On-line temperature-responsive restricted access material of sample preparation technique coupling with liquid chromatography for detection of tetracycline residues in milk

Abstract

Effort of sample preparation has been working towards the automation and improvement of accuracy. Development of on-line sample preparation coupled with liquid chromatography (LC) is a promising way. In this study, an on-line extraction-LC automated analysis was proposed based on a temperature-sensitive restricted access material (RAM) for the determination of tetracycline residues (TCs) in milk. The RAM was synthesized using silica as substrate by the two-step surface initiated-atom transfer polymerization (SI-ATRP), styrene (St), sodium 4-vinylbenzenesulfonate (Nass) and a lower critical solution temperature (LCST)-responsive n-vinylcaprolactam (VCl) were conjugated to fabricate multiple interaction-adsorption sites of P(St-co-Nass-co-VCl) on silica interior, while an upper critical solution temperature (UCST)-responsive sulfobetaine methacrylate (SBMA) was then block polymerized to form zwitterionic exclusion sites on exterior. Thereafter, the RAM was packed as extraction column to be fixed into LC ahead of C18 analytical column, establishing automatic analysis by rotation of switching valve. In conjunction with LCST and UCST dual-responsive properties to adjust the hydrophobic-hydrophilic transition of the grafted P(St-co-Nass-co-VCl)-b-PSBMA chains, the RAM column acquires the best extraction of TCs and meanwhile the furthest removal of proteins at about 30 ℃ of column temperature. And using the proposed automatic analysis for monitoring of TCs in milk, a wide linearity (20–900 ng mL−1), low limit of detection (1.0 ng mL−1) and limit of quantification (2.5 ng mL−1) as well as good inter-day (<3.9 % RSD) and intra-day precision (<4.8 % RSD) with satisfactory recoveries (97.2–101.9 %) show the superior efficiency. It is believed that the introduction of temperature-sensitive RAM to construct on-line extraction is a powerful tool for detecting trace substances in scientific analyses.