Determination of Diuretics in Urine Using Immobilized Multi-Walled Carbon Nanotubes in Hollow Fiber Liquid-Phase Microextraction Combined with Liquid Chromatography-Tandem Mass Spectrometry

A novel technique utilizing the adsorptive potential of immobilized multi-walled carbon nanotubes (I-MWCNT) in hollow fiber liquid-phase microextraction (HF-LPME) was developed for the determination of diuretics in urine. In this study, the potential of carbon nanotubes as a sorbent for three-phase liquid-phase microextraction of diuretics from urine samples was evaluated. Analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS). A novel method was applied to detect acetazolamide (AAA), chlorothiazide (CTA), hydrochlorothiazide (HCT), hydroflumethiazide (HFT), clopamide (CA), trichlormethiazide (TCM), althiazide (AT) and bendroflumethiazide (BFT) in urine. Two-step extractions using different times and temperatures for each step were adopted. Parameters influencing the extraction efficiency, including the extraction solvent, sample pH, salt concentration, extraction time and extraction temperature were systematically optimized. Under the resulting optimal extraction conditions, this method showed good linearity over an analytes concentration range of 1 to 1000 ng/mL, high extraction repeatability with relative standard deviations of less than 6%, and low detection limits (0.09 to 0.51 ng/mL). The application of the methods to the determination of diuretics in real samples was tested by analyzing urine samples of patient.