Off-line HPLC method combined to LC–MS for the determination of sildenafil and its active metabolite in post-mortem human blood according to confirmation criteria

Pistos, Constantinos; Papoutsis, Ioannis; Dona, Artemis; Stefanidou, Maria; Athanaselis, Sotiris; Maravelias, C; Spiliopoulou, Chara

doi:10.1016/j.forsciint.2008.03.018

Cited by 31 publications

(26 citation statements)

References 15 publications

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“…Tracqui 2003(Tracqui, et al, 2003 estimated SC using HPLC-MS Bupernorphine as an internal standard. Constantinos Pistos 2008(Pistos, et al, 2008 used LC-MS for estimation of SC in postmortem human blood samples. Daniela De Orsi 2009 (Daniela, et al, 2009) used HPLC-PDA and elctrospray MS and Benzoic acid as internal standard for estimation of SC.…”

Section: Introductionmentioning

confidence: 99%

RP-HPLC Method Development for Estimation of Sildenafil Citrate in Tablets and in Seminal Fluid

Sharma¹

2012

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A RP HPLC method for estimation of sildenafil citrate (SC) in tablet dosage form and seminal fluid was developed and validated. Best resolution was obtained with column Waters Spherisorb® C 18 bonded silica, (5 µm, 4.6 x 250 mm) at 230 nm with retention time of 5.01 min. The mobile phase used was TEA (0.2%) pH adjusted at 3 with OPA and ACN (60:40) with flow rate of 1.0 mL/min. The method for estimation of sildenafil citrate in tablet dosage form was found to be linear, accurate, precise, sensitive and selective. Whereas the bioanalytical estimation of SC in seminal fluid was found to be in the range of 100 ng/mL to 10µg/mL. Method was found to be highly sensitive as LOD and LOQ were found to 0.3 µg/ml and 0.9µg/ml. The repeatability and reproducibility were within the range i.e. less than 2%.The accuracy of the method was 99.3%. The percentage purity was calculated for market formulation was 102.8%. The internal standard used for bioanalytical methods was Diclofenac sodium. The drug was extracted from seminal fluid by protein precipitation using ACN as precipitating agent. The linearity range was from 100.0ng/mL to 2.0µg/mL. The LOD and LOQ were found to 0.03µg/mL and 0.1µg/mL. The repeatability and reproducibility were within the range i.e. % RSD less than 15%. The accuracy of the method was 90.36%.and the extraction efficiency was found to be 98.25%. The stability of drug was found to be within the range i.e. less the 15%. INTRODUCTIONSildenafil citrate marketed as Viagra TM (Pfizer) was approved as a drug for treating male erectile dysfunction (ED) by the US Food and Drug Administration on 27 March 1998 (Anonymous, 1998). SC is 1- [[3-(6,7-dihydro-1-methyl-7-oxo-3-propyl-1H pyrazolo[4,3-d] pyrimidin-5-yl) -4-ethoxyphenyl] sulfonyl]-4-methyl piperazine citrate is a PDE5 inhibitor used for treatment of ED (Anonymous, 2010). Nitric Oxide (NO) is released with sexual stimulation from nerve endings and endothelial cells in the spongy erectile tissue, the corpus cavernosum of the penis. This release of NO activates the enzyme guanylate cyclase. The enzyme guanylate cyclase then converts guanosine triphosphate (GTP) into cGMP causing the smooth muscle to relax, which causes an inflow of blood, which then leads to an erection. Cyclic guanosine monophosphate (cGMP) is then hydrolysed back to the inactive GMP by phosphodiesterase type 5 (PDE5). The level of cGMP is therefore controlled by the activation of cyclic nucleotide cyclase and the breakdown by PDE5.

