Sensitive and rapid HPLC-UV method with back-extraction step for the determination of sildenafil in human plasma. 2016

Hamza Al-Hroub, and Bayan Alkhawaja, and Eman Alkhawaja, and Tawfiq Arafat
Faculty of Pharmacy & Medical Sciences, Petra University, Amman-Jordan.

In this work we provided a selective, sensitive and rapid HPLC-UV method for quantification of sildenafil in human plasma. We have adopted a simple liquid-liquid extraction procedure followed a back-extraction in 5% perchloric acid solution. Chromatographic separation was achieved on a BDS C-18Column (150mm×4.6mm, 5μm) using a mobile phase consisted 63% water, 37% acetonitrile and 0.1% triethylamine (pH 7.7). The analysis was detected at 230nm. The achieved lower limit of quantification was 2.00ng/ml. The method showed linear calibration curve over the range of 2.00-200ng/ml. Intra- and inter day precision (CV%) were less than 6.80 and 5.19%, respectively. Whilst intra- and inter day accuracy% were ranged between (98.3 and 105%) and (99.4 and 103%), respectively. Tests confirmed the stability of sildenafil in plasma at room temperature for 24h, during three freeze-thaw cycles, after 24h in autosampler at 10°C and after 60 days in plasma at -30°C. The recovery of sildenafil was greater than 78.4%. The described simple UV method achieved very low limit of quantification and by using simple and inexpensive extraction procedure, complete separation was obtained within short run time. Having demonstrated the validity and novelty of our method, thus it is applicable for the clinical and pharmacokinetic studies of sildenafil in human volunteers especially in laboratories in countries where cost of modern techniques and instrumentation is prohibitive.

UI MeSH Term Description Entries
D002851 Chromatography, High Pressure Liquid Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed. Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000068677 Sildenafil Citrate A PHOSPHODIESTERASE TYPE-5 INHIBITOR; VASODILATOR AGENT and UROLOGICAL AGENT that is used in the treatment of ERECTILE DYSFUNCTION and PRIMARY PULMONARY HYPERTENSION. 1-((3-(6,7-Dihydro-1-methyl-7-oxo-3-propyl-1H-pyrazolo(4,3-d)pyrimidin-5-yl)-4-ethoxyphenyl)sulfonyl)-4-methylpiperazine citrate,Acetildenafil,Desmethyl Sildenafil,Desmethylsildenafil,Homosildenafil,Hydroxyhomosildenafil,NCX-911,Revatio,Sildenafil,Sildenafil Lactate,Sildenafil Nitrate,UK 92480-10,UK-92,480-10,Viagra,Citrate, Sildenafil,Lactate, Sildenafil,NCX 911,NCX911,Nitrate, Sildenafil,Sildenafil, Desmethyl,UK 92,480 10,UK 92480 10,UK 9248010
D014466 Ultraviolet Rays That portion of the electromagnetic spectrum immediately below the visible range and extending into the x-ray frequencies. The longer wavelengths (near-UV or biotic or vital rays) are necessary for the endogenous synthesis of vitamin D and are also called antirachitic rays; the shorter, ionizing wavelengths (far-UV or abiotic or extravital rays) are viricidal, bactericidal, mutagenic, and carcinogenic and are used as disinfectants. Actinic Rays,Black Light, Ultraviolet,UV Light,UV Radiation,Ultra-Violet Rays,Ultraviolet Light,Ultraviolet Radiation,Actinic Ray,Light, UV,Light, Ultraviolet,Radiation, UV,Radiation, Ultraviolet,Ray, Actinic,Ray, Ultra-Violet,Ray, Ultraviolet,Ultra Violet Rays,Ultra-Violet Ray,Ultraviolet Black Light,Ultraviolet Black Lights,Ultraviolet Radiations,Ultraviolet Ray
D015203 Reproducibility of Results The statistical reproducibility of measurements (often in a clinical context), including the testing of instrumentation or techniques to obtain reproducible results. The concept includes reproducibility of physiological measurements, which may be used to develop rules to assess probability or prognosis, or response to a stimulus; reproducibility of occurrence of a condition; and reproducibility of experimental results. Reliability and Validity,Reliability of Result,Reproducibility Of Result,Reproducibility of Finding,Validity of Result,Validity of Results,Face Validity,Reliability (Epidemiology),Reliability of Results,Reproducibility of Findings,Test-Retest Reliability,Validity (Epidemiology),Finding Reproducibilities,Finding Reproducibility,Of Result, Reproducibility,Of Results, Reproducibility,Reliabilities, Test-Retest,Reliability, Test-Retest,Result Reliabilities,Result Reliability,Result Validities,Result Validity,Result, Reproducibility Of,Results, Reproducibility Of,Test Retest Reliability,Validity and Reliability,Validity, Face
D057230 Limit of Detection Concentration or quantity that is derived from the smallest measure that can be detected with reasonable certainty for a given analytical procedure. Limits of Detection,Detection Limit,Detection Limits
D058986 Phosphodiesterase 5 Inhibitors Compounds that specifically inhibit PHOSPHODIESTERASE 5. PDE-5 Inhibitor,PDE5 Inhibitor,Phosphodiesterase 5 Inhibitor,Phosphodiesterase Type 5 Inhibitor,PDE-5 Inhibitors,PDE5 Inhibitors,Phosphodiesterase Type 5 Inhibitors,5 Inhibitor, Phosphodiesterase,Inhibitor, PDE-5,Inhibitor, PDE5,Inhibitor, Phosphodiesterase 5,Inhibitors, PDE-5,Inhibitors, PDE5,Inhibitors, Phosphodiesterase 5,PDE 5 Inhibitor,PDE 5 Inhibitors
D059625 Liquid-Liquid Extraction The removal of a soluble component from a liquid mixture by contact with a second liquid, immiscible with the carrier liquid, in which the component is preferentially soluble. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed) Liquid Phase Extraction,Extraction, Liquid Phase,Extraction, Liquid-Liquid,Extractions, Liquid Phase,Extractions, Liquid-Liquid,Liquid Liquid Extraction,Liquid Phase Extractions,Liquid-Liquid Extractions,Phase Extraction, Liquid,Phase Extractions, Liquid

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