Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard

Background and Objective: Homocysteine is an amino acid with a free thiol (Sulphydryl) group and is an intermediate formed during the metabolism of methionine to cysteine. Measurement of total plasma Homocysteine may be of value in several clinical conditions including homocysteinuria, atheroscleros...

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Autores principales:	F Darabi, B Jalali, M Rafiei, M Mahdavi, A Babaei
Formato:	article
Lenguaje:	EN FA
Publicado:	Babol University of Medical Sciences 2005
Materias:	homocysteine atherosclerosis high performance liquid chromatography plasma Medicine R Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/3c17bd6c77844c1bb03cecb9c0bc93fa
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spelling	oai:doaj.org-article:3c17bd6c77844c1bb03cecb9c0bc93fa2021-11-10T09:14:32ZMeasurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard1561-41072251-7170https://doaj.org/article/3c17bd6c77844c1bb03cecb9c0bc93fa2005-04-01T00:00:00Zhttp://jbums.org/article-1-2632-en.htmlhttps://doaj.org/toc/1561-4107https://doaj.org/toc/2251-7170Background and Objective: Homocysteine is an amino acid with a free thiol (Sulphydryl) group and is an intermediate formed during the metabolism of methionine to cysteine. Measurement of total plasma Homocysteine may be of value in several clinical conditions including homocysteinuria, atherosclerosis, thrombophilia and folate/vitamin B12 deficiency. The purpose of this study was to measure total plasma Homocysteine using RP-HPLC with fluorescence detection method. Methods: Total plasma Homocysteine was measured with reverse-phase HPLC with fluorescence detection after reduction of Homocysteine with tris (2-carboxyetile) phosphine (TCEP) and derivatization of plasma thiols with ammonium 7-fluorobenzo-2-oxa 1,3-diazole-4-sulfonate (SBD-F). Findings: The limit of detection for Homocysteine was 0.2 µmol/l. The within day coefficient of variation (CV) was 2.67% to 4.56% and the between day coefficient of variation was 5.43% to 8.17%. The mean recovery of Homocysteine was 93% to 103.4%. Conclusion: The results show that the HPLC method with fluorescence detection for measurement of total plasma Homocysteine, is sensitive (The limit of detection 0.2 µmol/l), accurate (CV between 2.67% and 8.17%) and it has acceptable recovery (93% to 103.4%). Therefore, it is confident method especially for investigate application.F DarabiB JalaliM RafieiM MahdaviA BabaeiBabol University of Medical Sciencesarticlehomocysteineatherosclerosishigh performance liquid chromatographyplasmaMedicineRMedicine (General)R5-920ENFAMajallah-i Dānishgāh-i ̒Ulūm-i Pizishkī-i Bābul, Vol 7, Iss 2, Pp 18-23 (2005)
institution	DOAJ
collection	DOAJ
language	EN FA
topic	homocysteine atherosclerosis high performance liquid chromatography plasma Medicine R Medicine (General) R5-920
spellingShingle	homocysteine atherosclerosis high performance liquid chromatography plasma Medicine R Medicine (General) R5-920 F Darabi B Jalali M Rafiei M Mahdavi A Babaei Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
description	Background and Objective: Homocysteine is an amino acid with a free thiol (Sulphydryl) group and is an intermediate formed during the metabolism of methionine to cysteine. Measurement of total plasma Homocysteine may be of value in several clinical conditions including homocysteinuria, atherosclerosis, thrombophilia and folate/vitamin B12 deficiency. The purpose of this study was to measure total plasma Homocysteine using RP-HPLC with fluorescence detection method. Methods: Total plasma Homocysteine was measured with reverse-phase HPLC with fluorescence detection after reduction of Homocysteine with tris (2-carboxyetile) phosphine (TCEP) and derivatization of plasma thiols with ammonium 7-fluorobenzo-2-oxa 1,3-diazole-4-sulfonate (SBD-F). Findings: The limit of detection for Homocysteine was 0.2 µmol/l. The within day coefficient of variation (CV) was 2.67% to 4.56% and the between day coefficient of variation was 5.43% to 8.17%. The mean recovery of Homocysteine was 93% to 103.4%. Conclusion: The results show that the HPLC method with fluorescence detection for measurement of total plasma Homocysteine, is sensitive (The limit of detection 0.2 µmol/l), accurate (CV between 2.67% and 8.17%) and it has acceptable recovery (93% to 103.4%). Therefore, it is confident method especially for investigate application.
format	article
author	F Darabi B Jalali M Rafiei M Mahdavi A Babaei
author_facet	F Darabi B Jalali M Rafiei M Mahdavi A Babaei
author_sort	F Darabi
title	Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
title_short	Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
title_full	Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
title_fullStr	Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
title_full_unstemmed	Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard
title_sort	measurement of total plasma homocysteine with reverse phase high performance liquid chromatography (rp-hplc) method without internal standard
publisher	Babol University of Medical Sciences
publishDate	2005
url	https://doaj.org/article/3c17bd6c77844c1bb03cecb9c0bc93fa
work_keys_str_mv	AT fdarabi measurementoftotalplasmahomocysteinewithreversephasehighperformanceliquidchromatographyrphplcmethodwithoutinternalstandard AT bjalali measurementoftotalplasmahomocysteinewithreversephasehighperformanceliquidchromatographyrphplcmethodwithoutinternalstandard AT mrafiei measurementoftotalplasmahomocysteinewithreversephasehighperformanceliquidchromatographyrphplcmethodwithoutinternalstandard AT mmahdavi measurementoftotalplasmahomocysteinewithreversephasehighperformanceliquidchromatographyrphplcmethodwithoutinternalstandard AT ababaei measurementoftotalplasmahomocysteinewithreversephasehighperformanceliquidchromatographyrphplcmethodwithoutinternalstandard
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Measurement of total plasma Homocysteine with reverse phase high performance liquid chromatography (RP-HPLC) method without internal standard

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