Determination of endothelin-1 in rats using a high-performance liquid chromatography coupled to electrospray tandem mass spectrometry.
A sensitive and specific liquid chromatography tandem mass spectrometry method with electrospray ionization for the determination of endothelin-1 in rat plasma and lung effluents has been developed and validated. Detection was achieved by an Applied Biosystems MDS Sciex API 2000 triple quadrupole mass spectrometer coupled to an Agilent 1100 LC system. The limit of detection and the limit of the quantification of ET-1 in matrix buffer was estimated at 40 pM and 1 nM, respectively. The precision and accuracy for both intra- and inter-day determination of the analyte ranged from 2.5% to 14.7% and from 104.2% to 113.3%, respectively. No significant relative matrix effect was observed. Stability of ET-1 established in a bench-top, autosampler, long-term storage stability as well as freeze/thaw cycles shown no significant degradation products in the samples. The results of the method validation indicated that this method is applicable for the determination of the ET-1 concentration in an effluent from the isolated lung preparation as well as in vivo in plasma samples to evaluate ET-1 as a potential biomarker of the progression of pulmonary endothelial dysfunction and pulmonary hypertension in rats induced by a monocrotaline injection.
Walczak
Maria
M
Department of Pharmacokinetics and Physical Pharmacy, Jagiellonian University Medical College, Kraków, Poland. mwalczak.farm@poczta.fm
Fedorowicz
Andrzej
A
Chłopicki
Stefan
S
Szymura-Oleksiak
Joanna
J
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004195:Disease Models, Animal; D019332:Endothelin-1 / Q000032:analysis / Q000097:blood*; D006976:Hypertension, Pulmonary / Q000503:physiopathology; D057230:Limit of Detection; D015394:Molecular Structure; D051381:Rats; D053719:Tandem Mass Spectrometry / Q000379:methods*
D019332:Endothelin-1