ABSTRACT
A sensitive and rapid ion chromatography (IC) method was developed for the low level determination of allylamine (AAM) in sevelamer (SVM) drug substances, i.e., sevelamer hydrochloride (SVH) and sevelamer carbonate (SVC). This method utilized a Dionex Ion Pack CS14 IC column, a mobile phase of 10mM methane sulfonic acid with conductivity detection. The total chromatographic run time was as short as 8 min. The various factors involved in the sample preparation such as, extraction solvent, extraction time and stirrer speed were evaluated. This method was validated as per United States Pharmacopoeia (USP) and International Conference on Harmonization (ICH) guidelines in terms of detection limit, quantitation limit, linearity, precision, accuracy, specificity and robustness. Linearity of the method was very good over the concentration range of 9-750 µg/mL with the coefficient of determination (r(2)) 0.999. The detection and quantitation limit of AAM were 2.7 and 9.0 µg/mL, respectively. The recovery data obtained for AAM were between 97% and 109%. Also, the specificity of the method was proved through IC coupled with mass spectrometer (IC-MS). The developed method was found to be robust and applied successfully to determine the content of AAM in Sevelamer bulk drugs.
Subject(s)
Allylamine/analysis , Chemistry, Pharmaceutical/methods , Chromatography, Ion Exchange/methods , Polyamines/analysis , Allylamine/chemistry , Calibration , Chemistry Techniques, Analytical , Drug Stability , Electric Conductivity , Humans , Mass Spectrometry/methods , Models, Chemical , Polyamines/chemistry , Reproducibility of Results , Sevelamer , Spectrometry, Mass, Electrospray Ionization/methods , Time FactorsABSTRACT
A new chiral purity method was developed for D-cycloserine (D-cys) by reverse phase HPLC and validated. Chiral derivatizing reagents, viz., o-phthalaldehyde and N-acetyl-L-cysteine were utilized in this method. The resultant diastereomers were resolved using Zorbax SB Phenyl HPLC column under isocratic elution. A mobile phase of 95:05 (v/v), 20mM Na(2)HPO(4) (pH 7), and acetonitrile, respectively, was used with the flow rate of 1.0 mL/min and UV detection at 335 nm. The method development with different chiral stationary phases and chiral derivatization reagents were also investigated. The stability of diastereomer derivative and influence of organic modifier and pH of the mobile phase were studied and optimized. The stability-indicating capability of the method was established by performing stress studies under acidic, basic, oxidation, light, humidity and thermal conditions. The detection and quantitation limit of L-cycloserine (L-cys) were 0.015 and 0.05% (w/w), respectively. A linear range from 0.05 to 0.30% (w/w) was obtained with the coefficient of determination (r(2)) 0.998. The recovery obtained for L-cys was between 92.9 and 100.2%. This method was applied successfully in pharmaceutical analysis to determine the content of L-cys in D-cys bulk drug.
