Liquid chromatographic high-throughput analysis of the new ultra-short acting hypnotic ‘HIE-124’ and its metabolite in mice serum using a monolithic silica column

Abstract

For the first time, a fast, high-performance liquid chromatography (HPLC) method was developed and validated for the simultaneous determination of the new ultra-short hypnotic HIE-124 and its metabolite in mice serum. Each compound, together with carbamazepine (internal standard) was extracted from the serum matrix using liquid-liquid extraction (LLE). Chromatographic resolution of the analytes was performed on a Chromolith Speed Rod monolithic silica column (100 mm × 4.6 mm i.d.) under isocratic conditions using a mobile phase of 65:35 (v/v), 20 mM phosphate buffer (pH 7.0 adjusted with phosphoric acid)-acetonitrile. The elution of the analytes were monitored at 240 nm and conducted at ambient temperature. Because of high column efficiency the mobile phase was pumped at a flow rate of 2.5 mL min(-1). The total run time of the assay was 2 min. The method was validated over the range of 60-2000 ng mL(-1) for HIE-124 and 200-1600 ng mL(-1) for the metabolite (r(2) = 0.99). The limit of detection (LOD) for HIE-124 and its metabolite were 20 ng mL(-1) and 65 ng mL(-1), respectively. The proposed method was validated in compliance with ICH guidelines, in terms of accuracy, precision, limits of detection and quantitation and other aspects of analytical validation. The developed method could be used for the trace analyses of HIE-124 and its metabolite in serum and was finally used for the pharmacokinetic study investigation of HIE-124 in mice serum.

Advances in Chemical and Biological Methods to Identify Microorganisms—From Past to Present

“…The detector creates a signal proportional to the quantity of sample component in the column and the digital microprocessor controls the instrument providing data analysis. Presently, it can be used in monolithic columns [83,84,85,86] or at higher temperatures—high temperature liquid chromatography [87,88,89]. Some concerns which have to be considered when choosing HPLC for microbial detection are that the equipment is expensive and it demands complex maintenance procedures, and therefore some caution is advisable for an adequate interpretation of results.…”

Monolithic Silica for Fast HPLC: Current Success and Promising Future

“…Monolthic columns have been successfully applied in the field of bioanalysis [105][106][107][108][109][110][111][112][113][114][115][116][117]. For example, they have been used in plasma analysis of drugs, which usually requires intensive sample preparation and filtration to avoid column blockage.…”

Development and validation of a LC method for the separation and determination of the anticancer-active FeIII(4-methoxy-salophene) using the new second-generation monolith

“…A successful application of first‐generation monolithic silica columns for fast LC analysis has recently been reviewed by El Deeb . The possibility of blockage after injection of biological samples is smaller on monolithic compared to particle‐packed LC columns because of the high column porosity . Analytical monolith columns have been successfully applied to control the quality of pharmaceutical products .…”