Liquid-liquid extraction combined with high performance liquid chromatography-diode array-ultra-violet for simultaneous determination of antineoplastic drugs in plasma

Abstract

A liquid-liquid extraction (LLE) combined with high-performance liquid chromatography-diode array detection method for simultaneous analysis of four chemically and structurally different antineoplastic drugs (cyclophosphamide, doxorubicin, 5-fluorouracil and ifosfamide) was developed. The assay was performed by isocratic elution, with a C18 column (5 µm, 250 x 4.6 mm) and mobile phase constituted by water pH 4.0-acetonitrile-methanol (68:19:13, v/v/v), which allowed satisfactory separation of the compounds of interest. LLE, with ethyl acetate, was used for sample clean-up with recoveries ranging from 60 to 98%. The linear ranges were from 0.5 to 100 µg mL -1 , for doxorubicin and 1 to 100 µg mL -1 , for the other compounds. The relative standard deviations ranged from 5.5 to 17.7%. This method is a fast and simple alternative that can be used, simultaneously, for the determination of the four drugs in plasma, with a range enabling quantification of the drugs in pharmacokinetics, bioequivalence and therapeutic drug-monitoring studies.Uniterms: Antineoplastic drugs/simultaneous determination in plasma. Liquid-liquid extraction. Highperformance liquid chromatography/diode array detector.Um método de extração líquido-líquido (ELL) combinado com cromatografia líquida de alta eficiência-detector de arranjo de diodos foi desenvolvido para análise simultânea de quatro fármacos antineoplásicos quimicamente e estruturalmente diferentes (ciclofosfamida, doxorrubicina, fluoruracila e ifosfamida). O estudo foi realizado sob condições isocráticas, com coluna C18 (5µm, 250 x 4.6 mm) e fase móvel constituída por água pH 4.0-acetonitrila-metanol (68:19:13, v/v/v), que permitiu separação satisfatória dos analitos de interesse. A ELL, com acetato de etila, foi utilizada para limpeza da amostra, com recuperação variando de 60 a 98%. As faixas foram lineares de 0,5 a 100 µg mL -1 para doxorrubicina e 1 a 100 µg mL -1 para os outros compostos. O desvio padrão relativo variou de 5,5 a 17,7%. Este método é uma alternativa rápida e simples que pode ser usado, simultaneamente, para a determinação dos quatro fármacos em plasma, com uma faixa que permite quantificá-los em estudos de farmacocinética, bioequivalência e monitorização terapêutica.Unitermos: Fármacos antineoplásicos/determinação simultânea em plasma. Extração líquido-líquido. Cromatografia líquida de alta eficiência/detector de arranjo de diodos.

Development and Validation of a Liquid Chromatographic Method for the Quantification of Related Compounds in Cyclophosphamide

“…Correlation coefficient for Cyclophosphamide and each related compounds was calculated for each calibration curve. ICH recommends that, for the establishment of linearity, a minimum of five concentration levels should be used .Acceptance criteria for correlation coefficient was used as not less than 0.99 [9].…”

Liquid Chromatography–Tandem Mass Spectrometry for the Simultaneous Determination of Doxorubicin and its Metabolites Doxorubicinol, Doxorubicinone, Doxorubicinolone, and 7-Deoxydoxorubicinone in Mouse Plasma

“…The Luna Omega C18 column exhibited better separation, an excellent peak shape, and good sensitivity for the analytes using a gradient elution of 0.1% formic acid in 95% methanol and 0.1% formic acid in 0.1% formic acid in 5% methanol compared with the Halo C18 and Acquity BEH C18 columns. Using methanol instead of acetonitrile [19][20][21][22][23][24][25][26][27][28][29][30][31][32][33] as the strong eluent of the mobile phase resulted in increased ionization efficiency and linearity range for the five analytes. Daunorubicin, an analog of doxorubicin, was selected as and the internal standard (IS).…”

“…Doxorubicinol, a major alcohol metabolite of doxorubicin formed by carbonyl reductases 1 and 3, is implicated in off-target cardiotoxicity of doxorubicin-treated patients [3,[7][8][9][10]. High-performance liquid chromatography (HPLC) with fluorescence [11][12][13][14][15][16][17][18][19] or ultraviolet (UV) [20,21] detection, LC with mass spectrometry (LC-MS) [22,23] or tandem mass spectrometry (LC-MS/MS) [24][25][26][27][28][29][30][31][32][33][34][35][36], and capillary electrophoresis [37][38][39] methods have been used to analyze doxorubicin alone or with its metabolite doxorubicinol in various biological matrices, such as blood, serum, plasma, cells, and tissues. Protein precipitation with methanol, acetonitrile, or acetone [13][14][15]18,19,21,22,[25][26][27]31,…”

Application of Charge Transfer Reactions for the Quantitative Spectrophotometric Determination of Cyclophosphamide in Pure and Pharmaceutical Formulation

“…1). This drug has been frequently administered in combination with other antineoplastic agents like Ifofhamide (IF) for the treatment of malignancies like lymphomas, leukemia and a variety of bone and soft tissue sarcomas [1,2]. CP is a prodrug that requires metabolic activation to phosphoramide mustard, the cytotoxic antineoplastic agent.…”