What should I order on the lab slip to test vitamin D levels?

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What to Order on Lab Slip to Test Vitamin D Levels

Order a serum 25-hydroxyvitamin D [25(OH)D] test that measures both 25(OH)D2 and 25(OH)D3 forms. This is the gold standard for assessing vitamin D status in clinical practice 1.

Specific Test Name and Requirements

  • Write "25-hydroxyvitamin D, total" or "25(OH)D" on the lab requisition 2
  • The assay must measure both 25(OH)D2 (ergocalciferol metabolite) and 25(OH)D3 (cholecalciferol metabolite) to capture total vitamin D status 1
  • If the assay separates D2 and D3, the lab should report the sum as the primary result 1
  • Request results in ng/mL rather than nmol/L for consistency with clinical guidelines 1

Sample Collection Specifications

  • Use serum as the sample of choice 1
  • Avoid gel separator tubes if possible, as they can interfere with certain measurement methods, particularly LC-MS/MS 1
  • 25(OH)D is very stable in serum, so timing of collection is not critical 1

Important Testing Considerations

  • Do NOT order 1,25-dihydroxyvitamin D [1,25(OH)2D] for routine vitamin D status assessment—this is the active hormone form but does not reflect body stores 2
  • Ensure your laboratory participates in external quality control programs like DEQAS (Vitamin D External Quality Assurance Scheme) or the CDC's Vitamin D Standardization Certification Program 1, 3
  • Be aware that significant inter-laboratory variability exists (4-32% variation in classification of deficiency depending on assay used) 1, 3

When to Order Testing

  • Population-wide screening is NOT recommended by the USPSTF or Endocrine Society 4, 3
  • Test selectively in high-risk populations: patients with osteoporosis, malabsorption syndromes, chronic kidney disease, chronic liver disease, darker skin pigmentation, limited sun exposure, obesity, elderly/institutionalized individuals, or those on medications affecting vitamin D metabolism 4, 3, 5
  • For dark-skinned, veiled, elderly (≥65 years), or institutionalized individuals, you may supplement with 800 IU daily without baseline testing 1, 6

Timing of Follow-Up Testing

  • Recheck levels at least 3 months after initiating supplementation to allow serum concentrations to plateau and accurately reflect treatment response 1, 6, 3
  • For intermittent dosing regimens (weekly or monthly), measure just prior to the next scheduled dose 1, 6
  • Once stable on maintenance therapy, annual monitoring is sufficient 3

References

Guideline

Guideline Directed Topic Overview

Dr.Oracle Medical Advisory Board & Editors, 2025

Guideline

Vitamin D Testing Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Guideline

Vitamin D Testing and Optimal Levels

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2025

Research

When should we measure vitamin D concentration in clinical practice?

Scandinavian journal of clinical and laboratory investigation. Supplementum, 2012

Guideline

Vitamin D Insufficiency Treatment Guidelines

Praxis Medical Insights: Practical Summaries of Clinical Guidelines, 2026

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Professional Medical Disclaimer

This information is intended for healthcare professionals. Any medical decision-making should rely on clinical judgment and independently verified information. The content provided herein does not replace professional discretion and should be considered supplementary to established clinical guidelines. Healthcare providers should verify all information against primary literature and current practice standards before application in patient care. Dr.Oracle assumes no liability for clinical decisions based on this content.

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