Determination of the Specific Radioactivity of [14C]Lactate by Enzymatic Decarboxylation and14CO2Collection
We present an enzymatic method for the determination ofl-[14C]lactate specific radioactivity in complex biological samples containing other radiolabeled compounds. The method is based on the conversion ofl-lactate tol-pyruvate by lactate oxidase (no EC number assigned) and the decarboxylation ofl-py...
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Veröffentlicht in: | Analytical biochemistry 1997-11, Vol.253 (2), p.190-195 |
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container_title | Analytical biochemistry |
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creator | Lehoux, Eric A. Svedružić, Željko Spivey, H.Olin |
description | We present an enzymatic method for the determination ofl-[14C]lactate specific radioactivity in complex biological samples containing other radiolabeled compounds. The method is based on the conversion ofl-lactate tol-pyruvate by lactate oxidase (no EC number assigned) and the decarboxylation ofl-pyruvate by pyruvate oxidase (EC 1.2.3.3). The14CO2produced by the enzymatic decarboxylation of pyruvate is quantitatively captured in a CO2trap and its radioactivity is measured. The method is simple, specific, and precise (2% relative SD). It can be conveniently used for routine multiple determinations ofl-[14C]lactate specific radioactivity in tracer metabolic studies. Under specified conditions, the method can also be used to determine the specific radioactivity ofl-[14C]pyruvate. |
doi_str_mv | 10.1006/abio.1997.2353 |
format | Article |
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The method is based on the conversion ofl-lactate tol-pyruvate by lactate oxidase (no EC number assigned) and the decarboxylation ofl-pyruvate by pyruvate oxidase (EC 1.2.3.3). The14CO2produced by the enzymatic decarboxylation of pyruvate is quantitatively captured in a CO2trap and its radioactivity is measured. The method is simple, specific, and precise (2% relative SD). It can be conveniently used for routine multiple determinations ofl-[14C]lactate specific radioactivity in tracer metabolic studies. Under specified conditions, the method can also be used to determine the specific radioactivity ofl-[14C]pyruvate.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1006/abio.1997.2353</identifier><language>eng</language><publisher>Elsevier Inc</publisher><ispartof>Analytical biochemistry, 1997-11, Vol.253 (2), p.190-195</ispartof><rights>1997 Academic Press</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c1333-9fd778716f63324f46ded9e7d51a8688d7354ff676f7ba45fba100b6277bb1373</citedby><cites>FETCH-LOGICAL-c1333-9fd778716f63324f46ded9e7d51a8688d7354ff676f7ba45fba100b6277bb1373</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0003269797923538$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids></links><search><creatorcontrib>Lehoux, Eric A.</creatorcontrib><creatorcontrib>Svedružić, Željko</creatorcontrib><creatorcontrib>Spivey, H.Olin</creatorcontrib><title>Determination of the Specific Radioactivity of [14C]Lactate by Enzymatic Decarboxylation and14CO2Collection</title><title>Analytical biochemistry</title><description>We present an enzymatic method for the determination ofl-[14C]lactate specific radioactivity in complex biological samples containing other radiolabeled compounds. The method is based on the conversion ofl-lactate tol-pyruvate by lactate oxidase (no EC number assigned) and the decarboxylation ofl-pyruvate by pyruvate oxidase (EC 1.2.3.3). The14CO2produced by the enzymatic decarboxylation of pyruvate is quantitatively captured in a CO2trap and its radioactivity is measured. The method is simple, specific, and precise (2% relative SD). It can be conveniently used for routine multiple determinations ofl-[14C]lactate specific radioactivity in tracer metabolic studies. 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title | Determination of the Specific Radioactivity of [14C]Lactate by Enzymatic Decarboxylation and14CO2Collection |
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