68000 HPLC kit coenzyme Q10 ubiquinone serum plasma whole blood

Coenzyme Q10 in Serum/Plasma/Whole Blood - HPLC

Order no.: 68000
100 Tests
  • Coenzyme Q10

No liquid-liquid extraction
No evaporation step
Stable standards and controls

Description

Clinical relevance

Coenzyme Q10 (ubiquinone, Q10 or UQ) is structurally related to vitamin E and is part of the mitochondrial respiratory chain in many organisms. It is essential for cellular energy production and also acts as an antioxidant in the body. Coenzyme Q10 is ingested with food, but it can also be produced inside the human body. Due to their high levels of energy turnover, heart, lungs and liver exhibit particularly high concentrations of Q10. Deficiencies are rare, but they can be caused by some muscular diseases (myopathy), and then require treatment. It is assumed that a Q10 deficiency may be a cause of cardiovascular disease.

 

Product advantages

  • No liquid-liquid extraction
  • No evaporation step
  • Stable calibrators and controls

 

This Chromsystems kit allows the reliable chromatographic determination of coenzyme Q10 in an isocratic HPLC run with UV detection. First, coenzyme Q10 is released by precipitating the proteins and then concentrated using solid phase extraction. Using this method, plasma extraction with neurotoxic hexane and evaporation of the supernatant is avoided. A stable internal standard is used to provide reliable quantification. The separately available quality controls are human matrix-based and produced according to strict standards.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

100

Parameter

Coenzyme Q10

Analysis Time

< 15 min

Limit of quantification

20 µg/l

Linearity

up to 20 000 μg/l

Recovery

90 %

Intraassay

CV = 5 %

Interassay

CV = 6 %

Gradient

Isocratic

Information

For the Chromsystems HPLC analysis of coenzyme Q 10 any isocratic HPLC system with UV detector is suitable.

Column temperature

ambient (~25 °C)

Flow rate

2.5 ml/min

Specimen

Plasma/Serum/Whole Blood

Pre-analytic Treatment

Keep cool for transport.

Sample Preparation

Plasma/Serum

  • Place 500 μl plasma/serum/whole blood and 250 μl Internal Standard into a labeled, light protected reaction vial and mix briefly.
  • Add 500 μl Precipitation Reagent 1 and mix 30 s (vortex).
  • Incubate 10 minutes at +2 to +8 °C.
  • Centrifuge 5 minutes at 15 000 x g. It is not necessary to transfer the supernatant into a new reaction vial!
  • Add 100 μl Precipitation Reagent 2 and mix 30 s (vortex).
  • Centrifuge 10 minutes at 15 000 x g.
  • Apply complete supernatant to Sample Clean Up Column and draw through completely by centrifugation (2 minutes at 700 x g) or suction, discard effluent.
  • Draw 500 μl Wash Buffer 1 through the Sample Clean Up Column (1 minute at 700 x g), followed by 160 μl Wash Buffer 2 (2 minutes at 700 x g), discard effluents.
  • Centrifuge 250 μl Elution Buffer through the Sample Clean up Column in a labeled, light protected reaction vial (2 min at 700 x g).
  • Inject 50 μl of the eluate into the HPLC system.

 

Whole Blood

  • Place 250 µl cold dist. water into a labelled, light protected reaction vial, add 250 µl whole blood and mix briefly. To make sure that the blood is transferred completely rinse the tip several times.
  • Add 250 µl Internal Standard and mix briefly.
  • Add 500 µl Precipitation Reagent 1 and mix for 30 sec (vortex).
  • Incubate 10 min at +2 to +8 °C.
  • Centrifuge 5 min at 15000 x g. It is not necessary to transfer the supernatant into a new reaction vial!
  • Add 100 µl Precipitation Reagent 2 and mix for 30 sec (vortex).
  • Centrifuge immediately for 10 min at 15000 x g.
  • Apply complete supernatant from the precipitation step to a labelled sample clean up column and draw through quickly by centrifugation (2 min at 700 x g) or suction, discard effluent.
  • Draw 500 µl Wash Buffer 1 through the column by centrifugation (1 min at 700 x g) or suction, discard effluent.
  • Draw 160 µl Wash Buffer 2 through the column by centrifugation (2 min at 700 x g) or suction, discard effluent.
  • Change the collection vial, apply 250 µl Elution Buffer to the column, and draw through completely by centrifugation (2 min at 700 x g) or suction.
  • Inject 50 µl eluate into the HPLC system.

Sample Stability

Storage life is up to 4 days at +2 to +8 °C. For longer storage, deep freeze samples below -18 °C.

Injection Volume

20 µl Plasma/Serum; 50 µl Whole Blood

Flow

2.5 ml/min

Additional Info

For the Chromsystems HPLC analysis of coenzyme Q 10 any isocratic HPLC system with UV detector is suitable.

Wavelength

275 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.: 68001

Internal Standard


Order no.: 68004

Precipitation Reagent 1


Order no.: 68005

Precipitation Reagent 2


Order no.: 68006

Wash Buffer 1


Order no.: 68007

Sample Clean Up Columns


Order no.: 68008

Wash Buffer 2


Order no.: 68009

Elution Buffer


Order no.: 68010

Reaction vials


Order no.: 33005

Calibrators

Calibrators available separately

Controls

Controls available separately

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