PNL Volume 15
Ludwicki, J.
Swiecicki, W. K.
Agricultural Academy, Warsaw, Poland
Plant Experiment Station, Wiatrowo, Poland
Elsewhere in this volume a new seed character is described, viz.
orange cotyledons. Because the difference in color in comparison to
typical yellow seeds was very clear we decided to analyze the pigment
extracts by chromatography. Seeds of two lines, Wt 11145 (orange
cotyledons) and Wt 3527 (typical yellow cotyledons), were analyzed.
The material was ground and hydrolyzed (KOH in alcohol) prior to
extraction with hexane.
Chromatographic separation of carotenoids was done by MgO column
chromatography using hexane and 20% solution of acetone in hexane as
solvents. For the calculation of carotenoid content, tabular values of
were used, viz: B-carotene 2592, B-cryptoxanthin 2386, lutein 2550.
For determination of the provitamin value the biological activity of
S-carotene was taken as 100%, 6-cryptoxanthin as 57% and lutein as 0.
Chromatographic analysis of pigment extracts from seeds of line
Wt 3527 (yellow cotyledons) showed the presence of two carotenoid frac-
tions (Fig. 1). Their elution was possible only after using a 20%
solution of acetone in hexane. Use of less polar solvents did not cause
visible migration of the pigments. Nor did the above conditions make
possible the removal of trace levels of blue-pink pigment (probably
anthocyanin) which formed an isolated band at the top of the column.
Chromatographic separation of pigments from seeds of line Wt 11145
(orange cotyledons) revealed three bands of carotenoids (Fig. 2). The
first, most mobile fraction which did not occur in the "typical" yellow
seeds was eluted by pure hexane. Two remaining fractions of carotenoids
were comparable with line Wt 3527. Again, the band of blue-pink pigment
was observed.
Spectral investigations showed (Fig. 3) that carotenoids in seeds
of line Wt 11145 had the maximum absorption in the following wave
lengths (nm):
I fraction - 425, 450, 477
II fraction - 425, 4 46, 475
III fraction - 422, 445, 475
On the basis of spectral data the respective fractions were identified
as I - 6-carotene; II - ft-cryptoxanthin; III - lutein.
Carotenoid pigments isolated from seeds of Wt 3527 (Fig. 4) had
maximum absorption in wave lengths (run):
I fraction - 425, 4 46, 477
II fraction - 422, 4 45, 475
The above values indicate the presence of 6-cryptoxanthin and lutein.
Table 1 gives the content of carotenoids and provitamin.
Thus it appears that seeds of Wt 11145 have more than double the
amount of biologically active f.-cryptoxanthin, plus a small amount of
B-carotene, in comparison with the yellow cotyledon line. The
provitamin activity was nearly tripled. The higher content of lutein in
line Wt 11145 is not significant from a nutritional point of view.
PNL Volume 15