Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection
A highly sensitive method for the determination of α-lipoic acid (LA) and dihydrolipoic acid (DHLA) in human plasma and urine has been developed. Samples were acidified and extracted with organic solvent, and the free sulfhydryls of DHLA protected as the dicarboxyethylate by treatment with ethylchlo...
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| Veröffentlicht in: | Journal of Chromatography A 2000-02, Vol.870 (1), p.381-388 |
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| creator | Haj-Yehia, Abdullah I Assaf, Peter Nassar, Taher Katzhendler, Jehoshua |
| description | A highly sensitive method for the determination of α-lipoic acid (LA) and dihydrolipoic acid (DHLA) in human plasma and urine has been developed. Samples were acidified and extracted with organic solvent, and the free sulfhydryls of DHLA protected as the dicarboxyethylate by treatment with ethylchloroformate. The free carboxylic function of LA and the SH-protected DHLA were converted into their amide derivatives with the strong fluorophore 2-(4-aminophenyl)-6-methylbenzothiazole in the presence of a coupling agent and a base catalyst. The resulting fluorescent amides of both LA and DHLA were separated on a reversed-phase column (Ultrasphere C
8) using simple isocratic elution with acetonitrile–water (80:20) and detected fluorimetrically (excitation 343, emission 423 nm). The method is highly sensitive, reproducible, and is easily applied for the simultaneous determination of LA and DHLA in biological samples. |
| doi_str_mv | 10.1016/S0021-9673(99)00857-2 |
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8) using simple isocratic elution with acetonitrile–water (80:20) and detected fluorimetrically (excitation 343, emission 423 nm). The method is highly sensitive, reproducible, and is easily applied for the simultaneous determination of LA and DHLA in biological samples.</description><identifier>ISSN: 0021-9673</identifier><identifier>DOI: 10.1016/S0021-9673(99)00857-2</identifier><identifier>PMID: 10722093</identifier><identifier>CODEN: JOCRAM</identifier><language>eng</language><publisher>Amsterdam: Elsevier B.V</publisher><subject>Aminophenylmethylbenzothiazole ; Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Chromatography, High Pressure Liquid - methods ; Coenzymes, vitamins ; Dihydrolipoic acid ; Fundamental and applied biological sciences. Psychology ; Humans ; Investigative techniques, diagnostic techniques (general aspects) ; Lipoic acid ; Medical sciences ; Miscellaneous. Technology ; Other biological molecules ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Fluorescence ; Thioctic Acid - analogs & derivatives ; Thioctic Acid - analysis ; Thioctic Acid - blood ; Thioctic Acid - urine ; Thiols</subject><ispartof>Journal of Chromatography A, 2000-02, Vol.870 (1), p.381-388</ispartof><rights>2000 Elsevier Science B.V.</rights><rights>2000 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c390t-6c5663dd13140b1a9130ec407c731631434091f35d3e5ad5336f7a6a7ffcdcf3</citedby><cites>FETCH-LOGICAL-c390t-6c5663dd13140b1a9130ec407c731631434091f35d3e5ad5336f7a6a7ffcdcf3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0021-9673(99)00857-2$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>309,310,314,780,784,789,790,3550,23930,23931,25140,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1320829$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10722093$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Haj-Yehia, Abdullah I</creatorcontrib><creatorcontrib>Assaf, Peter</creatorcontrib><creatorcontrib>Nassar, Taher</creatorcontrib><creatorcontrib>Katzhendler, Jehoshua</creatorcontrib><title>Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection</title><title>Journal of Chromatography A</title><addtitle>J Chromatogr A</addtitle><description>A highly sensitive method for the determination of α-lipoic acid (LA) and dihydrolipoic acid (DHLA) in human plasma and urine has been developed. Samples were acidified and extracted with organic solvent, and the free sulfhydryls of DHLA protected as the dicarboxyethylate by treatment with ethylchloroformate. The free carboxylic function of LA and the SH-protected DHLA were converted into their amide derivatives with the strong fluorophore 2-(4-aminophenyl)-6-methylbenzothiazole in the presence of a coupling agent and a base catalyst. The resulting fluorescent amides of both LA and DHLA were separated on a reversed-phase column (Ultrasphere C
