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Epic Code LAB1230925 Vitamin D 25-Hydroxyvitamin Level D2 and D3

Test Name Alias

D2 | D3 | Vitamin D Hydroxy | 25-Hydroxy | calcifediol | ergocalciferol

Interface Order Alias


Clinical Information

Useful for the assessment of vitamin D levels. 

Collection Instructions

Specimen Collection: Blood


Container(s): Gold top (Serum Separator SST) (Red top acceptable for add ons)

Preferred Volume to Collect: 5.0 mL

Minimum Volume to Collect: 1.0 mL

Neonate Volume to Collect: 0.5 mL

Capillary collect ok? Yes

Microtainer acceptable: Yes


Collection Instructions:

  • After collection, gently invert tube 8-10 times.
  • Specimen should be processed within 2 hours – See Processing Instructions below.


Processing Instructions (Laboratory, Outpatient or Off-site collection)

  • Processed Specimen: Serum
  • Allow blood to clot for 30 minutes and centrifuge within 2 hours.
  • Centrifuge/Spin: Yes
  • Aliquot: No
  • Minimum serum volume: 0.5 mL (neonates may submit less)
  • Transport Temperature: Refrigerated

Rejection Criteria

  • Not centrifuged promptly after collection.
  • Quantity not sufficient.

Specimen Stability

Ambient: 5 days

Refrigerate: 14 days

Frozen: 14 days

Laboratory Retention: 3 days

Test Frequency

Monday-Friday for patients >5 years of age.

For patients ≤5 years of age a modified assay is performed once a week on Friday to eliminate interference due to the C-3 epimer of 25-hydroxyvitamin D3.

Reference Range

≤20 ng/mL - Deficiency 
21 - 29 ng/mL - Insufficiency  
30 - 100 ng/mL - Optimal Level 
>100 ng/mL - Possible Toxicity 

Performing Department


Performing Department Laboratory Location

Corewell Health Reference Laboratory, Grand Rapids


Liquid Chromatography-Mass Spectrometry



CDM Code


Epic Test ID



62292-8 (Total), 49054-0 (D2), 1989-3 (D3)


  1. Holick MF, Binkley NC, Bischoff-Ferrari HA, Gordon CM, Hanley DA, Heaney RP, Murad MH, Weaver CM; Endocrine Society. Evaluation, treatment, and prevention of vitamin D deficiency: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2011 Jul;96(7):1911-30. doi: 10.1210/jc.2011-0385. Epub 2011 Jun 6. Erratum in: J Clin Endocrinol Metab. 2011 Dec;96(12):3908. PMID: 21646368. 
  2. Ross AC, Manson JE, Abrams SA, Aloia JF, Brannon PM, Clinton SK, Durazo-Arvizu RA, Gallagher JC, Gallo RL, Jones G, Kovacs CS, Mayne ST, Rosen CJ, Shapses SA. The 2011 report on dietary reference intakes for calcium and vitamin D from the Institute of Medicine: what clinicians need to know. J Clin Endocrinol Metab. 2011 Jan;96(1):53-8. doi: 10.1210/jc.2010-2704. Epub 2010 Nov 29. PMID: 21118827; PMCID: PMC3046611. 

Reviewed Date