Conventional method for determination of homocysteine

The conventional methods for determining homocysteine mainly include high-performance liquid chromatography and immunological methods (such as enzyme-linked immunosorbent assay and fluorescence polarization immunoassay). The following is a specific introduction:

High performance liquid chromatography (HPLC)

Principle: Through chromatographic separation technology, homocysteine in blood samples is separated from other substances, and then quantitatively determined by combining fluorescence detectors or ultraviolet detectors.

Features:

High accuracy: It can accurately determine the concentration of homocysteine and is one of the gold standards in the laboratory.

. Can distinguish homocysteine from other sulfur-containing amino acids: helpful for scientific research or diagnosis of difficult cases. The operation is relatively complex: it requires professional equipment and technical support, and the detection time is relatively long.

High cost: Some primary healthcare institutions may not be able to carry out.

. Immunological methods are commonly used detection methods in clinical practice, mainly including enzyme-linked immunosorbent assay (ELISA), fluorescence polarization immunoassay (FPIA), etc.

Principle: Based on the analysis method established by the binding of antigens and antibodies to generate precipitates, homocysteine levels are determined by measuring the concentration of immune complexes.

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Features:

Easy to operate: suitable for large-scale screening and rapid clinical diagnosis.

. Lower cost: Compared to high-performance liquid chromatography, immunological methods have lower costs. Less accurate: may be affected by cross reaction interference, and the detection accuracy for extremely high or low concentration samples is slightly lower than that of chromatography. High degree of automation: For example, fluorescence polarization immunoassay method uses fluorescently labeled antibodies to bind with homocysteine, and determines the content of homocysteine by detecting changes in fluorescence polarization, which is suitable for large-scale screening.

Other methods

Isotope method: using adenosine containing radioactive isotopes to react with homocysteine to determine the concentration of homocysteine in serum.

. This method has strong specificity, but it is contaminated with radiation and is currently not commonly used. Gas chromatography: Homocysteine is usually converted into derivatives that can be analyzed by gas chromatography, and quantified by gas chromatography. This method is complex to operate and requires specialized equipment, so its application is limited.

Urine testing: Collecting 24-hour urine for analysis of excretion can be used as an auxiliary method.

. However, this method is greatly affected by kidney function and urine concentration, and the fluctuation of results is higher than that of blood testing, so it is usually not used as an independent diagnostic basis.