PMID- 23526703
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20211021
IS  - 0009-5893 (Print)
IS  - 0009-5893 (Linking)
VI  - 76
IP  - 1-2
DP  - 2013 Jan 1
TI  - Development of On-line Liquid Chromatography-Biochemical Detection for Soluble 
      Epoxide Hydrolase Inhibitors in Mixtures.
PG  - 13-21
AB  - In this study, an end-point-based fluorescence assay for soluble epoxide 
      hydrolase (sEH) was transformed into an on-line continuous-flow format. The 
      on-line biochemical detection system (BCD) was coupled on-line to liquid 
      chromatography (LC) to allow mixture analysis. The on-line BCD was based on a 
      flow system wherein sEH activity was detected by competition of analytes with the 
      substrate hydrolysis. The reaction product was measured by fluorescence 
      detection. In parallel to the BCD data, UV and MS data were obtained through 
      post-column splitting of the LC effluent. The buffer system and reagent 
      concentrations were optimized resulting in a stable on-line BCD with a good assay 
      window and good sensitivity (S/N > 60). The potency of known sEH inhibitors 
      (sEHis) obtained by LC-BCD correlates well with published values. The LC-BCD 
      system was applied to test how oxidative microsomal metabolism affects the 
      potency of three sEHis. After incubation with pig liver microsomes, several 
      metabolites of sEHis were characterized by MS, while their individual potencies 
      were measured by BCD. For all compounds tested, active metabolites were observed. 
      The developed method allows for the first time the detection of sEHis in mixtures 
      providing new opportunities in the development of drug candidates.
FAU - Falck, David
AU  - Falck D
AD  - Department of BioMolecular Analysis, VU University Amsterdam, Amsterdam, The 
      Netherlands.
FAU - Schebb, Nils Helge
AU  - Schebb NH
FAU - Prihatiningtyas, Setyo
AU  - Prihatiningtyas S
FAU - Zhang, Jiawen
AU  - Zhang J
FAU - Heus, Ferry
AU  - Heus F
FAU - Morisseau, Christophe
AU  - Morisseau C
FAU - Kool, Jeroen
AU  - Kool J
FAU - Hammock, Bruce D
AU  - Hammock BD
FAU - Niessen, Wilfried M A
AU  - Niessen WM
LA  - eng
GR  - U50 OH007550/OH/NIOSH CDC HHS/United States
GR  - R01 ES002710/ES/NIEHS NIH HHS/United States
GR  - P42 ES004699/ES/NIEHS NIH HHS/United States
GR  - U54 OH007550/OH/NIOSH CDC HHS/United States
GR  - U54OH007550/ACL/ACL HHS/United States
PT  - Journal Article
DEP - 20121107
PL  - Germany
TA  - Chromatographia
JT  - Chromatographia
JID - 0316520
PMC - PMC3601784
MID - NIHMS443519
OTO - NOTNLM
OT  - Bioassays
OT  - Column liquid chromatography
OT  - Enzyme inhibition
OT  - Metabolism
OT  - On-line screening
OT  - Soluble epoxide hydrolase
EDAT- 2013/03/26 06:00
MHDA- 2013/03/26 06:01
PMCR- 2014/01/01
CRDT- 2013/03/26 06:00
PHST- 2013/03/26 06:00 [entrez]
PHST- 2013/03/26 06:00 [pubmed]
PHST- 2013/03/26 06:01 [medline]
PHST- 2014/01/01 00:00 [pmc-release]
AID - 10.1007/s10337-012-2343-0 [doi]
PST - ppublish
SO  - Chromatographia. 2013 Jan 1;76(1-2):13-21. doi: 10.1007/s10337-012-2343-0. Epub 
      2012 Nov 7.