Homocysteine in Serum/Plasma - HPLC

Order No.: 45000, for 200 tests
Parameters:
Homocysteine

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

CE-IVD validated product ready for IVDR within timeframes and transition periods specified by the IVDR 2017/746

Homocysteine

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 assay 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.

More Information
Method of Analysis HPLC
Number of Tests 200
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.
Lower Limit of Quantitation 0.4 µmol/l
Upper Limit of Quantification up to 400 µmol/l
Intraassay

CV = 0.9–2.2 %

Interassay

CV = 2.9–3.1 %

Recovery 96–99 %
Specimen Serum/Plasma
Sample Preparation
  • To 100 µl plasma/serum add 25 µl Internal Standard and 75 µl Reduction Reagent.
  • Shake for 2 s (vortex).
  • Incubate 10 min at ambient temperature.
  • Add 100 µl Precipitation Reagent.
  • Shake for 30 s (vortex).
  • Centrifuge 5 to 7 min at 9000 x g.
  • Pipette 100 µl derivatisation mix into a new light protected reaction vial.
  • Add 50 µl supernatant, mix well.
  • Incubate 10 min at +50 to +55 °C.
  • Cool down immediately.
  • Inject 20 to 50 µl into the HPLC system.
Run Time 5.5 min
Injection Volume 20–50 µl
Flow Rate 1.3–1.7 ml/min
Column Temperature ambient (~ 25 °C)
Wavelengths

EX 385 nm
EM 515 nm

Additional Info Any isocratic HPLC system with fluorescence detector is suitable.
Parameters Homocysteine
The following components are included in the kit:
As a customer please login or register to gain full access.

Homocysteine

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 assay 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.

More Information
Method of Analysis HPLC
Number of Tests 200
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.
Lower Limit of Quantitation 0.4 µmol/l
Upper Limit of Quantification up to 400 µmol/l
Intraassay

CV = 0.9–2.2 %

Interassay

CV = 2.9–3.1 %

Recovery 96–99 %
Specimen Serum/Plasma
Sample Preparation
  • To 100 µl plasma/serum add 25 µl Internal Standard and 75 µl Reduction Reagent.
  • Shake for 2 s (vortex).
  • Incubate 10 min at ambient temperature.
  • Add 100 µl Precipitation Reagent.
  • Shake for 30 s (vortex).
  • Centrifuge 5 to 7 min at 9000 x g.
  • Pipette 100 µl derivatisation mix into a new light protected reaction vial.
  • Add 50 µl supernatant, mix well.
  • Incubate 10 min at +50 to +55 °C.
  • Cool down immediately.
  • Inject 20 to 50 µl into the HPLC system.
Run Time 5.5 min
Injection Volume 20–50 µl
Flow Rate 1.3–1.7 ml/min
Column Temperature ambient (~ 25 °C)
Wavelengths

EX 385 nm
EM 515 nm

Additional Info Any isocratic HPLC system with fluorescence detector is suitable.
Parameters Homocysteine
The following components are included in the kit:
As a customer please login or register to gain full access.