Using a 2.1 mm x 50 mm C18 column and a flow rate of 300 uL/min the attached chromatograms were obtained using a time of flight mass spectrometer as a detector (Note the mass spectrometer data are not required to answer these questions). These 3 samples provide approximately 30 peaks (this is a high estimate) in ~3 minutes. Using these 3 chromatograms, answer the following questions about this chromatographic system: Define bed volume and determine this value for the column used across these 3 samples. Report your answer as an average value and include its standard deviation.
Using an intensity threshold of 50000 counts, and peak elution time window of +/- 0.02 minutes, integrate all peaks across the different spectra and report the area for all identified peaks. Be sure to include a brief description of the approach/method of peak integration. This should include the program/algorithm and settings used. To be clear, your answer should include a printed data sheet (e.g. Excel or Google Sheets) for each chromatogram that lists the peak location, area, and height.
⢠What is the average peak capacity for the separations (hint: you will need to calculate the number of theoretical plates)?
⢠If Chromatogram #3 represents an external standard with a concentration of 10 ng at an elution time of 2.73 min (+/- 0.02 min) what are the respective concentrations of the analyte in Chromatograms #1 and #2 assuming the same nominal retention time.?
Using a 2.1 mm x 50 mm C18 column and a flow rate of 300 uL/min the attached chromatograms were obtained using a time of flight mass spectrometer as a detector (Note the mass spectrometer data are not required to answer these questions). These 3 samples provide approximately 30 peaks (this is a high estimate) in ~3 minutes. Using these 3 chromatograms, answer the following questions about this chromatographic system: Define bed volume and determine this value for the column used across these 3 samples. Report your answer as an average value and include its standard deviation.
Using an intensity threshold of 50000 counts, and peak elution time window of +/- 0.02 minutes, integrate all peaks across the different spectra and report the area for all identified peaks. Be sure to include a brief description of the approach/method of peak integration. This should include the program/algorithm and settings used. To be clear, your answer should include a printed data sheet (e.g. Excel or Google Sheets) for each chromatogram that lists the peak location, area, and height.
⢠What is the average peak capacity for the separations (hint: you will need to calculate the number of theoretical plates)?
⢠If Chromatogram #3 represents an external standard with a concentration of 10 ng at an elution time of 2.73 min (+/- 0.02 min) what are the respective concentrations of the analyte in Chromatograms #1 and #2 assuming the same nominal retention time.?
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