Extraction and purification of squalene from Amaranthus grain

Abstract

Grain amaranth has been suggested as an alternative to marine animals as a natural source of squalene. Oil contents, squalene contents, and fatty acid profiles were determined in 11 genotypes of four grain amaranth species. Although the oil contents of grain amaranth were low (from 5.1% in Amaranthus tricolor to 7.7% in Amaranthus cruentus) as compared to other oil-containing grains, high concentrations of squalene were found in total lipids, ranging from 3.6% in Amaranthus hypochondriacus to 6.1% in A. tricolor. The major fatty acids in Amaranthus oil consisted of palmitic acid (19.1-23.4%), oleic acid (18.7-38.9%), and linoleic acid (36.7-55.9%). A high degree of unsaturation was observed in Amaranthus oils, with S/U ratios of 0.26-0.32. A method to isolate and purify the squalene from Amaranthus oil was developed. After the saponification of K112, the squalene content increased from 4.2% in the crude oil to 43.3% in the unsaponifiables by the removal of the saponifiables. The unsaponifiables were fractionated by silica gel column chromatography to get highly purified squalene. The squalene purity in certain fractions was as high as 98%. Combining the fractions rich in squalene gave a 94% squalene concentrate, with a yield of 90%. The structure of squalene in the purified sample was confirmed by comparison of its ultraviolet spectrum with a standard and from its nuclear magnetic resonance spectra.