Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC

Order No.: 52752/F, for 960 tests
Parameters:
Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Significantly reduced hands-on time for sample preparation
Accurate and fast results for both vitamins
Each parameter safeguarded by a specific internal standard

Vitamin B1 (TPP, thiamine pyrophosphate)

Vitamin B6 (PLP, pyridoxal 5’-phosphate)

Clinical relevance

Vitamin B1 (thiamine) plays an important role in deriving energy from carbohydrates and fats. The active form is thiamine pyrophosphate (TPP), a coenzyme for some enzymatic reactions in carbohydrate metabolism and for pyruvate decarboxylase. TPP is also significantly involved in the oxidative degradation of glucose. The determination of the TPP concentration is more significant than measurements of total thiamine, as by this method only the physiologically active form is taken into account.

Vitamin B6 comprises pyridoxine, pyridoxamine and pyridoxal, as well as their phosphate derivatives. It is ingested with food and transferred via several enzymatic conversions into its active form, pyridoxal-5’-phosphate (PLP). PLP is involved in nearly one hundred different enzymatic reactions, among other things, as co-factor in amino acid metabolism, and in the formation of haemoglobin or neurotransmitters in the brain.

 

Product advantages

  • Automated sample preparation for minimised workload in the laboratory
  • Reliable and reproducible results with a specific internal standard for each parameter

 

This HPLC kit facilitates the combined analysis of vitamin B1 in whole blood and vitamin B6 in whole blood or plasma. Sample preparation is automated in 96 well filter plates for maximum output and minimal workload. The internal standard for each of the two parameters and special quality controls ensure precise and reliable quantification.

Chromsystems also offers two kits that feature manual determination using HPLC: a standard version as well as a kit for simplified sample preparation with pre-mixed tubes.

Alternatively, an automated method using UHPLC is also available.

More Information
Number of Tests 960
Limit of quantification

PLP 2.0 μg/l
TPP 7.0 μg/l

Linearity

PLP 500 μg/l
TPP 750 μg/l

Intraassay

CV < 3 %

Interassay

CV < 5 %

Analysis Time 9 min
Sample Preparation
  • Add 100 μl whole blood/calibrator/control and 50 μl Internal Standard Mix in a 96 Well Collection Plate.
  • Add 150 μl Extraction Reagent.
  • Shake for 4 min.
  • Remove precipitate by centrifugation (3 min, 2000 x g).
  • Transfer 75 μl of supernatant in a 96 well filter plate.
  • Add 75 μl Prep Solution and 50 μl of finisher solution.
  • Mix for 30 s.
  • Remove precipitate by centrifugation (2.5 min, 1500 x g).
  • Incubate supernatant for 45 min at 60 °C.
  • Add 100 μl Dilution Solution, mix briefly (30 s).
  • Seal plate and transfer to autosampler.
Sample Stability Samples are stable up to 1 week at +2 to +8 °C and up to 2 weeks at below -18 °C. Protect from light.
Specimen Whole Blood
Injection Volume 25–50 μl
Flow rate 1.5–2.3 ml/min
Column temperature ambient (~ 25°C)
Wavelength

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

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.
Method of Analysis HPLC
Parameter Vitamin B1 (TPP, thiamine pyrophosphate), Vitamin B6 (PLP, pyridoxal 5’-phosphate)
  1. 52001 HPLC mobile phase A vitamin B1 vitamin B6 whole blood plasma
    Mobile Phase A
    Order no.: 52001

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

    Details
  2. 52022 HPLC mobile phase B vitamin B1 vitamin B6 whole blood plasma
    Mobile Phase B
    Order no.: 52022

    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

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

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  4. 52744 internal standard mix automation vitamin B1 vitamin B6 whole blood
    Internal Standard Mix
    Order no.: 52744

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  5. 52705 extraction reagent automation vitamin B1 vitamin B6 whole blood
    Extraction Reagent
    Order no.: 52705

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  6. 52706 prep solution automation vitamin B1 vitamin B6 whole blood
    Prep Solution
    Order no.: 52706

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  7. 52707 Finisher 1 automation vitamin B1 vitamin B6 whole blood
    Finisher 1
    Order no.: 52707

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  8. 52708 Finisher 2 automation vitamin B1 vitamin B6 whole blood
    Finisher 2
    Order no.: 52708

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  9. 52709 dilution buffer automation vitamin B1 vitamin B6 whole blood
    Dilution Buffer
    Order no.: 52709

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  10. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    96 Well Filter Plates
    Order no.: 52057

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  11. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    96 Well Collection Plates
    Order no.: 52058

    Vitamin B1/B6 in Whole Blood, fully automated analysis - UHPLC

    Details
  12. 52057 52058 52059 well filter plates vitamin B1 vitamin B6
    Pierceable adhesive seals for 96 Well Plates
    Order no.: 52059

    Vitamin B1/B6 in Whole Blood, fully automated analysis - HPLC/UHPLC

    Details
  13. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC column
    Order no.: 52100

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

    Details
  1. 52003 HPLC whole blood calibration standard vitamin B1 vitamin B6
    Vitamins B1/B6 Whole Blood Calibration Standard
    Order no.: 52003

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

    Details
  1. 0164 0165 0167 HPLC vitamin B1 vitamin B6 whole blood controls
    Vitamins B1/B6 Whole Blood Controls
    Order no.: 0164/0165/0167
    Details
  1. PEEK-encased prefilter, 2 µm
    Details
  2. PEEK prefilter housing
    PEEK prefilter housing
    Order no.: 15010
     
    Details
  3. 52100 HPLC column vitamin B1 vitamin B6 whole blood plasma
    HPLC column
    Order no.: 52100

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

    Details
  4. Precolumn cartridge holder 4/10
    Details
  5. Precolumn cartridge 4/10
    Precolumn cartridge 4/10
    Order no.: 18052
    Vitamins B1 and B6 - HPLC/UHPLC
    Details
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