The evaluation of the quality of beta-carotene derived from Azolla Filiculoides in theAnzali Wetland using the alkaline hydrolysis method in winter

Azolla is certainly a valuable laboratory plant that will thrive with very little care. The use of Azolla may be an important factor in the world's future food needs. Azolla is unique because it is one of the fastest growing plants on the planet – yet it does not need any soil to grow. Azolla is a small floating fern and is the only known pteridophyte that lives in symbiosis with a diazotrophic cyanobacterium. Azolla spp. are heterosporous free-floating freshwater ferns that live symbiotically with Anabaena Azollae, a nitrogen-fixing blue-green algae. There are six species of Azolla in the world. Azolla is a genus of aquatic ferns in the family Salviniaceae. The genus Azolla belongs to the single genus family Azollaceae. Azolla filiculoides is a species of Azolla. Azolla filiculoides is under the sub-genus Euazolla. It is native to warm temperate and tropical regions as well as most of the old world including Asia, Australia and Anzali wetland. This plant is dark green to reddish and float on the water surface, either individually or in mats, which can reach a thickness of up to 20 cm. When A. filiculoides plants are exposed to strong sunlight they obtain a red color. The same occurs in winter time. In shade they always remain green. A. filiculoides settles in ponds, ditches, water reservoirs, wetlands, channels and slow flowing rivers. A. filiculoides is very rich in protein, fat, ash and carotenoids. The chemical composition of Azolla species varies with ecotypes and with the ecological conditions and the phase of growth. The dry matter percentage of different Azolla species varies widely. Lysine and methionine contents in this species are moderate. But, essential amino acids in this species are poor. Carotenoids are the most common pigments in nature and are synthesized by all photosynthetic organisms. Carotenoids are considered key molecules for life. Biological properties of carotenoids allow for a wide range of commercial applications. Indeed, recent interest in the carotenoids has been mainly for their nutraceutical properties. Carotenoids as natural pigments, are used by the industry as pharmaceuticals, nutraceuticals, and animal feed additives, as well as colorants in cosmetics and special foods. Carotenoids have been studied for their ability to prevent chronic disease due to the free radical theory of aging in chronic disease etiology. The effect of carotenoids on biotechnology and the food industry is significantly attributed. Finally, carotenoids as fortified substances in foods and special aspects about carotenoids as health promoters are well presented along with a glance of carotenoids economics. β-carotene is one of the carotenoids. β-carotene is the main source of pro-vitamin A and is widely used as a food colorant. The majority of the β-carotene commercialized in the world is obtained by chemical synthesis from β-ionone. Therefore, research for the production of natural colors for use in food industry has particular importance.

The present project was aimed at determining the content, quality, and purity of β-carotene extracted from Azolla filiculoides in the Anzali Wetland, comparing it with synthetic β-carotene, and measuring its economic value.

One treatment had β-carotene derived from Azolla filiculoides in the Anzali Wetland through the alkaline hydrolysis method in the winter of 2014. Treatments were kept at 4 °C for one year. Synthetic β-carotene was used as the control. The quality of the treatments was assessed by applying some chemical tests, including the measurement of the content and quality of β-carotene, calorimetry using the Hunter-LAB method, determination of the purity and vitamin A employing high-performance liquid chromatography (HPLC), estimation of the dwell-time duration at 5 °C, and measurement of the solubility of β-carotene in water.

The results of the tests regarding the purity, concentration, calorimetry, vitamin compounds, dwell time, and solubility in the experimental β-carotene, compared with those in the control, revealed no significant difference (p>0.05). Moreover, the factors showed no significant difference between the control and experimental treatments during the dwell time (p>0.05). The natural β-carotene had a good quality during the storage period at 5 °C for one year.

Since there was no significant difference between the β-carotene derived from Azolla filiculoides and the synthetic one in terms of the chemical tests, purity, and dwell time, and since the natural β -carotene derived from Azolla filiculoides takes precedence over the synthetic one in terms of the food hygiene, it is recommended that natural β-carotene extracted from Azolla filiculoides be substituted for synthetic β-carotene in the food industry.