I have a method that asks me to take 300 mg of Active A and put it into 100 ml of solvent as stock solution A. From stock solution A, I take 10 ml of that solution and dilute again to working solution a, now containing 30 mg of active A. The purity of active A is actually only 97.4% pure. So, there are actually 29.22 mg of Active A in working solution a.
In the HPLC where I enter the injections(concentrations) I set it up as 2 uL(58.44), 4 uL(116.88), 6uL(175.32), 8uL(233.76), 10uL(292.2) injections of the working solution a. For the concentrations I entered the numbers in parenthesis. My units are in mg/100 ml. For some reason, my check standards are reading about 200 times high. I think I'm doing something wrong entering the math into the HPLC. Do you have any suggestions for reading material on entering calculations into HPLC or can you suggest a common error that beginners make when creating calibration curves?
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