62ª Reunião Anual da SBPC
D. Ciências da Saúde - 5. Farmácia - 5. Farmacognosia
HIGH-SPEED COUNTER-CURRENT CHROMATOGRAPHY AS A TOOL TO ISOLATE FLAVONOID-C-GLYCOSYDES FROM Passiflora edulis VAR. flavicarpa LEAVES ("MARACUJÁ AMARELO")  
Silvana Maria Zucolotto 1
Flávio Henrique Reginatto 2
Freddy Alejandro Ramos 3
Leonardo Castellanos 3
Carmenza Duque 3
Eloir Paulo Schenkel 2
1. Departamento de Farmácia, Universidade Federal do Rio Grande do Norte (UFRN)
2. Pós-Graduação em Farmácia, Universidade Federal de Santa Catarina (UFSC)
3. Departamento de Química, Universidad Nacional de Colombia (UNAL)
INTRODUÇÃO:
Passiflora edulis var. flavicarpa, known popularly as yellow passion fruit ("maracujá amarelo"), is widely cultivated in Brazil because their fruits are used in the juice preparation and also the tea of their leaves are used in the popular medicine mainly as sedative and tranquilizer. Recently reports have been attributed to flavonoid-C-glycosides the pharmacological effects of Passiflora species. Then an efficient method for the separation and purification of these compounds from P. edulis specie could be warranted. High-speed countercurrent chromatography (HSCCC) is a separation technique that uses a liquid stationary phase without solid support matrix, in conjunction with the use of centrifugal force. This technique eliminates the irreversible loss of sample onto the solid support matrix used in the conventional adsorption chromatographic column. In this context, the aim of this work was the development of an efficient method for the isolation and purification of flavonoid C-glycosides from P. edulis var. flavicarpa leaves by HSCCC.
METODOLOGIA:
The leaves of P. edulis var. flavicarpa were collected in Santa Catarina, Brazil and were air-dried, powdered, and extracted using hot water (90°C) by infusion (plant:solvent, 1:10,w/v) for 10 min. Thereafter, the aqueous extract was filtered and partitioned with ethyl acetate (3 × 300 mL) and n-butanol (3 × 300 mL), yielding ethyl acetate and butanol (BuOH) fractions. The BuOH fraction was selected to further analysis by HSCCC because it is rich in flavonoid C-glycosides. HSCCC analysis was performed on a SER 521, Merk- Hitachi coupled with L-6000Ap pump and an automatic fraction colector (Eldex U-200) using a two-phase solvent system composed of ethyl acetate: n-butanol: water (2:1:3, v/v/v), 750 rpm and flow=1.5 mL/min.
RESULTADOS:
Several two-phase solvent systems were tested and the solvent systems ethyl acetate: n-butanol: água (2:1:3, v/vv/v) was found to be satisfactory for the separation of these compounds. The separation time was 7 hours and retention of stationary phase was 63%. From 1.5 g of BuOH fraction were obtained two main fractions (fraction I and II), using the lower phase as stationary phase. Fraction I showed a main unknown compound (1) and fraction II showed isoorientin (2) as main compound. Both fractions were purified chromatographic column using Sephadex LH-20, as adsorbent and methanol as mobile phase, resulting in 46 mg of compound 1 and 30 mg of compound 2.  In addition, this solvent system allowed the retention of di-C-glycosyl flavonoids (vicenin-2, 6,8-di-C-glycosylchrysin, spinosin and an unknown component) in the stationary phase, facilitating the further purification of these compounds.
CONCLUSÃO:

The HSCCC procedure allowed to obtain two main components, 46 mg of an one unknown compound and 30 mg of the isoorientin and the di-C-glycosyl flavonoids were retained in stationary phase. HSCCC analysis showed be a powerful technique for preparative separation and purification of flavonoid C-glycosides from leaves of P. edulis.

Instituição de Fomento: CAPES, CNPq, DIB-Universidad Nacional de Colombia
Palavras-chave: Passiflora edulis var. flavicarpa, Counter Current Chromatography, flavonoid C-glycosides.