25-OH-VD

Getein Technical Team

 01
Introduction of Vitamin
D
 1.1 What is Vitamin
D?

Definitions

Vitamin D is a fat-soluble vitamin that is naturally present in very few

foods, added to others, and available as a dietary supplement.
It is also produced endogenously when ultraviolet rays from sunlight
strike the skin and trigger vitamin D synthesis.

Only 10% of the vitamin D our body needs is obtained from food. The
rest is made by our bodies when our skin is exposed to sunlight.

Vitamin D Structure
 1.2 Types of Vitamin
D
Several forms of vitamin D exist. The two major forms are:

VD2 Structure VD3 structure

Vitamin D2 (Ergocalciferol): Vitamin D3 (Cholecalciferol):

It is synthesized by plants and is not It is made in large quantities in the skin when
produced by the human body. sunlight strikes bare skin. It can also be
ingested from animal sources.
 1.3 25-OH-VD and 1,25-(OH)2-VD

Vitamin D obtained from sun exposure, food, and supplements is biologically inert and must undergo two
hydroxylations in the body for activation.
The first occurs in the liver and converts vitamin D to 25-hydroxyvitamin D [25(OH)D], also known as calcidiol.
The second occurs primarily in the kidney and forms the physiologically active 1,25-dihydroxyvitamin D
[1,25(OH)2D], also known as calcitriol.

Serum concentration of 25(OH)D is the best indicator of vitamin D status. It reflects vitamin D produced
cutaneously and that obtained from food and supplements and has a fairly long circulating half-life of 15 days.
Conversely, 1,25(OH)2D only lasts for a few hours in the body and levels are a thousand-fold less than those of
25(OH)D.
A blood test for 1,25(OH)2D may appear normal or elevated even when your actual levels of vitamin D are low.
1.4 Metabolism of Vitamin
D?

Vitamin D Metabolism Flowchart

 1.5 Vitamin D
Function
• Absorb calcium: Vitamin D, along with calcium, helps build bones and keep bones strong and healthy.
• Block the release of parathyroid hormone, which makes bones thin and brittle.
• Other roles: modulation of cell growth, neuromuscular and immune function, and reduction of inflammation,
etc.
 1.6 Food Source of Vitamin D

Very few foods in nature contain vitamin D.

Animal-based foods can provide some vitamin D in the
form of 25(OH)D, which appears to be approximately
five times more potent than the parent vitamin in
raising serum 25(OH)D concentrations.

Note: IUs = International Units, DV = Daily Value.

 02
Vitamin D Deficiency and Toxicity
 2.1 Vitamin D Deficiency

Vitamin D deficiency is when levels of vitamin D in your body fall below those recommended as necessary
to ensure all the processes in your body that rely on vitamin D can function properly.

Common Causes of Lack of Vitamin D

 Limited exposure to sunlight: Some of us live in northern latitudes, wear long clothes, or have a job that is taken place
mostly indoors. Also sunscreen inhibits vitamin D production.
 Dark skin: People with dark skin have higher levels of melanin, and this pigment reduces the skin’s ability to make vitamin
D when exposed to sunlight.
 Kidney and liver function: These organs play an important role in converting vitamin D to its active form, so kidney or liver
diseases can reduce the ability of these organs to create biologically active form of vitamin D in the body.
 Strict vegetarian diet: Food sources that contain vitamin D are mostly animal-based, such as fish and fish oils, egg yolks,
cheese, fortified milk and beef liver.
 Digestive problems: Certain medical conditions, such as Crohn’s disease, cystic fibrosis, and celiac disease can reduce the
ability of the intestines to absorb vitamin D from food.
 Obesity: Obesity may cause low vitamin D levels. Research suggests that vitamin D may become ‘trapped’ inside fat tissue
so less of it is available in our blood circulation.
2.2 Epidemiology

Prevalence Vitamin D Deficiency

2.3 Related Diseases
 2.3 Related Diseases

Rickets and osteomalacia are the classical vitamin D deficiency diseases.

In children aged between six months and 36 months, vitamin D

deficiency most commonly causes rickets, a disease characterized by a
failure of bone tissue to properly mineralize, resulting in soft bones and
skeletal deformities.
Rickets

In adults, vitamin D deficiency can lead to osteomalacia, resulting in

weak bones[1,5]. Symptoms of bone pain and muscle weakness can
indicate inadequate vitamin D levels, but such symptoms can be subtle
and go undetected in the initial stages.

Osteomalacia
2.4 Vitamin D Assessment

Schematic Representation of the Roadmap for Action

 2.5 Vitamin D Toxicity

Vitamin D toxicity (VDT) due to excess of vitamin D (hypervitaminosis D) is a clinical condition characterized by
severe hypercalcemia that may persist for a prolonged time, leading to serious health consequences.
You cannot get vitamin D toxicity from the sun or by eating foods that contain vitamin D.
Vitamin D toxicity is said to occur at blood levels above 150 ng/ml (375 nmol/L) of 25(OH)D.

