Development and Validation of a Novel LC-MS/MS Method for the Simultaneous Determination of Abemaciclib, Palbociclib, Ribociclib, Anastrozole, Letrozole, and Fulvestrant in Plasma Samples: A Prerequisite for Personalized Breast Cancer Treatment

Abstract

Palbociclib, ribociclib and abemaciclib were recently approved as chemotherapeutic agents and are currently in the post-marketing surveillance phase. They are used in combination with aromatase inhibitors anastrozole and letrozole or antiestrogen fulvestrant for HR+, HER2− breast cancer treatment. Here, a novel bioanalytical LC-ESI-MS/MS method was developed for the quantitation of these six drugs in human plasma. The samples were prepared by simple protein precipitation followed by solvent evaporation. A Kinetex biphenyl column (150 × 4.6 mm, 2.6 µm) used for chromatographic analysis adequately resolved even the closely eluting aromatase inhibitors’ peaks. The mobile phase consisted of 0.1% formic acid in water and in ACN, in a linear gradient. An additional gradient step was added to eliminate the observed carry-over. The proposed method was fully validated in the relevant linear ranges covering the expected plasma concentrations of all six drugs (correlation coefficients between 0.9996 and 0.9931). The intra-day method precision (CV) ranged from 3.1% to 15%, while intra-day accuracy (%bias) was between −1.5% and 15.0%. The inter-day precision ranged from 1.6% to 14.9%, with accuracy between −14.3% and 14.6%, which is in accordance with the EMA and ICH guidelines on bioanalytical method validation. The method was successfully applied to samples from patients treated for HR+, HER2− breast cancer.

Detection of abemaciclib, an anti-breast cancer agent, using a new electrochemical DNA biosensor

“…The limit of detection (LOD) of AMC was determined to be 2.7 nM (3S b /m). This value of detection limit and the linear dynamic range for AMC observed for the ds-DNA/PP/Ce-doped H-NiO-ND/PGE are comparable and better than those obtained for several other previous studies (Table 1) (Margaryan et al, 2022;Turkovic et al, 2022;Martínez-Chávez et al, 2021). The detection limit of only several LC-MS/MS method developed for detection of AMC was superior to our sensor.…”

Development and Validation of a Quantitative LC-MS/MS Method for CDK4/6 Inhibitors Palbociclib, Ribociclib, Abemaciclib, and Abemaciclib-M2 in Human Plasma

“…Although several bioanalytical assays for CDK4/6 inhibitors have been published, these assays do not allow for proper measurement of all said analytes simultaneously. [13][14][15][16][17][18] In particular, abemaciclib-M2 is not measured in conjunction with all CDK4/6 inhibitors by existing assays; thus, our assay, capable of measuring this active metabolite, represents an important and novel contribution. During clinical use, the possibility of monitoring abemaciclib-M2 plasma concentrations would be a major advantage for the following reasons:…”

“…Hence, both compounds are prone to interactions with CYP3A4-modulating drugs, warranting careful monitoring. 9 In addition to measuring all US Food and Drug Administration (FDA)-approved CDK4/6 inhibitors and abemaciclib-M2, we aimed to address issues of certain other CDK4/6 inhibitor assays, such as the lack of use of internal standards, 18 lengthy run times, 18 high plasma sample volumes, 13 or narrow working ranges for one or more analytes. 14,15 Here, we describe our efforts to develop and validate a novel liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) assay for the simultaneous quantitation of CDK4/6 inhibitors plus abemaciclib-M2 in human plasma that addresses these issues and can optimally support TDM, pharmacogenetic studies, and future clinical trials that require bioanalyses of these drugs.…”

Rapid Capillary Electrophoresis Method for Simultaneous Determination of Abemaciclib, Ribociclib, and Palbociclib in Pharmaceutical Dosage Forms: A Green Approach

“…Our proposed method is the first CE method for any CDK4/6 inhibitors as well as the first CE method for the simultaneous determination of ABE, RIB, and PAL. So far, there have been only a few methods reported for simultaneous determination of ABE, RIB, and PAL [ 16 , 17 , 18 ], but these are LC methods that include longer analyses and more organic solvents, making them less environmentally friendly. Although our method has lower sensitivity than LC-MS methods, this does not present any drawbacks for the determination of these drugs in pharmaceutical dosage forms.…”

“…However, this method is an RP-HPLC method applied only to standard solutions and not to pharmaceutical formulations. Currently, only a few methods are reported for the simultaneous determination of CDK4/6 inhibitors in biological samples [ 16 , 17 , 18 ]. Based on a literature search, there are no published CE methods for the analysis of CDK4/6 inhibitors at all.…”