Trace analysis of three antihistamines in human urine by on-line single drop liquid–liquid–liquid microextraction coupled to sweeping micellar electrokinetic chromatography and its application to pharmacokinetic study

A rapid and efficient dual preconcentration method of on-line single drop liquid–liquid–liquid microextraction (SD-LLLME) coupled to sweeping micellar electrokinetic chromatography (MEKC) was developed for trace analysis of three antihistamines (mizolastine, chlorpheniramine and pheniramine) in human urine. Three analytes were firstly extracted from donor phase (4 mL urine sample) adjusted to alkaline condition (0.5 M NaOH). The unionized analytes were subsequently extracted into a drop of n-octanol layered over the urine sample, and then into a microdrop of acceptor phase (100 mM H3PO4) suspended from a capillary inlet. The enriched acceptor phase was on-line injected into capillary with a height difference and then analyzed directly by sweeping MEKC. Good linear relationships were obtained for all analytes in a range of 6.25 × 10−6 to 2.5 × 10−4 g/L with correlation coefficients (r) higher than 0.987. The proposed method achieved limits of detections (LOD) varied from 1.2 × 10−7 to 9.5 × 10−7 g/L based on a signal-to-noise of 3 (S/N = 3) with 751- to 1372-fold increases in detection sensitivity for analytes, and it was successfully applied to the pharmacokinetic study of three antihistamines in human urine after an oral administration. The results demonstrated that this method was a promising combination for the rapid trace analysis of antihistamines in human urine with the advantages of operation simplicity, high enrichment factor and little solvent consumption.

► A dual preconcentration method combined in an on-line mode.
► Simple and rapid operation without time-consuming matrix-transfer steps.
► High enrichment efficiencies are got in trace analysis of analytes in urine matrix.
► The method is used for pharmacokinetic of chlorpheniramine in human urine.