Milan – The Italian company Indena has developed and validated an HPLC method that allows the identification and the direct quantification of all the bilberry anthocyanins both in plant material and in extracts. This innovative characterisation method provides an effective solution to a well-known issue in the analytical determination of the active constituents of bilberry extracts. These extracts are widely used in dietary botanicals and by the pharmaceutical market for the treatment of vascular and vision disorders.
The main actives, accounting for many of the beneficial properties of these extracts, are anthocyanins, natural pigments responsible for the red, pink, mauve, purple, blue or violet colouring of most flowers and fruits. Formed by a flavonoid molecule conjugated with a sugar, these compounds act as potent antioxidants and phosphodiesterase inhibitors, thus being particularly effective in the amelioration of capillary resistance and permeability.
Anthocyanidins, usually present in low quantity in the bilberry extract, are anthocyanins without the sugar moiety and should be considered anthocyanin degradation products occurring when there has been incorrect extract production and/or storage.
For the time being, the most common analytical method for evaluating anthocyanins involves UV-visible spectrophotometry, which allows the quantification of anthocyanins by detecting them in the visible region. Although widely used, this method lacks in specificity and it does not allow the identification of each anthocyanin. As a consequence, it is impossible to detect anthocyanins extracts produced from different plant materials (raspberry, blackberry, black currant, elder¬berry, etc.). The best technique for the standardization of anthocyanin extracts is High-performance Liquid Chromatography (HPLC), which allows the evaluation of individual anthocyanins, though this method is neither easily reproducible nor does it allow the complete separation of all the constituents.
In order to simplify the UV-visible and the HPLC procedures the original extract is often modified by acidic hydrolysis in order to convert all anthocyanins into anthocyanidins. As reported in the literature, these methods are far from being satisfactory as they form anthocyanidins by hydrolysis and provide an overestimation of anthocyanin content, if any proanthocyanidins are present in the extracts. Moreover, this procedure does not allow the quantification of the real content of free anthocyanidins, which are markers of extract degradation.
Recently introduced into the Italian Pharmacopoeia, the new HPLC method by Indena overcomes all of these analytical issues. As a result of its good reproducibility and its high specificity, the method is able to identify unequivocally the botanical raw materials used for manufacturing and evaluating the composition of the extracts, thus providing a high degree of product consistency and quality.
(Source: Mirtoselect® A new validated HPLC method of analysis - http://www.indena.com/pdf/mirtoselectlr.pdf)
Indena is the world's leading company dedicated to the identification, development and production of active principles derived from plants, for use in the pharmaceutical, health food and cosmetics industries.
Indena, a privately owned Italian company, reported € 140 million in consolidated turnover in 2004, mostly generated abroad. The company, with around 700 employees, including 10% dedicated to full-time research, manages cultivation, manufacturing, and distribution operations in more than 40 countries throughout the world.
The key to Indena's success is its research, covering: the screening of medicinal plants for their pharmacological benefits; the identification of new active principles; and the development of extraction and purification systems at the cutting-edge of industrial application.
The phyto-chemical research is carried out in Indena's own Settala Research Center, near Milan. Indena also co-operates with the world's most prestigious universities and private research institutions in the biological assessment of safety and effectiveness up to clinical phase I.
Backed up by over 80 years of botanical experience, the Italian company has developed a plantation network, managed by experts, to supply its research and production centers with officinal plants, while at the same time ensuring bio-diversity and protecting the ecosystem balance from uncontrolled harvesting. Today more than 60% of the raw material used in the manufacturing process comes from cultivation.
Indena's decades-long global presence in the plant extraction industry is a guarantee of its in-depth knowledge and understanding of the business, the market and the legal framework. The Company's experts communicate and interact constantly with the major international regulatory authorities such as WHO, EMEA, and ESCOP, and work with all the main pharmacopoeias.