Simultaneous determination of etoposide and a piperine analogue (PA-1) by UPLC–qTOF-MS: Evidence that PA-1 enhances the oral bioavailability of etoposide in mice

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2010-03, Vol.878 (9), p.823-830
• Main Authors: Sachin, B.S., Najar, I.A., Sharma, S.C., Verma, M.K., Reddy, M.V., Anand, R., Khajuria, R.K., Koul, S., Johri, R.K.
• Format: Article
• Language: English
• Subjects: Alkaloids - blood; Alkaloids - chemical synthesis; Alkaloids - chemistry; Analogue; Analysis; Analytical, structural and metabolic biochemistry; Animals; Antineoplastic Agents, Phytogenic - blood; Antineoplastic Agents, Phytogenic - pharmacokinetics; Benzodioxoles - blood; Benzodioxoles - chemical synthesis; Benzodioxoles - chemistry; Bias; Bioavailability; Biological and medical sciences; Biological Availability; Calibration; Chromatography, High Pressure Liquid - methods; Drug Carriers - analysis; Drug Carriers - chemical synthesis; Drug Carriers - chemistry; Etoposide; Etoposide - blood; Etoposide - pharmacokinetics; Fundamental and applied biological sciences. Psychology; General pharmacology; Medical sciences; Mice; Monitoring; Pharmacokinetics; Pharmacology. Drug treatments; Piperidines - blood; Piperidines - chemical synthesis; Piperidines - chemistry; Piperine analogue; Polarity; Polyunsaturated Alkamides - blood; Polyunsaturated Alkamides - chemical synthesis; Polyunsaturated Alkamides - chemistry; Solid phases; Tandem Mass Spectrometry - methods; UPLC–qTOF-MS/MS
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.01.048

In the present investigation, a UPLC–qTOF-MS/MS method has been developed for the simultaneous determination of etoposide and a piperine analogue, namely, 4-ethyl 5-(3,4-methylenedioxyphenyl)-2E,4E-pentadienoic acid piperidide (PA-1). The analytes were separated on a reverse phase C18 column using methanol–water (72:28, v/v) mobile phase with a flow rate of 250 μL/min. The qTOF-MS was operated under multiple reaction monitoring mode using electro-spray ionization (ESI) technique with positive ion polarity. The major product ions for etoposide and PA-1 were at m/ z 185.1350 and 164.1581, respectively. The recovery of the analytes from mouse plasma was optimized using solid phase extraction technique. The total run time was 6 min and the elution of etoposide and PA-1 occurred at 1.24 and 2.84 min, respectively. The calibration curves of etoposide as well as PA-1 were linear over the concentration range of 2–1000 ng/mL ( r 2, 0.9829), and 1–1000 ng/mL ( r 2, 0.9989), respectively. For etoposide intra-assay and inter-assay accuracy in terms of % bias was in between −7.65 to +6.26, and −7.83 to +5.99, respectively. For PA-1 intra-assay and inter-assay accuracy in terms of % bias was in between −7.01 to +9.10, and −7.36 to +6.71, respectively. The lower limit of quantitation for etoposide and PA-1 were 2.0 and 1.0 ng/mL, respectively. Analytes were stable under various conditions (in autosampler, during freeze–thaw, at room temperature, and under deep-freeze conditions). The method was used for a pharmacokinetic study which showed that PA-1 enhanced the oral bioavailability of etoposide in mice by 2.32-fold.