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Vitamin D Advanced (D2, D3 & D Total)

Also referred as

25-Hydroxy Vitamin D by LC-MS/MS

(Vitamin D2

Vitamin D3 & D Total)

5,139+ booked recently

For men & women

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72 hours

Contains

3 tests

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The Vitamin D Advanced (D2, D3 & D Total) test is a blood test that measures the levels of various forms of 25-hydroxy vitamin D (25-OH D2, 25-OH D3, and D total) in the body. This test is a valuable indicator of osteoporosis (bone weakness), rickets (bone malformation), and osteomalacia (bone softening). It can also help check the functioning of the parathyroid gland as this gland plays an important role in how the body uses vitamin D.

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Understanding Vitamin D Advanced (D2, D3 & D Total)

Vitamin D is a fat-soluble essential nutrient. It exists in two main forms, vitamin D3 and D2. Vitamin D3 (cholecalciferol) is derived mainly from animal sources. You can get it from fish, egg yolk, etc., and by spending time in sunlight, which provides ultraviolet B (UVB) rays. Vitamin D2 (ergocalciferol) comes from plant sources like mushrooms and some plant-based supplements. Both vitamin D2 and D3 are changed into 25-hydroxy D2 or 25-hydroxy D3 in the liver. Measuring the levels of 25-hydroxy vitamin D in the blood is the best way to check the vitamin D status.

As per a study by NCBI (National Center for Biotechnology Information), vitamin D deficiency is highly prevalent across all age groups in India. Low vitamin D levels can lead to an increased risk of fractures, muscle weakness and lethargy, dysfunction of the immune system, and an increased risk of chronic diseases like diabetes. The Vitamin D Advanced (D2, D3 & D Total) test can help find out if there is a vitamin D deficiency. It can also show if there is too much vitamin D in the body, usually from taking too many supplements.

The Vitamin D Advanced (D2, D3 & D Total) test offers a more detailed assessment of vitamin D status in the body. It evaluates Total 25-OH D as well as 25-OH metabolites of vitamin D2 and vitamin D3 using the highly specialized LC-MS/MS technique. On the other hand, a routine Vitamin D (25-OH) test measures only the Total 25-OH D vitamin (i.e., D2+D3) and does not differentiate between the two forms. The LC-MS/MS method is considered the gold standard for vitamin D evaluation. It is a highly specialized and sensitive method that can help differentiate between various forms of vitamin D, 25 (OH) D, and also substrates, D2 and D3. Additionally, LC-MS/MS can detect very small amounts of vitamin D and its byproducts.

The doctor may suggest Vitamin D Advanced (D2, D3 & D Total) test if there are symptoms suggestive of vitamin D deficiency or excess such as bone deformities, immune system dysfunction, muscle cramps and weakness, dental abnormalities, bone pain, fatigue, loss of appetite, increased thirst, nausea, frequent urination, etc.

No special preparation is required for this test. Consume a regular diet before taking the test. However, it is generally recommended to stop taking vitamin D supplements before this test, as they may affect the accuracy of the test results. Also, let the doctor know about all the other medications being taken, as some can affect the test results.

Lab test result ranges are approximate and may differ slightly between labs depending on the methodology and laboratory guidelines. Talk to the doctor about the specific test results. The test results will help them determine the medical condition, make recommendations for lifestyle modifications such as diet and exercise, prescribe medication if required to manage the condition and formulate an overall treatment plan.

What does Vitamin D Advanced (D2, D3 & D Total) measure?

Contains 3 tests

The Vitamin D Advanced (D2, D3 & D Total) test test checks the levels of all forms of 25-hydroxy vitamin D in the blood, which includes D2, D3, and the total amount.

Vitamin D is an important nutrient the body needs. It can be obtained in two ways: from sunlight or food. There are two main types of vitamin D:

Vitamin D2 (ergocalciferol): comes from plants, fortified foods, and some supplements.
Vitamin D3 (cholecalciferol): produced in the skin through sun exposure and also found in animal-based foods.

Before the body can use vitamin D, both D2 and D3 go through changes inside the body. They are absorbed in the small intestine and then processed in the liver and kidneys. They are turned into forms called 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D. This test measures 25-hydroxy vitamin D in your blood, which is the best way to check the body’s vitamin D levels.

