Measurement of homocysteine: a historical perspective

Abstract

Elevated plasma level of homocysteine is being increasingly associated with many diseases. There is a significant interest in the development of methods to determine the total homocysteine in biologically relevant tissues. Over the years, researchers use various methods to determine the exact concentrations of homo cysteine in these tissues. However, the precise method used in many studies earlier was questionable. We have reviewed various methodologies for the measurement of homocysteine. We list the commonly used methodologies currently in use to determine homocysteine levels. Through extensive literature search, we have come up with the most popular as well as the newest measurement modalities and listed them with a brief discussion of each of the methodology. In conclusion, we have presented the historical perspective of homocysteine measurement in biological fluids in this manuscript. Thus, the precise understanding of its concentration in biological fluids coupled with its importance in health and disease should justify a newer but reliable technique in the area of ongoing research in homocysteine.

Analysis of serum homocysteine in the laboratory practice - comparison of the direct chemiluminescence immunoassay and high performance liquid chromatography coupled with fluorescent detection

“…Homocysteine determination can be performed by many analytical techniques. The routinely used diagnostic methods are immunoassays, such as enzyme immunoassay (EIA), fluorescence polarization immunoassay (FPIA) and chemiluminescence immunoassay (CLIA) ( 6 , 9 ). However, recently the interest in the use of chromatographic methods in the diagnostic tests has increased ( 10 ).…”

“…These methods combine the separation power of chromatographic columns and the specific identity of detection techniques so they are regarded as the gold analytical standard suitable for very sensitive and selective analysis of organic compounds in the complex biological matrices. Compared to the immunological methods, they are distinguished not only by high specificity but also the ability to analyse various compounds at very different concentration levels as well as in a wide concentration range, often during the same analysis (9,11,12). For these reasons, chromatographic methods are becoming increasingly popular and even recommended as reference methods in routine clinical analysis (10)(11)(12)(13).…”

Ability of dietary factors to affect homocysteine levels in mice: a review

“…In addition, it must be kept in mind that the different methods used for quantification of homocysteine such as chromatography, immunoassays or capillary electrophoresis could have influenced the results [ 27 ]. In our review, the high-performance liquid chromatography (HPLC) was the most frequently used method to quantify plasma homocysteine (in 72 out of 113 studies, Additional file 1 : Table S1).…”

Hyperhomocysteinemia: Clinical Insights

“…5 For the estimation of the Hcy level, plasma tissue homogenate samples are analyzed using various methodologies such as immunoassay, capillary electrophoresis, enzymatic assay, liquid chromatography-mass spectrometry (LCMS) and high-pressure liquid chromatography (HPLC). 6 The normal Hcy levels is ranging between 5 and 15 μmol/L, while a mildly increased level is 15 to 30 μmol/L, moderate from 30 to 100 μmol/L, and a value >100 μmol/L is classified as severe hyperhomocysteinemia. 7,8 The levels of Hcy can be increased through a defective metabolism of Met, due to genetic defects of the transcription of enzymes responsible for Hcy metabolism or deficiencies of cofactors involved in these pathways such as vitamins B6, B12, and folate.…”