MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS

Order No.: 62000 & 62000/1000
Parameters:
25-OH-Vitamin D2, 25-OH-Vitamin D3

Online sample preparation
Isotopically labelled internal standard
Multilevel calibrators
Upgrade available for the determination of 3-epi-25-OH-Vitamin D3/D2

Grouped product items
Product Name Qty
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
Order No.: 62000
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma (integrated kit for 1000 tests) - LC-MS/MS
Order No.: 62000/1000

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

  • Online sample preparation
  • Isotopically labelled internal standard
  • 3PLUS1® and 6PLUS1® Multilevel Calibrator Set and MassCheck® Controls
  • Upgrade available for the determination of epimeric forms

 

This Chromsystems kit allows the specific determination of 25-OH-Vitamin D3 and 25-OH-Vitamin D2 in serum and plasma with LC-MS/MS. The manual sample preparation is limited to simple and effective protein precipitation. After this, the analytes are concentrated using an online trap column and interfering matrix components are separated. The trap column is connected by a simple switching system to the HPLC column, which provides further purification of the analytes. This method is characterised by the use of atmospheric pressure chemical ionisation (APCI). Isotopically labelled internal standards assure a high level of precision and robustness of the method and minimise matrix effects ("ion suppression"). The 3PLUS1® and 6PLUS1® multilevel calibrators are optionally available and guarantee a high level of precision of the results.

An upgrade for the determination of the epimeric forms of 25-OH-Vitamin D3 and 25-OH-Vitamin D2 is also available for this kit (see accessories), as well as a variant for automated sample preparation in 96 well filter plates (order no. 62000/1000/F).

 

Content for 200 tests (62000)
Mobile Phase A, 1000 ml
Mobile Phase B, 1000 ml
Precipitation Reagent, 5 ml
Internal Standard, 40 ml
Rinsing Solution, 1000 ml
Reaction Vials, 2 x 100 pcs. 

 

Content for 1000 tests (62000/1000)
Mobile Phase A, 5 x 1000 ml
Mobile Phase B, 5 x 1000 ml
Precipitation Reagent, 5 x 5 ml
Internal Standard, 5 x 40 ml
Rinsing Solution, 2 x 1000 ml
Reaction Vials, 10 x 100 pcs. 
3PLUS1® 25-OH-Vitamin D3/D2 Multilevel Serum Calibrator Set (lyoph.), 4 x 1 ml 
MassCheck® 25-OH-Vitamin D3/D2 Serum Control, Level I (lyoph.), 5 x 1 ml
MassCheck® 25-OH-Vitamin D3/D2 Serum Control, Level II (lyoph.), 5 x 1 ml 
Analytical Column, 1 pc. 
Trap Column, 2 pcs.

More Information
Method of Analysis LC-MS/MS
Limit of quantification

25-OH-Vitamin D2: 2.2 µg/l
25-OH-Vitamin D3: 3.0 µg/l

Linearity

up to 250 μg/l

Intraassay

CV < 5 %

Interassay

CV < 6 %

Analysis Time 5 min
Specimen Serum/Plasma
Pre-analytic Treatment

Patient samples are stable up to 3 days at ambient temperature and up to 1 week at +2 to +8 °C. For longer storage (maximum 1 month) deep-freeze the samples below -18 °C.

Sample Preparation
  • Place 100 µl sample into a reaction vial.
  • Add 25 µl Precipitation Reagent.
  • Add 200 µl Internal Standard and mix 20 s (vortex).
  • Incubate 10 min at +4 °C.
  • Centrifuge 5 min at 15000 x g.
  • Transfer 200 µl supernatant into an autosampler vial.
Sample Stability Samples are stable up to 1 week at +2 to +8 °C. For longer storage periods keep samples frozen below -18 °C.
Injection Volume 10 to 50 μl
Additional Info

HPLC parameters

Pump A delivers 100 % Mobile Phase A for loading the trap column.
Pump B delivers 100 % Mobile Phase A for separating analytes on the analytical column.

Ionisation APCI positive
MS/MS-Mode MRM
Information

The trap column and the analytical column are linked to the mass spectrometer through a 2-position 6-port valve.

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
Flow

Pump A: 0.02–1.00 ml/min
Pump B: 0.60–1.00 ml/min

  1. 62001 LCMS Vitamin D Mobile Phase A
    Mobile Phase A
    Order no.: 62001

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  2. 62002 LCMS Vitamin D Mobile Phase B
    Mobile Phase B
    Order no.: 62002

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  3. 62003 LCMS Vitamin D Precipitation Reagent
    Precipitation Reagent
    Order no.: 62003

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  4. 62004 LCMS Vitamin D Internal Standard
    Internal Standard
    Order no.: 62004

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  5. 62009 LCMS rinsing solution vitamin D3 vitamin D2 serum plasma
    Rinsing Solution
    Order no.: 62009

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  6. 3006 reaction vials
    Reaction Vials
    Order no.: 3006
    Details
  1. 0221 0222 0223 0256 Vitamin D Serum Controls
    Details
  1. 62100 LCMS Vitamin D Analytical Column
    Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Order no.: 62100

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS

    Details
  1. 62100 LCMS Vitamin D Analytical Column
    Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Order no.: 62100

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS

    Details
  2. 62110 LCMS Vitamin D Trap Column
    Trap Column
    Order no.: 62110

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  3. 62015 LCMS tuning mix vitamin D3 vitamin D2 serum plasma
    Tuning Mix 25-OH-Vitamin D3/D2
    Order no.: 62015

    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS

    Details
  4. Upgrade for 3-epi-25-OH-Vitamin D3/D2 - LC-MS/MS
    Upgrade for 3-epi-25-OH-Vitamin D3/D2 - LC-MS/MS
    Order no.: Epimer Upgrade

    Upgrade for order no. 62000
    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma

    Details
  5. PEEK-encased Prefilter, 2 µm
    Details
  6. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
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