Vitamins A and E in Serum/Plasma - HPLC

Order No.: 34000, for 100 tests
Parameters:
Vitamin A, Vitamin E

2 specific precipitation steps
Easy handling
Low costs per test

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

  • 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.

More Information
Number of Tests 100
Limit of quantification

Vitamin A: 0.02 mg/l
Vitamin E: 0.25 mg/l

Linearity

Vitamin A: 0.02-5.0 mg/l 
Vitamin E: 0.25-45.00 mg/l

Recovery Vitamin A: 106 %; Vitamin E: 101 %
Intraassay

Vitamin A: CV ≤ 2.5 %

Vitamin E: CV ≤ 2.4 %

Interassay

Vitamin A: CV ≤ 5.0 %

Vitamin E: CV ≤ 4.9 %

Analysis Time approx. 9 min
Sample Preparation
  • Pipette 200 µl serum/plasma + 20 µl Internal Standard + 25 µl Precipitation Reagent I into a light protected reaction vial.
  • Mix 30 s (vortex).
  • Add 400 µl Precipitation Reagent II, vortex again for 30 s.
  • Centrifuge 10 min at 9000 x g.
  • Inject 50 µl of the supernatant into the HPLC system.
Sample Stability Protect prepared samples from light, at +2 to +8°C stable up to 12 hours.
Specimen Serum/Plasma
Pre-analytic Treatment

Time of sample drawing

  • In the morning before breakfast
  • Prior to any medication

Collection and storage of patient specimens

  • Collect about 1 to 2 ml blood.
  • Centrifugate 5 min at 2000 x g.
  • Store serum/plasma at -18 °C and protect from light until analysed.
Injection Volume 50 µl
Flow rate 1.5 ml/min
Column temperature ambient (~ 25 °C)
Wavelength

325 nm, after 3.5 min switch to 295 nm

Gradient Isocratic
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.
Method of Analysis HPLC
Parameter Vitamin A, Vitamin E
Complete Specification 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..

Injection volume: 50 µl
Flow rate: 1.5 ml/min
Wavelengths: 325 nm,
after 3.5 min switch to 295 nm
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.

  1. 34001 HPLC vitamin A vitamin E serum plasma mobile phase
    Mobile Phase
    Order no.: 34001
    Vitamins A and E in Serum/Plasma - HPLC
    Details
  2. 34001 HPLC vitamin A vitamin E serum plasma mobile phase
    Mobile Phase, 10 x 1000 ml
    Order no.: 34002
    Vitamins A and E in Serum/Plasma - HPLC
    Details
  3. 34006 vitamin A vitamin E serum plasma precipitation reagent
    Precipitation Reagent I
    Order no.: 34006
    Vitamins A and E in Serum/Plasma - HPLC/UHPLC
    Details
  4. 34003 vitamin A vitamin E serum plasma precipitation reagent
    Precipitation Reagent II
    Order no.: 34003
    Vitamins A and E in Serum/Plasma - HPLC
    Details
  5. 34004 vitamin A vitamin E serum plasma calibration standard
    Vitamins A and E Serum Calibration Standard
    Order no.: 34004
    Vitamins A and E in Serum/Plasma- HPLC/UHPLC
    Details
  6. 34008 vitamin A vitamin E serum plasma internal standard
    Internal Standard
    Order no.: 34008
    Vitamins A and E in Serum/Plasma - HPLC
    Details
  7. Reaction Vials, 1.5 ml
    Reaction Vials, 1.5 ml
    Order no.: 33005
    Amber colour, light protected
    Details
  1. 34004 vitamin A vitamin E serum plasma calibration standard
    Vitamins A and E Serum Calibration Standard
    Order no.: 34004
    Vitamins A and E in Serum/Plasma- HPLC/UHPLC
    Details
  1. 0032 0036 0037 HPLC vitamin A vitamin E serum controls
    Vitamins A and E Serum Controls
    Order no.: 0032/0036/0037
    Details
  1. PEEK Prefilter Housing
    PEEK Prefilter Housing
    Order no.: 15010
    Details
  2. 34100 HPLC vitamin A vitamin E serum plasma analytical column
    HPLC Column Vitamins A and E in Serum/Plasma
    Order no.: 34100
    Vitamins A and E in Serum/Plasma - HPLC
    Details
  3. PEEK-encased Prefilter, 5 µm
    Details
  4. Precolumn Cartridge Holder 4/10
    Details
  5. Precolumn Cartridge 4/10
    Precolumn Cartridge 4/10
    Order no.: 18034
    Vitamins A and E - HPLC/UHPLC
    Details
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