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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 & 62062/1000
Parameters:
25-OH-Vitamin D2, 25-OH-Vitamin D3, 3-epi-25-OH-Vitamin D3

Isotopically labelled internal standard
Multilevel calibrators

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

Grouped product items
Product Name Qty
MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma (kit for 200 tests)
Order No.: 62062
MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma (complete kit for 1000 tests)
Order No.: 62062/1000
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 200 tests (Order no. 62062)
Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls (Order no. 62062/1000)

 

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.

 

Content for 200 tests (62062)
Mobile Phase A, 1000 ml (1x)
Mobile Phase B, 1000 ml (1x)
Precipitation Reagent, 5 ml (1x)
Internal Standard Mix, 40 ml (1x)
Rinsing Solution, 1000 ml (1x)
Reaction Vials, 100 pcs. (2x)

 

Content for 1000 tests (62062/1000)
Mobile Phase A, 1000 ml (5x)
Mobile Phase B, 1000 ml (5x)
Precipitation Reagent, 5 ml (5x)
Internal Standard Mix, 40 ml (5x)
Rinsing Solution, 1000 ml (2x)
Reaction Vials, 100 pcs. (10x)
3PLUS1® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level I (lyoph.), 1 ml (5x)
MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level II (lyoph.), 1 ml (5x)
Analytical Column, 1 pc. (1x)
Trap Column, 1 pc. (2x)

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. 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 reaction vials:

  • 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 of the supernatant into an autosampler vial.
  • Inject ≤ 50 µl of the supernatant into the LC-MS/MS system

For the sample preparation with 96 well filter plates, please refer to the instruction manual.
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
Order No. MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS
62062 Kit for 200 tests
62062/1000 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)
62011 Mobile Phase A, 1000 ml (1x)
62022 Mobile Phase B, 1000 ml (1x)
62003 Precipitation Reagent, 5 ml (1x)
62045 Internal Standard Mix, 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)
62130 Analytical Column, equilibrated, with test chromatogram, 1 pc.
62110/epi Trap Column, equilibrated, with test chromatogram, 1 pc.
62029 3PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3, 4 x 1 ml (lyoph.)
0311 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control Level I, 5 x 1 ml (lyoph.)
0312 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control Level II, 5 x 1 ml (lyoph.)
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
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:
  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. 3006 reaction vials
    Reaction Vials
    Order no.: 3006
    Details
The following products are not included in the kit but are required for the application of the method:
  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
  2. 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
  3. 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
  4. 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
  5. 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. 3006 reaction vials
    Reaction Vials
    Order no.: 3006
    Details
  2. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  3. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details
  4. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  5. 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 200 tests (Order no. 62062)
Kit for 1000 tests, including Analytical Column, Trap Column, 3PLUS1® Multilevel Calibrator Set and MassCheck® Controls (Order no. 62062/1000)

 

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.

 

Content for 200 tests (62062)
Mobile Phase A, 1000 ml (1x)
Mobile Phase B, 1000 ml (1x)
Precipitation Reagent, 5 ml (1x)
Internal Standard Mix, 40 ml (1x)
Rinsing Solution, 1000 ml (1x)
Reaction Vials, 100 pcs. (2x)

 

Content for 1000 tests (62062/1000)
Mobile Phase A, 1000 ml (5x)
Mobile Phase B, 1000 ml (5x)
Precipitation Reagent, 5 ml (5x)
Internal Standard Mix, 40 ml (5x)
Rinsing Solution, 1000 ml (2x)
Reaction Vials, 100 pcs. (10x)
3PLUS1® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level I (lyoph.), 1 ml (5x)
MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control, Level II (lyoph.), 1 ml (5x)
Analytical Column, 1 pc. (1x)
Trap Column, 1 pc. (2x)

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. 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 reaction vials:

  • 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 of the supernatant into an autosampler vial.
  • Inject ≤ 50 µl of the supernatant into the LC-MS/MS system

For the sample preparation with 96 well filter plates, please refer to the instruction manual.
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
Order No. MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in Serum/Plasma by LC-MS/MS
62062 Kit for 200 tests
62062/1000 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)
62011 Mobile Phase A, 1000 ml (1x)
62022 Mobile Phase B, 1000 ml (1x)
62003 Precipitation Reagent, 5 ml (1x)
62045 Internal Standard Mix, 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)
62130 Analytical Column, equilibrated, with test chromatogram, 1 pc.
62110/epi Trap Column, equilibrated, with test chromatogram, 1 pc.
62029 3PLUS1® Multilevel Serum Calibrator Set 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3, 4 x 1 ml (lyoph.)
0311 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control Level I, 5 x 1 ml (lyoph.)
0312 MassCheck® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 Serum Control Level II, 5 x 1 ml (lyoph.)
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
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:
  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. 3006 reaction vials
    Reaction Vials
    Order no.: 3006
    Details
The following products are not included in the kit but are required for the application of the method:
  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
  2. 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
  3. 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
  4. 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
  5. 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. 3006 reaction vials
    Reaction Vials
    Order no.: 3006
    Details
  2. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  3. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details
  4. 15070 15071 Prefilter and Housing
    Stainless Steel Prefilter Housing
    Order no.: 15070
    Details
  5. 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.