For the first time, a new version of the separation of 3-glucosides and 3,5-diglucosides of the five main anthocyanidins for grape fruits -delphinidin, cyanidin, petunidin, peonidin and mal-vidin under reverse-phase HPLC conditions was proposed. It has been shown that the replacement of the traditionally used acetonitrile with environmentally more acceptable ethanol upon acidifica-tion with ortho-phosphoric rather than formic acid makes it possible to significantly change the selectivity of the separation of two types of glucosides at their joint presence. The proposed separa-tion option allows differentiating between Vitis vinifera grapes, in the fruit peel of which only 3-glucosides of the above anthocyanidins are synthesized, and grapes of other species or hybrid grape varieties. For a complete review of the anthocyanin composition, it is necessary to use a gradient mode, since the acylation of anthocyanins with acetic and p-coumaric acids significantly changes the lipophilicity of anthocyanins. The type of anthocyanins was analyzed according to the previously proposed system, which takes into account the activity of three types of enzymes: 1) 5-O-glucosyltransferase that leads to 3,5-diglucosides synthesis; 2) 3',5'-hydroxylase responsible for ring B hydroxylation; 3) anthocyanin O-methyltransferase for converting cyanidin into peonidin derivatives as well as delphinidine into petunidin and malvidin glycosides. The method was used to determine anthocyanins in 43 grape varieties grown in Belgorod on farms and private farms. Among the studied grapes, varieties were found with grape fruits with the accumulation of only 3-glucosides, as well as 3-glucosides and 3,5-diglucosides simultaneously, with delphinidin and cyanidin types of anthocyanins, and with different degrees of methylation. The calculated parameters for all grape varieties are presented in two tables and a brief discussion of some varieties is given.

A group control procedure is developed, in which the anthocyanin complex of a grape extract is divided into six groups without differentiation by aglycons: 3-glucosides (Zone III); 3-glucosides acylated with p -coumaric acid (Zone I); 3-glucosides acylated with acetic acid (Zone II); 3,5-diglucosides (Zone IV); 3,5-diglucosides acylated with p -coumaric acid (Zone IV); and 3,5-diglucosides acylated with acetic acid (Zone V). The anthocyanins were separated by hydrophilic interaction chromatography in a column packed with a silica using the mobile phases of acetonitrile–water (20–8 vol %) containing 0.5 vol % of orthophosphoric acid. Due to the low viscosity of the mobile phase and the complete separation of anthocyanin groups, the procedure suitable for determining the inheritance of genetic traits in the selection of grapes is transferred to a format of microcolumn chromatography using MiLiChrome chromatographs.

В работе методом обращенно-фазовой ВЭЖХ с диодно-матричным и масс-спектроскопическим детектированиями исследован состав антоцианового комплекса плодов популярного местного сорта винограда неизвестной селекции. По появлению производных 3,5-диглюкозидов установлен гибридный характер данного сорта винограда. Показано, что вследствие сольватохромного эффекта количественная оценка видового состава по площадям пиков осложнена, однако при сопоставлении спектров, записанных при различном содержании в подвижной фазе ацетонитрила, найдена особая точка, при которой величина поглощения при СФ-детектировании не зависит от состава растворителя. Для выполнения анализа антоцианов винограда в изократических условиях предложен вариант флэш-хроматографии, позволяющий отделить неацилированные компоненты от ацилированных с использованием насадочных картриджей для твердофазной экстракции

Методом обращенно-фазовой ВЭЖХ c диодно-матричным, масс-спектрометрическим и амперометрическим детекторами исследованы комплексы антоцианов ряда сортов винограда с традиционным для Vitis vinifera составом, содержащие в качестве основных компонентов 3-глюкозиды пяти антоцианидинов – дельфинидина (Dp3G), цианидина (Cy3G), петунидина (Pt3G), пеонидина (Pn3G) и мальвидина (Mv3G). Разработана методика определения антиоксидантной активности индивидуальных антоцианов амперометрическим методом, выявлены особенности использования прибора Цвет Яуза-01-АА. Показано, что чувствительность амперометрического детектирования зависит не только от строения антоцианов, но и от напряжения на аноде. Предложена схема расчета антиоксидантной активности антоциановых комплексов по результатам амперометрического детектирования смеси индивидуальных антоцианов.

Anthocyan complexes of a series of grape varieties with the composition traditional for Vitis vinifera, containing 3-glucosides of five anthocyanidins, delphinidin (Dp3G), cyanidin (Cy3G), petunidin (Pt3G), peonidin (Pn3G), and malvidin (Mv3G) as the major components have been studied by reversed-phase HPLC with diode-array, mass spectrometric, and amperometric detectors. A procedure has been developed for the determination of the antioxidant activity of individual anthocyans using an amperometric method; the peculiarities of using a Tsvet Yauza-01-AA system have been evaluated. It has been shown that the sensitivity of amperometric detection depends not only on the structure of anthocyans, but also on the anode voltage. A scheme of the calculation of the antioxidant activity of anthocyan complexes by the results of amperometric detection of a mixture of individual anthocyans has been proposed.

Fruits of some cultivars of red-flesh apples derived from М alus niedzwetzkyana were found to accumulate up to 0,070 g per 100 g (for that of «Granatnoye» variety). The main component of all samples under investigation was found to be cyanindin-3-galactoside (more than 84% of anthocyanins sum) among some others cyanidin derivatives. The titrable acidity as well as antioxidant activity of «Granatnoye» fruits juice exceeded that for ordinary varieties (including red-skin ones); thus the former is of great interest for food industry.

Anthocyanins of two hawthorn species fruits: C. chlorosarca Maxim. and C. pentagyna Waldst. et Kit., were investigated by HPLC-DAD-ESI-MS method. Cyanidin-3-galactoside, cyanidine-3-glucoside, cyanidine-3-arabinoside and cyanidine-3-xiloside were found. Overall anthocyanins accumulation was the highest in the case of C. chlorosarca fruits: 0.460÷0.620 g per 100 g of fresh fruits as cyanidine-3-glucoside chloride equivalent. It has been found that according to anthocyanins accumulation as well as to the antioxidant activity C. chlorosarca fruits are rather comparable to dried hibiscus subdariffa calyces and may be used for another tea preparation at some acidification.