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MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma - LC-MS/MS

Order No.: 62062/1000/F, for 1000 tests
Parameters:
25-OH-Vitamin D2, 25-OH-Vitamin D3, 3-epi-25-OH-Vitamin D3

For sample preparation with 96 Well Filter Plates

Assay validated according to IVDR (=> Declaration of Conformity)

main product photo

25-OH-Vitamin D3

3-epi-25-OH-Vitamin D3

25-OH-Vitamin D2

MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS

Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls
For sample preparation with 96 Well Filter Plates

 

Clinical relevance

This assay allows the quantitative determination of 25-hydroxycholecalciferol (25-OH-vitamin D3) and 25-hydroxyergocalciferol (25-OH-vitamin D2), as well as 3-epi-25-OH-vitamin D3 in human plasma or serum samples.

It is intended to be used for screening and/or monitoring of 25-OH-vitamin D3/D2 levels where indicated:

  • in patients with suspected vitamin D deficiency,
  • in patients with suspected vitamin D insufficiency, or
  • in patients with suspected vitamin D excess, and/or
  • in patients under vitamin D supplementation therapy.

The test kit is further intended to be used as an aid to diagnosis of diseases for which determination of vitamin D levels is indicated.

This kit should be used when the levels of 3-epi-25-OH-vitamin D3 and 25-OH-vitamin D3 are to be determined separately.

 

Assay characteristics

Manual sample preparation is limited to simple and effective protein precipitation. The analytes are enriched with the aid of a trap column online and thus, interfering matrix components are separated. Chromatographic separation and further purification of all analytes is then performed by a special high‑resolution analytical column connected to the trap column by a valve. The method features the use of APCI (atmospheric pressure chemical ionisation) and the isotopically labelled internal standard adjusted to the epimeric forms. Matrix effects (“ion suppression”) are minimised and the precision and robustness of the method are optimised. The multilevel calibrator includes 25-hydroxyvitamin D3/D2 as well as the epimeric form 3-epi-25-hydroxyvitamin D3 and ensures high precision of results.

Detailed performance evaluation data for this assay can be found in Appendices II and III of the instruction manual.

More Information
Method of Analysis LC-MS/MS
Number of Tests 1000
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. Detailed performance evaluation data for this assay can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

25-OH-Vitamin D3: 1.5 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.5 µg/l – 250 µg/l
3-epi-25-OH-Vitamin D3: 1.5 µg/l – 250 µg/l

Different systems might show different performance data.
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.

Sample preparation with 96 well filter plates:

  • Pipette 200 µl Internal Standard into 96 well filter plate.
  • Add 25 µl Precipitation Reagent.
  • Add 100 µl sample/calibrator/MassCheck® control and shake 10 min.
  • Place 96 well filter plate on collection plate.
  • Centrifuge 5 min at 2000 x g.
  • Seal collection plate and transfer to autosampler.
  • Inject ≤ 50 µl eluate into the LC-MS/MS system
Run Time 3.25 or 3.9 min (depending on gradient system)
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature Analytical Column: +40 °C / Trap Column: +20 to +25 °C
Ionisation APCI positive
MS/MS Mode MRM
Parameters 25-OH-Vitamin D2, 25-OH-Vitamin D3, 3-epi-25-OH-Vitamin D3

MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS

Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls
For sample preparation with 96 Well Filter Plates

 

Order No. Kit components (Quantities in kit)
62011 Mobile Phase A, 1000 ml (5x)
62022 Mobile Phase B, 1000 ml (5x)
62003 Precipitation Reagent, 5 ml (5x)
62045 Internal Standard Mix, 40 ml (5x)
62009 Rinsing Solution, 1000 ml (2x)
62057 96 Well Filter Plates, 3 pcs. (4x)
62058 96 Well Collection Plates, 3 pcs. (4x)
62059 Pierceable Adhesive Seals for 96 Well Plates, 3 pcs. (4x)
62130 Analytical Column, equilibrated, with test chromatogram, 1 pc. (1x)
62110/epi Trap Column, equilibrated, with test chromatogram, 1 pc. (2x)
62029 3PLUS1® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
0311 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level I (lyoph.), 1 ml (5x)
0312 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level II (lyoph.), 1 ml (5x)

 

Order No. Required components (not included in the kit)
62017 Tuning Mix 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 (Analytes and Internal Standards), 1 ml

 

