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Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

Order No.: 52052/RUO, for 100 tests
Parameters:
Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Each analyte safeguarded by a specific internal standard
No post-column derivatisation required
One sample preparation and one run for both analytes

Vitamin B1 (TPP, thiamine pyrophosphate)

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

This Chromsystems kit allows for the HPLC determination of thiamine pyrophosphate and pyridoxal-5’-phosphate, analogues of Vitamin B1 and Vitamin B6, 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 in a reaction vial. The derivatised research sample is analysed using fluorescence detection with a wavelength switching, and the chromatographic separation is performed with 2 mobile phases in less than 9 minutes. The use of a tailor-made internal standard for each parameter ensures precise and accurate results.

More Information
Method of Analysis HPLC
Number of Tests 100
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.
Specimen Whole Blood and Whole Blood/Plasma
Sample Preparation
  • Mix 200 μl reserach sample/calibrator/control with 100 μl Internal Standard and 300 μl Precipitation Reagent in an amber vial (light protection) and shake 30 s (vortex).
  • Centrifuge for 5 min with at least 14000 x g.
  • Pipette 250 μl Neutralisation Reagent and 100 μl derivatisation mix into a new vial. Add 250 μl supernatant obtained above, mix briefly.
  • Incubate for 25 min at 60 °C (waterbath).
  • Cool sample for 10 min at +2 to +8 °C, then centrifuge for 2 min with at least 14000 x g.
  • Transfer supernatant into a light protected autosampler vial, inject 25–50 μl into the HPLC system.
Run Time 7 - 9 min
Injection Volume 25-50 µl
Flow Rate 1.5 - 2.3 ml/min
Column Temperature ambient (~ 25 °C)
Gradient

Gradient profile: specific binary gradients for whole blood and plasma
Wavelengths

Start EX 320 nm, EM 415 nm
after approx. 3.8 min switch to EX 367 nm, EM 435 nm
Additional Info

For the combined analysis of vitamins B1 and B6 in whole blood or plasma every common HPLC system with binary pump and programmable fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)
The following components are included in the kit:
  1. Mobile Phase A
    Mobile Phase A
    Order no.: 52001/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  2. Mobile Phase B
    Mobile Phase B
    Order no.: 52022/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  3. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003
     
    Details
  4. Internal Standard
    Internal Standard
    Order no.: 52044/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  5. Precipitation Reagent
    Precipitation Reagent
    Order no.: 52005/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  6. Neutralisation Reagent
    Neutralisation Reagent
    Order no.: 52006/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  7. Derivatisation Reagent 1
    Derivatisation Reagent 1
    Order no.: 52007/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  8. Derivatisation Reagent 2
    Derivatisation Reagent 2
    Order no.: 52008/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  9. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma
    Order no.: 52100

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  2. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  3. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005
     
    Details
  4. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  5. Precolumn Cartridge 4/10
    Precolumn Cartridge 4/10
    Order no.: 18052/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  1. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005
     
    Details
  2. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003
     
    Details
  1. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  2. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  1. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma
    Order no.: 52100

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  1. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  2. Precolumn Cartridge Holder 4/10
    Precolumn Cartridge Holder 4/10
    Order no.: 18001
    Details
  3. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  4. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details

Vitamin B1 (TPP, thiamine pyrophosphate)

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

This Chromsystems kit allows for the HPLC determination of thiamine pyrophosphate and pyridoxal-5’-phosphate, analogues of Vitamin B1 and Vitamin B6, 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 in a reaction vial. The derivatised research sample is analysed using fluorescence detection with a wavelength switching, and the chromatographic separation is performed with 2 mobile phases in less than 9 minutes. The use of a tailor-made internal standard for each parameter ensures precise and accurate results.

More Information
Method of Analysis HPLC
Number of Tests 100
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.
Specimen Whole Blood and Whole Blood/Plasma
Sample Preparation
  • Mix 200 μl reserach sample/calibrator/control with 100 μl Internal Standard and 300 μl Precipitation Reagent in an amber vial (light protection) and shake 30 s (vortex).
  • Centrifuge for 5 min with at least 14000 x g.
  • Pipette 250 μl Neutralisation Reagent and 100 μl derivatisation mix into a new vial. Add 250 μl supernatant obtained above, mix briefly.
  • Incubate for 25 min at 60 °C (waterbath).
  • Cool sample for 10 min at +2 to +8 °C, then centrifuge for 2 min with at least 14000 x g.
  • Transfer supernatant into a light protected autosampler vial, inject 25–50 μl into the HPLC system.
Run Time 7 - 9 min
Injection Volume 25-50 µl
Flow Rate 1.5 - 2.3 ml/min
Column Temperature ambient (~ 25 °C)
Gradient

Gradient profile: specific binary gradients for whole blood and plasma
Wavelengths

Start EX 320 nm, EM 415 nm
after approx. 3.8 min switch to EX 367 nm, EM 435 nm
Additional Info

For the combined analysis of vitamins B1 and B6 in whole blood or plasma every common HPLC system with binary pump and programmable fluorescence detector is suitable.
Parameters Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)
The following components are included in the kit:
  1. Mobile Phase A
    Mobile Phase A
    Order no.: 52001/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  2. Mobile Phase B
    Mobile Phase B
    Order no.: 52022/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  3. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003
     
    Details
  4. Internal Standard
    Internal Standard
    Order no.: 52044/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  5. Precipitation Reagent
    Precipitation Reagent
    Order no.: 52005/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  6. Neutralisation Reagent
    Neutralisation Reagent
    Order no.: 52006/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  7. Derivatisation Reagent 1
    Derivatisation Reagent 1
    Order no.: 52007/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  8. Derivatisation Reagent 2
    Derivatisation Reagent 2
    Order no.: 52008/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC/UHPLC

    Details
  9. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
The following products are not included in the kit but are required for the application of the method:
  1. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma
    Order no.: 52100

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  2. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  3. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005
     
    Details
  4. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  5. Precolumn Cartridge 4/10
    Precolumn Cartridge 4/10
    Order no.: 18052/RUO

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  1. 36005 HPLC plasma calibration standard vitamin B6
    Vitamin B6 Plasma Calibration Standard
    Order no.: 36005
     
    Details
  2. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003
     
    Details
  1. 0031 0038 0039 HPLC vitamin B6 plasma controls
    Vitamin B6 Plasma Controls
    Order no.: 0031/0038/0039
    Details
  2. 0164_0165_0167_hplc_vitamin_b1_vitamin_b6_whole_blood_controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  1. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC Column Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma
    Order no.: 52100

    Vitamin B1 in Whole Blood and Vitamin B6 in Whole Blood/Plasma - HPLC

    Details
  1. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  2. Precolumn Cartridge Holder 4/10
    Precolumn Cartridge Holder 4/10
    Order no.: 18001
    Details
  3. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  4. PEEK-encased Prefilter, 2 µm
    PEEK-encased Prefilter, 2 µm
    Order no.: 15011
    Details