PMID- 20934393
OWN - NLM
STAT- MEDLINE
DCOM- 20110113
LR  - 20191210
IS  - 1873-376X (Electronic)
IS  - 1570-0232 (Linking)
VI  - 878
IP  - 29
DP  - 2010 Nov 1
TI  - Biogenic amines and their metabolites in mouse brain tissue: development, 
      optimization and validation of an analytical HPLC method.
PG  - 3003-14
LID - 10.1016/j.jchromb.2010.09.019 [doi]
AB  - 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 degrees C on a 
      microbore ALF-115 column (150 mm x 1.0 mm, 3 mum particle size) filled with porous 
      C(18) silica stationary phase. In this study, a two-level fractional factorial 
      experimental design ((1/2) 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 (Societe Francaise 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.
CI  - Copyright (c) 2010 Elsevier B.V. All rights reserved.
FAU - Nguyen, A T
AU  - Nguyen AT
AD  - Laboratory of Neurochemistry and Behaviour, Institute Born-Bunge, University of 
      Antwerp, Department of Biomedical Sciences, Universiteitsplein 1, B-2610 Wilrijk, 
      Belgium.
FAU - Aerts, T
AU  - Aerts T
FAU - Van Dam, D
AU  - Van Dam D
FAU - De Deyn, P P
AU  - De Deyn PP
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20100929
PL  - Netherlands
TA  - J Chromatogr B Analyt Technol Biomed Life Sci
JT  - Journal of chromatography. B, Analytical technologies in the biomedical and life 
      sciences
JID - 101139554
RN  - 0 (Biogenic Amines)
SB  - IM
MH  - Animals
MH  - Biogenic Amines/*analysis/*metabolism
MH  - Brain/*metabolism
MH  - *Brain Chemistry
MH  - Chromatography, High Pressure Liquid/*methods
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
EDAT- 2010/10/12 06:00
MHDA- 2011/01/14 06:00
CRDT- 2010/10/12 06:00
PHST- 2010/07/12 00:00 [received]
PHST- 2010/08/28 00:00 [revised]
PHST- 2010/09/18 00:00 [accepted]
PHST- 2010/10/12 06:00 [entrez]
PHST- 2010/10/12 06:00 [pubmed]
PHST- 2011/01/14 06:00 [medline]
AID - S1570-0232(10)00601-X [pii]
AID - 10.1016/j.jchromb.2010.09.019 [doi]
PST - ppublish
SO  - J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Nov 1;878(29):3003-14. doi: 
      10.1016/j.jchromb.2010.09.019. Epub 2010 Sep 29.