Determination of Total Homocysteine in Human Plasma by Isocratic High-Performance Liquid Chromatography

A simple, sensitive and precise isocratic HPLC method for the determination of total homocysteine in human plasma is described. The thiol compounds were liberated from plasma proteins by reduction with tri-n-butylphosphine and derivatized with a thiol-specific fluorogenic marker, 7-fluoro-benzo-2-oxa-1,3-diazole-4-sulphonate. The derivatives were separated isocratically within 7 min by reversed-phase HPLC using a Superspher 100 RP-18 column as stationary phase. By using this approach more than 200 samples a day can be assayed for total homocysteine. The method was linear up to 100 mumol/l and proved to be sensitive with a detection limit of 0.1 mumol/l and the lowest limit of reliable quantification of 0.5 mumol/l for homocysteine in buffer. Intra- and inter-assay coefficients of variation were both < 4% at a concentration of 10 mumol/l homocysteine. Similar results were obtained for homocysteine concentrations between 0.5 and 100 mumol/l. The analytical recovery for these concentrations ranged from 94.9 to 117.0%. As compared to other protocols published so far, this modified method is less complicated but equally sensitive and reproducible and allows a rapid determination of total homocysteine and cysteine in human plasma under routine conditions.

• Publication year

  1997

• Venue

  European journal of clinical chemistry and clinical biochemistry : journal of the Forum of European Clinical Chemistry Societies

• Publication date

  Unknown publication date

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1515/cclm.1997.35.9.687PMID 9352231
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Post-methionine load hyperhomocysteinemia in persons with normal fasting total plasma homocysteine: initial results from the NHLBI Family Heart Study.

  1995cited by this paper
• Mutated methylenetetrahydrofolate reductase as a risk factor for spina bifida.

  1995cited by this paper
• Homocysteine in plasma: stabilization of blood samples with fluoride.

  1995cited by this paper
• Rapid, fully automated measurement of plasma homocyst(e)ine with the Abbott IMx analyzer.

  1995cited by this paper
• Homocyst(e)ine and arterial occlusive diseases

  1994cited by this paper
• Rapid HPLC determination of total homocysteine and other thiols in serum and plasma: sex differences and correlation with cobalamin and folate concentrations in healthy subjects.

  1994cited by this paper
• Effects of age, lipoproteins, and hemostatic parameters on the role of homocyst(e)inemia as a cardiovascular risk factor in men.

  1994cited by this paper
• Homocysteine and other thiols in plasma and urine: automated determination and sample stability.

  1993cited by this paper
• Modification of a high-performance liquid chromatographic method for assay of homocysteine in human plasma.

  1993cited by this paper
• Vitamin status and intake as primary determinants of homocysteinemia in an elderly population.

  1993cited by this paper
• A prospective study of plasma homocyst(e)ine and risk of myocardial infarction in US physicians.

  1992cited by this paper
• Hyperhomocysteinemia: an independent risk factor for vascular disease.

  1991cited by this paper
• The association of elevated plasma homocyst(e)ine with progression of symptomatic peripheral arterial disease.

  1991cited by this paper
• Rapid high-performance liquid chromatographic assay for total homocysteine levels in human serum.

  1991cited by this paper
• Impaired homocysteine metabolism in early-onset cerebral and peripheral occlusive arterial disease. Effects of pyridoxine and folic acid treatment.

  1990cited by this paper
• Prevalence of hyperhomocyst(e)inemia in patients with peripheral arterial occlusive disease.

  1989cited by this paper
• Fully automated fluorescence assay for determining total homocysteine in plasma.

  1989cited by this paper
• Reduction of plasma lipid and homocysteine levels by pyridoxine, folate, cobalamin, choline, riboflavin, and troxerutin in atherosclerosis.

  1989cited by this paper
• Determination of free and total homocysteine in human plasma by high-performance liquid chromatography with fluorescence detection.

  1987cited by this paper
• Quantitation of total homocysteine, total cysteine, and methionine in normal serum and urine using capillary gas chromatography-mass spectrometry.

  1987cited by this paper
• Heterozygosity for homocystinuria in premature peripheral and cerebral occlusive arterial disease.

  1985cited by this paper
• Radioenzymic determination of homocysteine in plasma and urine.

  1985cited by this paper
• Protein-Bound Homocyst(e)ine in Normal Subjects and in Patients with Homocystinuria

  1979cited by this paper
• Technical pitfalls leading to errors in the quantitation of plasma amino acids.

