Vitamin C in Plasma/Serum - HPLC
Order no.: 65065
100 Tests 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
HPLC Number of Tests
100 Parameter
Vitamin C Analysis Time
5 min Limit of quantification
0.4 mg/l
Linearity
0.4–100 mg/l
Recovery
97–103 % Intraassay
CV ≤ 3.8 %
Interassay
CV ≤ 4.8 %
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.
Sample Preparation
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Pipette 100 μl sample/calibrator/control into a labelled light protected reaction vial.
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Add 100 μl Internal Standard and mix 30 s (vortex).
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Centrifuge 5 min at at least 13000 x g.
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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. Additional Info
Any isocratic HPLC system with UV detection is suitable.
Injection Volume
20 µl Flow rate
1.5 ml/min Gradient
Isocratic Column temperature
ambient (~25 °C) 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
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