Application of ICH Guidelines for Studying the Degradation Behavior of Rocuronium Bromide Coupled with Stability-Indicating RP-LC Method.

Non-depolarizing neuromuscular blocking agent Rocuronium bromide was quantified in drug substance and drug product using reversed-phase liquid chromatographic method. Forced degradation studies were conducted for Rocuronium bromide in drug substance under acidic (2MHCl), basic (2MNaOH), oxidative (3% H2O2), thermal (135°C) and photolytic (254 nm) stress conditions. An Agilent H12 C18 column was used for separation using diammonium hydrogen phosphate buffer (pH 8; 0.04M)- acetonitrile (50:50; v/v) as mobile phase at flow rate of 1 mL/min. The quantification was done using UV detection at 210 nm. The limit of quantification and detection was 11.1 and 3.66 µg/mL, respectively, and the recovery percentage was 99% in drug substance and drug product. ICH guidelines were adopted for method validation. The proposed LC method monitored the degradation profile for Rocuronium bromide under various stress conditions and provided a specific LC method for its routine analysis. Besides, the MS data were used to identify all Rocuronium bromide degradation products and the possible degradation pathway was designated.

Stability indicating liquid chromatography method for the analysis of Vecuronium bromide: study of the degradation profile.

Neuromuscular blocker agent namely; Vecuronium bromide (VEC) was quantified through developing a simple reversed phase liquid chromatographic (RP-LC) method, in drug substance and in drug product. The proposed method could quantify VEC in the presence of its degradation products produced from exposing VEC to different stress conditions as recommended by the International Conference on harmonization (ICH) guidelines. Acidic (2M HCl), basic (2MNaOH) hydrolysis, oxidation (3% H2O2), photolysis (UV light at 254nm), and thermal (135 °C) degradation were estimated by exposing the drug substances to different stress conditions. The separation of the drug from its degradation products was successfully conducted on Tracer Extrasil CN (150 × 4.6mm; 5µm) column using O-phosphoric acid (pH6; 0.05M)-acetonitrile (50:50v/v) as mobile phase. The detection and quantification was done with UV detection at210nm.The validation data were found to be acceptable over a concentration range10-120 µg/ml. The limit of quantification (LOQ) and detection (LOD) were 8.10 and 2.67 µg/ml, respectively. The proposed method met all criteria for validation in accordance with the International Conference on harmonization (ICH) guidelines. The presented work monitored the degradation profile for VEC under various stress conditions and provided a simple LC method for its routine analysis. The structures of the forced degradation products had been described in details using the MS data and the possible degradation pathways were outlined. Besides, the results obtained from the developed method compared statistically with that of the official method indicting high accuracy and precision.

Screen printed potentiometric sensor for therapeutic monitoring of rocuronium at the point of care.

Rocuronium bromide (ROC) is currently regarded as the 'gold-standard' in emergency medicine and anesthesia. Globally, millions of human beings are daily administered ROC at emergency settings where it is favored among all the neuromuscular blockers, particularly succinylcholine, for both its fast onset of action and short duration. However, it has been reported that 45% of patients in the post-anesthesia care unit are susceptible to residual postoperative paralysis, undesired ventilator effects and incomplete recovery after ROC administration. From an analytical chemistry perspective, direct determination of ROC is a difficult approach due to the complexity in isolation from biological specimens as well as the lack of a sensitive detection techniques and detectable chromophore. This contribution describes the development of a calix[6]arene-based screen-printed electrode (SPE) that is capable of ROC detection in biological samples at the point of care. This fabricated SPE (sensor 1) exhibited superior performance characteristics (slope, LOD and life time) with respect to an ionophore-free liquid-contact electrode, LCE, (sensor 2). The proposed SPE showed a linear response over a concentration from 1 µM to 10 mM, with a Nernstian slope of 57.9 mV/decade and a detection limit of 0.39 µM. Moreover, this sensor showed a considerable selectivity towards ROC in presence of the anticipated interfering ions. To investigate the ability of the SPE to detect ROC in real biological specimens, ROC has been spiked at a concentration comparable to its anticipated level in human plasma (Cmax~ 40 µM) and the proposed SPE displayed an excellent platform for therapeutic drug monitoring (TDM) of ROC with respect to UV-spectrophotometry and LC/MS. Finally, the developed SPE was used for the determination of ROC in its commercial pharmaceutical formulation.