Combination of hollow‐fiber liquid‐phase microextraction and capillary electrophoresis for pioglitazone and its main metabolites determination in rat liver microsomal fraction

Abstract

Pioglitazone (PGZ), a thiazolidinedione antidiabetic agent, is reported as a potent and selective activator of peroxisome proliferator-activated receptor γ (PPAR γ). This drug has been widely prescribed for the treatment of Type 2 diabetes mellitus. In this regard, this manuscript presents, for the first time, an alternative electrophoretic method for PGZ and its main metabolites determination in rat liver microsomal fraction. The electrophoretic analyses were performed using an uncoated fused-silica capillary of 50 μm id, 48 cm in total length and 40 cm in effective length, and 50 mmol/L sodium phosphate buffer solution (pH 2.5). All experiments were carried out under the normal mode. The capillary temperature was set at 35°C and a constant voltage of +30 kV was applied during the analyses. Samples were introduced into the capillary by hydrodynamic injection (50 mbar, 15 s) and detection was performed at 190 nm. The sample preparation procedure, based on hollow-fiber liquid-phase microextraction, was optimized using multifactorial experiments. Next, the following optimal condition was established: sample agitation at 1500 rpm, extraction for 15 min, 0.01 mol/L hydrochloric acid as acceptor phase, 1-octanol as organic phase, and donor phase pH adjustment to 6.0. The method demonstrated LOQs of 200 ng/mL. Additionally, it was linear over the concentration range of 200-25,000 ng/mL for PGZ and 200-2000 ng/mL for the metabolites. Finally, the validated method was employed to study the in vitro metabolism of PGZ using rat liver microsomal fraction.

Pioglitazone: A review of analytical methods

“…Calixto et al [36] proposed an alternative electrophoretic method for PIO and its main metabolites determination in rat liver microsomal fraction. It was carried out on an uncoated fused-silica capillary (48 cm×50 µm i.d.)…”

Development of Capillary Zone Electrophoresis Method for the Simultaneous Separation and Quantification of Metformin and Pioglitazone in Dosage Forms; and Comparison with HPLC Method

“…In addition, the preliminary investigation also showed that phosphate buffer recorded better electropherograms, in terms of resolution, migration time, peak shape, peak height, baseline noise, and the electric current produced, than borate and acetate buffers. The literature also reported that the use of phosphate buffer was appropriate for MET [ 31 , 36 ] and PIO [ 34 ], with other analytes, assay methods.…”

“…The acidity of the buffer may affect mobility and electroosmosis flow (EOF) by altering the dissociation constant of analyte and Si-OH groups on the capillary [ 37 ]. Based on the pKa values of MET (12.4) [ 38 ] and PIO (5.6) [ 34 ], both compounds are positively charged under acidic conditions and can be analyzed by CZE. Notably, atenolol was chosen as an internal standard (IS) because it has a pKa value of 9.6.…”

“…In this context, CZE was exploited for the assay of MET [ 26 , 31 , 32 , 33 ] and PIO [ 34 ] and with other drugs or metabolites. Nevertheless, to our knowledge, no CZE method for the assay of PIO and MET in combination dosage form has been reported.…”

Spectrophotometric assay of pioglitazone hydrochloride using permanganate in acidic and basic media

“…reviewed. 4 Various techniques such as uv-spectrophotometry, [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23] potentiometry, voltammetry, [27][28] high-performance liquid chromatography, [29][30][31][32][33][34][35][36][37][38][39] ultra-performance liquid chromatography, 40 highperformance thin layer chromatography [41][42][43][44][45][46] and capillary electrophoresis 47 have been reported for the determination of PGH in bulk sampleand tablets.…”