Hi everyone. I am currently working on HPLC assay of glucosamine, in accordance to USP method. Below are some of the specs of the method:
There are several problems I am facing currently, such as the retention times are not at around 10min (as stated in USP). Also, I am having a problem of delaying of retention time after each injection. For example, a sequence of 6 injections that I have ran gave such retention times, from injection 1 to 6: 5.265, 5.279, 5.294, 5.303, 5.313, 5.322.
The equilibration of the column took about 2hour before the initiation of sequence.
Did anyone encounter such situation before? Your advice is much appreciated.
Keng HONG Tee Here i am attaching File .it might help you. According to USP method mobile phase ratio is 30:70 (buffer : ACN) & flow rate 1.1 ml/min.
Did you condition the column? And do you have freshly prepared mobile phase each day? If so, what is the impact of it?
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