Linked Life Data

20934393

Source:http://linkedlifedata.com/resource/pubmed/id/20934393

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
29
pubmed:dateCreated
2010-10-18
pubmed:abstractText
A simple and fast HPLC method based on an isocratic, reversed-phased ion-pair with amperometric end-point detection for simultaneous measurement of noradrenergic (MHPG/NA and A), dopaminergic (DOPAC, HVA/DA) and serotonergic (5-HIAA/5-HT) compounds in mouse brain tissue was developed. In order to improve the chromatographic resolution (Rs) with an acceptable total analysis time, experimental designs for multivariate optimization of the experimental conditions were applied. The optimal conditions for the separation of the eight neurotransmitters and metabolites, as well as two internal standards, i.e., DHBA and 5-HMT, were obtained using a mixture of methanol-phosphate-citric buffer (pH 3.2, 50 mM) (9:91, v/v) containing 2 mM OSA as mobile phase at 32°C on a microbore ALF-115 column (150 mm × 1.0 mm, 3 ?m particle size) filled with porous C(18) silica stationary phase. In this study, a two-level fractional factorial experimental design (½ 2(K)) was employed to optimize the separation and capacity factor (k') of each molecule, leading to a good separation of all biogenic amines and their metabolites in brain tissue. A simple method for the preparation of different bio-analytical samples in phosphate-citric buffer was also developed. Results show that all molecules of interest were stabilized for at least 24 h in the matrix conditions without any antioxidants. The method was fully validated according to the requirements of SFSTP (Société Française des Sciences et Techniques Pharmaceutiques). The acceptance limits were set at ±15% of the nominal concentration. The method was found accurate over a concentration range of 4-2000 ng/ml for MHPG, 1-450 ng/ml for NA, 1-700 ng/ml for A, 1-300 ng/ml for DOPAC, 1-300 ng/ml for 5-HIAA, 1-700 ng/ml for DA, 4-2800 ng/ml for HVA and 1-350 ng/ml for 5-HT. The assay limits of detection for MHPG, NA, A, DOPAC, 5-HIAA, DA, HVA and 5-HT were 2.6, 2.8, 4.1, 0.7, 0.6, 0.8, 4.2 and 1.4 pg, respectively. It was found that the mean inter- and intra-assay relative standard deviations (RSDs) over the range of standard curve were less than 3%, the absolute and the relative recoveries were around 100%, demonstrating the high precision and accuracy, and reliability of the analytical method described to apply in routine analysis of biogenic amines and their metabolites in brain tissue.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
1873-376X
pubmed:author
pubmed:copyrightInfo
Copyright © 2010 Elsevier B.V. All rights reserved.
pubmed:issnType
Electronic
pubmed:day
1
pubmed:volume
878
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3003-14
pubmed:meshHeading
pubmed:year
2010
pubmed:articleTitle
Biogenic amines and their metabolites in mouse brain tissue: development, optimization and validation of an analytical HPLC method.
pubmed:affiliation
Laboratory of Neurochemistry and Behaviour, Institute Born-Bunge, University of Antwerp, Department of Biomedical Sciences, Universiteitsplein 1, B-2610 Wilrijk, Belgium.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Evaluation Studies, Validation Studies