Order Code MTHX Methotrexate, Serum
Additional Codes
MAYO CODE MTHX
Useful For
Determining whether methotrexate is being cleared appropriately and verifying that a nontoxic concentration has been attained following therapy
Method Name
Enzyme-Immunoassay (EIA)
Reporting Name
Methotrexate, SSpecimen Type
SerumOrdering Guidance
For patients that have received glucarpidase (carboxypeptidase G2) as a high-dose methotrexate rescue therapy, order MTXSG / Methotrexate Post Glucarpidase, Serum.
Shipping Instructions
Ship specimen in amber vial to protect from light.
Specimen Required
Supplies: Amber Frosted Tube, 5 mL (T915)
Collection Container/Tube:
Preferred: Serum gel
Acceptable: Red top
Submission Container/Tube: Amber vial
Specimen Volume: 0.5 mL
Collection Instructions:
1. Methotrexate is sensitive to fluorescent light; avoid prolonged exposure of specimen to direct light.
2. Within 2 hours of collection, centrifuge the specimen.
3. For serum gel tubes, aliquot serum into a plastic vial within 2 hours of collection. Keep protected from light.
4. For red-top tubes, aliquot serum into a plastic vial immediately. Keep protected from light.
Specimen Minimum Volume
0.25 mL
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Serum | Refrigerated (preferred) | 14 days | LIGHT PROTECTED |
Frozen | 28 days | LIGHT PROTECTED | |
Ambient | 72 hours | LIGHT PROTECTED |
Reject Due To
Gross hemolysis | Reject |
Reference Values
Nontoxic drug concentration after 72 hours: <0.1 mcmol/L
Method Description
The Roche ONLINE TDM Methotrexate assay is a homogeneous enzyme immunoassay based on competition between drug in the specimen and methotrexate labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PD) for binding to the antibody reagent. As the latter binds antibody, enzyme activity decreases. In the presence of drug from the specimen, enzyme activity increases and is directly proportional to the drug concentration. Active enzyme converts the coenzyme nicotinamide adenine dinucleotide (NAD) to NADH, which is measured spectrophotometrically as a rate of change in absorbance. Endogenous serum G6PD does not interfere with the results because the coenzyme NAD functions only with the bacterial enzyme used in the assay.(Package insert: MTX: ONLINE TDM Methotrexate. Roche Diagnostics; V 1.0, 03/2024)
Performing Laboratory

CPT Code Information
80204
Day(s) Performed
Monday through Sunday
Forms
If not ordering electronically, complete, print, and send a Therapeutics Test Request (T831) with the specimen.