  1969cited by this paper
• Enzymic transpeptidation reactions involving γ-glutamyl peptides and α-amino-acyl peptides

  1952cited by this paper
• Enzymic transpeptidation reactions involving gamma-glutamyl peptides and alpha-amino-acyl peptides.

  1952cited by this paper

• Association between high BMI and high homocysteine levels in Chinese patients with bipolar disorder.

  2021cites this paper
• Hyperhomocysteinemia Induced by Methionine Excess is Effectively Suppressed by Betaine in Geese

  2020cites this paper
• Sex differences in the prevalence and clinical correlates of hyperhomocysteinemia in patients with bipolar disorder

  2020cites this paper
• Prevalence and clinical-demographic correlates of hyperhomocysteinemia in inpatients with bipolar disorder in a Han Chinese population.

  2018cites this paper
• Therapeutic impact of grape leaves polyphenols on certain biochemical and neurological markers in AlCl3-induced Alzheimer's disease.

  2017cites this paper
• Quantitative Analysis of Total Plasma Homocysteine by LC‐MS/MS

  2016cites this paper
• Niacin Reverses Migratory Macrophage Foam Cell Arrest Mediated by oxLDL In Vitro

  2014cites this paper
• Aged garlic extract restores nitric oxide bioavailability in cultured human endothelial cells even under conditions of homocysteine elevation.

  2013cites this paper
• Thiol Reactive Probes and Chemosensors

  2012cites this paper
• Loss of mitochondrial thioredoxin reductase delays angiogenic switch and impairs tumour growth

  2011cites this paper
• Addressing the Role of Mitochondrial Thioredoxin Reductase and xCT in the Maintenance of Redox Homeostasis

  2010cites this paper
• Complex Redundancy between the Mammalian Thioredoxin and Glutathione Systems in Cell Proliferation and Tumorigenesis Thesis

  2009cites this paper
• The impact of homocysteine levels on cognition in euthymic bipolar patients: a cross-sectional study.

  2008cites this paper
• Acute hyperhomocysteinemia induces microvascular and macrovascular endothelial dysfunction.

  2007cites this paper
• Association of cognitive deficits with elevated homocysteine levels in euthymic bipolar patients and its impact on psychosocial functioning: preliminary results.

  2007cites this paper
• The cystine/cysteine cycle: a redox cycle regulating susceptibility versus resistance to cell death

  2006cites this paper
• Cystine-Import and regulation of apoptosis n B-Lymphocytes

  2005cites this paper
• Relationship of aging and tobacco use with the development of aberrant crypt foci in a predominantly African-American population.

  2005cites this paper
• In vivo quantification of intracellular amino acids and intermediates of the methionine pathway in Corynebacterium glutamicum.

  2005cites this paper
• Homocysteine and carotid atherosclerosis in chronic renal failure--the confounding effect of renal function.

  2004cites this paper
• Heme oxygenase 1 expression in young uremic patients on hemodialysis

  2004cites this paper
• Bioanalytical considerations for compounds containing free sulfhydryl groups.

  2003cites this paper
• Refinement and delineation of the breakpoint regions of a chromosome 1;22 translocation in a patient with Costello syndrome.

  2002cites this paper
• Hydroxocobalamin reduces hyperhomocysteinemia in end-stage renal disease.

  2002cites this paper
• Methods for homocysteine analysis and biological relevance of the results.

  2002cites this paper
• Methylenetetrahydrofolate reductase gene polymorphism, hyperhomocysteinemia and occlusive retinal vascular disease in type 2 diabetic and non-diabetic subjects.

  2002cites this paper
• Critical role of pH for derivatization of homocysteine with benzofurazanes.

  2001cites this paper
• Parenteral vitamin B12 reduces hyperhomocysteinemia in end-stage renal disease.

  2001cites this paper
• Données récentes sur l'homocystéine

  2001cites this paper
• Animal models of diet-induced atherosclerosis.

  2001cites this paper
• Parenteral vitamin B12 reduces hyperhomocysteinemia in end-stage renal disease.

  2001cites this paper
• Total homocysteine measurement in clinical practice

  2000cites this paper
• A tale of two homocysteines--and two hemodialysis units.

  2000cites this paper
• Determination of homocysteine in plasma by liquid chromatography with fluorescence detection.

  1999cites this paper
• Determination of retinol, antioxidant vitamins and homocysteine in skin puncture blood.

  1999cites this paper
• Plasma homocysteine determined by capillary electrophoresis with laser-induced fluorescence detection.

  1999cites this paper
• Validation of a point-of-care assay for the urinary albumin:creatinine ratio.

  1999cites this paper
• High-performance liquid chromatographic method for measuring total plasma homocysteine levels.

  1998cites this paper