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Vitamin D (25-Hydroxy) & Vitamin B12

Vitamin D (25-Hydroxy) & Vitamin B12

Also referred as
25-OH vitamin D
Calcidiol
Cyanocobalamin
+2
Methylcobalamin
Cobalamin
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The Vitamin D (25-Hydroxy) & Vitamin B12 test measures the level of vitamin D and vitamin B12 in your body. This test helps determine deficiencies related to vitamin D and B12. It also helps assess if you have too much of these vitamins in your body (most likely from excessive supplements). Vitamin D helps keep bones, teeth, and muscles healthy, whereas vitamin B12 maintains blood, brain, and nerve functions.

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Understanding Vitamin D (25-Hydroxy) & Vitamin B12

Vitamins, although required in small amounts, are significant in maintaining overall health and well-being.  Vitamin D (25-hydroxy) plays a critical role in various body functions, including:

  • Supports immune system functioning
  • Helps maintain healthy bones
  • Promotes cell growth

Vitamin D also acts as a hormone because it is synthesized in the skin (via sunlight) and then converted to its active form (known as calcitriol) in the kidneys. This active form helps regulate the levels of calcium and phosphorus in the body. Low vitamin D levels can result in an increased risk of fractures, muscle weakness and lethargy, dysfunction of the immune system, and an increased risk of diabetes.

Vitamin B12, also known as cobalamin, belongs to the B complex group of vitamins. It is vital for various body functions such as:

  • Aids in the production of red blood cells (RBCs)
  • Supports repair and regeneration of cells and tissues
  • Promotes optimal brain function and nerve health

Some common symptoms of vitamin B12 deficiency include fatigue, mouth ulcers, loss of appetite, burning sensation in feet, hands, arms, and legs, muscle weakness, etc.

The Vitamin D (25-Hydroxy) & Vitamin B12 checks for the levels of Vitamin D and Vitamin B12 in your body. The vitamin D test is important for assessing bone health and calcium metabolism. It helps detect deficiencies that can lead to rickets in children and osteomalacia or osteoporosis in adults. The vitamin B12 test helps determine the response to vitamin B12 therapy in conditions like anemia. Additionally, it is important to monitor parameters to maintain a healthy pregnancy. Low vitamin B12 levels have been associated with adverse pregnancy outcomes like anemia, low birth weight, and growth retardation of the baby.  The Vitamin D (25-Hydroxy) & Vitamin B12 test is also done to check for excessive vitamin D and B12, particularly due to overuse of supplementation.

Usually, no special preparation is required for this test. Eat and drink as per the daily routine. However, do let the doctor know about all the medications being taken, as some of them may affect the accuracy of the test results.

Test result ranges are approximate and may differ slightly between labs depending on the methodology and laboratory guidelines. Talk to the doctor about specific test results. They can help you understand what the results mean, suggest lifestyle changes like diet or exercise, give medicine if needed, and plan the right treatment for you.

Lifestyle Tips to Support Healthy Vitamin D and Vitamin B12 Levels:

🥤 Hydration: Drink enough water daily to support proper digestion and vitamin absorption.

🌞 Exposure to the sun: Get regular morning sunlight for natural vitamin D synthesis.  Avoid smoking and limit alcohol, as these can interfere with both vitamin D metabolism and vitamin B12 absorption.

🏃 Activity (Exercise): Regular physical activity, especially outdoors, supports vitamin D production and may enhance B12 metabolism.

🍔 Lifestyle (Balanced Diet): Include vitamin D-rich foods (like eggs, mushrooms, salmon, fortified milk) and vitamin B12-rich foods (like meat, fish, dairy, or fortified cereals for vegetarians/vegans).

👩‍⚕️ Talk to Your Doctor (Consultation): Avoid self-supplementation.  Consult your doctor for proper guidance on vitamin D or B12 levels and safe supplementation if needed.

🚭 Habits (Smoking and Alcohol): These habits can interfere with the absorption and activation of vitamins B12 and D.

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What does Vitamin D (25-Hydroxy) & Vitamin B12 measure?

Contains 2 tests

The Vitamin D (25-Hydroxy) & Vitamin B12 test is tailored to determine the levels of two important vitamins in your body: vitamin D (25-OH) and vitamin B12. Vitamin D is an essential nutrient that can be synthesized in the body upon healthy exposure to sunlight or absorbed from dietary sources. On the other hand, vitamin B12 is vital for maintaining a healthy nervous system, making red blood cells, and creating the genetic material of our cells.

