25-OH-Vitamin D3/D2 in Serum/Plasma - HPLC
Vitamins D3 and D2 with one analysis
Improved separation
Quality controls and calibrator available


25-OH-Vitamin D2
25-OH-Vitamin D3
Clinical relevance
Vitamin D (cholecalciferol) is a recognised clinical chemical determinant associated with osteoporosis and osteopenia (reduction in bone density). It also plays a role in the body's immune response, cardiovascular diseases and some types of cancer. Therefore, vitamin D is highly important from a number of diagnostic perspectives.
25-OH-vitamin D3 is the primary metabolite in the human body and the direct precursor of the physiologically active form 1.25-(OH)2-vitamin D3. First-line treatment for vitamin D deficiency is nutritional supplements with vitamin D3 or vitamin D2. Vitamin D2 is easier to synthesise and is widely used as a supplement outside of the EU, and especially in the United States. Therefore, its determination in serum/plasma is also relevant.
Product advantages
- Combined analysis of vitamins D3 and D2
- Reliable separation
- Reliable quantification with a stable internal standard
This Chromsystems kit allows the simultaneous and reliable chromatographic determination of 25-OH-Vitamin D3 and 25-OH-Vitamin D2 on an isocratic HPLC system with UV detection. By means of efficient protein precipitation and selective solid phase extraction interfering components are removed. A non-physiological derivative of vitamin D is used as a stable internal standard to produce reliable quantification. This way, reproducible and reliable results can be produced at a low expenditure of cost and time.
Alternatively, for high sample throughput Chromsystems offers an online method with selective online solid phase extraction (order no. 38900/1000) as well as methods for analysis using mass spectrometry (order no. 62000).
Number of Tests | 100 | ||||||||||||||||||||||
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Limit of quantification | 1.1 µg/l - 1.4 µg/l |
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Linearity | up to at least 250 µg/l | ||||||||||||||||||||||
Recovery | 25-OH-Vitamin D3: 86 %; 25-OH-Vitamin D2: 87 % | ||||||||||||||||||||||
Intraassay | 25-OH-Vitamin D3: CV 3.0 % |
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Interassay | 25-OH-Vitamin D3: CV 3.3 % |
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Analysis Time | 10 min | ||||||||||||||||||||||
Sample Preparation |
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Sample Stability | At ambient temperature up to 3 days, at +2 to +8 °C up to 1 week. It is recommended to keep samples cool for transport. Freeze for longer storage (up to 1 year) at -20 °C. | ||||||||||||||||||||||
Specimen | Serum/Plasma | ||||||||||||||||||||||
Injection Volume | 25 µl (10-50 µl) | ||||||||||||||||||||||
Flow rate | 0.7 ml/min | ||||||||||||||||||||||
Column temperature | approx. 25 °C | ||||||||||||||||||||||
Wavelength | 265 nm | ||||||||||||||||||||||
Please note | The freely available information on this website, in particular on the sample preparation, are not sufficient to work with our products. Please read instructions and warning notices on products and/or instruction manuals. | ||||||||||||||||||||||
Method of Analysis | HPLC | ||||||||||||||||||||||
Parameter | 25-OH-Vitamin D2, 25-OH-Vitamin D3 | ||||||||||||||||||||||
Complete Specification |
HPLC parametersIsocratic HPLC system with UV detector.
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Additional Info | Isocratic HPLC system with UV detector. | ||||||||||||||||||||||
Injection interval | 12 min | ||||||||||||||||||||||
Autosampler rinsing solution | Water:acetonitrile = 50:50 |
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Vitamin D3/D2 Serum Calibration StandardOrder no.: 3803325-OH-Vitamin D3/D2 in Serum/Plasma - HPLC
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Vitamin D3/D2 Serum Calibration StandardOrder no.: 3803325-OH-Vitamin D3/D2 in Serum/Plasma - HPLC
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HPLC Column 25-OH-Vitamin D3/D2 in Serum/PlasmaOrder no.: 3813025-OH-Vitamin D3/D2 in Serum/Plasma - HPLC