Vitamin C in Plasma/Serum - HPLC

Order No.: 65065, for 100 tests
Parameters:
Vitamin C

Simplified sample preparation
Enhanced column stability
High stability of standards and controls

Vitamin C

Clinical relevance

Vitamin C (ascorbic acid, ascorbate) is a water-soluble vitamin that is an essential cofactor for the human body, for instance in the catabolism of tyrosine and the synthesis of catecholamines. Regular consumption of fruit and vegetables is usually sufficient to cover the body’s vitamin C needs.

In the case of vitamin C deficiency, procollagen chains are produced insufficiently and stable fibrils cannot be formed in the affected cells. Without functioning collagen, the blood vessels and tendons become fragile and the typical skin-related symptoms associated with scurvy start to develop. In addition to the collagen damages described above, further diseases caused by vitamin C deficiency include bleeding of uncertain origin, loss of teeth and overactive thyroid gland (hyperthyroidism). Vitamin C deficiency is also observed in dialysis patients, in chronic alcohol abuse and during pregnancy.

 

Product advantages

  • High stability of processed samples and controls
  • Enhanced column stability
  • Reliable quantification with a stable internal standard

 

This Chromsystems kit allows the reliable chromatographic determination of vitamin C in one isocratic HPLC run with UV detection. An efficient protein precipitation step removes interfering components and stabilises Vitamin C simultaneously. A stable internal standard is used for reliable quantification. Due to a special stabilisation of the prepared samples, it is also possible to reliably process and measure large sample batches.

 

More Information
Number of Tests 100
Limit of quantification

0.4 mg/l

Linearity

0.4–100 mg/l

Recovery 97–103 %
Intraassay

CV ≤ 3.8 %

Interassay

CV ≤ 4.8 %

Analysis Time 5 min
Sample Preparation
  • Pipette 100 μl sample/calibrator/control into a labelled light protected reaction vial.

  • Add 100 μl Internal Standard and mix 30 s (vortex).

  • Centrifuge 5 min at at least 13000 x g.

  • Inject 20 μl of the supernatant into the HPLC system.

Sample Stability The patient samples are stable for 12 h at room temperatur, at +2 to +8 °C for at least 2 weeks. For longer storage (up to 6 weeks) keep samples below -18 °C.
Specimen Plasma/Serum
Pre-analytic Treatment

Fresh lithium heparin plasma is used for analysis. To minimise the oxidation of vitamin C the whole blood sample should be centrifuged immediately after collection and the plasma samples must then be transported deep frozen. Start sample preparation immediately after thawing. The stability for vitamin C in heparin plasma is 5 days at -20 °C. At room temperature this is only up to two hours. If whole blood is used for transportation, store at +2 to +8 °C , do not freeze.

Injection Volume 20 µl
Flow rate 1.5 ml/min
Column temperature ambient (~25 °C)
Wavelength 245 nm
Gradient Isocratic
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 C
Additional Info Any isocratic HPLC system with UV detection is suitable.

  1. 65001 HPLC mobile phase vitamin C plasma serum
    Mobile Phase
    Order no.: 65001
    Vitamin C in Plasma/Serum - HPLC
    Details
  2. 65001 HPLC mobile phase vitamin C plasma serum
    Mobile Phase, 10 x 1000ml
    Order no.: 65002
    Vitamin C in Plasma/Serum - HPLC
    Details
  3. 65044 HPLC internal standard vitamin C plasma serum
    Internal Standard
    Order no.: 65044
    Vitamin C in Plasma/Serum - HPLC
    Details
  4. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  5. 65003 HPLC plasma calibration standard vitamin C
    Vitamin C Plasma Calibration Standard
    Order no.: 65003
    Vitamin C in Plasma/Serum - HPLC
    Details
  1. 65003 HPLC plasma calibration standard vitamin C
    Vitamin C Plasma Calibration Standard
    Order no.: 65003
    Vitamin C in Plasma/Serum - HPLC
    Details
  1. 0074 0075 0076 HPLC vitamin C plasma controls
    Vitamin C Plasma Controls
    Order no.: 0074/0075/0076
    Details
  1. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. 65100 HPLC column vitamin C plasma serum
    HPLC column
    Order no.: 65100
    Vitamin C in Plasma/Serum - HPLC
    Details
  3. PEEK-encased Prefilter, 5 µm
    Details
  4. Precolumn cartridge 4/10
    Precolumn cartridge 4/10
    Order no.: 18065
    Vitamin C in Plasma/Serum - HPLC
    Details
  5. Precolumn Cartridge Holder 4/10
    Details
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