This test is generally recommended as a part of routine health checkups, nutritional assessments, or to monitor vitamin levels in individuals with dietary restrictions or those who are on vitamin supplements.

The Vitamin B12 measures the amount of vitamin B12 in your blood. Vitamin B12 is important for a healthy nervous system, making red blood cells, and creating DNA in your cells. Low B12 levels are common in older adults, children, vegans or vegetarians, people with diabetes, those who had gastric bypass surgery, breastfeeding women, and people with conditions like Crohn’s disease that affect vitamin absorption. High B12 levels can occur in liver diseases, certain blood disorders, or from taking vitamin supplements. Usually, high levels from supplements are not harmful, as extra B12 is removed through urine.

What Do the Results Mean?

Pathological Factors 

Test Parameter

Interpretation (possible causes)

Low levels

  • Anaemia

  • Term pregnancy

  • Intrinsic factor deficiency

  • Partial gastrectomy/ileal damage

  • Celiac disease

  • Parasitic infestation

  • Pancreatic deficiency

  • Treated epilepsy

  • Hemodialysis 

High levels

  • Renal failure

  • Hepatocellular disorders

  • Myeloproliferative disorders and at times with excess

  • Supplementation of vitamin pills.

Physiological Factors

🧒 Age: Older adults may absorb less vitamin B12 due to lower stomach acid.

🫄 Pregnancy: Higher nutritional needs during pregnancy can temporarily lower B12 levels.

💊 Hormones: Hormone therapy or birth control pills can slightly affect B12 absorption.

Lifestyle Factors

🥗 Diet: Vegetarian or vegan diets without animal products may lead to low B12 intake.

🍷 Alcohol: Excessive alcohol can reduce B12 absorption and storage in the liver.

🚬 Smoking: Smoking cigarettes may reduce vitamin B12 activity.

Disclaimer: This information is for educational purposes only and should not replace professional medical advice, diagnosis, or treatment. Always consult your healthcare provider for personalized guidance.

What Do Normal Vitamin B12 Levels Mean?

Normal B12 levels show that your body has enough vitamin B12 for healthy red blood cells, nerve function, and metabolism. This helps maintain energy, mood, and brain health.

What Do High Vitamin B12 Levels Mean? 

High B12 levels are uncommon. They can happen due to too many supplements or health conditions like liver or kidney problems, or certain blood disorders. Your doctor may order extra tests to find the cause.

What to Do If Your Vitamin B12 Is High? 

Do not stop supplements suddenly. Your doctor will review your medical history, medications, and may suggest tests to check liver, kidney, or blood health. Often, adjusting supplements is enough to bring levels back to normal.

What Do Low Levels of Vitamin B12 Indicate?

Low vitamin B12 levels suggest a deficiency, which can lead to anemia, fatigue, and nerve damage if left untreated. Common causes include:

  • Poor dietary intake (especially in vegetarians or vegans)
  • Malabsorption due to gastric or intestinal issues
  • Pernicious anemia
  • Certain medications (e.g., metformin, proton pump inhibitors)
  • Treatment usually involves vitamin B12 supplements or injections based on deficiency severity.

Can Vitamin B12 Results Be Inaccurate?

Yes. While the test is reliable, results may sometimes be inaccurate due to recent vitamin B12 injections or supplements, pregnancy or hormonal therapy, liver or kidney diseases affecting vitamin metabolism, or laboratory variations in sample handling.

What Factors Can Affect Vitamin B12 Results?

Though it is a reliable test, several factors can influence accuracy:

  • Recent intake of vitamin B12 supplements or injections
  • Liver or kidney disorders
  • Alcohol consumption or chronic illness
  • Use of medications like metformin or oral contraceptives
  • Pregnancy or hormonal therapy

What Follow-Up Tests May Be Required?

Based on your Vitamin B12 results, your doctor may recommend additional tests to get a clearer picture of your health or to monitor possible complications. These may include: 

Table: Tests Commonly Ordered Alongside Vitamin B12

Test Name

Primary Purpose

Folate (Vitamin B9) Test

Check folate levels, since low folate can also cause anemia similar to B12 deficiency.

Homocysteine Test

Measure homocysteine in the blood, which can rise when B12 is low.

Methylmalonic Acid (MMA) Test

Detect early B12 deficiency by measuring MMA, which increases when B12 is low.

Complete Blood Count (CBC) Test

Provide details about red blood cells and hemoglobin to indicate type of anemia.

Intrinsic Factor Antibody Test

Detects autoimmune causes of B12 deficiency.

