Development of New Liquid Chromatographic Method for Mitotane and Its Metabolites Determination in Human Plasma Employing Design of Experiments Methodology

A simple and reliable new HPLC method with UV detection has been developed and validated for simultaneous determination of mitotane and its two metabolites DDA and DDE. Method development was carried out utilizing systematical approach of the design of experiments (DoE) methodology. For estimation of factors influence on selected chromatographic responses and definition of the optimal chromatographic conditions, Box–Behnken experimental design was applied. The defined optimal separation conditions were: column Restek Ultra Aqua C18 with pre-column Restek Ultra Aqua C18 operating at temperature 35°C; mixture of acetonitrile and 0.5% formic acid as mobile phase with 1.2 mL min−1 flow rate and detection at 230 nm. As sample preparation method, liquid–liquid extraction was chosen. Method was fully validated and LOQ and LOD were experimentally determined. Finally, method was successfully applied for determination of mitotane and its metabolites in plasma samples of patients with adrenocortical carcinoma.

[1]  Sai Sandeep Mannemala,et al.  Statistical design in optimization and robustness testing of a RP-HPLC method for determination of warfarin and its process-related impurities , 2015, Journal of the Iranian Chemical Society.

[2]  Sai Sandeep Mannemala,et al.  Multiple Response Optimization of a HPLC Method for the Determination of Enantiomeric Purity of S-Ofloxacin , 2014, Chromatographia.

[3]  M. Jovanović,et al.  Robust optimization of psychotropic drug mixture separation in hydrophilic interaction liquid chromatography. , 2013, Acta chimica Slovenica.

[4]  M. Jovanović,et al.  Chemometrically assissted optimization and validation of RP-HPLC method for the analysis of itraconazole and its impurities , 2013, Acta pharmaceutica.

[5]  B. Nigović,et al.  Simultaneous analysis of mitotane and its main metabolites in human blood and urine samples by SPE-HPLC technique. , 2012, Biomedical chromatography : BMC.

[6]  S. Ackland,et al.  A simple HPLC method for plasma level monitoring of mitotane and its two main metabolites in adrenocortical cancer patients. , 2011, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[7]  Cristina Maria Quintella,et al.  Statistical designs and response surface techniques for the optimization of chromatographic systems. , 2007, Journal of chromatography. A.

[8]  M. Papotti,et al.  Adjuvant mitotane treatment for adrenocortical carcinoma. , 2007, The New England journal of medicine.

[9]  F. Ghezzo,et al.  A new simple HPLC method for measuring mitotane and its two principal metabolites Tests in animals and mitotane-treated patients. , 2006, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[10]  Martin Fassnacht,et al.  Adrenocortical Carcinoma: Clinical Update , 2006 .

[11]  Robert L. Mason,et al.  Statistical Design and Analysis of Experiments , 2003 .

[12]  C. Cordon-Cardo,et al.  Adrenocortical carcinoma: clinical, morphologic, and molecular characterization. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[13]  E. Baudin,et al.  Impact of monitoring plasma 1,1‐dichlorodiphenildichloroethane (o,p′DDD) levels on the treatment of patients with adrenocortical carcinoma , 2001, Cancer.

[14]  A. Latronico,et al.  Adrenocortical carcinoma , 2000, Cancer.

[15]  M. Madera,et al.  Adrenal Cortical Carcinoma , 1999 .

[16]  T. Lundstedt,et al.  Experimental design and optimization , 1998 .

[17]  M. Brennan,et al.  An eleven-year experience with adrenocortical carcinoma. , 1992, Surgery.