Validation of a Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Method for Determination of All-Trans Retinoic Acid in Human Plasma and Its Application to a Bioequivalence Study

Abstract

A sensitive, reliable and specific LC-MS-MS method was developed and validated for the identification and quantitation of all-trans retinoic acid (ATRA) in human plasma. Acitretin was used as the internal standard (IS). After liquid-liquid extraction of 500 μL plasma with methyl tert-butyl ether (MTBE), ATRA and the IS were chromatographed on a HyPURITY C18 column (150 mm × 2.1 mm, 5 μm) with the column temperature set at 40 °C. The mobile phase was consisted of 40% phase A (MTBE-methanol-acetic acid, 50:50:0.5, v/v) and 60% phase B (water-methanol-acetic acid, 50:50:0.5, v/v) with a flow rate of 0.3 mL/min. The API 4000 triple quadrupole mass spectrometer was operated in multiple reaction monitoring (MRM) mode via the positive electrospray ionization interface using the transition m/z 301.4 → 123.1 for ATRA and m/z 326.9 → 177.1 for IS, respectively. The calibration curve was linear over the range of 0.45-217.00 ng/mL (r ≥ 0.999) with a lower limit of quantitation (LLOQ) of 0.45 ng/mL. The intra-and inter-day precisions values were below 8% relative standard deviation and the accuracy was from

Physiologically Based Pharmacokinetic Model of All-trans-Retinoic Acid with Application to Cancer Populations and Drug Interactions

“…(A) Administration of 10-mg atRA PO (as described in Thudi et al, 2011). (B) Administration of 20 mg atRA PO (as described in Peng et al, 2014). (C) Administration of 22.5 mg/m 2 atRA PO BID (as described in Ozpolat et al, 2003).…”

“…In brief, the fraction absorbed (F a ) and a nominal flow in gut model (Q gut ) were predicted using atRA Caco-2 cell permeability data within Simcyp. The k a and lag time were estimated using the observed concentration-time profiles from three studies in healthy volunteers administered with a single oral dose of atRA (Ozpolat et al, 2003;Thudi et al, 2011;Peng et al, 2014). The reported concentration-time profiles were digitized using Plot Digitizer software (http://plotdigitizer.sourceforge.net), and one compartment pharmacokinetic model with first-order absorption was fitted to the data using Phoenix WinNonlin 6.4 (Pharsight Corporation, Cary, NC).…”

“…For one study (Peng et al, 2014) that was reported in Chinese, the Simcyp Chinese healthy volunteer profile was used with the modifications of the population as described earlier. The population profile was also modified to match the clinical study population for the age range and proportion of females.…”

“…a Data fromThudi et al, 2011. b Data fromPeng et al, 2014. c Data fromOzpolat et al, 2003. d Unpaired t test P , 0.05 between the observed and predicted AUC.PBPK Model of atRA Disposition and Interaction in Humans 253 at ASPET Journals on September 5, 2020 jpet.aspetjournals.org Downloaded from…”

Mass spectrometry in the pharmacokinetic studies of anticancer natural products

“…Separation of the analytes was achieved applying chromatographic conditions quite similar over the different methods developed: the column commonly used was the C 18 (Lehman & Franz, 1996; Gundersen et al, ; Peng et al, ) (C 30 in the case of the method proposed by Ruhl () and an amide column in the case of the assay developed by Arnold et al ()) with gradients formed from mobile phases composed of water and solvents (generally acetonitrile or methanol, or mixture of both) acidified with 0.1% formic acid (Gundersen et al, ; Arnold et al, ) or 0.1% acetic acid (Lehman & Franz, 1996). Total run times were in the range of 7 min (Gundersen et al, ) to 30 min (Lehman & Franz, 1996).…”