title: A highly efficient and sensitive LC-MS/MS method for the determination of afatinib in human plasma: application to a metabolic stability study.
abstract: Afatinib (AFT) is a new tyrosine kinase inhibitor approved for the treatment of nonsmall cell lung cancer. In the present study, a simple, specific, rapid and sensitive liquid chromatography tandem mass-spectrometric method for the quantification of AFT in human plasma, was developed and validated. Chromatographic separation of the analytes was accomplished on a reversed-phase Luna(®) -PFP 100 Å column (50 × 2.0 mm; 3.0 μm) maintained at ambient temperature. Isocratic elution was carried out using acetonitrile-water (40:60, v/v) containing 10 mm ammonium formate buffer (pH 4.5) adjusted with formic acid at a flow rate of 0.4 mL min(-1) . The analytes were monitored by electrospray ionization in positive ion multiple reaction monitoring mode. The method yields a linear calibration plot (r(2)  = 0.9997) from a quantification range of 0.5-500 ng mL(-1) with the lower limit of quantification and lower limit of detection of 1.29 and 0.42 ng mL(-1) , respectively. The intra- and inter-day precision and accuracy were estimated and found to be in the ranges of 1.53-4.11% for precision and -2.80-0.38% for accuracy. Finally, quantification of afatinib in a metabolic stability study in rat liver microsomes was achieved through the proposed method. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.
authors.forename: Adnan A
authors.initials: AA
authors.lastname: Kadi
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.
authors.forename: Ali S
authors.initials: AS
authors.lastname: Abdelhameed
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.|Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Kasr El-Aini St, Cairo, 11562, Egypt.
authors.forename: Hany W
authors.initials: HW
authors.lastname: Darwish
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.
authors.forename: Mohamed W
authors.initials: MW
authors.lastname: Attwa
authors.affiliation: Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, PO Box 2457, Riyadh, 11451, Kingdom of Saudi Arabia.
authors.forename: Nasser S
authors.initials: NS
authors.lastname: Al-Shakliah
mesh_terms: D000077716:Afatinib; D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D047428:Protein Kinase Inhibitors / Q000097:blood*; D011799:Quinazolines / Q000097:blood*; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D047428:Protein Kinase Inhibitors; D011799:Quinazolines; D000077716:Afatinib
keywords: LC-MS/MS; afatinib; human plasma; metabolic stability study