Vitamin D3

The Vitamin D3 test measures the amount of vitamin D3, also known as cholecalciferol, a type of vitamin D. Vitamin D3 is naturally found in animal-derived products like milk and may be consumed in certain fortified foods or dietary supplements. It is directly synthesized in the skin when exposed to the sun. The Vitamin D3 test determines whether you have enough vitamin D3 in your body to support normal bodily functions.

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Vitamin D Total

The Vitamin D Total test measures the levels of vitamin D2 and D3 in your body. This test is an appropriate indicator of the total Vitamin D storage in the body. A vitamin D total test is also recommended in case of abnormal calcium, phosphorus, and parathyroid hormone levels. Individuals with abnormal vitamin D levels become prone to heart diseases, high blood pressure, immune system disorders, different types of cancers (prostate, breast, and colon), and various infectious diseases like influenza, tuberculosis, sepsis, etc. All these reasons make it more important for individuals to check their vitamin D levels.

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Vitamin D2

The Vitamin D2 test measures the amount of vitamin D2, also known as ergocalciferol, which is a type of vitamin D. It is usually found in plant-based sources such as mushrooms and in some fortified foods. The Vitamin D2 test is ordered to determine if a deficiency, insufficiency, or abnormally high level of vitamin D is present in the body, or to monitor ongoing treatment for a previously diagnosed deficiency.

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FAQs related to Vitamin D Advanced (D2, D3 & D Total)

What is the Vitamin D Advanced (D2, D3 & D Total) test, and why is it important?

The Vitamin D Advanced (D2, D3 & D Total) test measures your vitamin D levels, which are crucial for bone health, immune function, and overall well-being. It helps find vitamin D deficiency, which can cause bone pain, joint and muscle aches, tiredness, muscle twitching, and weakness. If not treated, it can lead to weak bones, fractures, or osteoporosis.

What are the signs and symptoms of vitamin D deficiency?

Signs and symptoms of vitamin D deficiency may include fatigue, bone pain, depression, hair loss, muscle weakness, appetite loss, pale skin, and sleep disturbances.

What can cause vitamin D deficiency?

The primary causes of vitamin D deficiency are insufficient sunlight exposure or a vitamin D-deficient diet.

Who should get the Vitamin D Advanced (D2, D3 & D Total) test done?

Anyone at risk of vitamin D deficiency should think about getting this test. This includes people over 45, those who don’t get much sunlight, have darker skin, or have digestive problems like celiac disease or Crohn’s disease, as well as those with certain other health conditions.

How is the Vitamin D Advanced (D2, D3 & D Total) test performed?

A phlebotomist will clean your skin using an antiseptic alcohol cotton swab or wipe and take a blood sample from your vein using a needle. The blood sample will be stored safely and transported to the laboratory for analysis.

What is the right time to get the Vitamin D Advanced (D2, D3 & D Total) test done?

This test can be done at any time of the day. However, one should stop taking vitamin D supplements before giving the test sample to get accurate results.

Are there any risks associated with this test?

No, the Vitamin D Advanced (D2, D3 & D Total) test is generally safe with minimal risks. You might feel a little discomfort when the blood sample is taken, but serious problems are very rare.

What happens if my vitamin D is low?

Low vitamin D can have adverse effects on the whole body. It can increase the risk of heart disease, high blood pressure, diabetes, infections, immune system disorders, multiple sclerosis, and colon, prostate, and breast cancers.

Can vitamin D deficiency be corrected through diet alone?

Though some food items such as fortified foods, green vegetables, mushrooms, egg yolks, and fatty fish contain vitamin D, supplements are often necessary to reach adequate levels.

How much sun exposure is sufficient to get adequate vitamin D?

Regular sun exposure is the most natural way to get enough vitamin D. Usually, you need the afternoon sun for about 10 to 30 minutes several times weekly. However, people with darker skin may need a little longer than this. Your exposure time depends on how sensitive your skin is to sunlight.

How much vitamin D is needed daily?

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).

What other tests can be ordered along with a vitamin D profile?

Serum calcium, phosphorus, and parathyroid hormone can be ordered in addition to vitamin D profile.

What are the differences between Vitamin D2 and Vitamin D3?

Vitamin 25-OH D2 (ergocalciferol) is found in some plant-based foods and is used in some supplements. Vitamin 25-OH D3 (cholecalciferol) is produced by the skin in response to sunlight and is also found in animal-based foods and supplements. Vitamin 25-OH D3 is generally considered more effective in raising blood levels of Vitamin D.

Content created by

Dr. Swati Gupta (Anand)

MD Lab Medicine, MBBS

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Last Updated

6th Oct 2025