Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – HPLC Reagent Kit
Each parameter safeguarded by a specific internal standard
No post-column derivatisation required
One sample preparation and one run for both parameters
Assay validated according to IVDR (=> Declaration of Conformity)
Chromatogram of a whole blood calibrator
Chromatogram of a plasma calibrator
Vitamin B1 (TPP, thiamine pyrophosphate)
Vitamin B6 (PLP, pyridoxal 5’-phosphate)
Clinical relevance
This assay allows the quantitative determination of the physiologically active forms of vitamin B1, thiamine pyrophosphate, in human whole blood samples and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood or plasma samples.
It is intended to be used for for screening and/or monitoring of vitamin B1 and/or B6 levels where indicated
• in patients with suspected vitamin B1 and/or B6 deficiency,
• in patients with suspected vitamin B1 and/or B6 excess,
• in patients under vitamin B1 and/or B6 supplementation therapy.
Product advantages
• Specific internal standard for each parameter
• No post-column derivatisation required
• One sample preparation and one run for both parameters
Assay characteristics
Thiamine pyrophosphate (TPP) and pyridoxal 5’-phosphate (PLP) can be determined in one run, with one sample preparation and without complex post-column derivatisation. Sample preparation is restricted to a single precipitation step and subsequent derivatisation and is possible with Reaction Vials (assays 52052 and 52052/Premix) or 96 Well Filter Plates (assay 52752/F). Pre-mixed Neutralisation Tubes simplify sample preparation when performing the 52052/Premix assay. Chromatographic separation is carried out with two Mobile Phases by HPLC. The derivatised analytes are determined by fluorescence detection with wavelength switching. The use of two internal standards assures high precision and safety in quantifying the analytes.
Two types of kits available
Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators
|
52052-BK Basic Kit |
|
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x) |
Reagent Kit
• Mobile phase and calibrator included
|
52052 Reagent Kit |
|
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x) |
| 52001 | Mobile Phase A, 1000 ml (1x) |
| 52022 | Mobile Phase B, 1000 ml (1x) |
| 52003 | Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (1x) |
| Method of Analysis | HPLC |
|---|---|
| Number of Tests | 100 |
| 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 the appendix of the instruction manual. |
| Lower and Upper Limit of Quantitation | TPP 3.5 – 750 µg/l (whole blood) |
| Specimen | Whole Blood / 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.
|
| Run Time | 7 min (plasma), 9 min (whole blood) |
| Injection Volume | 25 μl |
| Flow Rate | 1.5–2.3 ml/min |
| Column Temperature | +20 to +25 °C |
| Gradient | binary |
| Wavelengths | Start EX 320 nm, EM 415 nm, |
| Additional Info | Any common HPLC system with binary pump and programmable fluorescence detector is suitable. |
| Parameters | Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate) |
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – HPLC Reagent Kit (52052)
For 100 tests
| Order No. | Reagent Kit components (Quantities in kit) |
| 52001 | Mobile Phase A, 1000 ml (1x) |
| 52022 | Mobile Phase B, 1000 ml (1x) |
| 52003 | Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (1x) |
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour, 100 pcs. (2x) |
| Order No. | Required components (not included in the Reagent Kit) |
| 52100 | HPLC Column, equilibrated, with test chromatogram, 1 pc. |
| Required for the analysis of vitamin B1 and vitamin B6 in whole blood: | |
| 0165 | Vitamin B1/B6 Whole Blood Control, Level I, 5 x 2.0 ml (lyoph.) |
| 0167 | Vitamin B1/B6 Whole Blood Control, Level II, 5 x 2.0 ml (lyoph.) |
| Required for the analysis of vitamin B6 in plasma: | |
| 36005 | Plasma Calibration Standard, 5 x 1.0 ml (lyoph.) |
| 0038 | Vitamin B6 Plasma Control, Level I, 5 x 2.0 ml (lyoph.) |
| 0039 | Vitamin B6 Plasma Control, Level II, 5 x 2.0 ml (lyoph.) |
| Order No. | Lab equipment |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. |
| 15010 | PEEK Prefilter Housing, 1 pc. |
| 15011 | PEEK-encased Prefilter, 2 µm, 5 pcs. |
| 18001 | Precolumn Cartridge Holder 4/10, 1 pc. |
| 18010 | Precolumn Cartridge 4/10, 1 pc. |
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Mobile Phase AOrder no.: 52001Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
Mobile Phase BOrder no.: 52022Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
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Vitamins B1/B6 Whole Blood Calibration StandardOrder no.: 52003Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Internal StandardOrder no.: 52044Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
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Precipitation ReagentOrder no.: 52005Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Neutralisation ReagentOrder no.: 52006Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
Derivatisation Reagent 1 (lyoph.)Order no.: 52007Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC -
Derivatisation Reagent 2Order no.: 52008Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/PlasmaOrder no.: 52100Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
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-
-
Vitamins B1/B6 Whole Blood Calibration StandardOrder no.: 52003
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/PlasmaOrder no.: 52100
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
Chromatogram of a whole blood calibrator
Chromatogram of a plasma calibrator
Vitamin B1 (TPP, thiamine pyrophosphate)
Vitamin B6 (PLP, pyridoxal 5’-phosphate)
Clinical relevance
This assay allows the quantitative determination of the physiologically active forms of vitamin B1, thiamine pyrophosphate, in human whole blood samples and of vitamin B6, pyridoxal 5‘-phosphate, in human whole blood or plasma samples.
