Development of a HPLC method for the analysis of carotenoids in vegetables and fruits

Tee E.S., . and Lim C.L., . (1990) Development of a HPLC method for the analysis of carotenoids in vegetables and fruits. [Proceedings Paper]

Preliminary studies were carried out to apply a HPLC method developed for the analysis of carotenoids in plant foods. The eight vegetables selected were prepared by two pre-treatment procedures namely with and without prior saponification and chromatographed by the reverse-phase HPLC method developed. Carotenoids peaks obtained were tentatively identified using eight reference standards similar chromatographed. To assist in the confirmation of the pigments obtained samples were also chromatographed by open-column chromatography employing a mixture of magnesia and diatomaceous earth. Fractions were eluted with an acetone-hexane mixture using a stepwise increase in the proportion of acetone in hexane studied by UV-vis absorption spectrophotometry and re-chromatographed on HPLC. The major carotenoids alpha- and beta- carotenes lutein and lycopene were thus identified. Saponification removed various non-carotenoid pigments in green vegetables resulting in HPLC profiles which were easier to quantitate as well as prolonging the life of the HPLC columns. Total carotenoid content of the saponified vegetables could be calculated by taking the absorbance reading of the extracts at 450 nm. The saponification process did not result in a significant loss of beta-carotene although there appeared to be some loss of lutein which however possesses no pro-vitamin A activity.

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Abstract

Preliminary studies were carried out to apply a HPLC method developed for the analysis of carotenoids in plant foods. The eight vegetables selected were prepared by two pre-treatment procedures namely with and without prior saponification and chromatographed by the reverse-phase HPLC method developed. Carotenoids peaks obtained were tentatively identified using eight reference standards similar chromatographed. To assist in the confirmation of the pigments obtained samples were also chromatographed by open-column chromatography employing a mixture of magnesia and diatomaceous earth. Fractions were eluted with an acetone-hexane mixture using a stepwise increase in the proportion of acetone in hexane studied by UV-vis absorption spectrophotometry and re-chromatographed on HPLC. The major carotenoids alpha- and beta- carotenes lutein and lycopene were thus identified. Saponification removed various non-carotenoid pigments in green vegetables resulting in HPLC profiles which were easier to quantitate as well as prolonging the life of the HPLC columns. Total carotenoid content of the saponified vegetables could be calculated by taking the absorbance reading of the extracts at 450 nm. The saponification process did not result in a significant loss of beta-carotene although there appeared to be some loss of lutein which however possesses no pro-vitamin A activity.

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Item Type:	Proceedings Paper
Additional Information:	1 ill.; 10 graphs; 2 tables; 16 ref. Summary En
AGROVOC Term:	HORTALIZAS
AGROVOC Term:	FRUTAS
AGROVOC Term:	CROMATOGRAFIA LIQUIDA ALTA PRESION
AGROVOC Term:	CAROTENOIDES/ SAPONIFICACION
Geographical Term:	MALAYSIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:26
URI:	http://webagris.upm.edu.my/id/eprint/14719

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