High Performance Liquid

Chromatography Method
for Rapid and Accurate
Determination of Homocysteine
in Plasma and Serum

By Birte Vester and K. Rasmussen

Klinisk-kemisk afdeling, Aarhus Kommunehospital,
Aarhus, Denmark.
Eur. J. Clin. Chem. Clin. Biochem.
Vol. 29, 1991,pp. 549-554
© 1991 Walter de Gruyter Ä Co.
Berlin · New York
Group 6 :
- Shafira Azzahra (P17334115423)
- Anisa Safanah (P17334115425)
- Yoga Romi Anggara (P17334115428)
 This research used Liquid Chromatography.
And the procedures are :
1. a. Blood no addictive put into the tube (for serum)
b. Blood with EDTA put into the tube (for plasma)
2. Take blood to the centrifuge for separate between
erythrocyte and serum
3. Take specimen to Liquid Chromatography
Instrument for measure homocysteine
The Aim of the study
For determining the normal
value of homocysteine in
plasma or serum in a certain
area.
 Clarifying the terms
• Liquid Chromatography = is the collective
term for a set of laboratory techniques for the
separation of mixtures. The mixture is dissolved
in a fluid called the mobile phase, which carries it
through a structure holding another material
called the stationary phase.
• The different of Plasma and Serum =
Plasma, blood specimen with addictive
Serum, blood specimen no-addictive
• Cobalamin and Folat = are important
substances for
 Background of the study
The residents of every region has a
different normal value of homocysteine
because the residents are influenced by
the weather, air temperature, air
pressure, and environmental factors.
Therefore, the normal value of
homocysteine in a region is needed.
The sample of the study
Blood
( plasma or serum )
Data Collection Techniques
The researchers use Liquid
Chromatography to measure
the homocysteine from plasma
or serum
 Data Collection Analysis
•Blood samples must be cooled or centrifuged
immediately after venipuncture. The method is
useful for evaluation of cobalamin or folate
deficiency, especially in patients with normal or
moderately depressed cobalamin or folate
concentrations in blood.

•Blood samples take into Chromatography for

measure homocysteine
 Finding of the research
Mean SD Range

Plasma

Men 9.8 1.6 7.8 – 11.9

Woman 6.8 1.2 5.5 – 8.4

Serum

Men 10.8 1.6 8.7 – 13.0

Woman 7.6 1.2 5.8 – 9.1

 The conclusion of the research
The method uses only commercially
available reagents and requires only a
Standard HPLC apparatus equipped with a
fluorescence detector in addition to normal
laboratory equipment. The addition of an
internal Standard ensures good precision,
accuracy and performance. The analysis is
robust, fast and inexpensive and is now
suitable for routine use in the clinical
laboratory.
• suitable for routine use in the clinical
laboratory.
• Plasma/serum homocysteine concentrations
were measured in blood samples from 16
apparently healthy volunteers.