Abstract
Mitotane is a cytotoxic drug used in the treatment of inoperable adrenocortical carcinoma, it inhibits steroidogenesis as well, and therefore monitoring the level of steroid hormones in patients treated with mitotane is a crucial point of therapy. Hence, we have developed a simple, fast, and efficient electrophoretic method combined with reverse polarity sweeping as online preconcentration technique and dispersive liquid-liquid microextraction for the simultaneous determination of mitotane, its main metabolite DDA, and five steroid hormones (progesterone, testosterone, epitestosterone, cortisol, and corticosterone) in urine samples. In addition, a new sample matrix consisting of β-CD2 SDS1 complexes for a high hydrophobic compounds solubilization was developed. Approach based on the application of β-cyclodextrin and SDS complex of a ratio 2:1 allowed for hydrodynamic injection into the capillary of a solution containing both mitotane and other analytes. The detection limits of the analytes for the reverse polarity sweeping-dispersive liquid-liquid microextraction method were found to be in the range of 1.5-3ng/mL, which were approximately 1000 times lower than in the conventional hydrodynamic injection (5 s, 0.5psi) without any preconcentration procedure. All analytes were completely resolved in less than 13 min by uncoated silica capillary with an inner diameter of 75μm (ID) × 60cm. Electrophoretic separation was performed in reverse polarity with a voltage of -25kV with a background electrolyte (BGE) consisting of 100mM SDS, 25% ACN, 25mM phosphate buffer (pH 2.5), and 7 mMβ-cyclodextrin.
Published Version
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