Stereoselective determination of hydroxychloroquine and its metabolites in human urine by liquid‐phase microextraction and CE

Abstract

Liquid-phase microextraction based on polypropylene hollow fibers and CE were applied for the chiral determination of hydroxychloroquine (HCQ) and its metabolites (desethylchloroquine, DCQ; desethylhydroxychloroquine, DHCQ; bisdesethylchloroquine, BDCQ) in human urine. The analytes were extracted from 3 mL of urine spiked with the internal standard (metoprolol) and alkalinized with 250 muL of 2 M NaOH. The analytes were extracted into 1-octanol impregnated in the pores of the hollow fiber, and into an acid acceptor solution inside the hollow fiber. The electrophoretic separations were carried out in 100 mmol/L Tris buffer (pH adjusted to 9.0 with phosphoric acid) containing 1% w/v S-beta-CD and 30 mg/mL HP-beta-CD with a constant voltage of +18 kV. The method was linear over the concentration range of 10-1000 ng/mL for each HCQ stereoisomer and 21-333 ng/mL for each metabolite stereoisomer. Within-day and between-day assay precision and accuracy for the analytes were studied at three concentration levels for each stereoisomer and were lower than 15%. The developed method was applied for the determination of the cumulative urinary excretion of HCQ, DCQ, and DHCQ after oral administration of rac-HCQ to a health volunteer. The results obtained are in agreement with previous literature data.

Stereoselective determination of hydroxychloro‐quine and its major metabolites in human urine by solid‐phase microextraction and HPLC

“…BDCQ concentration found in human urine is very low, so, most of the published papers quantify just the two major metabolites in human urine, DCQ and DHCQ [8 -11]. The excretion of HCQ and its metabolites has shown to be stereoselective and studies show that, in a short period, (+)-(S)-enantiomer is excreted in a higher amount than ( -)-(R)-enantiomer [9,11]. Nevertheless, when the excretion of HCQ was studied in a longer period, the difference between (+)-(S)-and ( -)-(R)-tended to decrease, the ratio being (+)/( -) equal to 50:50 in a period of 360 h [10].…”

“…The most common methods employed to analyze these compounds in human biological fluids is HPLC using chiral stationary phases [8 -10, 12, 13] and CE [11]. Almost all of these mentioned papers are based on traditional liquid -liquid extraction [8 -10, 12, 13] and just one method, recently developed by our group, is based on liquid -liquid microextraction [11].…”

The role of cyclodextrins in chiral capillary electrophoresis

“…In this way, it was possible to establish the enantiomer ratio of metoprolol, and its ortho and meta by-products during one analysis [31]. The separation of the enantiomers of pharmaceuticals and their metabolites can also be managed during one CE analysis [15,21,22]. CDs are excellent to chiraly analyze main and by-products simultaneously [31,56].…”

“…Capillaries are washed with concentrated bases or acids resulting in elimination of endogenous dirt among analytes [3,15,21,22].…”

Microextração em fase líquida (LPME): fundamentos da técnica e aplicações na análise de fármacos em fluidos biológicos

“…Diversos trabalhos vêm sendo realizados utilizando urina como matriz (Tabela 1). Recentemente, De Oliveira et al 61 desenvolveram um método para análise enantiosseletiva da hidroxicloroquina e metabólitos em urina empregando n-octanol como solvente de extração e posterior análise por CE. Neste trabalho, valores de recuperação em torno de 80% foram observados e o método foi aplicado em um estudo piloto de disposição cinética, mostrando potencial para aplicação em estudos farmacocinéticos de excreção destes analitos.…”