PNL Volume 17 1985

RESEARCH REPORTS 51

CAROTENOID COMPOSITION AND PROVITAMIN A VALUES OF F₂ SEEDS FROM THE

CROSS OF WL 3527 (orc) x Wt 11145 (Orc)

Ludwicki J., . Agricultural University/SGGW, Warsaw, Poland

and W. K. Swiecicki Plant Experiment Station, Wiatrowo, Poland

The composition and content of carotenoids was investigated in dry

and fresh seeds of lines Wt 3527 (orc I- yellow cotyledons), Wt 11145

Orc 1 - orange cotyledons), and in dry F₂ seeds of the hybrid Wt 3527 x

Wt 11145. F2 seeds were divided into five groups: yellow, dark yellow,

light orange, orange, brick (see [1]).

The seeds were ground and hydrolyzed (KOH in alcohol) prior to ex-

traction with hexane. Chromatographic separation of carotenoids was

done on an MgO column using hexane as the mobile phase followed by 10

and 20% solutions of acetone in hexane.

Moreover, the separation of pigments was investigated by TLC on

silica gel using solvent mixture of hexane-ethylacetate-methanol

(75:20:7) as the mobile phase.

The content of a given carotenoid was estimated on the basis of a

spectrophotometric assay using tabular values of E^1%_1cm, viz: beta-carotene

2592; beta-cryptoxanthin 2386; lutein 2550.

For the determination of the provitamin A value, the biological

activity of beta-carotene was taken as 100% beta-cryptoxanthin as 57%, and

lutein as 0%.

PNL Volume 17

1985

RESEARCH REPORTS

Column chromatography of pigment extracts from dry seeds of line

Wt 3527 showed the presence of two fractions migrating in 10% and 20%

solution of acetone in hexane. But carotenoids from dry seeds of line

Wt 11145, fresh seeds of line Wt 3527, and all F₂ recombinants had three

fractions, eluted in turn by hexane, 10% and 20% solution of acetone in

hexane. Fresh seeds of line Wt 11145 did not contain the carotenoid

eluted from the column by 20% solution of acetone.

The TLC method showed that the fraction eluted from the column by

20% solution of acetone contained not only lutein but also two addi-

tional carotenoid pigments (Fig. 1). The following substances were iden-

tified in solutions obtained by separation of carotenoids by column

chromatography:

Further examination of the pigment in the 20% fraction which migra-

ted below lutein on the TLC chromatogram (Fig. Id) had absorption maxima

in the following wave lengths (nm): 420, 445, 472, corresponding to

taraxanthin. The lowest migrating carotenoid was not identified (Fig.

le).

Wt 11145 had two to four times more total carotenoids than the

other lines, but 90% of this was lutein (Table 1). Lutein was the pre-

dominant carotenoid in dry seeds. Fresh seeds contained greater amounts

of 6-carotene and beta-cryptoxanthin. The amount of beta-cryptoxanthin in dry

seeds was the least variable constituent: 94-110 mkg/lOOg.

The highest provitamin value was found in dry seeds of an F₂ from

cross Wt 3527 x Wt 11145 having dark yellow cotyledons (Table 2). Other

segregants had less than half the provitamin A.

Fresh

Dry

1. Blixt, S., and W. K. Swiecicki. 1983. PNL 15:9-10.

Fig. 1. Carotenoid separation

extracted from seeds of

hybrids WT 3527 and WT 11145.

a) beta-carotene;

b) beta-cryptoxanthin;

c) lutein;

d) taraxanthin;

e) unidentified;

s) standard all-trans-

beta-carotene.



	PNL Volume 17 1985	RESEARCH REPORTS 53

	Table 1. Carotenoid content isolated from pea seeds by column chromatography. Fresh and dry seeds of parents and dry seeds of selected F₂'s.



	Table 2. Provitamin A value of pea seed (in g of -carotene/lOOg)