65065 HPLC kit vitamin C plasma serum

Vitamin C in Plasma/Serum - HPLC

Order no.: 65065
100 Tests
  • Vitamin C

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

Description

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.

 

Kit content

Mobile Phase, 1000 ml
Vitamin C Plasma Calibration Standard (lyoph.), 5 x 0.5 ml
Internal Standard, 10 ml
Reaction vials, amber coloured (light protection), 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Vitamin C

Analysis Time

5 min

Limit of quantification

0.4 mg/l

Linearity

1-100 mg/l

Recovery

103 %

Intraassay

CV < 4 %

Interassay

CV < 5 %

Gradient

Isocratic

Information

Any isocratic HPLC system with UV detection is suitable.

Column temperature

ambient (~25 °C)

Flow rate

1.5 ml/min

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. If whole blood is used for transportation, store at +2 to +8 °C , do not freeze.

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 15 000 x g.

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

  • Storage of prepared samples: The resulting supernatant is stable up to 12 h at ambient temperature and at least 1 week at +2 to +8 °C. For longer storage, keep

    below -18 °C.

Sample Stability

The stability for vitamin C in heparin plasma is 5 days at -20 °C. At room temperature this is only up to two hours.

Injection Volume

20 µl

Flow

1.5 ml/min

Additional Info

For the Chromsystems HPLC analysis of vitamin C any isocratic HPLC system with UV detection is suitable.

Wavelength

245 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.

Kit Components

Components available separately

Mobile Phase


Order no.: 65001

Internal Standard


Order no.: 65044

Reaction vials


Order no.: 33005

Calibrators

Calibrators available separately

Controls

Controls available separately

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