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A rapid and convenient preparation of [4-3H]NADP and stereospecifically tritiated NADP3H

✍ Scribed by Richard G. Moran; Pamela Sartori; Valenrie Reich

Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
731 KB
Volume
138
Category
Article
ISSN
0003-2697

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✦ Synopsis

The enzymatic preparation and chromatographic purification of [4-'H]NADP and NADPH stereospecifically labeled with 'H on either the A or B faces at position 4 have been simplified. Commercially available [ l-3H]glucose was used as a starting material for the sequential synthesis of [4B-"HINADPH, [4-'HINADP, and [4A-'HINADPH. These products were rapidly purified by step elution of DEAE-cellulose minicolumns so that [4B-3H]NADPH was produced and purified from [ l-3H]glucose in 2 h, [4-'HINADP in 5 h, and [4A-'H]NADPH in 8 h. Yields of these products were 65 to 88% starting with [ l-3H]glucose. Analysis of the products by highperformance liquid chromatography indicated radiochemical purities of 82-95% for these compounds and specific activities equivalent to that of the starting material (lo-15 Ci/mmol).

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