Vitamin C in Plasma/Serum - Automated HPLC

• Vitamin C

Automated sample preparation with 96 well filter plates

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
96 Well filter plates, 5 pcs.
Collection plates, 5 pcs.
Pierceable adhesive seals for 96 well plates, 5 pcs.
 

CHROMATOGRAM

TECHNICAL DATA

Method of Analysis

Number of Tests

Parameter

Analysis Time

Limit of quantification

0.5 mg/l

Linearity

Intraassay

CV <= 3 %

Interassay

Information

Specimen

Pre-analytic Treatment

Specimens: Fresh lithium heparin plasma

Sample stability: 5 days at -20 °C, up to 2 hours at room temperature
Detailed information on stability and transportation can be obtained from the instruction manual.

Sample Preparation

• Place 100 µl Internal Standard in a 96 well filter plate.
• Add 100 µl sample/calibrator/control.
• Vortex for 30 s at 1100 rpm (orbit: 2 mm).
• Centrifuge for 5 min at 2000 x g.
• Close the 96 well filter plate.
• Vortex for 30 s at 800 rpm (orbit: 2 mm).
• Inject 20 µl of the prepared sample into the HPLC system.

Additional Info

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

Injection Volume

Flow rate

Flow

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

Vitamin C Plasma Calibration Standard

Order no.: 65003

DETAILS

Internal Standard

Order no.: 65044

DETAILS

96 Well Filter Plates

Order no.: 65757

DETAILS

Collection Plates

Order no.: 65758

DETAILS

Pierceable adhesive seals for 96 Well Plates

Order no.: 65759

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