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Metabolism and Enzymology

Comparison of Tissue Preparation Methods for Assay of Nicotinamide Coenzymes

Department of Botany, Colorado State University, Fort Collins, Colorado 80523

To prepare tissues for analysis of NAD⁺, NADH, NADP⁺, and NADPH, common practice is to freeze samples in liquid nitrogen, often followed by freeze-drying, before extraction in HCl or NaOH. With cucumber (Cucumis sativus L.) cotyledons, prefreezing in liquid nitrogen or slower freezing to –20°C yielded substantially lower values for NADH and NADPH than obtained from samples homogenized immediately in acid or base. Freeze-drying after freezing in liquid nitrogen generally caused even lower values of those coenzymes. We suggest that direct extraction is more likely to yield accurate results with cotyledons and other plant parts.

² Permanent address: Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China.

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