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Shin, Young G.,Lee, Hamm,Murakami, Stanley,Buirst, Kenji,Buonarati, Michael H.,Cox, April,Regal, Kelly,Hunt, Kevin W.,Scearce-Levie, Kimberly,Watts, Ryan J.,Liu, Xingrong 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.5
A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed and applied for the determination of human $A{\beta}1$-40 and $A{\beta}1$-42 peptides in transgenic mouse plasma to support preclinical pharmacodynamics studies. The method consisted of micro-elution solid phase extraction for sample preparation and LC-MS/MS analysis in the negative ion mode using electrospray ionization for analysis. $^{15}N_{53}-A{\beta}1$-40 and $^{15}N_{55}-A{\beta}1$-42 peptides were used as internal standards. A quadratic regression (weighted 1/concentrations), with an equation $y=ax^2+bx+c$, was used to fit calibration curves over the concentration range of 0.500-100 ng/mL for both $A{\beta}1$-40 and $A{\beta}1$-42 peptides. For quality control samples at 6.00, 40.0 and 80.0 ng/mL from the qualification experiment, the within-run accuracy ranged from -2.69 to 0.583 % with precision values ${\geq}8.23%$ for $A{\beta}1$-40. Within-run accuracy ranged from -4.83 to 10.1 % with precision values ${\geq}8.87%$ for $A{\beta}1$-42. Samples from a pharmacodynamics study using Tg2576 transgenic mice were analyzed by this qualified LC-MS/MS method and concentrations were compared to those generated by ELISA. The two methods were shown to be comparable for $A{\beta}1$-40 quantification of samples from the Tg2576 amyloid precursor protein transgenic mouse model, but varied slightly for $A{\beta}1$-42.
Young G. Shin,Lee Hamm,Stanley Murakami,Kenji Buirst,Michael H. Buonarati,April Cox,Kelly Regal,Kevin W. Hunt,Kimberly Scearce-Levie,Ryan J. Watts,Xingrong Liu 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.5
A liquid chromatographic–tandem mass spectrometric(LC–MS/MS) method was developed and appliedfor the determination of human Ab1-40 and Ab1-42 peptidesin transgenic mouse plasma to support preclinicalpharmacodynamics studies. The method consisted ofmicro-elution solid phase extraction for sample preparationand LC–MS/MS analysis in the negative ion mode usingelectrospray ionization for analysis. 15N53-Ab1-40 and15N55-Ab1-42 peptides were used as internal standards. A quadratic regression (weighted 1/concentrations), withan equation y = ax2 ? bx ? c, was used to fit calibrationcurves over the concentration range of 0.500–100 ng/mLfor both Ab1-40 and Ab1-42 peptides. For quality controlsamples at 6.00, 40.0 and 80.0 ng/mL from the qualificationexperiment, the within-run accuracy ranged from-2.69 to 0.583 % with precision values B8.23 % for Ab1-40. Within-run accuracy ranged from -4.83 to 10.1 % with precision values B8.87 % for Ab1-42. Samples from apharmacodynamics study using Tg2576 transgenic micewere analyzed by this qualified LC–MS/MS method andconcentrations were compared to those generated byELISA. The two methods were shown to be comparable forAb1-40 quantification of samples from the Tg2576 amyloidprecursor protein transgenic mouse model, but variedslightly for Ab1-42.