Determination of bupropion hydrochloride in rat plasma by LC-MS/MS and Its application to pharmacokinetic study

A selective and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for quantitation of bupropion hydrochloride in rat plasma using triazolam as an internal standard. Chromatographic separation was achieved on a SB-C18 column at 30 °C, with 50: 50 (v/v) acetonitrile-0.1 % formic acid in water as mobile phase. The flow rate was 0.3 mL/min. The determination of bupropion was performed in MRM mode, m/z 239.9 → 183.7 for bupropion and m/z 343.0 → 308.0 for triazolam (IS) and positive ion electrospray ionization interface. Calibration curve was linear over range of 1.2 to 480 ng/mL. The intra- and inter-run relative standard deviations of the assay were less than 10 %. The mean absolute recoveries determined at the concentrations of 2.4, 48 and 360 ng/mLwere 91.00%, 92.06%, 91.71%, respectively. The validated method is successfully used to analyze the drug in samples of rat plasma for pharmacokinetic study.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Determination of bupropion hydrochloride in rat plasma by LC-MS/MS and Its application to pharmacokinetic study

A selective and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for quantitation of bupropion hydrochloride in rat plasma using triazolam as an internal standard. Chromatographic separation was achieved on a SB-C18 column at 30 °C, with 50: 50 (v/v) acetonitrile-0.1 % formic acid in water as mobile phase. The flow rate was 0.3 mL/min. The determination of bupropion was performed in MRM mode, m/z 239.9 → 183.7 for bupropion and m/z 343.0 → 308.0 for triazolam (IS) and positive ion electrospray ionization interface. Calibration curve was linear over range of 1.2 to 480 ng/mL. The intra- and inter-run relative standard deviations of the assay were less than 10 %. The mean absolute recoveries determined at the concentrations of 2.4, 48 and 360 ng/mLwere 91.00%, 92.06%, 91.71%, respectively. The validated method is successfully used to analyze the drug in samples of rat plasma for pharmacokinetic study.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Case Report: A novel heterozygous nonsense mutation in KRIT1 cause hereditary cerebral cavernous malformation

Cerebral cavernous malformation (CCM) is a vascular malformation of the central nervous system and mainly characterized by enlarged capillary cavities without intervening brain parenchyma. Genetic studies have identified three disease-causing genes (CCM1/KRIT1, CCM2/MGC4607 and CCM3/PDCD10) responsible for CCM. Here, we characterized a four-generation family diagnosed with CCM and identified a novel heterozygous mutation c.1159C>T, p.Q387X in KRIT1 gene by whole exome sequencing and Sanger sequencing. The Q387X mutation resulted in premature termination of KRIT1 protein, which was predicted to be deleterious by the ACMG/AMP 2015 guideline. Our results provide novel genetic evidence support that KRIT1 mutations cause CCM, and are helpful to the treatment and genetic diagnosis of CCM

Determination of bupropion hydrochloride in rat plasma by LC-MS/MS and Its application to pharmacokinetic study

A selective and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for quantitation of bupropion hydrochloride in rat plasma using triazolam as an internal standard. Chromatographic separation was achieved on a SB-C18 column at 30 °C, with 50: 50 (v/v) acetonitrile-0.1 % formic acid in water as mobile phase. The flow rate was 0.3 mL/min. The determination of bupropion was performed in MRM mode, m/z 239.9 → 183.7 for bupropion and m/z 343.0 → 308.0 for triazolam (IS) and positive ion electrospray ionization interface. Calibration curve was linear over range of 1.2 to 480 ng/mL. The intra- and inter-run relative standard deviations of the assay were less than 10 %. The mean absolute recoveries determined at the concentrations of 2.4, 48 and 360 ng/mLwere 91.00%, 92.06%, 91.71%, respectively. The validated method is successfully used to analyze the drug in samples of rat plasma for pharmacokinetic study.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Determination of troxerutin in rabbit plasma by LC-ESI-MS and its application to a pharmacokinetic study

