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

CE-IVD validated product ready for IVDR within timeframes and transition periods specified by the IVDR 2017/746

Grouped product items
Product Name Qty
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma (kit for 200 tests)
Order No.: 62000
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma (complete kit for 1000 tests)
Order No.: 62000/1000

25-OH-Vitamin D2

25-OH-Vitamin D3

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

Kit for 200 tests (Order no. 62000)
Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls (Order no. 62000/1000)

 

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
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Different systems might show different performance data.
Lower and Upper Limit of Quantitation

25-OH-Vitamin D3: 1.0 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.0 µg/l – 250 µg/l

Specimen Serum/Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

  • Pipette 100 µl sample/calibrator/MassCheck® control into a reaction vial.
  • Add 25 µl Precipitation Reagent.
  • Add 200 µl Internal Standard and mix 20 s (vortex).
  • Incubate 10 min at +2 to +8 °C.
  • Centrifuge 5 min at 15000 x g.
  • Transfer 200 µl supernatant into an autosampler vial.
  • Inject ≤ 50 µL of the supernatant into the LC-MS/MS system
Run Time 5 min
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature +20 to +25 °C
Ionisation APCI positive
MS/MS Mode MRM
Parameters 25-OH-Vitamin D2, 25-OH-Vitamin D3
Order No. MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma by LC-MS/MS
62000 Kit for 200 tests
62000/1000 Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls

 

Order No. Kit components (Quantities in kit for 200 tests)
62001 Mobile Phase A, 1000 ml (1x)
62002 Mobile Phase B, 1000 ml (1x)
62003 Precipitation Reagent, 5 ml (1x)
62004 Internal Standard, 40 ml (1x)
62009 Rinsing Solution, 1000 ml (1x)
3006 Reaction Vials, 100 pcs. (2x)

 

Order No. Required components (not included in the kit for 200 tests)
62100 Analytical Column (equilibrated, with test chromatogram), 1 pc.
62110 Trap Column, equilibrated, with test chromatogram, 1 pc.
62028 3PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2, 4 x 1 ml (lyoph.)
62039 6PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2, 7 x 1 ml (lyoph.)
0222 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level I, 5 x 1 ml (lyoph.)
0223 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level II, 5 x 1 ml (lyoph.)
0256 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level III, 5 x 1 ml (lyoph.)
62015 Tuning Mix 25-OH-Vitamin D3/D2, Analytes and Internal Standards, 2 ml

 

Order No. General lab equipment (non-CE/IVD products)
3006 Reaction Vials, 100 pcs.
15010 PEEK Prefilter Housing, 1 pc.
15011 PEEK-encased Prefilter, 2 µm, 5 pcs.
15070 Stainless Steel Prefilter Housing, 1 pc.
15071 Stainless Steel Prefilter, 0,5 µm, 5 pcs.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
General lab equipment (non-CE-IVD products), available from Chromsystems:
As a customer please login or register to gain full access.

25-OH-Vitamin D2

25-OH-Vitamin D3

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

Kit for 200 tests (Order no. 62000)
Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls (Order no. 62000/1000)

 

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
Please note The information listed here, including the sample preparation, is not sufficient for using the product. Please read the information provided in the instruction manual, which includes detailed information on limitations associated with the use of the product in line with its intended purpose. Different systems might show different performance data.
Lower and Upper Limit of Quantitation

25-OH-Vitamin D3: 1.0 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.0 µg/l – 250 µg/l

Specimen Serum/Plasma
Sample Preparation

The information on the sample preparation presented here is not sufficient for use in the laboratory. For a detailed step by step description, please refer to the instruction manual.

  • Pipette 100 µl sample/calibrator/MassCheck® control into a reaction vial.
  • Add 25 µl Precipitation Reagent.
  • Add 200 µl Internal Standard and mix 20 s (vortex).
  • Incubate 10 min at +2 to +8 °C.
  • Centrifuge 5 min at 15000 x g.
  • Transfer 200 µl supernatant into an autosampler vial.
  • Inject ≤ 50 µL of the supernatant into the LC-MS/MS system
Run Time 5 min
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature +20 to +25 °C
Ionisation APCI positive
MS/MS Mode MRM
Parameters 25-OH-Vitamin D2, 25-OH-Vitamin D3
Order No. MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma by LC-MS/MS
62000 Kit for 200 tests
62000/1000 Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls

 

Order No. Kit components (Quantities in kit for 200 tests)
62001 Mobile Phase A, 1000 ml (1x)
62002 Mobile Phase B, 1000 ml (1x)
62003 Precipitation Reagent, 5 ml (1x)
62004 Internal Standard, 40 ml (1x)
62009 Rinsing Solution, 1000 ml (1x)
3006 Reaction Vials, 100 pcs. (2x)

 

Order No. Required components (not included in the kit for 200 tests)
62100 Analytical Column (equilibrated, with test chromatogram), 1 pc.
62110 Trap Column, equilibrated, with test chromatogram, 1 pc.
62028 3PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2, 4 x 1 ml (lyoph.)
62039 6PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2, 7 x 1 ml (lyoph.)
0222 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level I, 5 x 1 ml (lyoph.)
0223 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level II, 5 x 1 ml (lyoph.)
0256 MassCheck® 25-OH-Vitamin D3/D2 Serum Control Level III, 5 x 1 ml (lyoph.)
62015 Tuning Mix 25-OH-Vitamin D3/D2, Analytes and Internal Standards, 2 ml

 

Order No. General lab equipment (non-CE/IVD products)
3006 Reaction Vials, 100 pcs.
15010 PEEK Prefilter Housing, 1 pc.
15011 PEEK-encased Prefilter, 2 µm, 5 pcs.
15070 Stainless Steel Prefilter Housing, 1 pc.
15071 Stainless Steel Prefilter, 0,5 µm, 5 pcs.
The following components are included in the kit:
The following products are not included in the kit but are required for the application of the method:
General lab equipment (non-CE-IVD products), available from Chromsystems:
As a customer please login or register to gain full access.