Gastrointestinal Absorption Tests

Checks if the stomach or intestines are absorbing B12 properly, e.g., after gastric surgery or in conditions like Crohn’s disease.

How Does the Vitamin B12 Test Differ from Other Blood Tests?

It is important to understand how the Vitamin B12 test differs from other common blood tests:

Complete Blood Count (CBC): Checks overall blood health and red blood cells. CBC can suggest anemia but doesn’t show the cause. Vitamin B12 specifically measures the nutrient needed to produce healthy red blood cells.

Folate (Vitamin B9) Test: Folate levels also affect red blood cell production. Unlike B12, folate deficiency alone does not directly indicate nerve or brain-related issues.

Iron Studies: Tests like serum iron or ferritin detect iron-related anemia. Vitamin B12 deficiency anemia occurs even if iron levels are normal.

Homocysteine & Methylmalonic Acid (MMA): These are functional tests that rise when B12 is low. The Vitamin B12 test directly measures the B12 level, making it the first step to detect deficiency.

Know more about Vitamin B12

The Vitamin D (25-Hydroxy) test measures the levels of vitamin D in the body. Vitamin D is an essential nutrient that can be synthesized in the body upon healthy exposure to sunlight or absorbed from dietary sources. It exists in two forms: vitamin D2 (ergocalciferol) and vitamin D3 (cholecalciferol). Vitamin D2 is present in plants, such as yeast or mushrooms, and is available as a supplement in fortified foods. Vitamin D3 is found in foods like cheese, green vegetables, mushrooms, egg yolks, and fatty fish.

Both forms of vitamin D need to be converted in the liver and kidneys before the body can use them effectively. This test shows the total level of vitamin D (D2 + D3) circulating in the body and guides your doctor in recommending the right treatment or lifestyle changes if needed.

What Do the Results Mean?

Pathological Factors 

Test Parameter

Interpretation (possible causes)

Normal / Optimal

  • Sufficiency

Possible causes: HPT (Hyperparathyroidism)

Humoral hypercalcemia of malignancy

Low

  • Deficiency 

Possible causes can be renal failure, hyperphosphatemia, vitamin D–dependent rickets, types 1 and 2

Borderline / Moderate Low

  • Insufficiency (due to inadequate dietary intake or low sunlight exposure)

High / Excess

  • Hypervitaminosis

Possible causes can be sarcoidosis, non-Hodgkin lymphoma 

Key: ⇧: Indicates High Levels , ⇩: Indicates Low Levels

Physiological Factors

🧒 Age: Older adults synthesize less vitamin D from sunlight primarily because of age-related changes in their skin.

🫄 Pregnancy: Vitamin D demand increases during pregnancy for fetal bone development.

💊 Hormonal Influence: Hormonal changes or contraceptive use may alter vitamin D metabolism.

Lifestyle Factors

🌤️ Sun Exposure: Limited sunlight reduces vitamin D synthesis.

🥗 Diet: Low intake of vitamin D-rich foods (like fish, egg yolks, or fortified dairy) can cause deficiency.

🍷 Alcohol Consumption: Chronic alcohol use affects liver function, impacting vitamin D metabolism.

🚬 Smoking: Smoking is closely associated with reduced bone density and altered vitamin D activity.

Disclaimer: This information is for educational purposes only and should not replace professional medical advice, diagnosis, or treatment. Always consult your healthcare provider for personalized guidance.

What Do Normal Vitamin D (25-Hydroxy) Levels Mean?

Normal vitamin D levels indicate your body has sufficient vitamin D for bone health, muscle strength, and immune function.

What Do High Levels of Vitamin D (25-Hydroxy) Indicate?

High vitamin D levels may occur due to excessive supplementation or conditions like sarcoidosis or hyperparathyroidism. Persistently high levels can cause vitamin D toxicity, leading to high blood calcium (hypercalcemia), which may result in nausea, vomiting, muscle weakness, and, in severe cases, kidney stones or kidney damage.

What to Do if Your Vitamin D (25-Hydroxy) Is Higher Than Normal?

If your Vitamin D (25-Hydroxy) level is high, consult your doctor for further guidance. They may review your dosage, identify causes of high calcium like excess supplement intake, or check for conditions such as hypercalcemia. Do not stop or adjust supplements without your doctor’s advice.

What Do Low Levels of Vitamin D (25-Hydroxy) Indicate?

