Combination of a sub‐3 μm superficially porous particle packed column with charged aerosol detection for the simple and sensitive measurement of nine macrolides in human urine

Abstract

Due to the lack of chromophores in many macrolides, analytical methods based on mass spectrometry and electrochemical detection coupled to liquid chromatography have been suggested to be suitable for the quantification of macrolides in complex matrices. In this study, a simple and sensitive analytical method was established for the simultaneous measurement of nine macrolides in human urine by combining a sub-3 μm superficially porous particle packed column with charged aerosol detection. After thorough investigation of various sample preparation methods, including two liquid-liquid extraction methods and four solid-phase extraction methods, HLB solid-phase extraction was selected and further optimized. Absolute recovery of the optimized sample preparation method ranged from 99.5-110.2%, indicating its very high extraction/clean-up efficiency. For chromatography, parameters influencing macrolide separation were systematically optimized, and the resulting conditions allowed baseline separation of nine macrolides within 24 min using a very simple mobile phase. The established method was validated for linearity, limit of detection, limit of quantification, absolute recovery, and precision. Based on its limit of detection (0.025-0.100 μg/mL), the method had similar or greater sensitivity than most methods based on electrochemical detection. It was found that the current method was appropriate for application to real human urine samples after drug administration.

Development and validation of high‐performance liquid chromatography method for determination of clarithromycin in pharmaceutical tablets

“…The analytical methods reported previously for the quantification of CLA in biological fluids as well as in formulations include microbiological bioassay [5] and chemical methods, such as ultraviolet (UV) spectrophotometry [6], spectrofluorimetry [7], thin layer chromatography [8], voltammetry [9,10], and capillary zone electrophoresis [11,12]. A survey of literature revealed that most techniques used for the determination of this drug are highperformance liquid chromatography with electrochemical detection [13][14][15][16][17], mass spectrometry [18][19][20][21], fluorescence detection after pre-column derivatization [22], UV detection [23][24][25], and diode array detection [26]. These methods were time-consuming, tedious, and mainly devoted to the determination of CLA in biological fluids.…”

Development and validation of a stability‐indicating HPLC method for topiramate using a mixed‐mode column and charged aerosol detector

“…Charged aerosol detector (CAD) is a relatively new type of HPLC detector introduced by Dixon and Peterson and represents an alternative detection method for non‐volatile analytes that lack chromophore groups, such as topiramate . CAD has been described as advantageous over other universal detectors due to its high sensitivity, consistent response, and broad dynamic range . In a system employing a CAD, the HPLC eluent is nebulized by nitrogen carrier gas that transforms the liquid phase into small droplets.…”