Geeking out here with a story from today's experiments for developing a bioanalytical method for measuring fluoxetine (Prozac) and metabolite in human plasma.
I was puzzled why I could not see the signal response for my analyte that I was expecting. I checked, rechecked, and checked again my method to ensure that the HPLC was set up properly. I calculated, recalculated, and calculated again my procedures to ensure that I had spiked in the correct amounts of drug. I confirmed, reconfirmed, and confirmed again that all of the parameters were being properly transferred from the computer to the HPLC. Then I noticed that a pool of mobile phase was collecting in the column heater. After connecting the end of my column to the detector, I saw my analyte peak.
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