A sensitive liquid chromatography–electrospray ionization–tandem mass spectrometry (LC–ESI–MS–MS) method was developed and
validated for the quantification of cepharanthine (CEP) in beagle dog plasma. The chromatographic separation was performed
on an Agilent-C18 column and the mobile phase was composed of methanol:water with 10mM ammonium acetate (20:80, v/v). Detection was operated in the positive ion mode and the tandem mass spectrometer was tuned in the multiple reactions monitoring
mode (MRM) to monitor m/z transitions 607→365 for CEP and 285→193 for the internal standard (IS) diazepam. This method exhibited a linear range
of 5–2,500ngmL−1. The precision (RSD%) and accuracy (RME%) of the assay were <8.7 and 2.4%, respectively. The limit of quantification was
5ngmL−1 and no significant matrix effect was observed. The validated method has been successfully applied to pharmacokinetic study
of CEP in beagle dog.
KeywordsColumn liquid chromatography–tandem mass spectrometry–Pharmacokinetic study–Cepharanthine
"The present work involves the development of a semiautomated microextraction by packed sorbent (MEPS) for the extraction of cepharanthine in plasma sample. Previous works have reported analytical methods for the extraction and quantification of cepharanthine in plasma sample from beagle dog and human  . However, no analytical methods have been reported for the quantification of cepharanthine in mouse, which is a classic malaria model. "
[Show abstract][Hide abstract] ABSTRACT: The spread of Plasmodium falciparum resistance toward most of the used drugs requires new antimalarial compounds. Taking advantage of the biodiversity, the ethnopharmacological approach opens the way for the discovery and the characterization of potent original molecules. Previous works led to the selection of a bisbenzylisoquinoline, cepharanthine, extracted from Stephania rotunda, which is mainly present in Cambodia. A sensitive and selective liquid chromatography method has been developed for the determination of cepharanthine in mouse plasma. The method involved a semiautomated microextraction by packed sorbent (MEPS) using 4 mg of solid phase silica-C8 sorbent. LC separation was performed on a Kinetex XB-C18 column (2.6 µm) with a mobile phase of acetonitrile containing formic acid and 10 mM ammonium formate buffer pH 3.5. Data were acquired at 282 nm with a diode array detector. The drug/internal standard peak area ratios were linked via linear relationships to plasma concentrations (75-2,000 ng/mL). Precision was below 5% and accuracy was 99.0-102%. Extraction recovery of cepharanthine was 56-58%. The method was successfully used to determine the pharmacokinetic profile of cepharanthine in healthy and Plasmodium berghei infected mice. The infection did not impact pharmacokinetic parameters of cepharanthine.
Journal of Analytical Methods in Chemistry 02/2014; 2014(1):695231. DOI:10.1155/2014/695231 · 0.79 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: The leaf hydroalcoholic extract of Cissampelos sympodialis Eichl. (Menispermaceae) has shown promising activity in different animal models of asthma. Several alkaloids have been identified in the extract, including warifteine and methylwarifteine (bisbenzylisoquinoline), as well as milonine (morphinandienone).
To develop and validate an analytical method for the simultaneous quantitation of the bioactive markers of C. sympodialis hydroalcoholic leaf extract and to apply the method to a seasonal (phenological) study of the concentration of the alkaloid markers.
The method used reversed phase high performance liquid chromatography with UV detection and calibration by standard addition. Separation was achieved using a C₁₈-column (250 × 4.6 mm, 5 µm) and a mobile phase consisting of a mixture of 0.05% aqueous (Et)₃NH₂ (A):MeOH(B) in gradient mode at a flow rate of 1.0 mL/min.
The method proved to be linear in the concentration range tested (2-100 µg/mL, r² > 0,99), precise (RSD ≤ 15%), accurate (85-115%), selective and robust. Detection limits for warifteine, methyl-warifteine and milonine were 0.39, 1.10 and 1.77 µg/mL respectively. The highest concentration of total alkaloids (determined as the sum of the three alkaloids) in the hydroalcoholic extract of the leaves was 2.9 ± 0.2 mg/g extract (n = 3), prior to fruit development. Both warifteine and methylwarifteine were detected in the total alkaloid fraction of the ripened fruits.
The results demonstrated that significant variations in the concentration of the biomarkers occurred throughout the vegetative cycle. The lowest concentration of the alkaloids in the leaves coincided with their appearance in the ripened fruits.
[Show abstract][Hide abstract] ABSTRACT: Although traditional Chinese medicines (TCMs) play important role in drug discovery and human health, the actual value of TCMs has not been fully recognized worldwide due to its complex components and uncontrollable quality. For the modernization and globalization of TCMs, it is important to establish selective, sensitive and feasible analytical methods for determination and quantification of bioactive components of TCMs in body fluids primarily due to the low concentration, the complex nature of the biological matrices, and multi-components and their metabolites present in biological fluids. The present review summarizes the current extraction techniques, chromatographic separation and spectroscopic (especially mass spectrometric) analysis methods and new trends on the analysis of bioactive components and metabolites of TCMs in biological fluids. In addition, the importance of establishment of pharmacokinetics and bioavailability profiles and simultaneous determination of multi-active components in TCMs is discussed to provide proper examples of analytical methods for pharmacological and clinical studies of TCMs.
Journal of pharmaceutical and biomedical analysis 04/2013; 87. DOI:10.1016/j.jpba.2013.04.006 · 2.98 Impact Factor
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