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Determining vitamin D content in foods is difficult because in natural foods of highest vitamin D activity, and even in vitamin D-fortified foods, only small quantities are present, and many other compounds are extracted along with vitamin D that cause difficulties in purifying the extract or in the spectrophotometry or colorimetry that follows. Several physicochemical methods--such as spectrophotometric, colorimetric, thin-layer chromatographic, adsorption, partition, gas-liquid, and high-performance column chromatographic--have been tried for assay foods for vitamin D, but none of them have been accepted for official or routine use; they are time consuming and expensive, or lack the required sensitivity, precision, or accuracy. Curative biological assays, based on degree of healing of a leg bone of rats previously made rachitic, is the generally accepted method to determine vitamin D content of foods. However, that method also requires too much time and is expensive. The recently developed high-performance liquid chromatographic method may offer the most for establishing a satisfactory physicochemical method for determining vitamin D in foods. Many of the difficulties and problems in assaying foods for vitamin D are discussed.

The vitamin E group includes tocopherols and tocotrienols and their isomers, esters, and derivatives. They differ not only in biopotencies as antisterility agents but also in activities in other physiological and chemical relationships. Unlike vitamins A and D, foods (vegetable oils) are among the richest sources of vitamin E, and assay methods for vitamin E include food applications more often than for the former vitamins. Physicochemical methods are replacing bioassays for vitamin E and tocopherol wherever possible because of greater specificity and less variability, time, and, sometimes, expense. Unless careful purifications and isolations are carried out and some of the relative vitamin E activities of components are calculated, bioassays are still required for total vitamin E activity. The vitamin E group is separated by column, paper, thin-layer, gas-liquid, and high-pressure liquid chromatography (HPLC). Gas-liquid chromatography has been more successfully used for vitamin E than for other fat-soluble vitamins. Recently developed HPLC methods for vitamin E are sensitive and apparently require less cleanup of extracts and less time than former methods; HPLC may prove to be the most useful technique for vitamin E in foods, especially if other fat-soluble vitamins can be determined simultaneously on the same sample extract.

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