Determination of the serum concentration of spironolactone and its metabolites by high-performance liquid chromatography

Development and validation of a dried blood spot—LC–APCI–MS assay for estimation of canrenone in paediatric samples

“…Several HPLC methods have been reported for estimation of CAN in various biomatrices such as serum, plasma and urine [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26], however there are no published methods which have employed whole blood in liquid form or in the form of DBS. Many of the methods have several limitations such as lack of selectivity [10,11], low sensitivity [13,17] or large volume of biomatrix required (1 ml plasma) [18,25] and as such are not suitable for estimation of CAN in very low volume paediatric blood samples.…”

“…Many of the methods have several limitations such as lack of selectivity [10,11], low sensitivity [13,17] or large volume of biomatrix required (1 ml plasma) [18,25] and as such are not suitable for estimation of CAN in very low volume paediatric blood samples. We have previously reported a sensitive HPLC method for estimation of spironolactone and its metabolites, including CAN, in paediatric plasma [24] employing 200 l plasma samples however, the same method cannot be extrapolated to DBS samples as the volume of biomatrix to be processed is approximately 20 times lower than that utilised for plasma.…”

Simultaneous determination of spironolactone and its active metabolite canrenone in human plasma by HPLC‐APCI‐MS

“…7,8 With the introduction of high-performance liquid chromatography (HPLC) methods to measure canrenone concentration, it became clear that the fluorimetric method was not specific for canrenone but measured other fluorescigenic metabolites as well. Afterward, Overdiek et al 9,10 developed an HPLC method with an ultraviolet detector to determine spironolactone and its metabolites in human and animal serums. In these studies, only one wavelength (240 nm) was used, although the UV absorption maximum for canrenone appears at 280 nm.…”

“…The limits of quantitation (LOQ), as derived from their calibration curves, were 12.5 ng/ml in guinea pig plasma 10 and 50 ng/ml in human plasma. 9 Varin et al 11 determined spironolactone and its metabolites in human biological fluids by HPLC based on two sequential solid-phase extractions, which is an expensive and timeconsuming method. Then, Jankowski et al 12 modified the HPLC method, which used a one-step liquid-liquid extraction and a programmed switchover of the UV wavelength.…”

Development and validation of an HPLC method for the determination of spironolactone and its metabolites in paediatric plasma samples

“…In addition, sample preparation should be quick and efficient, facilitating use of the assay for routine analysis. Numerous HPLC methods [21][22][23][24][25][26][27][28][29][30][31][32][33] have been developed for the determination of spironolactone and/or its metabolites in plasma or serum. A number of these methods are unsuitable for the determination of spironolactone, 7␣-thiomethylspirolactone and canrenone in paediatric plasma samples either because not all three compounds were determined or the plasma sample size was at least 1 ml.…”