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Microsoft powerpoint - rosuvastatin_asms_2008

Simultaneous Determination of Rosuvastatin and N-Desmethylrosuvastatin in Human Plasma using Negative Ion ESI-LC/MS/MS
Linge Li, William Mylott, Bruce Hidy, and Rand Jenkins • PPD, Richmond, Virginia
Figure 2. Plasma Blank with Internal Standard
A bioanalytical method was developed for the Validation Samples
Calibration curves were constructed using the analysis of rosuvastatin, N-desmethylrosuvastatin, chromatographic peak area ratios of the analyte and and their respective isotope-labeled internal Calibration standards were prepared in human plasma internal standard from the calibration standards by applying at eight concentration levels spanning the quantitation a quadratic, 1/concentration squared (1/c2) weighted dipotassium EDTA. Samples were processed ranges. Quality control (QC) pools containing the regression algorithm. Analyte concentrations in unknown through mixed-mode solid phase extraction using analytes were prepared in human plasma at the lower samples were then calculated from their peak area ratios a Waters Oasis MAX SPE 10-mg, 96-well plate. limit of quantitation and five concentrations spanning Analysis was performed with a Sciex API4000, triple the quantitation range for the evaluation of assay performance, stability, and recovery.
TurboIonSpray source. Negative ions, formed by Table 1. Regression Equations
deprotonation of the analyte molecules, were measured using multiple reaction monitoring (MRM) Extraction Method
mode. The mass transitions monitored were m/z 480Æ340 (rosuvastatin) and m/z 466Æ362 (N- Sample aliquots (250 µL) were diluted with 75 µL of water and 350 µL of 2% ammonium hydroxide and approximately 6.0 minutes. The assay was validated fortified with 25 µL of working internal standard solution. Analytes were isolated through mixed-mode solid phase extraction using a Waters Oasis MAX SPE 0.0500 to 25.0 ng/mL for N-desmethylrosuvastatin 10-mg, 96-well plate and eluted with 500 µL of 2:98 and requires a 250-µL sample aliquot. Samples are formic acid/acetonitrile, v/v. The eluate was kept frozen at -70 °C prior to analysis. Figure 3. Lower Limit of Quantitation Standard
evaporated under a nitrogen stream at 45 °C, and the A validation was performed to evaluate the remaining residue was reconstituted with 250 µL of specificity, precision, accuracy, recovery, and stability characteristics of the assay. The validation Table 2. Inter-assay Precision and Accuracy
data presented demonstrate that the assay meets the performance requirements needed to support its Chromatographic and
Mass Spectrometric Conditions
Figure 1. Chemical Structures
Sample extracts (25 µL) were injected onto a Waters ACQUITY UPLC BEH HILIC analytical column (2.1 mm x 50 mm, 1.7 µm) connected to an LC/MS/MS system. Mobile Phase A was 2.5 mM ammonium bicarbonate in 98:2:0.01 water/acetonitrile/ammonium hydroxide, v/v/v. Mobile Phase B was 2.5 mM ammonium bicarbonate in 2:98:0.01 water/acetonitrile/ ammonium hydroxide, v/v/v. Mobile phase composition was controlled The analytes were detected by multiple reaction CONCLUSIONS
monitoring on a Sciex API 4000 triple quadrupole mass spectrometer using negative ion TurboIonSpray. Table 3. Stability Summary
An LC/MS/MS assay has been successfully developed and validated for the Retention times for the analytes and their labeled determination of rosuvastatin and N-desmethylrosuvastatin in human plasma internal standards were approximately 2 min.
containing dipotassium EDTA. The assay is suitable for the analysis of clinical study samples as demonstrated by its specificity, precision, accuracy, recovery, and stability characteristics.
The assay was validated over three separate ACKNOWLEDGMENTS
validation runs. Representative results of the validation are summarized in the following tables.
Margaret L. Ware’s contribution to the poster


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EPIDEMIOLOGY THE DIOXIN POLLUTION AS A RISK OF DEVELOPMENT FEMALE BREAST CANCER. CHAPAEVSK STUDY, RUSSIA B.Revich1, T.Ushakova2, O.Sergeev,V.Zeilert31Center for Demography and Human Ecology of Institute for Forecasting of Russian Academy ofSciences. Nakhimovsky prosp., 47. 117 418, Moscow, Russia. 2Cancer Scientific Research Center of Russian Academy of Medicine Sciences. Kashira Drive

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