title: A sensitive quantitative assay for the determination of propafenone and two metabolites, 5-hydroxypropafenone and N-depropylpropafenone, in human K2EDTA plasma using LC-MS/MS with ESI operated in positive mode.
abstract: Propafenone is a potent antiarrhythmic agent; clinically propafenone has been used for a number of cardiac arrhythmias because it possesses multiple modes of action, via beta adrenergic receptor blockade and calcium antagonistic activity. Propafenone (PPF) exhibits extensive saturable presystemic biotransformation (first-pass effect) resulting in two active metabolites: 5-hydroxypropafenone (5-OH PPF) formed by CYP2D6 and N-depropylpropafenone (NDP) formed by both CYP3A4 and CYP1A2 enzymes. A specific and sensitive LC-MS/MS method was developed and validated for quantitation of PPF, 5-OH PPF and NDP using turboion spray in a positive ion mode. A solid-phase extraction was employed for the extraction from human plasma. Chromatographic separation of analytes was achieved using an ACE-5 C8(50 × 4.6 mm) column with a gradient mobile phase comprising ammonium acetate containing 0.01% TFA in purified water and acetonitrile. The retention times achieved were 1.36, 1.23, 1.24 min and 1.34 min for PPF, 5-OH PPF, NDP and IS (carbamazepine), respectively. Quantitation was performed by monitoring multiple reaction monitoring transition pairs of m/z 342.30 to m/z 116.20, m/z 358.30 to m/z 116.20, m/z 300.30 to m/z 74.20 and m/z 237.20 to m/z 194.10, respectively. The developed method was validated for various parameters. The calibration curves of PPF and 5-OH PPF showed linearity from 1 to 500 ng/mL, with a lower limit of quantitation of 1.0 ng/mL and for NDP linearity from 0.1 to 25 ng/mL with a lower limit of quantitation of 0.1 ng/mL. The bias and precision for intra- and-inter batch assays were <10 and 5%, respectively. The developed assay was used to evaluate pharmacokinetic properties of propafenone and its major metabolites in healthy human subjects.
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.|Department of Chemistry, St Xavier's College (Autonomous), Ahmedabad, India.
authors.forename: Harilal
authors.initials: H
authors.lastname: Patel
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.
authors.forename: Ashok
authors.initials: A
authors.lastname: Ghoghari
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.
authors.forename: Chandrakant
authors.initials: C
authors.lastname: Bhatt
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.
authors.forename: Shaival
authors.initials: S
authors.lastname: Shah
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.
authors.forename: Anilkumar
authors.initials: A
authors.lastname: Jha
authors.affiliation: Department of Chemistry, St Xavier's College (Autonomous), Ahmedabad, India.
authors.forename: Nirmal
authors.initials: N
authors.lastname: Desai
authors.affiliation: Zydus Research Centre, Bioanalytical Laboratory, Ahmedabad, India.
authors.forename: Nuggehally R
authors.identifier: http://orcid.org/0000-0002-5768-1332
authors.initials: NR
authors.lastname: Srinivas
mesh_terms: D000284:Administration, Oral; D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D004492:Edetic Acid; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D011405:Propafenone / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011405:Propafenone; C044307:5-hydroxypropafenone; D004492:Edetic Acid; C045806:N-depropylpropafenone
keywords: LC-MS/MS; drug interaction; drug monitoring; pharmacokinetics; plasma samples; propafenone