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The composition of common dogwood fruits anthocyanins was determined using reverse-phase HPLC with spectrophotometric and mass spectrometric detection. A procedure for determining the structure of individual components of extracts including 3-galactosides of delphinidin, cyanidin and pelargonidin and 3-robinobiosides (3-rhamnosylgalactosides) of cyanidin and pelargonidin has been described. It was found that the content of delphinidin-3-galactoside, as well as both 3-robinobiosides, may be insignificant when cyanidin and pelargonidin 3-galactosides predominate. For the studied fruits, the level of accumulation of anthocyanins can reach (depending on color intensity) 46 mg per 100 g of fresh fruit, while the concentration of anthocyanins in the peel of fruits is significantly higher - more than 390 mg per 100 g. When developing an exhaustive extraction of anthocyanins, a peculiarity of dogwood fruits was established, which consists in the fact that extraction of anthocyanins by 0.1 M aqueous hydrochloric acid solution, unlike a large number of other natural sources studied, is practically impossible. The problem can be solved only by extraction with acidified aqueous-alcoholic mixtures when the ethanol content in the extractant is more than 30 vol.%, which indicates the high affinity of fruit pulp components to anthocyanins., В работе с использованием обращенно-фазовой ВЭЖХ со спектрофотометрическим и масс-спектрометрическим детектированием определен состав антоцианов плодов кизила обыкновенного. Описана процедура установления строения индивидуальных компонентов экстрактов, включающих 3-галактозиды дельфинидина, цианидина и пеларгонидина и 3-робинобиозиды (3-рамнозилгалактозиды) цианидина и пеларгонидина. Установлено, что содержание дельфинидин-3-галактозида, как и обоих 3-робинобиозидов, может быть незначительным при превалировании в составе 3-галактозидов цианидина и пеларгонидина. Для исследованных плодов уровень накопления антоцианов может достигать (в зависимости от интенсивности окраски) 46 мг на 100 г свежих плодов, при этом концентрация антоцианов в кожуре плодов существенно выше – более 390 мг на 100 г. При разработке исчерпывающей экстракции антоцианов установлена особенность плодов кизила, состоящая в том, что экстракция антоцианов 0.1 М водным раствором соляной кислоты, в отличие от большого числа других исследованных природных источников, практически невозможна. Задача может быть решена только при экстракции подкисленными водно-спиртовыми смесями при содержании этанола в экстрагенте более 30 об.%, что указывает на высокое сродство компонентов мякоти плодов к антоцианам.

The anthocyanin composition of some species of gooseberries with colored fruits was studied in comparison with anthocyanins of currant fruits from the collection of the Tsitsin Main Botanical garden. In fruits of currants Ribes nigrum var. sibiricum E. Wolf, R. nigrum L., R. americanum Mill., as well as in dark-colored fruits R. aureum Pursh., 3-glucosides and 3-rutinosides of cyanidin and delphinidin were found as major components in species-dependent ratios. It was found that in the fruits of gooseberry species Ribes oxyacanthoides L., Ribes missouriensis (Nutt.) Coville & Britton, Ribes rusticum Jancz. and Ribes cynosbati (L.) Mill. and in the fruits of one of the currants (R. alpinum L.) there is practically no biosynthesis of delphinidin derivatives, and 3-glucoside and 3-rutinoside of cyanidin are detected. Meanwhile, the biosynthesis of acylated by substituted cinnamic acids (mainly – p-coumaric acid) anthocyanins were found in the fruits of Ribes oxyacanthoides and Ribes cynosbati (about 10%), Ribes rusticum (about 20%). The inheritance of characteristic for the fruit of some species of gooseberry biosynthesis of acylated mainly p-coumaric acid, was installed in the fruits of jostaberry - Ribes x nidigrolaria Rud. Bauer & A. Bauer (about 10%). The species composition of anthocyanins was established using traditional reversed-phase HPLC in gradient elution mode using diode-array and mass spectrometric detection options. In addition, a method for the separation of anthocyanins of the same objects in the isocratic mode under hydrophilic interaction chromatography on diol stationary phases, allowing the use of simpler equipment. The variants of sample preparation of extract samples for the subsequent HPLC determination of anthocyanins by two different methods to avoid the appearance of artifacts are proposed., Исследован антоциановый состав некоторых видов крыжовника с окрашенными плодами в сопоставлении с антоцианами плодов смородин из коллекции Главного ботанического сада им. Н.В. Цицина. В плодах смородин Ribes nigrum var. sibiricum E.Wolf, Ribes nigrum L., Ribes americanum Mill., как и в темноокрашенных плодах Ribes aureum Pursh., обнаружены 3-глюкозиды и 3-рутинозиды цианидина и дельфинидина в качестве основных компонентов взависящих от видов соотношениях. Установлено, что в плодах крыжовника видов Ribes oxyacanthoides L., Ribes missouriensis (Nutt.) Coville & Britton, Ribes rusticum Jancz. и Ribes cynosbati (L.) Mill. и в плодах одной из смородин (Ribes alpinum L.) практически отсутствует биосинтез производных дельфинидина, а также детектируются 3-глюкозид и 3-рутинозид цианидина. При этом биосинтез ацилированных замещенными коричными кислотами (главным образом – пара-кумаровой) антоцианов найден в плодах Ribes oxyacanthoides и Ribes cynosbati (около 10%), Ribes rusticum (около 20%). Наследование характерного для плодов некоторых видов крыжовника признака – биосинтеза ацилированных восновном пара-кумаровой кислотой, – установлено в плодах йошты – Ribes x nidigrolaria Rud. Bauer & A. Bauer (около 10%). Видовой состав антоцианов устанавливали с использованием традиционной обращенно-фазовой ВЭЖХ вградиентном режиме элюирования с применением диодно-матричного и масс-спектрометрического вариантов детектирования. Кроме того, разработан способ разделения антоцианов этих же объектов в изократическом режиме в условиях гидрофильной хроматографии на диольных стационарных фазах, позволяющий использовать более простое оборудование. Предложены варианты пробоподготовки образцов экстрактов для последующего ВЭЖХ определения антоцианов двумя различными методами, позволяющими избежать появления артефактов.

