One Step Vitamins A and E in Serum/Plasma - UHPLC

Order No.: 34900/UHPLC, for 1000 tests
Parameters:
Vitamin A, Vitamin E

High throughput UHPLC methode
One step sample preparation with pre-mixed reaction tubes
Fully automated workflow available

Vitamin A

Vitamin E

Clinical relevance

Vitamins A and E are fat-soluble vitamins, which perform important functions in the body.

Vitamin A plays an important role in vision, bone metabolism, and the synthesis of steroid hormones. Deficiency of vitamin A can lead to night blindness, skin dehydration, and hair loss.

Vitamin E protects, among others, unsaturated fatty acids in the cell membranes and the LDL cholesterol from attacks by reactive radicals, which can occur as a result of increased oxidative stress in the body. Deficiency of vitamin E can lead to permanent joint pain, making the determination of vitamin E levels essential in this case.

However, overdoses of these vitamins may cause hypervitaminosis with intoxication symptoms. In addition, chemotherapies should not be "accompanied" by extra vitamin supplements – especially high doses of vitamins A and E may limit the success of treatment.

 

Product advantages

  • High throughput UHPLC method
  • One-step sample preparation with pre-mixed tubes
  • Automated version available

 

The new Chromsystems UHPLC kit for the analysis of vitamins A and E in plasma/serum combines the reliability of the Chromsystems method with the throughput and speed of UHPLC technology. Sample preparation is reduced to a single pipetting step using pre-mixed reaction tubes, which contain all necessary reagents and the customized internal standard, so all that is left to do is add the sample. Separation of the analytes is carried out in less than 4 minutes. For large numbers of samples there is also a method using 96-well filter plates available for semi-automated or automated sample preparation with a pipetting robot.

 

Content for 1000 Tests
Mobile Phase (for UHPLC), 2 x 500 ml
Pre-mixed Reaction Tubes, 10 x 100 pcs.
Serum Calibration Standard Vitamins A and E (lyoph.), 5 x 1 ml
UHPLC Column, 1 pc.
UHPLC Precolumn Cartridge, 3 x 1 pc.

More Information
Method of Analysis UHPLC
Number of Tests 1000
Limit of quantification Vitamin A: 0.005 mg/l
Vitamin E: 0.100 mg/l
Linearity Vitamin A: up to 4.5 mg/l
Vitamin E: up to 38 mg/l
Recovery Vitamin A: 104 %; Vitamin E: 101 %
Intraassay

Vitamin A: CV = 0.6-1.3 %
Vitamin E: CV = 0.5-1.4 %

Interassay

Vitamin A: CV = 2.9-3.1 %
Vitamin E: CV = 2.3-3.0 %

Analysis Time 3.5 min
Specimen Serum/Plasma
Pre-analytic Treatment

Time of sample drawing

  • In the morning before breakfast
  • Prior to any medication

Collection and storage of specimens

  • Collect about 1 to 2 ml blood.
  • Centrifugate 5 min at 2000 x g.
  • Store serum/plasma below -18 °C and protect from light until analysed.
Sample Preparation
  • Pipette 100 µl serum/plasma into a Pre-mixed Reaction Tube.
  • Mix 30 s (vortex).
  • Centrifuge 10 min at at least 9000 x g.
  • Transfer supernatant into a light protected autosampler vial.
  • Inject 5 µl into the UHPLC system.
Sample Stability The prepared, lightprotected samples are stable at ambient temperature for at least 24 h and at +2 to +8°C up to 48 h. For longer storage (up to 3 weeks) keep samples below -18 °C.
Injection Volume 5 µl
Flow rate 0.6 ml/min
Column temperature 35 °C
Wavelength 325 nm, after 1.30 min switch to 295 nm
Gradient Isocratic
Additional Info

For the analysis of vitamins A and E in serum/plasma, any major UHPLC system with isocratic pump, programmable UV detector and column oven is suitable.

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.
Parameter Vitamin A, Vitamin E
Pressure approx. 400 bar
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