25-OH-Vitamin D3/D2 in Serum/Plasma (Online Method) - HPLC
Small sample volume: 100 µl
Online sample preparation
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.
- Combined analysis of vitamins D3 and D2
- 100 µl sample volume is sufficient
- Selective online sample preparation
- Up to 100 samples per day
This Chromsystems kit allows simple and fast determination of 25-OH-Vitamin D3 and 25-OH-Vitamin D2 in a single HPLC run with UV detection. Following an efficient protein precipitation step, selective online solid phase extraction enables up to 100 analyses per day. Requiring only small sample volume, this analysis is also suitable for measuring paediatric samples. A stable internal standard ensures reliable quantification. This allows reproducible and reliable results while keeping costs and required laboratory time low.
Alternatively, there is also a standard method for HPLC (order no. 38038) as well as methods for determination using mass spectrometry (order no. 62000).
Content for 1000 tests
Mobile Phase A (online method), 10 x 1000 ml
Mobile Phase B (online method), 10 x 1000 ml
Internal Standard (online method), 10 x 25 ml
25-OH-Vitamin D3/D2 Serum Calibration Standard (lyoph.), 2 x 2 ml
25-OH-Vitamin D3/D2 Serum Control, Level I, 2 x 2 ml
25-OH-Vitamin D3/D2 Serum Control, Level II, 2 x 2 ml
HPLC Column, 1 pc.
Trap Column, 1 pc.
PEEK-encased Prefilter, 2 μm, 5 pcs.
Reaction Vials, transparent, 100 pcs.
|Method of Analysis||HPLC|
|Number of Tests||1000|
|Limit of quantification||5 µg/l|
|Linearity||5 - 250 µg/l|
|Recovery||25-OH-Vitamin D3: 99 %; 25-OH-Vitamin D2: 93 %|
25-OH-Vitamin D3: CV ≤ 3.2 %
25-OH-Vitamin D3: CV ≤ 5.6 %
|Analysis Time||10 min|
|Sample Stability||At ambient temperature: 3 days, at +2 to +8 °C: 1 week and below -18 °C: 1 year. Keep samples light protected and cool for transport.|
|Injection Volume||75 µl|
|Flow rate||Mobile Phase A (trap column): 1.0 ml/min; Mobile Phase B (analytical column): 0.7 ml/min|
|Column temperature||ambient (~25 °C)|
The HPLC analysis of 25-OH-vitamin D3/D2 (online method) requires two HPLC pumps, a 6-port-valve and a UV detector.
|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.|
|Parameter||25-OH-Vitamin D2, 25-OH-Vitamin D3|
3PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2Order no.: 62028