Vitis vinifera L. cv. Shiraz
(syn. Syrah) grape berries grown during the 2001 harvest
season on the Waite Campus at the University of Adelaide
were used as the source material for proanthocyanidins.
Samples were collected at three specific times during berry
development: three weeks after fruit set, ve´raison (the commencement
of fruit ripening), and commercial maturity. The
sample collected at ve´raison was divided into unpigmented and
pigmented berries. All samples (5 kg) consisted of randomly
collected fruit clusters within the same vineyard block.
Briefly, proanthocyanidin purification was carried out as
follows. Isolated skins were extracted in 2:1 acetone/water.
Afterward, acetone was removed under reduced pressure, and
the aqueous portion was extracted with chloroform, freezedried
to a powder, and purified by adsorption chromatography
using Toyopearl TSK HW 40-F size exclusion media (Sigma;
Castle Hill, NSW, Australia). Adsorbed proanthocyanidins
were rinsed (3 column volumes) with 1:1 methanol/water
containing 0.1% v/v trifluoroacetic acid to remove low molecular
weight contaminants. The sample collected at commercial
maturity was rinsed with an additional 7 column volumes of
this solvent in an attempt to remove apparent anthocyanin
impurities. After rinsing the samples, proanthocyanidins were
eluted with 2:1 acetone/water with 0.1% v/v trifluoroacetic acid
added. The acetone was evaporated, and the aqueous residue
was freeze-dried to a powder. With the exception of the
chloroform extraction, proanthocyanidin isolation and purification
has been previously described (19).
Vitis vinifera L. cv. Shiraz(syn. Syrah) grape berries grown during the 2001 harvestseason on the Waite Campus at the University of Adelaidewere used as the source material for proanthocyanidins.Samples were collected at three specific times during berrydevelopment: three weeks after fruit set, ve´raison (the commencementof fruit ripening), and commercial maturity. Thesample collected at ve´raison was divided into unpigmented andpigmented berries. All samples (5 kg) consisted of randomlycollected fruit clusters within the same vineyard block.Briefly, proanthocyanidin purification was carried out asfollows. Isolated skins were extracted in 2:1 acetone/water.Afterward, acetone was removed under reduced pressure, andthe aqueous portion was extracted with chloroform, freezedriedto a powder, and purified by adsorption chromatographyusing Toyopearl TSK HW 40-F size exclusion media (Sigma;Castle Hill, NSW, Australia). Adsorbed proanthocyanidinswere rinsed (3 column volumes) with 1:1 methanol/watercontaining 0.1% v/v trifluoroacetic acid to remove low molecularweight contaminants. The sample collected at commercialmaturity was rinsed with an additional 7 column volumes ofthis solvent in an attempt to remove apparent anthocyaninimpurities. After rinsing the samples, proanthocyanidins wereeluted with 2:1 acetone/water with 0.1% v/v trifluoroacetic acidadded. The acetone was evaporated, and the aqueous residuewas freeze-dried to a powder. With the exception of the
chloroform extraction, proanthocyanidin isolation and purification
has been previously described (19).
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