ABSTRACT
A liquid chromatography (HPLC) method with UV detection was developed for determination of sodium hyaluronate in pharmaceutical formulation. Sodium hyaluronate is a polymer of disaccharides, composed of D-glucuronic acid and D-N-acetylglucosamine, linked via alternating β-1, 4 and β-1, 3 glycosidic bonds. Being a polymer compound it lacks a UV absorbing chromophore. In the absence of a UV absorbing chromophore and highly polar nature of compound, the analysis becomes a major challenge. To overcome these problems a novel method for the determination of sodium hyaluronate was developed and validated based on size exclusion liquid chromatography (SEC) with UV detection. An isocratic mobile phase consisting of buffer 0.05 M potassium dihydrogen phosphate, pH adjusted to 7.0 using potassium hydroxide (10%) was used. Chromatography was carried out at 25 1C on a BioSep SEC S2000, 300 mm ? 7.8 mm column. The detection was carried out using variable wavelength UV-vis detector set at 205 nm. The compounds were eluted isocratically at a steady flow rate of 1.0 mL/min. Sodium hyaluronate retention time was about 4.9 min with an asymmetry factor of 1.93. A calibration curve was obtained from 1 to 38 g/mL (r40.9998). Within-day%RSD was 1.0 and between-day%RSD was 1.10. Specificity/selectivity experiments revealed the absence of interference from excipients, recovery from spiked samples for sodium hyaluronate was 99-102. The developed method was applied to the determination of sodium hyaluronate in pharmaceutical drug substance and product.
ABSTRACT
Methanolic extract of leaves of A. squamosa was tested for mosquitocidal effect against C. quinquefasciatus. A liquid mosquito insecticide formulation was prepared with the extract (1, 3 and 5 %w/w) using deodorized kerosene as solvent and investigated for its knock-down and 24 hr mortality. The extract formulation produced dose dependent activity, exhibited significantly shorter knock down KD50 and KD90 values and produced significant mortality. The results suggest the potential mosquitocidal effect of A. squamosa on C. quinquefasciatus.