MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
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-2022 regulation


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.
Method of Analysis | LC-MS/MS |
---|---|
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. |
Lower Limit of Quantification | 25-OH-Vitamin D2: 2.2 µg/l |
Upper Limit of Quantification | up to 250 μg/l |
Intraassay | CV < 5 % |
Interassay | CV < 6 % |
Run 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 |
|
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. |
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. |
Parameter | 25-OH-Vitamin D2, 25-OH-Vitamin D3 |
Flow | Pump A: 0.02–1.00 ml/min |
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. |
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Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MSOrder no.: 62100
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
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Tuning Mix 25-OH-Vitamin D3/D2Order no.: 62015
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
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Upgrade for 3-epi-25-OH-Vitamin D3/D2 - LC-MS/MSOrder no.: Epimer Upgrade
Upgrade for order no. 62000
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma
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Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MSOrder no.: 62100
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS


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.
Method of Analysis | LC-MS/MS |
---|---|
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. |
Lower Limit of Quantification | 25-OH-Vitamin D2: 2.2 µg/l |
Upper Limit of Quantification | up to 250 μg/l |
Intraassay | CV < 5 % |
Interassay | CV < 6 % |
Run 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 |
|
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. |
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. |
Parameter | 25-OH-Vitamin D2, 25-OH-Vitamin D3 |
Flow | Pump A: 0.02–1.00 ml/min |
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. |
-
Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MSOrder no.: 62100
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
-
Tuning Mix 25-OH-Vitamin D3/D2Order no.: 62015
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS
-
Upgrade for 3-epi-25-OH-Vitamin D3/D2 - LC-MS/MSOrder no.: Epimer Upgrade
Upgrade for order no. 62000
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma
-
Analytical Column 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MSOrder no.: 62100
MassChrom® 25-OH-Vitamin D3/D2 in Serum/Plasma - LC-MS/MS