Development and validation of highly sensitive method for determination of misoprostol free acid in human plasma by liquid chromatography-electrospray ionization tandem mass spectrometry: application to a clinical pharmacokinetic study.
ABSTRACT A highly sensitive, selective and evaporation free SPE extraction, ESI-LC-MS/MS method has been developed for estimation of misoprostol free acid in human plasma using misoprostol acid-d(5) as an internal standard (IS). The analyte was separated using isocratic mobile phase on reverse phase column and analyzed by MS/MS in the multiple reaction monitoring mode using the respective [M-H] anions, m/z 367-249 for misoprostol acid and m/z 372-249 for the IS. The total run time was 5.0 min and the elution of misoprostol acid and misoprostol acid-d(5) (IS) occurred at 3.6 min. The developed method was validated in human plasma with a lower limit of quantification of 2.5 pg/mL. A linear response function was established for the range of concentrations 2.5-1200 pg/mL (r>0.998) for misoprostol acid in human plasma. The intra and inter-day precision values for misoprostol acid met the acceptance as per FDA guidelines. Misoprostol acid was stable in the battery of stability studies viz., bench-top, auto-sampler and freeze/thaw cycles. The developed assay method was applied to an oral pharmacokinetic study in humans.
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ABSTRACT: Efforts to prosecute women for induced abortion have included allegations that misoprostol was found in body fluids. These claims, however, are questionable owing to the timing of specimen collection for accurate results, the scarcity and expense of validated assays, and the onerous lab procedures required to determine the presence of the substance. Adequate scrutiny should be applied each time such a claim is made. (C) 2014 Published by Elsevier Ireland Ltd. on behalf of International Federation of Gynecology and Obstetrics.International Journal of Gynecology & Obstetrics 08/2014; 127(2). DOI:10.1016/j.ijgo.2014.08.006 · 1.56 Impact Factor
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ABSTRACT: Misoprostol is a pharmaceutical synthetic compound, analog of prostaglandin E1, frequently used as an abortifacient in not medically supervised or self-induced abortions, particularly in countries with restrictive abortion laws representing a serious public health problem. The aim of this study was to develop and validate a sensitive analytical method for the determination of misoprostol acid in whole blood samples. The samples were prepared by SPE and the chromatographic separation was performed by UPLC-MS/MS using ESI- and MRM mode with an Acquity UPLC(®) BEH C18 (50mm×2.1mm i.d., 1.7μm) column using a methanol-ammonium 0.1% solution gradient in a total run time of 7.0min. The method showed to be selective and linear in range 25-2000ng/L. The LOD and LOQ were 10ng/L and 25ng/L, respectively. The recovery ranged from 89 to 97%. No carryover and significant matrix effect were observed. The intra- and inter-assay precisions and the inter-assay accuracy results were 4.0% and 5.4%, 5.5% and 4.1%, and -1.4% and -2.8%, for the concentrations 50 and 500ng/L, respectively. The method developed allows the analysis of misoprostol acid in whole blood samples with adequate sensitivity to the concentration range obtained from therapeutic doses. The method was successfully used in a controlled misoprostol administration study and has been applied in our laboratory in the forensic toxicology field.Journal of pharmaceutical and biomedical analysis 08/2012; 71:198-201. DOI:10.1016/j.jpba.2012.08.005 · 2.83 Impact Factor
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ABSTRACT: Enzyme- and free radical-catalyzed oxidation of polyunsaturated fatty acids (PUFAs) produces the eicosanoids, docosanoids and octadecanoids. This large family of potent bioactive lipids is involved in many biochemical and signaling pathways which are implicated in physiological and pathophysiological processes and can be viable therapeutic targets. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) offers selectivity, sensitivity, robustness and high resolution and is able to analyze a large number of eicosanoids in biological samples in a short time. The present article reviews and discusses reported LC-MS/MS methods and the results obtained from their application in cell models. Reliable analytical outcomes are critically important for understanding physiological and pathophysiological cellular processes, such as inflammation, diseases with inflammatory components (e.g., cardiovascular disease, diabetes, metabolic syndrome), as well as cancer. Reported findings obtained by using the LC-MS/MS methodology in cell systems may have important predictive as well as nutritional and pharmacological implications. We conclude that the LC-MS/MS methodology is a versatile and reliable analytical tool for the simultaneous analysis of multiple PUFA-derived metabolites including the eicosanoids in cell culture samples at concentrations on the pM/nM threshold, i.e. at baseline and after stimulation.Journal of chromatography. B, Analytical technologies in the biomedical and life sciences 05/2014; 964. DOI:10.1016/j.jchromb.2014.05.024 · 2.69 Impact Factor