45000 HPLC kit homocysteine plasma serum

Homocysteine in Serum/Plasma - HPLC

Order no.: 45000
200 Tests
  • Homocysteine

Fast sample preparation
Short analysis time
Column life-time more than 2000 runs
Suitable for all blood collection systems

Description

Clinical relevance

Homocysteine (Hcy) is a non-proteinogenic amino acid, synthesized during the demethylation of the amino acid methionine. Some enzymes are involved in this process, which require cofactors such as vitamin B6 and vitamin B12. A deficiency of these cofactors can lead to an increase in the Hcy concentration, but inherited genetic mutations may also be the cause.

Even moderately increased Hcy levels are a risk factor for cardiovascular diseases, arteriosclerosis and coronary heart diseases. Increased Hcy levels are also detected in the case of homocystinuria, an inherited genetic disorder in which the enzymes required for metabolism are not present or at levels that are too low. In this case, already newborns have high levels of Hcy, however, the disease is treatable if detected in time. Screening of newborns (NBS) for homocystinuria has therefore become mandatory in some European countries such as Ireland. In Germany it is only performed as part of more intensive screening or in cases where the disease is suspected.

 

Product advantages

  • Fast sample preparation
  • Short analysis time
  • Column lifetime of more than 2000 runs
  • For all blood collection systems

 

This reagent kit allows the simple and reliable determination of total homocysteine in serum/plasma. Sample preparation is simply a reduction step for releasing homocysteine from its protein binding, followed by precipitation and subsequent precolumn derivatisation. Chromatographic separation is performed on an isocratic HPLC system with fluorescence detection. The optimised reagents ensure the accurate and reliable quantification of total homocysteine for all blood collection systems within a brief analysis period of less than 6 minutes.

Chromatogram

Technical Data

Method of Analysis

HPLC

Number of Tests

200

Parameter

Homocysteine

Analysis Time

< 6 min

Limit of quantification

0.4 µmol/l

Linearity

up to 400 µmol/l

Recovery

> 96 %

Intraassay

CV < 2 %

Interassay

CV < 3.2 %

Column temperature

ambient (~25 °C)

Flow rate

1.3 to 1.7 ml/min

Specimen

Serum/Plasma

Pre-analytic Treatment

Time of sample drawing

Blood should be taken from the fasted patient. Prepare the plasma or serum within 45 min after blood collection. In case special homocysteine blood collection systems are used, please contact the respective manufacturer.

Sample Preparation

  • To 100 µl plasma/serum add 25 µl Internal Standard and 75 µl Reduction Reagent.
  • Vortex 2 s.
  • Incubate 10 min at ambient temperature (~25 °C).
  • Add 100 µl Precipitation Reagent.
  • Mix 30 s.
  • Centrifuge 5 to 7 min at 9000 x g.
  • Pipette 100 µl derivatisation mix (= Derivatisation Reagent 1 + Derivatisation Reagent 2) into a separate reaction vial.
  • Add 50 µl supernatant, mix well.
  • Incubate 10 min at +50 to +55 °C in a water bath.
  • Cool down immediately.
  • Inject 20 to 50 µl into the HPLC system.

Sample Stability

At ambient temperature approx. 4 days, at +2 to +8 °C approx. 4 weeks and below -18 °C several months.

Injection Volume

20 - 50 µl

Additional Info

For the analysis of homocysteine any isocratic HPLC system with fluorescence detector is suitable.

Wavelength

EX 385 nm
EM 515 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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