Rapid and convenient preparation of (4-/sup 3/H)NADP and stereospecifically tritiated NADP/sup 3/H
The enzymatic preparation and chromatographic purification of (4-/sup 3/H)NADP and NADPH stereospecifically labeled with /sup 3/H on either the A or B faces at position 4 have been simplified. Commercially available (1-/sup 3/H)glucose was used as a starting material for the sequential synthesis of (4B-/sup 3/H)NADPH, (4-/sup 3/H)NADP, and (4A-/sup 3/H)NADPH. These products were rapidly purified by step elution of DEAE-cellulose minicolumns so that (4B-/sup 3/H)NADPH was produced and purified from (1-/sup 3/H)glucose in 2 h, (4-/sup 3/H)NADP in 5 h, and (4A-/sup 3/H)NADPH in 8 h. Yields of these products were 65 to 88% starting with (1-/sup 3/H)glucose. Analysis of the products by high-performance liquid chromatography indicated radiochemical purities of 82-95% for these compounds and specific activities equivalent to that of the starting material (10-15 Ci/mmol). 15 references.
- Research Organization:
- Childrens Hospital of Los Angeles, CA
- OSTI ID:
- 7025085
- Journal Information:
- Anal. Biochem.; (United States), Vol. 138:1
- Country of Publication:
- United States
- Language:
- English
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ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
COENZYMES
STEREOCHEMISTRY
SYNTHESIS
GLUCOSE
LABELLED COMPOUNDS
LIQUID COLUMN CHROMATOGRAPHY
PURIFICATION
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ALDEHYDES
CARBOHYDRATES
CHROMATOGRAPHY
HEXOSES
ISOTOPE APPLICATIONS
MONOSACCHARIDES
ORGANIC COMPOUNDS
SACCHARIDES
SEPARATION PROCESSES
400102* - Chemical & Spectral Procedures