34000 HPLC kit vitamin A vitamin E serum plasma

Vitamins A and E in Serum/Plasma - HPLC

Order no.: 34000
100 Tests
  • Vitamin A
  • Vitamin E

2 specific precipitation steps
Easy handling
Low costs per test

Description

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

  • Specific method
  • Easy sample preparation
  • Low analysis costs

 

This established kit allows the simultaneous analysis of vitamins A and E in serum/plasma in a single isocratic HPLC run with UV detection. An easy sample preparation is initiating the effective protein precipitation step. A stable internal standard is used for precise quantification.

Chromsystems offers additional kits for the fast and efficient determination of vitamins A and E. You can choose between several options depending on the device capabilities (HPLC or UHPLC) and the sample quantities in your laboratory. Alternatively, the kit 34000/Premix with simplified sample preparation in premixed tubes is available, as well as the 34900/UHPLC kit for higher sample throughput and using UHPLC determination. Our kits are also available with automated workflow (34700/F and 34900/F).

 

Kit content

Mobile Phase, 1000 ml
Precipitation Reagent I, 2,5 ml
Precipitation Reagent II, 40 ml
Vitamins A and E Serum Calibration Standard, 5 x 1 ml (lyoph.)
Internal Standard, 2 ml
Reaction vials, amber coloured (light protection), 100 pcs.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Vitamin A, Vitamin E

Analysis Time

ca. 9 min

Limit of quantification

Vitamin A: 0.02 mg/l
Vitamin E: 0.50 mg/l

Linearity

Vitamin A: 0.02-2.25 mg/l 
Vitamin E: 0.50-45.00 mg/l

Recovery

Vitamin A: 103.9 %; Vitamin E: 97.5 %

Intraassay

CV ≤ 3.1 %

Interassay

CV ≤ 2.5 %

Column temperature

ambient (~25 °C)

Flow rate

1.5 ml/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 rpm.
  • Store serum/plasma at -20 °C and protect from light until analysed.

Sample Preparation

  • Pipette 200 µl serum/plasma + 20 µl Internal Standard + 25 µl Precipitation Reagent I into a light protected reaction vial.
  • Vortex 30 s.
  • Add 400 µl Precipitation Reagent II.
  • Vortex again 30 s.
  • Centrifuge 10 min at 9000 x g.
  • Inject 50 µl of the supernatant into the HPLC system.

Sample Stability

Protect samples from light, at +2 to +8°C stable up to 12 hours. For longer storage periods (up to 3 weeks) keep samples frozen below -18 °C.

Injection Volume

50 µl

Additional Info

For the analysis any isocratic HPLC system with UV detector is suitable. For simultaneous analysis of the vitamins A and E, a detector with programmable wavelength switching is very useful as these vitamins are detected at different wavelengths.

Wavelength

325 nm, after 3.5 min switch to 295 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

Calibrators

Calibrators available separately

Controls

Controls available separately

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