Development, Validation and Application of an LC-MS/MS Bioanalytical Method for the Quantification of GF449, A Novel 5-HT<sub>1A</sub> Agonist, in Rat Plasma and Brain

ISSN: 1875-6638 (Online)
ISSN: 1573-4064 (Print)

Volume 12, 8 Issues, 2016

Download PDF Flyer

Medicinal Chemistry

Aims & ScopeAbstracted/Indexed in

Submit Abstracts Online Submit Manuscripts Online

Honorary Life Fellow
Kings College
University of Cambridge

View Full Editorial Board

Subscribe Purchase Articles Order Reprints

Current: 1.363
5 - Year: 1.388

Development, Validation and Application of an LC-MS/MS Bioanalytical Method for the Quantification of GF449, A Novel 5-HT1A Agonist, in Rat Plasma and Brain

Medicinal Chemistry, 10(5): 449-459.

Author(s): Silvia Franchini, Laura Taddia, Diego Pinetti, Gianluca Carnevale and Livio Brasili.

Affiliation: Dipartimento di Scienze Farmaceutiche, Università degli Studi di Modena e Reggio Emilia, Via Campi 183, 41100 Modena, Italy.


We have recently reported a novel class of selective 5-HT1A agonists among which GF449 emerged for its high potency and almost full agonist activity (pKi 5-HT1A = 8.8; pD2 = 9.22, %Emax = 91.6). In order to quantify GF449 in rat plasma and brain, a sensitive LC-MS/MS method was developed and validated.

Solid phase extraction (SPE) or a combined protein precipitation SPE permitted an efficient analyte recovery and sample clean-up. Multiple reaction monitoring (MRM) was used to track both GF449 and its internal standard (IS), MM189. GF449 was determined and quantitated to nanomolar concentrations in both plasma and brain matrix (LOQs = 0.0025 nmol/mL). Specificity was ensured using three further MRM qualifier transitions for both analyte and IS. Linearity was found in the range of 0.0025 nmol/mL to 1.00 nmol/mL (R2 = 0.9965) and from 0.0025 nmol/mL to 50 nmol/mL (R2 = 0.9999) for plasma and brain respectively. Intraday trueness ranged from 94.0% to 117.5% for brain and from 93.7% to 108.1% for plasma, while precision values were within 3.0% - 6.7% and 2.5% - 9.2% for plasma and brain respectively. The interday trueness of plasma ranged from 89.6% to 107.7% and the precision values (CV%) ranged from 4.6% to 7.5%. Interday trueness and precision (CV%) of the brain ranged from 94.3% to 101.2% and from 1.6% to 11.5% respectively.

The method was validated in accordance with the EMEA guidelines and was successfully applied to plasma and brain samples obtained from rats treated with a 10 mg/kg single oral dose of GF449, thus demonstrating its applicability to preclinical pharmacokinetic studies.


5-HT<sub>1A</sub> receptors agonist, rat plasma, rat brain, LC-MS/MS, quantitative analysis, pharmacokinetics.

Purchase Online Order Reprints Order Eprints Rights and Permissions

Article Details

Volume: 10
Issue Number: 5
First Page: 449
Last Page: 459
Page Count: 11
DOI: 10.2174/1573406409666131128150507

Related Journals

Webmaster Contact: Copyright © 2016 Bentham Science