Ecofriendly and biocompatible biochars derived from waste-branches for direct and efficient solid-phase extraction of benzodiazepines in crude urine sample prior to LC-MS/MS.

Biochars (BCs) derived from waste-branches of apple tree, grape tree, and oak were developed for direct solid-phase extraction (SPE) of five benzodiazepines (BZDs) in crude urine samples prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS) determination. Scanning electron microscopy, elemental analyzer, X-ray diffractometry, N adsorption/desorption experiments, and Fourier transform infrared spectrometry characterizations revealed the existence of their mesoporous structure and numerous oxygen-containing functional groups. The obtained BCs not only possessed high affinity towards BZDs via π-π and hydrogen bond interactions, but also afforded the great biocompatibility of excluding interfering components from undiluted urine samples when using SPE adsorbents. Variables affecting SPE of target analytes were systematically optimized including pH, ionic strength, dilution ratio, washing solution, desorption solvent, and its volume. The method of BC-based SPE combined with LC-MS/MS exhibited a wide linear range of 0.03-100 ng/mL, a low detection limit of 0.01-0.08 ng/mL, and satisfactory recovery of 77.6-106% for the studied BZDs. Notably, this method allowed the possibility of direct loading of undiluted urine samples and avoided tedious filtration and dilution steps, which significantly simplified the pretreatment process. Additionally, these BC sorbents derived from waste-branches were ecofriendly and cost-effective, providing a sustainable alternative for the traditional SPE sorbents. Thus, the proposed method has promising application for ecofriendly, simple, efficient, and reliable monitoring of BZDs in urine samples.