Order No. Lab equipment
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:
  1. 62011 Mobile Phase A 25-OH-Vitamin-D3-D2
    Mobile Phase A
    Order no.: 62011

    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma

    Details
  2. 62022 Mobile Phase B 25-OH-Vitamin-D3-D2
    Mobile Phase B
    Order no.: 62022
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 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. 62045 Internal Standard Mix
    Internal Standard Mix
    Order no.: 62045
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 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. 96 Well Filter Plates
    96 Well Filter Plates
    Order no.: 62057
    Details
  7. 96 Well Collection Plate
    Collection Plates
    Order no.: 62058
    Details
  8. Pierceable Adhesive Seals for 96 Well Plates
    Pierceable Adhesive Seals for 96 Well Plates
    Order no.: 62059
    Details
  9. 62130 Analytical Column 25-OH-Vitamin D3-D2
    Analytical Column
    Order no.: 62130
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma
    Details
  10. 62110_epi trap column epimers vitamin D
    Trap Column
    Order no.: 62110/Epi
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma – LC-MS/MS
    Details
  11. 62029 3PLUS1 Calibrator 3-epi Vitamin D - LCMS
    3PLUS1® Multilevel Serum Calibrator Set (including epimeric form of Vitamin D3)
    Order no.: 62029
    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Details
  12. 0310 0311 0312 3-epi Vitamin D Controls
    MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Controls
    Order no.: 0310/0311/0312

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

    Details
The following products are not included in the kit but are required for the application of the method:
  1. 62017 Tuning Mix D3-D2
    Tuning Mix 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3
    Order no.: 62017
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma - LC-MS/MS

    Details
  1. 62029 3PLUS1 Calibrator 3-epi Vitamin D - LCMS
    3PLUS1® Multilevel Serum Calibrator Set (including epimeric form of Vitamin D3)
    Order no.: 62029
    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Details
  1. 0310 0311 0312 3-epi Vitamin D Controls
    MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Controls
    Order no.: 0310/0311/0312

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

    Details
  1. 62130 Analytical Column 25-OH-Vitamin D3-D2
    Analytical Column
    Order no.: 62130
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma
    Details
  1. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details
  3. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  4. Stainless Steel Prefilter - 0.5 µm
    Stainless Steel Prefilter - 0.5 µm
    Order no.: 15071
    Details
As a customer please login or register to gain full access.

25-OH-Vitamin D3

3-epi-25-OH-Vitamin D3

25-OH-Vitamin D2

MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS

Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls
For sample preparation with 96 Well Filter Plates

 

Clinical relevance

This assay allows the quantitative determination of 25-hydroxycholecalciferol (25-OH-vitamin D3) and 25-hydroxyergocalciferol (25-OH-vitamin D2), as well as 3-epi-25-OH-vitamin D3 in human plasma or serum samples.

It is intended to be used for screening and/or monitoring of 25-OH-vitamin D3/D2 levels where indicated:

  • in patients with suspected vitamin D deficiency,
  • in patients with suspected vitamin D insufficiency, or
  • in patients with suspected vitamin D excess, and/or
  • in patients under vitamin D supplementation therapy.

The test kit is further intended to be used as an aid to diagnosis of diseases for which determination of vitamin D levels is indicated.

This kit should be used when the levels of 3-epi-25-OH-vitamin D3 and 25-OH-vitamin D3 are to be determined separately.

 

Assay characteristics

Manual sample preparation is limited to simple and effective protein precipitation. The analytes are enriched with the aid of a trap column online and thus, interfering matrix components are separated. Chromatographic separation and further purification of all analytes is then performed by a special high‑resolution analytical column connected to the trap column by a valve. The method features the use of APCI (atmospheric pressure chemical ionisation) and the isotopically labelled internal standard adjusted to the epimeric forms. Matrix effects (“ion suppression”) are minimised and the precision and robustness of the method are optimised. The multilevel calibrator includes 25-hydroxyvitamin D3/D2 as well as the epimeric form 3-epi-25-hydroxyvitamin D3 and ensures high precision of results.

Detailed performance evaluation data for this assay can be found in Appendices II and III of the instruction manual.

More Information
Method of Analysis LC-MS/MS
Number of Tests 1000
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. Detailed performance evaluation data for this assay can be found in Appendices II and III of the instruction manual.
Lower and Upper Limit of Quantitation

25-OH-Vitamin D3: 1.5 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.5 µg/l – 250 µg/l
3-epi-25-OH-Vitamin D3: 1.5 µg/l – 250 µg/l

Different systems might show different performance data.
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.