The method of reversed-phase HPLC with diode-array and mass-spectrometric detection was explored to determine the qualitative composition of red cabbage anthocyanins. Mono- and diacylated derivatives of cyanidin-3,5-diglycosides with a complex structure of the substituent in position were found, as well as non-esterified compounds. The acylation acids were paracoumaric, ferulic and sinapic. Quantitative analysis was performed according to a simplified spectrophotometric method: absorbance measurement at pH 4.5 was excluded. The measurement showed that the level of anthocyanins accumulation is as high as 0.100 g of anthocyanins per 100 g of raw plant material (as cyanidin-3-glucoside chloride equivalent). Analysis of electronic spectra of the extract at different pH revealed the existence of some basic structures with absorbance maxima shifted bathochromically as compared to the spectrum of flavylium structure. The existence of one basic form at pH 4.5 forced us to exclude absorbance measurement at pH 4.5 during quantitative anthocyanin determination. Thus, the possibility of obtaining some differently colored encapsulated forms was proved. Because of relative instability of non-flavylium forms of anthocyanins the freeze drying technique was used to prepare six differently colored forms of 3% anthocyanins encapsulated in maltodextrin. The decrease of anthocyanins contents at drying did not exceed 30%, while no statistically proved loss of anthocyanins was found at storage in a refrigerator during more than six months.

Reversed-phase HPLC with mass spectrometric and diode array detection as well as some literature data were used to reveal the individual types of solutes in anthocyanin complexes of tulip flower petals that are responsible for tulip flower petals coloration of the samples available in the local flower market. It has been found that the main components of the complexes are 3-rutinosides and their 2”’ and 3”’ acylated with acetic acid derivatives of the three anthocyanidins - delphinidin, cyanidin and pelargonidin in the color dependent ratios, though trace quantities of 3-glucosides were found in some cases. For the anthocyanin structure confirmation a correlation analysis of solute retentions of cyanidin or pelargonidin derivatives vs that of delphinidin was proposed based upon equivalence of structures alteration in the solute pairs for each series. The specificity of solutes retention modes was revealed by relative retention analysis, the trend parameters reflected particularities of chromatographic behavior as well as that of electron spectra of the solutes. The difference of acylated anthocyanins retentions was proposed to disclose the conformation states of solutes in the sorbent interface.

Reversed-phase HPLC with mass spectrometric and diode array detection as well as some literature data were used to reveal the individual types of solutes in anthocyanin complexes of tulip flower petals that are responsible for tulip flower petals coloration of the samples available in the local flower market. It has been found that the main components of the complexes are 3-rutinosides and their 2”’ and 3”’ acylated with acetic acid derivatives of the three anthocyanidins - delphinidin, cyanidin and pelargonidin in the color dependent ratios, though trace quantities of 3-glucosides were found in some cases. For the anthocyanin structure confirmation a correlation analysis of solute retentions of cyanidin or pelargonidin derivatives vs that of delphinidin was proposed based upon equivalence of structures alteration in the solute pairs for each series. The specificity of solutes retention modes was revealed by relative retention analysis, the trend parameters reflected particularities of chromatographic behavior as well as that of electron spectra of the solutes. The difference of acylated anthocyanins retentions was proposed to disclose the conformation states of solutes in the sorbent interface.