Low levels suggest deficiency, which may cause weak bones, muscle pain, or fatigue. Common causes include:

  • Lack of sunlight exposure
  • Poor dietary intake
  • Malabsorption syndromes (e.g., celiac disease)
  • Obesity or liver/kidney disease

Treatment involves lifestyle changes, dietary improvement, or vitamin D supplements.

Can Vitamin D (25-Hydroxy) Results Be Inaccurate?

Yes. Results may vary due to recent supplement intake, laboratory variations, or conditions affecting vitamin D metabolism, such as liver or kidney disease. In some cases, additional tests may be advised to confirm the deficiency.

What Factors Can Affect Vitamin D (25-Hydroxy) Results?

Though it is a reliable test, some circumstances can influence accuracy:

  • Recent vitamin D or calcium supplement intake
  • Limited sun exposure
  • Chronic illness or liver/kidney disorders
  • Malabsorption syndromes
  • Obesity or hormonal imbalances

Know more about Vitamin D (25-Hydroxy)

FAQs related to Vitamin D (25-Hydroxy) & Vitamin B12

The Vitamin D (25-Hydroxy) & Vitamin B12 test can be done at any time of the day as no fasting is required for getting tested.
No, the Vitamin D (25-Hydroxy) & Vitamin B12 test is generally a safe procedure. While rare, bruising, bleeding, and infection at the puncture site may occur. In very uncommon cases, there can be swelling of the vein after the blood withdrawal.
Bone pain, depression, hair loss, muscle weakness, appetite loss, pale skin, and sleep disturbances are some common signs and symptoms of vitamin D deficiency.
Insufficient sunlight exposure or a vitamin D-deficient diet are the primary cause of vitamin D deficiency.
Always refer to your doctor in case your test results indicate low levels of vitamin D. Although consuming oily fish such as salmon or sardines, red meat, egg yolks, and fortified cereals may help improve vitamin D levels. Additionally, sufficient exposure to the sun can assist you in naturally increasing your levels of vitamin D.
Muscle weakness, loss of appetite, vision problems, a painful tongue, mouth ulcers, a fast and irregular heartbeat, shortness of breath, tingling or numbness, and pins and needles or burning sensations in the feet, hands, arms, and legs are common signs and symptoms of vitamin B12 deficiency.
Deficiency of vitamin B12 could be due to poor dietary intake, pernicious anemia, and certain types of stomach-related surgeries, like weight loss surgery or intestinal surgery.  Some medicines and immune system problems like lupus or Graves' disease can also cause it. Talk to your doctor if you have any of these issues.
Low vitamin B12 levels can lead to symptoms like fatigue, weakness, pale skin, appetite loss, weight loss, infertility, numbness or tingling in hands and feet, balance issues, depression, confusion, memory problems, and mouth soreness. In infants, it may cause growth delays and anemia. Early treatment is crucial to prevent nerve damage.
Always consult your doctor if your vitamin B12 levels are lower than normal. Incorporating food like meat, fish, eggs, milk, cheese, soy products, and fortified cereals may also help boost your vitamin B12 levels.
Regular sun exposure is the most natural way to get enough vitamin D. Usually, you need midday sunlight for about 10 to 30 minutes several times weekly. However, darker-skinned people may need a little longer than this. Exposure time also depends on how sensitive your skin is to sunlight.
a) 0–12 months: 400 IU/day; b). Ages 1–70: 600 IU/day; c) Over 70: 800 IU/day. For adults with low levels (<30 ng/mL), 1,500–2,000 IU/day is advised to restore balance (as per Endocrine Society).
Yes, low Vitamin B12 levels can lead to mental health problems like depression, anxiety, confusion, and memory issues. Vitamin B12, along with other B vitamins, helps control brain chemicals that affect mood and behavior.
Pregnant women need enough nutrients to help the baby grow and develop well. Low Vitamin B12 levels can lead to problems like anemia, low birth weight, and slow growth in the baby. It may also cause serious issues like spina bifida (a problem with the baby’s spine) and poor brain development.
Content created by
Written by:
Shweta Prakash
MSc, BSc
Reviewed by:
Dr. Naziya Maner
DNB, DCP
Want to know moreRead our editorial policy
Last Updated29th Nov 2025

Vitamin D (25-Hydroxy) & Vitamin B12 test price for other cities

Price inBangaloreRs. 1149
Price inHyderabadRs. 649
Price inPuneRs. 1149
Price inMumbaiRs. 1149
Price inNew DelhiRs. 1149

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Contains 2 tests
Vitamin B12
Vitamin D (25-Hydroxy)