Greener and whiter analytical method development and validation for determining the presence of zolpidem tartrate infused in apple juice using RP-HPLC via magnetic solid-phase extraction followed by LC-MS confirmatory analysis

Abstract

The research work entails a newly developed RP-HPLC method, aimed at analyzing the modern date rape drug, zolpidem tartrate (ZT), infused in apple juice matrix. The work relies on dispersive solid-phase extraction (DSPE) with polyethylene imine (PEI)-coated magnetic nanoparticles to preconcentrate zolpidem from the matrix, in the presence of trifluoroacetic acid (TFA) for matrix isolation, for the first time. The optimized conditions emphasize the use of an environmentally preferable mobile phase [methanol: 0.5% acetic acid (60 : 40% v/v; pH 2.50)] at a 1 ml min⁻¹ flow rate, employed with a Platisil Octa-Decyl Silane (ODS) column (250 × 4.6 mm; 5 μm). Further, the validated results were confirmed to be within the ICH guidelines, marking the method demonstrated to be linear (R² = 0.9988; 0.9957), robust (% RSD below 1), sensitive (LOD = 1.8 μg ml; LOQ = 6 μg ml⁻¹), precise and accurate (% recovery = 92–120%). Following the same conditions, a confirmatory analysis of zolpidem was accomplished using LC-MS, verifying the method's suitability notably, with good peak resolution, less matrix interference and a confirmation of the presence of zolpidem using mass spectrometry. The recycling ability of the PEI@SiO₂@Fe₃O₄ nanoparticles was also assessed. To determine the sustainability of the proposed work, a greener and whiter assessment has been carried out in a comparative mode with previous similar works. For green tools, the recently developed AGREE software was utilized for assessing the method's greeness and it demonstrated a good green score of 0.68, supported by method assessment using ComplexGAPI software. For the assessment of the method's blue principles, the latest software utilizing the blue applicability grade index (BAGI) was applied, resulting in a decent score of 62.5. To consider sustainability, the RGB methodical software in its latest version the RGBfast model, was incorporated in the study for furnishing a balance of the three different major principles (Red–Green–Blue) and for assessing a check on sustainability of the current method compared to similar previously established proposed works.