Simultaneous determination of spironolactone and its active metabolite canrenone in human plasma by HPLC-APCI-MS.
A sensitive and specific liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) method for the simultaneous determination of spironolactone and its active metabolite canrenone in human plasma has been developed and validated. After the addition of estazolam as the internal standard (IS), plasma samples were extracted with methylene chloride : ethyl acetate mixture (20 : 80, v/v) and separated by high-performance liquid chromatography (HPLC) on a reversed-phase C18 column with a mobile phase of methanol-water (57 : 43, v/v). Analytes were determined in a single quadrupole mass spectrometer using an atmospheric pressure chemical ionization (APCI) source. LC-APCI-MS was performed in the selected-ion monitoring (SIM) mode using target ions at m/z 341.25 for spironolactone and canrenone, m/z 295.05 for estazolam. The method was proved to be sensitive and specific by testing six different plasma batches. Calibration curves of spironolactone and canrenone were linear over the range 2-300 ng/ml. The within- and between-batch precisions (relative standard deviation (RSD)%) were lower than 10% and the accuracy ranged from 85 to 115%. The lower limit of quantification (LLOQ) was identifiable and reproducible at 2 ng/ml. The proposed method was successfully applied to study the pharmacokinetics of spironolactone and its major metabolite in healthy male Chinese volunteers.
Center for Instrumental Analysis, China Pharmaceutical University, Nanjing 210009, PR China.
Haijuan
H
Dong
Fengguo
F
Xu
Zunjian
Z
Zhang
Yuan
Y
Tian
Yun
Y
Chen
D001774:Blood Chemical Analysis / Q000379:methods*; D002192:Canrenone / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D045424:Complex Mixtures / Q000097:blood; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013148:Spironolactone / Q000097:blood*
D045424:Complex Mixtures; D013148:Spironolactone; D002192:Canrenone