS | 51776-3 |
608261 | Diazepam by LC-MS/MS | 16227-1 |
608262 | Nordiazepam by LC-MS/MS | 16228-9 |
608263 | Midazolam by LC-MS/MS | 16233-9 |
608264 | Alpha-Hydroxy Midazolam by LC-MS/MS | 59590-0 |
608265 | Oxazepam by LC-MS/MS | 16201-6 |
608266 | Temazepam by LC-MS/MS | 20559-1 |
608267 | Clobazam by LC-MS/MS | 59635-3 |
608268 | N-Desmethylclobazam by LC-MS/MS | 97160-6 |
608269 | Flunitrazepam by LC-MS/MS | 20528-6 |
608270 | 7-aminoflunitrazepam by LC-MS/MS | 51777-1 |
608271 | Flurazepam by LC-MS/MS | 16231-3 |
608272 | 2-Hydroxy Ethyl Flurazepam by LC-MS/MS | 97159-8 |
608273 | Lorazepam by LC-MS/MS | 17088-6 |
608274 | Prazepam by LC-MS/MS | 17479-7 |
608275 | Triazolam by LC-MS/MS | 16232-1 |
608276 | Alpha-Hydroxy Triazolam by LC-MS/MS | 49876-6 |
608277 | Zolpidem by LC-MS/MS | 72770-1 |
608278 | Zolpidem Phenyl-4-Carboxylic acid by LC-MS/MS | 72768-5 |
608448 | Benzodiazepines Interpretation | 69050-3 |