Using the reversed phase high performance liquid chromatography with diode-array and massspectrometric (in ESI mode) detection, a composition of Catharanthus roseum petals was established. After the acid hydrolysis, all five anthocyanidins were found to be different comparing to the ordinary anthocyanidins from Vitis vinifera fruits. The anthocyanins were elucidated to be 7-O-methyl derivatives of delphinidin, cyanidin, petunidin, peonidin and malvidin by the analysis of retention in RP HPLC, mass- and UV-visible spectra.The anthocyanins were characterized with UV-visible spectra, having the same fixtures as the set of nonmethylated (in position 7) anthocyanin with hypsochromic (4 nm) of spectral maxima. The absorption bands for 7-methylcyanidin and 7-methylpeonidin aglycons and derivatives were indistinguishable while for the set of 7-methydelphinidin, 7-methypetunidin and 7-methylmalvidine (hirsutidin) a consecutive shift of absorption maximaby approximately 1 – 1.5 nm was found. The same was true for non-methylates at position 7 derivatives. The analysis of retention of anthocyanins of the flowers including the comparison with the retention of Mangifera indica skin anthocyanins, mass- and UV-visible spectra indicated that a minor set of anthocyanins included two sets of derivatives. The minor compounds were found to be 3-galactosides for samples under investigation, while the set of the major anthocyanins was represented by 3-rhamnosylgalactosides. Indeed, though through mass-spectra it was not possible to differentiate 3-rhamnosylgalatosides and 3-(p-coumaroylgalactosides) because of m/z coincidence, the retention difference between the two found anthocyanins sets as well as UV-visible spectra excluded the latter type of derivatives.
The anthocyanin composition of five purple leaves cultivars of Ocimum basilicum L. was investigated by reversed-phase HPLC with mass-spectrometric detection by ESI mode with ion partial fragmentation as well as preparation of dried differently colored forms of anthocyanins encapsulated into maltodextrinmatrix. Analysis of the mass spectra revealed that according to the chromatographic profile the set of basil cultivar anthocyanins under investigation may be divided into two groups with the common feature being ahigh level of acylation with (mainly) p-coumaric, ferulic and malonic acids of the same base: cyanidin-3-dihexoside-5-hexoside. The presence of acylation with substituted cinnamic acids permits us to obtain solutions not only with a red color (the property of the flavylium form) but also with blue shades of coloration due to quinonoid and negatively charged quinonoid forms. All forms except that of flavylium are not stable in solution but stable enough to prepare dried encapsulated forms by lyophilization. Although the loss of anthocyaninswith drying is not negligible, the final product is characterized with high stability for storage in a refrigerator.
The selectivity of the separation of some anthocyanins on Diasphere-11-C10CN stationary phase (phase I) is compared with the traditional reversed Symmetry C18 phase (phase II). It is found that, in contrast to phase II, phase I is effective in the separation of isomeric pairs of anthocyanins of 6-hydroxycyanidin-3-rutinoside and delphinidin-3-rutinoside, 6-hydroxypelargonidin-3-rutinoside and cyanidin-3-rutinoside, which ensures the determination of anthocyanins of Alstroemeria flowers. A comparison of separation maps shows that, on phase I, as compared with phase II, retention does not decrease so much, when OH groups are added to the anthocyanidin structure; trend lines for 3-mono-, di-, and triglucosides have a higher slope, and the addition of a glucosidic substituent at position 5 results in a more significant decrease in the retentionof anthocyanins. Different selectivity of the separation of anthocyanins on phase I makes this separation version a good alternative to traditional reversed phase chromatography.
Chemical preservation is an important process that prevents foods, personal care products, woods and household products, such as paints and coatings, from undesirable change or decomposition by microbial growth. To date, many different chemical preservatives are commercially available, but they are also associated with health threats and severe negative environmental impact. The demand for novel, safe, and green chemical preservatives is growing, and this process is further accelerated by the European Green Deal. It is expected that by the year of 2050 (or even as soon as 2035), all preservatives that do not meet the ‘safe-by-design’ and ‘biodegradability’ criteria are banned from production and use. To meet these European goals, there is a large need for the development of green, circular, and bio-degradable antimicrobial compounds that can serve as alternatives for the currently available biocidals/ preservatives. Anthocyanins, derived from fruits and flowers, meet these sustainability goals. Furthermore, preliminary research at the Hanze University of Applied Science has confirmed the antimicrobial efficacy of rose and tulip anthocyanin extracts against an array of microbial species. Therefore, these molecules have the potential to serve as novel, sustainable chemical preservatives. In the current project we develop a strategy consisting of fractionation and state-of-the-art characterization methods of individual anthocyanins and subsequent in vitro screening to identify anthocyanin-molecules with potent antimicrobial efficacy for application in paints, coatings and other products. To our knowledge this is the first attempt that combines in-depth chemical characterization of individual anthocyanins in relation to their antimicrobial efficacy. Once developed, this strategy will allow us to single out anthocyanin molecules with antimicrobial properties and give us insight in structure-activity relations of individual anthocyanins. Our approach is the first step towards the development of anthocyanin molecules as novel, circular and biodegradable non-toxic plant-based preservatives.