It is intended to be used for for screening and/or monitoring of vitamin B1 and/or B6 levels where indicated
• in patients with suspected vitamin B1 and/or B6 deficiency,
• in patients with suspected vitamin B1 and/or B6 excess,
• in patients under vitamin B1 and/or B6 supplementation therapy.
Product advantages
• Specific internal standard for each parameter
• No post-column derivatisation required
• One sample preparation and one run for both parameters
Assay characteristics
Thiamine pyrophosphate (TPP) and pyridoxal 5’-phosphate (PLP) can be determined in one run, with one sample preparation and without complex post-column derivatisation. Sample preparation is restricted to a single precipitation step and subsequent derivatisation and is possible with Reaction Vials (assays 52052 and 52052/Premix) or 96 Well Filter Plates (assay 52752/F). Pre-mixed Neutralisation Tubes simplify sample preparation when performing the 52052/Premix assay. Chromatographic separation is carried out with two Mobile Phases by HPLC. The derivatised analytes are determined by fluorescence detection with wavelength switching. The use of two internal standards assures high precision and safety in quantifying the analytes.
Two types of kits available
Basic Kit
• Only sample preparation reagents and vials included
• No excess quantities of mobile phase and calibrators
|
52052-BK Basic Kit |
|
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x) |
Reagent Kit
• Mobile phase and calibrator included
|
52052 Reagent Kit |
|
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. (2x) |
| 52001 | Mobile Phase A, 1000 ml (1x) |
| 52022 | Mobile Phase B, 1000 ml (1x) |
| 52003 | Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (1x) |
| Method of Analysis | HPLC |
|---|---|
| Number of Tests | 100 |
| 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 the appendix of the instruction manual. |
| Lower and Upper Limit of Quantitation | TPP 3.5 – 750 µg/l (whole blood) |
| Specimen | Whole Blood / 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.
|
| Run Time | 7 min (plasma), 9 min (whole blood) |
| Injection Volume | 25 μl |
| Flow Rate | 1.5–2.3 ml/min |
| Column Temperature | +20 to +25 °C |
| Gradient | binary |
| Wavelengths | Start EX 320 nm, EM 415 nm, |
| Additional Info | Any common HPLC system with binary pump and programmable fluorescence detector is suitable. |
| Parameters | Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate) |
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood / Plasma – HPLC Reagent Kit (52052)
For 100 tests
| Order No. | Reagent Kit components (Quantities in kit) |
| 52001 | Mobile Phase A, 1000 ml (1x) |
| 52022 | Mobile Phase B, 1000 ml (1x) |
| 52003 | Whole Blood Calibration Standard, 5 x 1.0 ml (lyoph.) (1x) |
| 52044 | Internal Standard, 10 ml (1x) |
| 52005 | Precipitation Reagent, 30 ml (1x) |
| 52006 | Neutralisation Buffer, 25 ml (1x) |
| 52007 | Derivatisation Reagent 1, 2 x 0.3 ml (1x) |
| 52008 | Derivatisation Reagent 2, 15 ml (1x) |
| 33005 | Reaction Vials, 1.5 ml, amber colour, 100 pcs. (2x) |
| Order No. | Required components (not included in the Reagent Kit) |
| 52100 | HPLC Column, equilibrated, with test chromatogram, 1 pc. |
| Required for the analysis of vitamin B1 and vitamin B6 in whole blood: | |
| 0165 | Vitamin B1/B6 Whole Blood Control, Level I, 5 x 2.0 ml (lyoph.) |
| 0167 | Vitamin B1/B6 Whole Blood Control, Level II, 5 x 2.0 ml (lyoph.) |
| Required for the analysis of vitamin B6 in plasma: | |
| 36005 | Plasma Calibration Standard, 5 x 1.0 ml (lyoph.) |
| 0038 | Vitamin B6 Plasma Control, Level I, 5 x 2.0 ml (lyoph.) |
| 0039 | Vitamin B6 Plasma Control, Level II, 5 x 2.0 ml (lyoph.) |
| Order No. | Lab equipment |
| 33005 | Reaction Vials, 1.5 ml, amber colour (light protection), 100 pcs. |
| 15010 | PEEK Prefilter Housing, 1 pc. |
| 15011 | PEEK-encased Prefilter, 2 µm, 5 pcs. |
| 18001 | Precolumn Cartridge Holder 4/10, 1 pc. |
| 18010 | Precolumn Cartridge 4/10, 1 pc. |
-
Mobile Phase AOrder no.: 52001
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
Mobile Phase BOrder no.: 52022
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
Vitamins B1/B6 Whole Blood Calibration StandardOrder no.: 52003
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Internal StandardOrder no.: 52044
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Precipitation ReagentOrder no.: 52005
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Neutralisation ReagentOrder no.: 52006
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
Derivatisation Reagent 1 (lyoph.)Order no.: 52007Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
Derivatisation Reagent 2Order no.: 52008
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/PlasmaOrder no.: 52100
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC
-
-
-
Vitamins B1/B6 Whole Blood Calibration StandardOrder no.: 52003
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC
-
HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/PlasmaOrder no.: 52100
Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC