Health Guide

Urine methylmalonic acid level

What is this test?

This test measures the level of methylmalonic acid in urine. Vitamin B12 helps to transform methylmalonic acid into another substance in the body. This test is used to assess vitamin B12 (cobalamin) deficiency[1][2][3][4][5][6][7][8].

What are other names for this test?

  • Urine 2-Methylpropanedioic acid level
  • Urine 2-Methylpropanedioic acid measurement
  • Urine methylmalonic acid measurement

What are related tests?

  • Homocysteine measurement
  • Serum methylmalonic acid level
  • Vitamin B12 deficiency screening

Why do I need this test?

Laboratory tests may be done for many reasons. Tests are performed for routine health screenings or if a disease or toxicity is suspected. Lab tests may be used to determine if a medical condition is improving or worsening. Lab tests may also be used to measure the success or failure of a medication or treatment plan. Lab tests may be ordered for professional or legal reasons. You may need this test if you have:

  • Not enough vitamin B12

When and how often should I have this test?

When and how often laboratory tests are done may depend on many factors. The timing of laboratory tests may rely on the results or completion of other tests, procedures, or treatments. Lab tests may be performed immediately in an emergency, or tests may be delayed as a condition is treated or monitored. A test may be suggested or become necessary when certain signs or symptoms appear.

Due to changes in the way your body naturally functions through the course of a day, lab tests may need to be performed at a certain time of day. If you have prepared for a test by changing your food or fluid intake, lab tests may be timed in accordance with those changes. Timing of tests may be based on increased and decreased levels of medications, drugs or other substances in the body.

The age or gender of the person being tested may affect when and how often a lab test is required. Chronic or progressive conditions may need ongoing monitoring through the use of lab tests. Conditions that worsen and improve may also need frequent monitoring. Certain tests may be repeated to obtain a series of results, or tests may need to be repeated to confirm or disprove results. Timing and frequency of lab tests may vary if they are performed for professional or legal reasons.

How should I get ready for the test?

24 hour urine collection:

During a 24-hour urine collection, follow your usual diet and drink fluids as you ordinarily would, unless healthcare workers give you other instructions. Avoid drinking alcohol before and during the urine collection.

You may be asked to take a drug before this test[9]. Ask your healthcare worker the details of how to prepare for this test.

Random urine:

To prepare for giving a urine sample, be sure to drink enough fluids before the test, unless you have been given other instructions. Try not to empty your bladder before the test.

Fast overnight before the test[9].

A sample of your second urination in the morning is collected for this test[9].

How is the test done?

A sample of urine collected over 24 hours or a randomly collected urine may be collected for this test.

24 hour urine collection:

For a 24-hour urine collection, all of the urine that you pass over a 24-hour time period must be collected. If you are in the hospital, a healthcare worker will collect your urine. You will receive a special container to collect the sample in if you are doing the collection at home. The following are directions for collecting a 24-hour urine sample while at home:In the morning scheduled to begin the urine collection, urinate in the toilet and flush away the first urine you pass. Write down the date and time. That is the start date and time for the collection.Collect all urine you pass, day and night, for 24 hours. Use the container given to you to collect the urine. Avoid using other containers. The urine sample must include the last urine that you pass 24 hours after starting the collection. Do not allow toilet paper, stool, or anything else to be added to the urine sample.Write down the date and time that the last sample is collected. The urine sample may need to be kept cool during the 24-hour collection period. If so, keep the closed container in a pan on ice. Do not put ice in the container with the urine.

Random urine:

To provide a sample of urine, you will be asked to urinate into a container. Fill the container as much as you can, but do not overfill it. Urine samples may also be taken from a catheter.

How will the test feel?

The amount of discomfort you feel will depend on many factors, including your sensitivity to pain. Communicate how you are feeling with the person doing the test. Inform the person doing the test if you feel that you cannot continue with the test.

