Your lab uses the BioRad HPLC system for hemoglobin analysis.As you set up for the analysis, you power up the instrument to allow it to reach operational temperature and prepare the mobile phase buffer.When the instrument and mobile phase are ready, you begin pumping the mobile phase through the system to allow the instrument to equilibrate.You next inject your calibrator.The resulting chromatogram shows nothing but a flat baseline.You inject a larger volume of calibrator and get the same result.Which of the following would explain these results?

Question

Quizplus · Accepted Answer

The detector set at the wrong wavelength is the likely cause for the flat baseline.Even though the calibrator is passing through the stationary phase, when it elutes it would not be detected if the wavelength of the detector was set to the wrong wavelength.The column temperature and the ionic strength of the mobile phase would influence the retention time of the components of the calibrator but would not prevent the elution of these components.Chromatographic windows are retention-time measures and have no influence on chromatographic elution or detection.

Your Lab Uses the BioRad HPLC System for Hemoglobin Analysis