Vitamin C in Plasma/Serum - HPLC

• 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.

CHROMATOGRAM

TECHNICAL DATA

Method of Analysis

Number of Tests

Parameter

Analysis Time

Limit of quantification

0.4 mg/l

Linearity

0.4–100 mg/l

Recovery

Intraassay

CV ≤ 3.8 %

Interassay

CV ≤ 4.8 %

Specimen

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.

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

Additional Info

Injection Volume

Flow rate

Gradient

Column temperature

Wavelength

Please note

KIT COMPONENTS

Components available separately

Mobile Phase

Order no.: 65001

DETAILS

Mobile Phase, 10 x 1000ml

Order no.: 65002

DETAILS

Internal Standard

Order no.: 65044

DETAILS

Reaction vials

Order no.: 33005

DETAILS

Vitamin C Plasma Calibration Standard

Order no.: 65003

DETAILS

ACCESSORIES

PEEK prefilter housing

Order no.: 15010

DETAILS

HPLC column

Order no.: 65100

DETAILS

PEEK-encased prefilter, 5 µm

Order no.: 15009

DETAILS

Precolumn cartridge 4/10

Order no.: 18065

DETAILS

Precolumn cartridge holder 4/10

Order no.: 18001

DETAILS

CALIBRATORS

Calibrators available separately

Vitamin C Plasma Calibration Standard

Order no.: 65003

DETAILS

CONTROLS

Controls available separately

Vitamin C Plasma Controls

DETAILS