Estimation of plasma homocysteine level in young healthy population of Saudi Arabia


  1. Santosh Kumar
  2. Sreyoshi Fatima Alam
  3. Heba Musallam
  4. Ghazal Muteb Binsultan
  5. Hajar A Alreefi
  6. Ghadah Nasser Binzuman
  7. Zahraa Mhd Imad Qamhieh
  8. Sara M Fakher Maaz
  9. Ahmed Yaqinuddin
  10. Paul Ganguly


Clinical Sciences 2


Alfaisal University



  • A sulphur containing amino acid.
  • An intermediate product in biosynthesis of methionine and cysteine, with B-complex vitamins affecting its metabolism as co-enzymes.


  • Elevated plasma level of homocysteine is being increasingly associated with many diseases:Cardiovascular diseases (atherosclerosis, congestive heart failure, etc), stroke, Alzheimer’s disease, Parkinson’s disease, diabetes, non-alcoholic fatty liver disease (NAFLD), hearing loss, macular degeneration, and migraine.
  • However, there are no studies reporting plasma homocysteine levels for normal Saudi population.

Aim of the Study: 

  • Establishment of the methodology protocol for determining plasma homocysteine level using high pressure liquid chromatography (HPLC) at Alfaisal University campus.
  • To determine plasma homocysteine levels in young (18 – 25 years old) healthy Saudi population (who have no known diseases which are associated with elevated plasma homocysteine levels). 

Location of the Study:

  • The entire study and all the experiments have been conducted on campus at the research labs of Alfaisal University.
  • The study is partially funded by Internal Research Grant (IRG2015) from Alfaisal University.
Summary of Work


  • 5 ml of blood via venipuncture from the consenting individual.
  • The first and foremost component is establishment of the protocols and optimized methods for determining serum/plasma homocysteine level. 

We have established the protocols and optimized methods for determining serum/plasma homocysteine level using:

  • The Waters High performance liquid chromatography system, 
  • The Waters fluorescence detector.
  • The Empower pro software for data generation.
  • Chromsystems Reagent kits to process and run serum/plasma samples through HPLC system for homocysteine determination. 

Statistical Analysis: Data analysis will be performed using IBM SPSS Statistics version 22 software. The means and standard deviations of quantitative variables will be recorded. T-test and Mann-Whitney U test will be used in variables with normal and without normal distribution respectively.

figure: Simplified Illustration of HPLC


figure: HPLC System at the research lab at Alfaisal University


Figure: Autosampler and HPLC Pump


Summary of Results

We have established the protocols and optimized methods for determining serum/plasma homocysteine level using high pressure liquid chromatography (HPLC).

The following image shows a typical chromatogram of a plasma calibration standard, acquired during one of our recent experiments done to calibrate and optimize the instrument. 



Since we have completed the first and foremost step of this crucial project, that is establishment and optimization of the methodology protocol, we shall now proceed and determine plasma homocysteine levels in young (18 – 25 years old) healthy Saudi population. Our initial focus is on the young population, because increasing age is also known to be associated with increasing levels of homocysteine. This is a pioneering study in the Kingdom which will not only benefit the population in general but also add to the much needed research in the field of homocysteine. 

Future Goals:

  • To investigate the co-factors (any of folate, vitamin B6, vitamin B12, and H2S levels and MTHFR C677T/A1298C polymorphisms) which may affect plasma homocysteine levels.
  • Follow-up of the consenting individuals on yearly basis for the next 6 years to study changes in plasma homocysteine levels over this period.


Take-home Messages

Plasma levels of homocysteine are potentially amenable to therapeutic (with B-complex vitamins) and dietary interventions. Better understanding of homocysteine level and its contributing factors in the normal Saudi population will help design interventional therapeutic and preventive studies, leading to possible simple but important health care recommendations which may reduce burden of the above mentioned chronic diseases in the Kingdom.


Ganguly, P. and S. F. Alam (2015). "Role of homocysteine in the development of cardiovascular disease." Nutr J 14: 6.

Summary of Work
Summary of Results
Take-home Messages
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