24 hour urine or random urine collection:

This test usually causes no discomfort.

What should I do after the test?

24 hour urine collection:

When 24-hour urine collection is complete, close the container and seal the lid tightly. Return the sample in the urine container to the facility or healthcare worker as instructed. If you had the sample in an ice bath, return the sample within two hours after removing the container from the ice bath.

Random urine:

After collecting a urine sample, close the container if it has a lid. Place the container where the healthcare worker asked you to put it. Clean your hands with soap and water.

What are the risks?

Urine: A urine test is generally considered safe. Talk to your healthcare worker if you have questions or concerns about this test.

What are normal results for this test?

Laboratory test results may vary depending on your age, gender, health history, the method used for the test, and many other factors. If your results are different from the results suggested below, this may not mean that you have a disease. Contact your healthcare worker if you have any questions. The following are considered to be normal results for this test:

  • Normal ranges are technique specific [9]:
    • Colorimetry (24-hour urine): less than or equal to 11.2 mg/d (less than or equal to 95 micromol/d)
    • Gas-liquid chromatography (24-hour urine): less than or equal to 9.0 mg/d (less than or equal to 76 micromol/d)
    • Gas chromatography-mass spectrometry (random urine): less than 3.76 mg/g creatinine (less than 3.6 micromol/mol creatinine)

What follow up should I do after this test?

Ask your healthcare worker how you will be informed of the test results. You may be asked to call for results, schedule an appointment to discuss results, or notified of results by mail. Follow up care varies depending on many factors related to your test. Sometimes there is no follow up after you have been notified of test results. At other times follow up may be suggested or necessary. Some examples of follow up care include changes to medication or treatment plans, referral to a specialist, more or less frequent monitoring, and additional tests or procedures. Talk with your healthcare worker about any concerns or questions you have regarding follow up care or instructions.

Where can I get more information?

Related Companies

  • Linus Pauling Institute's Micronutrient Information Center -
  • Food and Nutrition Information Center, National Agricultural Library -
  • American Academy of Family Physicians -


[1] Bjorke Monsen AL & Ueland PM: Homocysteine and methylmalonic acid in diagnosis and risk assessment from infancy to adolescence. Am J Clin Nutr 2003; 78:7-21.

[2] Gultepe M, Ozcan O, Avsar K, et al: Urine methylmalonic acid measurements for the assessment of cobalamin deficiency related to neuropsychiatric disorders. Clin Biochem 2003; 36:275-282.

[3] Elin RJ & Winter WE: Methylmalonic acid: a test whose time has come. Arch Pathol Lab Med 2001; 125(6):824-827.

[4] Magera MJ, Helgeson JK, Matern D, et al: Methylmalonic acid measured in plasma and urine by stable-isotope dilution and electrospray tandem mass spectrometry. Clin Chem 2000; 46:1804-1810.

[5] Klee GG: Cobalamin and folate evaluation: measurement of methylmalonic acid and homocysteine vs vitamin B(12) and folate. Clin Chem 2000; 46(8 Pt 2):1277-1283.

[6] Snow CF: Laboratory diagnosis of vitamin B12 and folate deficiency: a guide for the primary care physician. Arch Intern Med 1999; 159(12):1289-1298.

[7] Norman EJ & Morrison JA: Screening elderly populations for cobalamin (vitamin B12) deficiency using the urinary methylmalonic acid assay by gas chromatography mass spectrometry. Am J Med 1993; 94(6):589-594.

[8] Matchar DB, Feussner JR, Millington DS, et al: Isotope-dilution assay for urinary methylmalonic acid in the diagnosis of vitamin B12 deficiency. A prospective clinical evaluation. Ann Intern Med 1987; 106(5):707-710.

[9] Tietz NW (Ed): Clinical Guide to Laboratory Tests, 3rd ed. W. B. Saunders, Philadelphia, PA, 1995.

Last Updated: 7/4/2018
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