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

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 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

This assay allows the quantitative determination of 25-hydroxycholecalciferol (25-OH-vitamin D3) and 25-hydroxyergocalciferol (25-OH-vitamin D2) 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 assay is further intended to be used as an aid to diagnosis of diseases for which determination of vitamin D levels is indicated.

 

Assay characteristics

Manual sample preparation is limited to simple and effective protein precipitation. A trap column concentrates the analytes and separates interfering substances. A two-position-six-port-valve connects the trap column to an HPLC column where the chromatographic separation takes place. The use of atmospheric pressure chemical ionisation (APCI) and a deuterated internal standard assures precision and robustness and minimises ion suppression effects.

This kit does not separate 3-epi-25-OH-vitamin D3 and 3-epi-25-OH-vitamin D2 from 25-OH-vitamin D3 and 25-OH-vitamin D2, respectively, and should therefore only be used to analyse samples that contain none or just negligible amounts of these metabolites.
We recommend the use of the assay MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in serum/plasma (order no. 62062) for chromatographic separation of the diastereomers and quantitation of 3-epi-25-OH-vitamin D3 to ensure correct assessment of the vitamin D status in patient samples that might contain 3-epi-25-OH-vitamin D3/D2.

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

 

Content for 200 tests (62000)
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 (62000/1000)
Mobile Phase A, 1000 ml (5x)
Mobile Phase B, 1000 ml (5x)
Precipitation Reagent, 5 ml (5x)
Internal Standard, 40 ml (5x)
Rinsing Solution, 1000 ml (2x)
Reaction Vials, 100 pcs. (10x)
3PLUS1® 25-OH-Vitamin D3/D2 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
MassCheck® 25-OH-Vitamin D3/D2 Serum Control, Level I (lyoph.), 1 ml (5x)
MassCheck® 25-OH-Vitamin D3/D2 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: 1.0 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.0 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 5.0 or 6.0 min (depending on gradient system)
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature Analytical Column and Trap Column: +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

This assay allows the quantitative determination of 25-hydroxycholecalciferol (25-OH-vitamin D3) and 25-hydroxyergocalciferol (25-OH-vitamin D2) 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 assay is further intended to be used as an aid to diagnosis of diseases for which determination of vitamin D levels is indicated.

 

Assay characteristics

Manual sample preparation is limited to simple and effective protein precipitation. A trap column concentrates the analytes and separates interfering substances. A two-position-six-port-valve connects the trap column to an HPLC column where the chromatographic separation takes place. The use of atmospheric pressure chemical ionisation (APCI) and a deuterated internal standard assures precision and robustness and minimises ion suppression effects.

This kit does not separate 3-epi-25-OH-vitamin D3 and 3-epi-25-OH-vitamin D2 from 25-OH-vitamin D3 and 25-OH-vitamin D2, respectively, and should therefore only be used to analyse samples that contain none or just negligible amounts of these metabolites.
We recommend the use of the assay MassChrom® 25-OH-Vitamin D3/D2 and 3-epi-25-OH-Vitamin D3 in serum/plasma (order no. 62062) for chromatographic separation of the diastereomers and quantitation of 3-epi-25-OH-vitamin D3 to ensure correct assessment of the vitamin D status in patient samples that might contain 3-epi-25-OH-vitamin D3/D2.

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

 

Content for 200 tests (62000)
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 (62000/1000)
Mobile Phase A, 1000 ml (5x)
Mobile Phase B, 1000 ml (5x)
Precipitation Reagent, 5 ml (5x)
Internal Standard, 40 ml (5x)
Rinsing Solution, 1000 ml (2x)
Reaction Vials, 100 pcs. (10x)
3PLUS1® 25-OH-Vitamin D3/D2 Multilevel Serum Calibrator Set (lyoph.), 1 ml (4x)
MassCheck® 25-OH-Vitamin D3/D2 Serum Control, Level I (lyoph.), 1 ml (5x)
MassCheck® 25-OH-Vitamin D3/D2 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: 1.0 µg/l – 250 µg/l
25-OH-Vitamin D2: 1.0 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 5.0 or 6.0 min (depending on gradient system)
Injection Volume ≤ 50 µl (mass spectrometer dependent)
Column Temperature Analytical Column and Trap Column: +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.