8) using simple isocratic elution with acetonitrile–water (80:20) and detected fluorimetrically (excitation 343, emission 423 nm). The method is highly sensitive, reproducible, and is easily applied for the simultaneous determination of LA and DHLA in biological samples.</description><subject>Aminophenylmethylbenzothiazole</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>Coenzymes, vitamins</subject><subject>Dihydrolipoic acid</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Lipoic acid</subject><subject>Medical sciences</subject><subject>Miscellaneous. Technology</subject><subject>Other biological molecules</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><subject>Spectrometry, Fluorescence</subject><subject>Thioctic Acid - analogs & derivatives</subject><subject>Thioctic Acid - analysis</subject><subject>Thioctic Acid - blood</subject><subject>Thioctic Acid - urine</subject><subject>Thiols</subject><issn>0021-9673</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc1O3TAQhb1oBZT2EVp5gaqySBnHN_b1ClW0FCQkFrC35vqHGCVxsJNW9zl4YXx_VNh1NdLRNzNH5xDymcF3Bkyc3QHUrFJC8m9KnQIsG1nV78jRP_mQfMj5EYBJkPUBOWRl1KD4EXn-6SaX-jDgFOJAo6ddGGMwFE2wFAdLbWjXNsW3chhoO_c40LHD3OMWm1MYHF2taRse2mp0ycdUEOPKwae5LJk2xR6n-JBwbNf0b5ha6rs5ptC7KZXTtjgxGxcfyXuPXXaf9vOY3F_-ur-4qm5uf19f_LipDFcwVcI0QnBrGWcLWDFUjIMzC5BGciaKyBegmOeN5a5B23AuvESB0ntjjefH5Ovu7Jji0-zypPuQjes6HFycs5aghFrCsoDNDjQp5pyc12MxjWmtGehNAXpbgN4krZXS2wJ0Xfa-7B_Mq97ZN1u79AtwsgcwG-x8KnmF_MrxGpa1Ktj5DnMljD_BJZ1NcCVaG1JJTNsY_uPkBehhp2U</recordid><startdate>20000218</startdate><enddate>20000218</enddate><creator>Haj-Yehia, Abdullah I</creator><creator>Assaf, Peter</creator><creator>Nassar, Taher</creator><creator>Katzhendler, Jehoshua</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20000218</creationdate><title>Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection</title><author>Haj-Yehia, Abdullah I ; Assaf, Peter ; Nassar, Taher ; Katzhendler, Jehoshua</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c390t-6c5663dd13140b1a9130ec407c731631434091f35d3e5ad5336f7a6a7ffcdcf3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Aminophenylmethylbenzothiazole</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>Coenzymes, vitamins</topic><topic>Dihydrolipoic acid</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Lipoic acid</topic><topic>Medical sciences</topic><topic>Miscellaneous. Technology</topic><topic>Other biological molecules</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><topic>Spectrometry, Fluorescence</topic><topic>Thioctic Acid - analogs & derivatives</topic><topic>Thioctic Acid - analysis</topic><topic>Thioctic Acid - blood</topic><topic>Thioctic Acid - urine</topic><topic>Thiols</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Haj-Yehia, Abdullah I</creatorcontrib><creatorcontrib>Assaf, Peter</creatorcontrib><creatorcontrib>Nassar, Taher</creatorcontrib><creatorcontrib>Katzhendler, Jehoshua</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of Chromatography A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Haj-Yehia, Abdullah I</au><au>Assaf, Peter</au><au>Nassar, Taher</au><au>Katzhendler, Jehoshua</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection</atitle><jtitle>Journal of Chromatography A</jtitle><addtitle>J Chromatogr A</addtitle><date>2000-02-18</date><risdate>2000</risdate><volume>870</volume><issue>1</issue><spage>381</spage><epage>388</epage><pages>381-388</pages><issn>0021-9673</issn><coden>JOCRAM</coden><abstract>A highly sensitive method for the determination of α-lipoic acid (LA) and dihydrolipoic acid (DHLA) in human plasma and urine has been developed. Samples were acidified and extracted with organic solvent, and the free sulfhydryls of DHLA protected as the dicarboxyethylate by treatment with ethylchloroformate. The free carboxylic function of LA and the SH-protected DHLA were converted into their amide derivatives with the strong fluorophore 2-(4-aminophenyl)-6-methylbenzothiazole in the presence of a coupling agent and a base catalyst. The resulting fluorescent amides of both LA and DHLA were separated on a reversed-phase column (Ultrasphere C
8) using simple isocratic elution with acetonitrile–water (80:20) and detected fluorimetrically (excitation 343, emission 423 nm). The method is highly sensitive, reproducible, and is easily applied for the simultaneous determination of LA and DHLA in biological samples.</abstract><cop>Amsterdam</cop><pub>Elsevier B.V</pub><pmid>10722093</pmid><doi>10.1016/S0021-9673(99)00857-2</doi><tpages>8</tpages></addata></record> |
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| subjects | Aminophenylmethylbenzothiazole Analytical, structural and metabolic biochemistry Biological and medical sciences Chromatography, High Pressure Liquid - methods Coenzymes, vitamins Dihydrolipoic acid Fundamental and applied biological sciences. Psychology Humans Investigative techniques, diagnostic techniques (general aspects) Lipoic acid Medical sciences Miscellaneous. Technology Other biological molecules Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Reproducibility of Results Sensitivity and Specificity Spectrometry, Fluorescence Thioctic Acid - analogs & derivatives Thioctic Acid - analysis Thioctic Acid - blood Thioctic Acid - urine Thiols |
| title | Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection |
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