title: Application of the dried spot sampling technique for rat cerebrospinal fluid sample collection and analysis.
abstract: Dried blood spotting (DBS) sample collection is gaining favor in the pharmaceutical industry due to benefits that include reduced animal usage and easier sample shipment and storage when compared to traditional plasma collection/analysis. The applicability of the DBS card to alternate, limited-volume, matrices has not been as fully characterized as their use with whole blood. In this paper we explored the application of the DBS sample collection technique to rat cerebrospinal fluid (CSF). A reverse phase HPLC-MS/MS method was developed and characterized for the quantitative bioanalysis of the α7 neutonal nicotinic acetylcholine receptor agonist PHA-00543613 in CSF using the dried spot sampling technique. The characterized assay and dried spot sampling technique was employed to analyze serially collected in vivo rat CSF samples after a single 4mg/kg dose of PHA-00543613 in CSF-cannulated rats. The DBS strategy enabled the collection of more timepoints and produced comparable exposure results to those obtained by the collection and analysis of liquid CSF samples but notably with eight less animals.
authors.affiliation: Department of Pharmacokinetics, Dynamics and Metabolism, Pfizer Worldwide Research and Development, Eastern Point Rd, Groton, CT 06340, USA. brian.rago@pfizer.com
authors.forename: Brian
authors.initials: B
authors.lastname: Rago
authors.forename: Jianhua
authors.initials: J
authors.lastname: Liu
authors.forename: Beijing
authors.initials: B
authors.lastname: Tan
authors.forename: Christopher
authors.initials: C
authors.lastname: Holliman
mesh_terms: D053000:Analytic Sample Preparation Methods / Q000379:methods*; D000818:Animals; D001774:Blood Chemical Analysis / Q000379:methods*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000134:cerebrospinal fluid; D002138:Calibration; D002555:Cerebrospinal Fluid / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods; D002626:Chemistry, Pharmaceutical / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods*; D008297:Male; D008956:Models, Chemical; D011812:Quinuclidines / Q000134:cerebrospinal fluid; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000378:metabolism*; D015203:Reproducibility of Results; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; C579165:Chrna7 protein, rat; C513163:N-(1-azabicyclo(2.2.2)oct-3-yl)furo(2,3-c)pyridine-5-carboxamide; D011812:Quinuclidines; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor