Total homocysteine measurement in clinical practice

Rasmussen, Karsten; Møller, Jan

doi:10.1258/0004563001899915

Cited by 133 publications

(119 citation statements)

References 143 publications

(203 reference statements)

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“…Recovery: A serum sample, whose homocysteine was previously measured, was split into two aliquots, the first was enriched with known amount of homocystine and homocystine-d 8 and treated according to the extraction procedure. The second aliquot was reduced, deproteinized and after centrifugation was spiked with the same amount of homocystine and homocystine-d 8 previously reduced in order to have the same concentration of homocysteine and homocystine-d 4 as the first aliquot.…”

Section: Methods Validationmentioning

confidence: 99%

“…Homocysteine measurements have been performed by different methods based on immunoassay, chromatographic and enzymatic techniques [7,8].…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, these techniques such as fluorescence polarization immunoassays, chemiluminescent microparticle immunoassays and enzyme-linked immunoassays, even showing a good correlation with reference methods, need expensive reagents [7,8].…”

Section: Introductionmentioning

confidence: 99%

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A simplified method for the determination of total homocysteine in plasma by electrospray tandem mass spectrometry

Persichilli

Gervasoni

Iavarone

et al. 2010

J of Separation Science

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A simplified method for the determination of total homocysteine in plasma by electrospray tandem mass spectrometry Hyperhomocysteinemia is a risk factor for different diseases. Several methods have been developed to analyze homocysteine and the immunometric ones, although expensive, they are in widespread use. A rapid LC-MS/MS method for homocysteine assay has been developed for the application of large clinical chemistry routines. Selected reaction monitoring was performed through the transitions m/z 136.0-90.1 for homocysteine and m/z 140.0-94.0 for the internal standard. ESI was used to generate [H1] adduct ions. Chromatographic isocratic separation was achieved using a strong cation exchange column. The mobile phase was methanol/water (20:80 v/v, containing 0.1% formic acid and 1.5 mmol/L ammonium formate in the water phase) at a flow rate of 0.250 mL/min (351C). Samples treatment consisted in the reduction with DTT and deproteinization with methanol. Recovery, linearity, LOD, LOQ and total imprecision were evaluated to validate the method. Homocysteine values on 100 serum samples were compared with those obtained by HPLC and immunometric methods. The method is robust, selective and precise in the whole range of values studied. Moreover, low reagent cost and easiness of sample treatment make this method useful, not only for research, but also for routine work.

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Section: Methods Validationmentioning

confidence: 99%

“…Homocysteine measurements have been performed by different methods based on immunoassay, chromatographic and enzymatic techniques [7,8].…”

Section: Introductionmentioning

confidence: 99%

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A simplified method for the determination of total homocysteine in plasma by electrospray tandem mass spectrometry

Persichilli

Gervasoni

Iavarone

et al. 2010

J of Separation Science

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“…Blood were collected into EDTA-treated evacuated tubes for whole blood and into untreated evacuated tubes for serum preparation. Blood samples for tHcy, folate, MMA and B12 were refrigerated immediately, and plasma and serum was separated by centrifugation at 1500 g for 15 min at 41C within 30 min of sampling (Rasmussen and Moller, 2000). Plasma and serum were stored at À201C until analysed.…”

Section: Methodsmentioning

confidence: 99%

Folate and vitamin B12 in relation to lactation: a 9-month postpartum follow-up study

et al. 2005

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Objective: To investigate the relation between lactation and markers of folate and vitamin B12 (B12) deficiency in women with and without vitamin supplementation. Design: A 9-month follow-up study. Subjects and methods: Blood samples from 91 women, who gave birth to a single healthy child, were collected 3 weeks, 4 and 9 months postpartum and analysed for circulating level of homocysteine (tHcy), methylmalonic acid (MMA), folate and B12. The participants were categorized as exclusively, partly or not breast-feeding dependent on the degree of lactation 4 months postpartum. During follow-up, lifestyle factors were recorded by structured interviews. Results: Among 72 exclusively breast-feeding women, the median (10-90% percentile) tHcy was 5.8 (3.1-8.3) mmol/l 3 weeks postpartum, 6.1 (4.1-10.3) mmol/l 4 months postpartum and 5.3 (3.6-8.7) mmol/l 9 months postpartum. At 9 months postpartum, none of the women breast-fed exclusively. No significant change occurred in the concentration of B12 and folate. Exclusively breast-feeding women without vitamin supplementation had higher median tHcy than supplemented exclusively breast-feeding women 4 and 9 months postpartum (7.0 vs 5.4 mmol/l (Po0.001) and 5.8 vs 4.5 mmol/l (P ¼ 0.003), respectively). Six women had increased (415 mmol/l) tHcy; four of these were unsupplemented and exclusively breast-feeding. Conclusion: We found no overall indication of depletion of the folate and B12 stores during the lactation period in this population. However, folate-supplemented women had lower tHcy and higher folate levels, suggesting a beneficial effect of supplementation with folate throughout lactation. Sponsorship: The Biomedical Laboratory Scientist Education and Research Fund and LEO Pharma Research Foundation supported this study. DPC Scandinavia, Denmark donated reagents for the folate and tHcy analysis.

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“…In the recent years, it has emerged as a risk factor for cardiovascular disease independent of known other factors (1)(2)(3). The first methods developed for total plasma HCy measurements were HPLC, with or without derivatization, and gas chromatography.…”

mentioning

confidence: 99%

Reference Intervals for Plasma Homocysteine by the AxSYM Immunoassay after Collection in Fluoride Tubes

Cotton¹,

Wautrecht²,

Léchevin³

et al. 2003

Clinical Chemistry

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Total homocysteine measurement in clinical practice

Abstract: This article was prepared at the invitation of the Analytical Investigations Standing Committee of the Association of Clinical Biochemists.

Cited by 133 publications

References 143 publications

A simplified method for the determination of total homocysteine in plasma by electrospray tandem mass spectrometry

A simplified method for the determination of total homocysteine in plasma by electrospray tandem mass spectrometry

Folate and vitamin B12 in relation to lactation: a 9-month postpartum follow-up study

Reference Intervals for Plasma Homocysteine by the AxSYM Immunoassay after Collection in Fluoride Tubes

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