Levels of B-carotene and-tocopherol present in the Spirulina (Arthrospira) grown in different photoperiods Platensis

A microalga cianofÃcea Spirulina platensis tem sido comercializada e estudada devido ao seu alto valor nutricional, possuindo vitaminas, minerais, proteÃnas, Ãcidos graxos poli-insaturados, carotenÃides, ficobilinas e polissacarÃdeos com propriedades terapÃuticas. O objetivo do presente trabalho foi avaliar os teores de -caroteno e -tocoferol presentes na microalga S. platensis cultivada em laboratÃrio com diferentes fotoperÃodos. A microalga S. platensis foi cultivada em seis aquÃrios de 20 L sob quatro condiÃÃes diferentes de iluminaÃÃo. Um cultivo foi submetido a um fotoperÃodo de 8 horas de luz (L) e 16 de escuro (E), o segundo a 12 L e 12 E, o terceiro a 16 L e 8 E e o Ãltimo exposto a 24 h de iluminaÃÃo constante. Os aquÃrios foram dispostos lado a lado em duas incubadoras de madeira, com temperatura ambiente de mantida em 28  2ÂC com auxÃlio de um âcoolerâ (220 V) e iluminÃncia na superfÃcie da Ãgua em torno de 78,1 μ E cm-2 s-1 fornecida por duas lÃmpadas fluorescentes de 40 W. Os cultivos foram monitorados por espectrofotometria a 680 nm e pela contagem dos filamentos (tricomas) microalgais utilizando um microscÃpio Ãptico comum. A anÃlise simultÃnea dos carotenos e tocoferÃis foi realizada por cromatografia lÃquida de alta eficiÃncia em uma coluna Waters Spherisorb-Hichrom S5 ODS-2 (4,6 x 250 mm) com uma fase mÃvel de MeOH: THF (90:10, v/v), fluxo de 1,5 mL min-1 e detecÃÃo a 450 nm e 292 nm, respectivamente. A extraÃÃo de -caroteno foi realizada suspendendo-se 0,1 g de Spirulina liofilizada em 10 mL de metanol:Ãgua Milli-Q (90:10, v/v). Para a saponificaÃÃo dos lipÃdios, a mistura foi homogeneizada com 7% de hidrÃxido de potÃssio, levada ao banhomaria a 70ÂC por 30 min e submetida à partiÃÃo em n-hexano que foi evaporado. O resÃduo foi suspenso em 1 mL de metanol e 100 μL foram injetados manualmente, em duplicata, no sistema cromatogrÃfico. O -caroteno foi detectado na biomassa obtida de todos os cultivos e o -tocoferol apenas nos cultivos com fotoperÃodos de 8 L e 16 E e de 12 L e 12 E, enquanto o -tocoferol nÃo foi detectado. O teor de -caroteno variou de acordo com a intensidade luminosa e quando as culturas foram expostas a 24 h de iluminaÃÃo constante, atingiram mais rapidamente a produtividade mÃxima e apresentaram o maior teor de -carotenoThe microalga cyanophyceae Spirulina platensis has been commercialized and studied due to its high nutritional value. It has shown to be a rich source of a great variety of vitamins, minerals, proteins, polyunsaturated fatty acids, carotenoids, phycobilins, and polysaccharides that exhibit potential therapeutic properties. The aim of the present work was to evaluate the content of -carotene and -tocopherol in S. platensis cultivated in laboratory under different conditions of photoperiods. S. platensis was cultivated within six 20-liter reservoirs under four different illumination conditions. The first culture was grown under 8- hour light (L) and 16-hour dark (D) photoperiod; the second, 12 L and 12 D photoperiod; the third, 16 L and 8 D photoperiod, and the forth cultivation condition was constant illumination. The reservoirs were placed side by side in two wood chambers, where room temperature was kept at 28  2ÂC with a cooler (220 V). The water surface illumination of 78.1 μE cm-2 s-1 was supplied by two 40-Watt fluorescent lamps. The cultures were monitored by spectrometry at 680 nm and by counting microalga filaments (trichomes) under an optical microscope. The simultaneous analysis of carotenoids and tocopherols was performed by high performance liquid chromatography in a Waters Spherisorb-Hichrom S5 ODS-2 (4.6 x 250 mm) column using MeOH:THF (90:10, v/v) as mobile phase, delivered at 1.5 mL min-1. The detector was set at 450 nm and 292 nm for carotenes and tocopherols, respectively. For the extraction of both compounds, three portions of 0.1 g lyophilized Spirulina were suspended in 10 mL methanol:MiliQ water (90:10, v/v). The mixture was homogenized with a 7% potassium hydroxide aqueous solution, brought to a water bath at 70ÂC for 30 min, for saponification, followed of partitioning into n-hexane which was evaporated to dryness. The residues were suspended in 1 mL methanol, and 100 μL aliquots were manually injected in the chromatographic system. -Carotene was detected in the biomass obtained from all the cultures and a-tocopherol was detected only in 8 L and 16 D and 12 L and 12 D photoperiods cultures. -Tocopherol, on the other hand, was not detected. The content of -carotene varied with illumination intensity and in the cultivation performed under constant illumination both maximum productivity and highest -carotene content were reached rapidl