Sample preparation with 96 well filter plates:

  • Pipette 200 µl Internal Standard into 96 well filter plate.
  • Add 25 µl Precipitation Reagent.
  • Add 100 µl sample/calibrator/MassCheck® control and shake 10 min.
  • Place 96 well filter plate on collection plate.
  • Centrifuge 5 min at 2000 x g.
  • Seal collection plate and transfer to autosampler.
  • Inject ≤ 50 µl eluate into the LC-MS/MS system
Run Time 3.25 or 3.9 min (depending on gradient system)
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature Analytical Column: +40 °C / Trap Column: +20 to +25 °C
Ionisation APCI positive
MS/MS Mode MRM
Parameters 25-OH-Vitamin D2, 25-OH-Vitamin D3, 3-epi-25-OH-Vitamin D3

MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS

Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls
For sample preparation with 96 Well Filter Plates

 

Order No. Kit components (Quantities in kit)
62011 Mobile Phase A, 1000 ml (5x)
62022 Mobile Phase B, 1000 ml (5x)
62003 Precipitation Reagent, 5 ml (5x)
62045 Internal Standard Mix, 40 ml (5x)
62009 Rinsing Solution, 1000 ml (2x)
62057 96 Well Filter Plates, 3 pcs. (4x)
62058 96 Well Collection Plates, 3 pcs. (4x)
62059 Pierceable Adhesive Seals for 96 Well Plates, 3 pcs. (4x)
62130 Analytical Column, equilibrated, with test chromatogram, 1 pc. (1x)
62110/epi Trap Column, equilibrated, with test chromatogram, 1 pc. (2x)
62029 3PLUS1® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
0311 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level I (lyoph.), 1 ml (5x)
0312 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level II (lyoph.), 1 ml (5x)

 

Order No. Required components (not included in the kit)
62017 Tuning Mix 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 (Analytes and Internal Standards), 1 ml

 

Order No. Lab equipment
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:
  1. 62011 Mobile Phase A 25-OH-Vitamin-D3-D2
    Mobile Phase A
    Order no.: 62011

    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma

    Details
  2. 62022 Mobile Phase B 25-OH-Vitamin-D3-D2
    Mobile Phase B
    Order no.: 62022
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 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. 62045 Internal Standard Mix
    Internal Standard Mix
    Order no.: 62045
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 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. 96 Well Filter Plates
    96 Well Filter Plates
    Order no.: 62057
    Details
  7. 96 Well Collection Plate
    Collection Plates
    Order no.: 62058
    Details
  8. Pierceable Adhesive Seals for 96 Well Plates
    Pierceable Adhesive Seals for 96 Well Plates
    Order no.: 62059
    Details
  9. 62130 Analytical Column 25-OH-Vitamin D3-D2
    Analytical Column
    Order no.: 62130
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma
    Details
  10. 62110_epi trap column epimers vitamin D
    Trap Column
    Order no.: 62110/Epi
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma – LC-MS/MS
    Details
  11. 62029 3PLUS1 Calibrator 3-epi Vitamin D - LCMS
    3PLUS1® Multilevel Serum Calibrator Set (including epimeric form of Vitamin D3)
    Order no.: 62029
    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Details
  12. 0310 0311 0312 3-epi Vitamin D Controls
    MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Controls
    Order no.: 0310/0311/0312

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

    Details
The following products are not included in the kit but are required for the application of the method:
  1. 62017 Tuning Mix D3-D2
    Tuning Mix 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3
    Order no.: 62017
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma - LC-MS/MS

    Details
  1. 62029 3PLUS1 Calibrator 3-epi Vitamin D - LCMS
    3PLUS1® Multilevel Serum Calibrator Set (including epimeric form of Vitamin D3)
    Order no.: 62029
    MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
    Details
  1. 0310 0311 0312 3-epi Vitamin D Controls
    MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Controls
    Order no.: 0310/0311/0312

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

    Details
  1. 62130 Analytical Column 25-OH-Vitamin D3-D2
    Analytical Column
    Order no.: 62130
    MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma
    Details
  1. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details
  3. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  4. Stainless Steel Prefilter - 0.5 µm
    Stainless Steel Prefilter - 0.5 µm
    Order no.: 15071
    Details
As a customer please login or register to gain full access.