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 ß-1, 4 and ß-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 °C 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 (r>0.9998). 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
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
A simple, specific, and precise high-performance liquid chromatographic method is developed and validated for the simultaneous determination of chlorocresol (CC), mometasone furoate (MF), and fusidic acid (FA) in a cream formulation. The isocratic mobile phase consists of 1.5% w/v aqueous ammonium acetate buffer-acetonitrile, 55:45 (v/v) of pH 3.8. The column contains octylsilyl chemically bonded to porous silica particle (Symmetry C8, 150x3.9 mm, 5 microm). The detection is carried out using variable wavelength UV-vis detector set at 240 nm. The solutions are chromatographed at a steady flow rate of 1.0 mL/min. The current method separates CC, MF, and FA in less than 8 min with good resolution and peak shapes, minimal tailing, and with retention factors between approximately 1 and 5. Linearity range and percent recoveries for CC, MF, and FA are 10-30, 10-30, and 200-600 microg/mL; and 100.31%, 100.38%, and 100.34%, respectively. The method is validated according to International Conference on Harmonization guidelines and proven to be suitable for stability testing, content uniformity testing, and quality control of these compounds in pharmaceutical preparations.
Subject(s)
Chromatography, High Pressure Liquid/methods , Cresols/analysis , Fusidic Acid/analysis , Pregnadienediols/analysis , Dosage Forms , Drug Contamination , Drug Stability , Mometasone Furoate , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
A simple and precise high performance liquid chromatographic method has been developed and validated for the simultaneous determination of bisoprolol fumarate (BF), and hydrochlorothiazide (HCTZ) in a tablet formulation. Chromatography was carried out at 25 degrees C on a 4.6 mm x 250 mm, 5 microm cyano column with the isocratic mobile phase of 0.1M aqueous phosphate buffer, acetonitrile and tetrahydrofuran (85:10:5, v/v/v) at a flow rate of 1.0 ml/min. The UV detection was carried out at 225 nm. HCTZ and BF were separated in less than 10 min with good resolution and minimal tailing, without interference of excipients. The method was validated according to ICH guidelines and the acceptance criteria for accuracy, precision, linearity, specificity and system suitability were met in all cases. The method was linear in the range of 50-150 microg/ml for BF and 125-375 microg/ml for HCTZ.
Subject(s)
Adrenergic beta-Antagonists/analysis , Bisoprolol/analysis , Chromatography, High Pressure Liquid/methods , Diuretics/analysis , Hydrochlorothiazide/analysis , Acetonitriles/chemistry , Buffers , Chlorothiazide/chemistry , Chromatography, High Pressure Liquid/instrumentation , Dosage Forms , Drug Contamination , Furans/chemistry , Guidelines as Topic , Phosphates/chemistry , Reference Standards , Reproducibility of Results , Sensitivity and Specificity , Tablets , Temperature , Time Factors , Water/chemistryABSTRACT
A new specific, accurate, precise, and reproducible selective online dissolution method for rosiglitazone maleate is developed and validated for the dissolution of rosiglitazone maleate in pharmaceutical formulations. The rationale of the method is based on the direct measurement of the absorbance of the analyte in the buffer medium at 242 nm using buffer as blank. Dissolution is achieved on a dissolution test apparatus consisting of photo diode array spectrophotometer, peristaltic pump, and temperature controller, using 0.01N HCl and 0.05M potassium chloride as the dissolution medium. The proposed method is developed, optimized, and validated in terms of linearity, reproducibility, accuracy, and selectivity for the dissolution of rosiglitazone maleate in pharmaceutical formulations. The method is found to be linear in the range of 1 to 14 microg/mL of rosiglitazone maleate with a correlation coefficient of 0.999. The dissolution studies of rosiglitazone maleate tablets obtained by the proposed method are in good agreement with those by high-performance liquid chromatography.