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Section: Introductionmentioning

confidence: 99%

RP-HPLC Method Development for Estimation of Sildenafil Citrate in Tablets and in Seminal Fluid

Sharma¹

2012

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“…A few analytical methods have been published for the quantification of SDL alone [3,12,13] or with DSDL [1,[14][15][16] in human plasma [1,3,[12][13][14], blood [15] and urine [16]. Most of them are based on liquid chromatography and the only published GC/MS [16] is applied in urine samples, which are not suitable for therapeutic drug monitoring.…”

Section: Introductionmentioning

confidence: 99%

Development and validation of a gas chromatography‐mass spectrometric method for the determination of sildenafil and desmethyl‐sildenafil in whole blood

Papoutsis

Νικολάου

Athanaselis

et al. 2011

J of Separation Science

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Sildenafil (SDL) is a phosphodiesterase type 5 inhibitor and it is approved for the treatment of erectile dysfunction and pulmonary hypertension. SDL is extensively metabolized to its pharmacologically active metabolite, desmethyl-sildenafil (DSDL). A sensitive and specific GC/MS method for the determination of SDL and DSDL in whole blood was developed and validated to support therapeutic drug monitoring of SDL patients. The combination of solid-phase extraction with derivatization using BSTFA with 1% TMCS in acetonitrile efficiently reduced matrix effect and improved sensitivity of the method. In this assay, protriptyline was used as internal standard for both analytes. The LODs were 1.50 and 5.00 ng/mL for SDL and DSDL, respectively, whereas the respective LOQs were 5.00 and 15.0 ng/mL. The calibration curves were linear up to 500.0 ng/mL (SDL: R(2) 0.992, DSDL: R(2) 0.990). Absolute recovery values for both analytes ranged from 83.1 to 93.2%. Within- and between-batch accuracy was less than 11.8 and 10.2%, respectively, whereas within- and between-batch precision was less than 8.1 and 10.8%, correspondingly. The developed method is suitable for the determination of SDL and DSDL concentrations in blood samples obtained from patients under Viagra(®) treatment, for pharmacokinetic studies or for the investigation of related forensic cases.

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“…The MRM transition used for pregabalin and UK-103320 were the same as previously reported in literature [14, 15,26]. However, the transition monitored for sildenafil was different from the one previously reported [15,26] since on our system better sensitivity was achieved using the optimised MRM transition. The selectivity of the method was tested by using six different batches of plasma.…”

Section: Methods Developmentmentioning

confidence: 79%

Simultaneous Determination of Pregabalin, Sildenafil and Its Active Metabolite in Rat Plasma Utilising SPE Followed by LC–MS–MS

Dżygiel

Fraier

2011

Chromatographia

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A sensitive and selective analytical method for the quantification of pregabalin, sildenafil and the active desmethyl metabolite of sildenafil (UK-103320) has been developed. The method can simultaneously quantify the three analytes within the expected in vivo concentration ranges using 50 lL of rat plasma. It utilises solid-phase extraction followed by high performance liquid chromatography coupled with tandem mass spectrometry. Quantitation in rat plasma demonstrated good accuracy and precision over the following dynamic ranges for each analyte: pregabalin (70-10,000 ng mL -1 ), sildenafil (1-2,000 ng mL -1 ) and UK-103320 (1-2,000 ng mL -1 ). For each analyte, the following lower limits of quantitation were obtained: 70 ng mL -1 for pregabalin and 1 ng mL -1 for sildenafil and UK-103320, respectively. The method was successfully used to analyse plasma samples from rats when pregabalin and sildenafil were administered in combination.Keywords LC-MS-MS Á Monolithic-based chromatography Á Solid-phase extraction Á Pregabalin Á Sildenafil Experimental Chemicals, Reagents, SolutionsPregabalin, sildenafil, UK-103320, as well as UK-464273 and UK-343664 which were used for pregabalin and P. D_ zygiel (

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Off-line HPLC method combined to LC–MS for the determination of sildenafil and its active metabolite in post-mortem human blood according to confirmation criteria

Cited by 31 publications

References 15 publications

RP-HPLC Method Development for Estimation of Sildenafil Citrate in Tablets and in Seminal Fluid

RP-HPLC Method Development for Estimation of Sildenafil Citrate in Tablets and in Seminal Fluid

Development and validation of a gas chromatography‐mass spectrometric method for the determination of sildenafil and desmethyl‐sildenafil in whole blood

Simultaneous Determination of Pregabalin, Sildenafil and Its Active Metabolite in Rat Plasma Utilising SPE Followed by LC–MS–MS

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