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Use of Nitrobenzene Oxidation for Study of Lignin Composition with an Improved Method for Product Extraction¹

Ruminant Nutrition Laboratory, Animal Science Institute, Agricultural Research Service, US Department of Agriculture, Beltsville, MD 20705

ABSTRACT

Sample stability, extraction procedures and solvents and various gas chromatographic procedures were studied in an attempt to improve procedures for the determination of lignin by the nitrobenzene oxidation technique. Extraction of oxidation products with ether, although needed to extract p-coumaric acid, resulted in a marked decrease in sample stability and increased extraneous material. Studies using a variety of phases and sample preparations showed that with 5% OV-25 on Gas Chrom Q the peaks corresponding to caffeic and ferulic acids were largely or entirely azoxybenzene and p-phenylazophenol. The introduction of disposable diatomaceous earth columns for extracting oxidation products resulted in a faster procedure and the removal of the by-products azoxybenzene and p-phenylazophenol from the extracted products.

¹ Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the US Department of Agriculture and does not imply its approval to the exclusion of other products that may be suitable.