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HPLC Method Development for Naringenin and its Glucoside in Rat Serum and their Bioavailibilty Studies

Varsha Gupta, Madhumita Srivastava, Rakesh Maurya, Paliwal SK and Anil Kumar Dwivedi

This study was aimed at finding a potent derivative of Naringenin (N) with osteogenic action. In CDRI, we have isolated Naringenin-6- C -Glucoside (NCG). This was found more active than naringenin. This paper reports a bioanalytical HPLC method for comparative bioavailability studies of N and NCG. In this method, separation was achieved on a Lichrosphere Lichrocart RP 18 (250 mm, 4 mm, 5 μm, Merck) column, with the mobile phase consisting of a mixture of 0.5% phosphoric acid in triple distilled water & acetonitrile (75:25). The flow rate was kept at 1.5 ml/ minute and the column effluents were monitored at 290 nm and 325 nm. The retention time of NCG was about 2.5 min, whereas naringenin eluted at about 14.5 min. There is no interference of serum impurities at these retention times. Validation parameters were checked and were found within limits. The highest serum concentrations (C max ) of naringenin (5 mg/kg dose) was recorded at 4 h after dosing and reached 1584 ± 439 ng/ml, followed by a marked decrease between 6 and 24 h. In case of NCG (5 mg/kg dose) highest concentration 738 ± 300 ng/ml was found at 3 hours (C max ). These data indicate that N & NCG are efficiently absorbed after feeding to rats and that their bioavailability is related to the glucoside moiety.