The analysis of proanthocyanidin cleavage products after acid-catalysis in the presence of excess phloroglucinol was investigated. In the developed analytical method, a solution of 0.1 N HCl in methanol, containing 50 g/L phloroglucinol and 10 g/L ascorbic acid was prepared. The proanthocyanidin of interest was reacted in this solution (5 g/L) at 50 degrees C for 20 min, and afterward combined with 5 volumes of 40 mM aqueous sodium acetate before analysis by reversed-phase HPLC using an aqueous acetic acid and methanol gradient. This procedure was used to investigate the composition of proanthocyanidins isolated from the seed and skin tissue of Vitis vinifera L. berries. The results compared favorably to results obtained when benzyl mercaptan was used as the trapping nucleophile, indicating that phloroglucinol is an effective reagent for this analysis.

The composition of grape (Vitis vinifera L. cv. Shiraz) skin proanthocyanidins has been determined at different stages of berry development. Beginning approximately 3 weeks after fruit set and concluding at commercial ripeness, the composition of isolated skin proanthocyanidins was determined using the following analytical techniques: elemental analysis, UV-Vis absorption spectroscopy, reversed-phase HPLC after acid-catalysis in the presence of excess phloroglucinol, gel permeation chromatography, electrospray ionization mass spectrometry (ESI-MS), and (13)C NMR. On the basis of these analyses, berry development was correlated with an increase in proanthocyanidin mean degree of polymerization, an increase in the proportion of (-)-epigallocatechin extension subunits, and increases in the level of anthocyanins associated with the proanthocyanidin fraction. Additionally, data acquired from ESI-MS of the isolates following acid-catalysis in the presence of excess phloroglucinol is consistent with pectin-bound proanthocyanidins.

Polyphenols extracted from the seeds of Vitis vinifera L. cv. Shiraz berries were monitored during berry development. Initially seeds were green, plump and had pliable seed coats, but beginning at veraison the seeds browned in colour, became desiccated and the seed coats hardened. Isolated polyphenols consisted of flavan-3-ol monomers ((+)-catechin, (-)-epicatechin and (-)-epicatechin-3-O-gallate) and procyanidins. The procyanidins were maximal in the 3 weeks prior to veraison, increasing little during this period. The amounts of flavan-3-ol monomers increased 5-fold during this same period of time, indicating that the procyanidins and the flavan-3-ol monomers accumulate at different stages. Beginning at veraison, amounts of all polyphenols declined and changed in composition. The decrease in amount followed second-order kinetics. Polyphenol changes after veraison could be explained by oxidation and therefore, electron paramagnetic resonance (EPR) spectroscopy was used to follow the potential development of radical species in the developing seeds. Spectra consistent with a phenoxyl radical were observed in the developing seeds. The concentration of radicals remained low until veraison but then increased, reaching a maximum three weeks later, declining slowly thereafter. Changes in radical intensity together with other documented changes in the seed are consistent with an oxidative event occurring during fruit ripening. Abbreviations Kennedy et al. Development of seed polyphenols 245Development of seed polyphenols

Organic materials responsible for water-repellency in some Australian soils were extracted with an amphiphilic mixture of iso-propanol/l5.7 M ammonia (7 : 3, v : v) in a Soxhlet apparatus, after which the water-repellent soils were rendered wettable. The successful extraction by an organic solvent system indicates that the bulk of hydrophobicity in these soils is not covalently linked to the surface of the sand. The extracted materials restored hydrophobicity on acid washed sands or ignited sands at levels comparable to the original soils.Spectroscopic and chromatographic examination of the extracted materials indicated that both free and esterified long-chain, 1G32 carbon atom, fatty acids were present with a bimodal distribution showing maxima at C,, and CZ2. The I3C-NMR and infrared spectra of the most hydrophobic extract suggest that hydrophobicity is caused by molecules with extensive polymethylene chains. Calculations with model compounds indicate that at least a close packed monolayer is required before measurable hydrophobicity can be detected with the molarity of ethanol droplet penetration test.