Reasons For HPLC Column Peak Fork And No Indication
1.HPLC column no indication
(1) The pump seal gasket is worn;
(2) A large number of air bubbles enter the pump body. Treatment For this case, replace the gasket; for the second case, use a 50 ml glass syringe to help extract air at the pump outlet while the pump is acting.
2. HPLC column peak fork
(1) The HPLC column is contaminated;
(2) The stigma packing collapses. Treatment For this case, first flush the column with pure water, then replace it with methanol, then rinse the column with methanol + isopropanol (4+6) (the length of the rinse time is determined by the contamination of the sample), and then Rinse with methanol, rinse with pure water, and finally rinse with methanol for more than 30 minutes.
If the peak is still poor after rinsing, consider the second case. For the second case, unscrew the column head and check that the HPLC column packing is indented or collapsed.
Remove the indurated part (contaminated packing), fill in the new packing, drop a drop of methanol, fill the packing, refill, press with a smooth stainless steel rod with the same inner diameter of the column, fill it again, drop the methanol, and then press it again. Once, until it is filled and filled. Rinse the HPLC column head with methanol, wipe the packing on the outer wall of the column, tighten the column head, and rinse with pure methanol for more than 30 minutes.