Symptoms of vitamin D toxicity may include the following:

 Dehydration  Muscle weakness

 Vomiting  Metastatic calcification of the soft tissues
 Decreased appetite  …
 Irritability
 Constipation
 Fatigue
 03
Vitamin D Test
 3.1 Clinical Significances

A vitamin D test is used to:

• Determine if bone weakness, bone malformation, or abnormal metabolism

of calcium is occurring as a result of a deficiency or excess of vitamin D;

• Help diagnose or monitor problems with parathyroid gland functioning since

PTH is essential for vitamin D activation;

• Screen people who are at high risk of deficiency;

• Help monitor the health status of individuals with diseases that interfere
with fat absorption;

• Monitor people who have had gastric bypass surgery and may not be able to
absorb enough vitamin D;

• Help determine the effectiveness of treatment when vitamin D, calcium,

 3.2 Reference Range

Optimal serum concentrations of 25(OH)D for bone and general health have not been established.

General Recommends for 25-OH-VD

Serum 25-Hydroxyvitamin D [25(OH)D] Concentrations and Health

 3.3 Testing Suitable
People

 Low calcium or phosphate levels or an unexplained increase

in serum alkaline phosphatase atypical osteoporosis;

 Unexplained limb pain near a joint in older people;

 Unusual fractures, unexplained bone pain, or other

symptoms suggesting metabolic bone disease.

Most experts recommend that babies, children, and adults who are at high risk of vitamin D deficiency should
be prescribed supplements without testing for deficiency, even if they don’t have any symptoms.
 3.4 Comparison of VD Measurement

 Chemiluminescentmicroparticle immunoassay (CMIA)

 Electrochemiluminescence (ECL) method
Immunoassay
 Homogenous competitive immunoassay
 Enzyme immunoassay

High-performance liquid
chromatography (HPLC)

Liquid chromatography–
tandem mass spectrometry Gold standard
(LC-MS/MS)
 Good accuracy
 Low susceptibility to interferences

Wallace, A. M., et al. "Measurement of 25-hydroxyvitamin D in the clinical laboratory: current procedures, Results with small uncertainties
performance characteristics and limitations." Steroids 75.7 (2010): 477-488.
 • IMMUNOASSAYS VS LC-MS/MS

Box-and-whisker plot showing the distribution of results for all assays tested.

The central boxes represent the 25th to 75th percentile range. The lines inside the boxes show the median value for each method.
The whiskers extend from the minimum to the maximum value, excluding outliers. An outlier value is defined as a value that
exceeds the upper or lower quartile plus or minus 1.5 times the interquartile range. The dashed horizontal line shows the median
value as measured by LC-MS/MS.

Farrell CJ . State-of-the-art vitamin D assays: a comparison of automated immunoassays with liquid chromatography–tandem mass spectrometry methods. Clin Chem . 2012;58(3):531–542.
• RESULTS
• DISCUSSIONS
 3.5 Interactions with Medications

Vitamin D supplements have the potential to interact with several types of medications. A few examples are provided below.
Individuals taking these medications on a regular basis should discuss vitamin D intakes with their healthcare providers.

Steroids
Corticosteroid medications such as prednisone, often prescribed to reduce inflammation, can reduce calcium absorption and impair
vitamin D metabolism. These effects can further contribute to the loss of bone and the development of osteoporosis associated with
their long-term use.

Other medications
Both the weight-loss drug orlistat (brand names Xenical® and alli TM) and the cholesterol-lowering drug cholestyramine (brand names
Questran®, LoCholest®, and Prevalite®) can reduce the absorption of vitamin D and other fat-soluble vitamins.
Both phenobarbital and phenytoin (brand name Dilantin®), used to prevent and control epileptic seizures, increase the hepatic
metabolism of vitamin D to inactive compounds and reduce calcium absorption.
 04
Getein Product
 4.1 Getein Product

Measurement Sample Reference Reaction

Test Method Item Volume
Range Type Value Time
Immunofluorescence 25-OH-VD 8-70 ng/mL S/P 30-50 ng/mL 40 uL 15 min
 4.2 Dietary Reference Intakes

• Determining intake levels for vitamin D is somewhat

complicated. Vitamin D levels in the body may come
from not only vitamin D in the diet but also from
synthesis in the skin through sunlight exposure.

• In 2010, American Institute of Medicine (IOM)

established the DRIs for vitamin D (refer to the right
table) based on the minimal sun exposure.

*For infants, Adequate Intake is 400 IU/day for 0 to 6 months of age and
400 IU/day for 6 to 12 months of age.
“Dietary Reference Intakes for Calcium and Vitamin D”, 2010, Institute of Medicine
Thanks for Your Attention

POCT