A sensitive and selective liquid chromatography/electrospray mass spectrometry (LC-ESIMS) method for determination of troxerutin in rabbit plasma was developed. After addition of paeoniflorin as internal standard (IS), protein precipitation by methanol: acetonitrile (3:1, v/v) was used as sample preparation. Chromatographic separation was achieved on an Allure (TM) PFP Propyl (2.1 mm × 100 mm, 5 μm) column with methanol-water as mobile phase with gradient elution. Electrospray ionization (ESI) source was applied and operated in positive ion mode; selected ion monitoring (SIM) mode was used to quantification using target fragment ions m/z 765 for troxerutin and m/z 503 for the IS. Calibration plots were linear over the range of 10-5000 ng/mL for troxerutin in rabbit plasma. Lower limit of quantitation (LLOQ) for troxerutin was 10 ng/mL. Mean recovery of troxerutin from plasma was in the range 92.6-98.1 %. RSD of intra-day and inter-day precision were both less than 11 %. This method is simple and sensitive enough to be used in pharmacokinetic research for determination of troxerutin in rabbit plasma.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Genetic Identification and Molecular Modeling Characterization Reveal a Novel PROM1 Mutation in Stargardt4-like Macular Dystrophy

Stargardt disease-4 (STGD4) is an autosomal dominant complex, genetically heterogeneous macular degeneration/dystrophy (MD) disorder. In this paper, we used targeted next generation sequencing and multiple molecular dynamics analyses to identify and characterize a disease-causing genetic variant in four generations of a Chinese family with STGD4-like MD. We found a novel heterozygous missense mutation, c.734T\u3eC (p.L245P) in the PROM1 gene. Structurally, this mutation most likely impairs PROM1 protein stability, flexibility, and amino acid interaction network after changing the amino acid residue Leucine into Proline in the basic helix-loop-helix leucine zipper domain. Molecular dynamic simulation and principal component analysis provide compelling evidence that this PROM1 mutation contributes to disease causativeness or susceptibility variants in patients with STGD4-like MD. Thus, this finding defines new approaches in genetic characterization, accurate diagnosis, and prevention of STGD4-like MD

Determination of sulpiride in rabbit plasma by LC-ESI-MS and its application to a pharmacokinetic study

A sensitive and selective liquid chromatography-mass spectrometry (LC-MS) method for determination of sulpiride in rabbit plasma was developed and validated. The analyte and internal standard (IS) were extracted from plasma by liquid-liquid extraction using ethyl acetate, and chromatography involved Agilent Extend-C18 column (2.1 mm x 50 mm, 3.5 μm) using 0.2 % formic acid in water and acetonitrile (60: 40, v/v) as a mobile phase. Detection involved positive ion mode electrospray ionization (ESI), and selective ion monitoring (SIM) mode was used for quantification of target fragmentions m/z 342.0 for sulpiride and m/z 294.8 for estazolam (internal standard, IS). The assay was linear over the range of 10-2000 ng/mL for sulpiride, with a lower limit of quantitation (LLOQ) of 10 ng/mL for sulpiride. Intraand inter-day precisions were less than 12 % and the accuracies were in the range of 94.1-108.7 % for sulpiride. This developed method was successfully applied for the determination of sulpiride in rabbit plasma for pharmacokinetic study.Colegio de Farmacéuticos de la Provincia de Buenos Aire

Genetic analysis of Penthorum chinense Pursh by improved RAPD and ISSR in China

Background: Penthorum chinense Pursh (P. chinense) is a well-known traditional Chinese medicine (TCM) plant, which has long been used for the prevention and treatment of hepatic diseases. This study aimed to genetically characterize the varieties of P. chinense from different geographic localities of China by random amplification of polymorphic DNA (RAPD)-PCR technique and verified with inter-simple sequence repeat (ISSR) markers. Results: The P. chinense samples were collected from nine different geographic localities. Previously improved RAPD and ISSR markers were utilized for genetic analysis using DNA amplification. The genetic relationship dendrogram was obtained by conducting cluster analysis to the similarity coefficient of improved RAPD and ISSR markers. Improved RAPD yielded 185 scorable amplified products, of which 68.6% of the bands were polymorphic, with an average amplification of 9.25 bands per primer. The ISSR markers revealed 156 alleles with 7.8 bands per primers, where 59.7% bands were polymorphic. Furthermore, the similarity coefficient ranges of RAPD and ISSR markers were 0.71\u20130.91 and 0.66\u20130.89, respectively. Conclusions: This study indicated that improved RAPD and ISSR methods are useful tools for evaluating the genetic diversity and characterizing P. chinense. Our findings can provide the theoretical basis for cultivar identification, standardization, and molecular-assisted breeding of P. chinense for medicinal use