This is an Application Brief and does not contain a detailed Experimental section.
This application brief demonstrates the gradient separation of morphine and its metabolites on Atlantis HILIC Silica column.
The compounds used in this study are-
            
        | 
                     Column:  | 
                     Atlantis HILIC Silica, 2.1 x 50 mm, 3 μm  | 
            
| 
                     Part Number:  | 
                     186002011  | 
            
| 
                     Mobile Phase A:  | 
                     10 mM NH4COOH in H2O, 0.125% HCOOH in 50:50 ACN:H2O  | 
            
| 
                     Mobile Phase B:  | 
                     10 mM NH4COOH in H2O, 0.125% HCOOH in 90:10 ACN:H2O  | 
            
| 
                     Flow Rate:  | 
                     0.6 mL/min  | 
            
| 
                     Injection Volume:  | 
                     5 μL  | 
            
| 
                     Sample Concentration:  | 
                     25 μg/mL each  | 
            
| 
                     Sample Diluent:  | 
                     75:25 ACN:MeOH with 0.2% HCOOH  | 
            
| 
                     Column Temperature:  | 
                     30 °C  | 
            
| 
                     Detection:  | 
                     UV @ 280 nm  | 
            
| 
                     Sampling Rate:  | 
                     20 points/sec  | 
            
| 
                     Time Constant:  | 
                     0.1  | 
            
| 
                     Instrument:  | 
                     Waters ACQUITY UPLC with ACQUITY PDA  | 
            
| 
                     Time(min)  | 
                     Profile  | 
            |
|---|---|---|
| 
                     %A  | 
                     %B  | 
            |
| 
                     0.00  | 
                     0.1  | 
                     99.9  | 
            
| 
                     1.05  | 
                     0.1  | 
                     99.9  | 
            
| 
                     4.35  | 
                     99.9  | 
                     0.1  | 
            
| 
                     4.50  | 
                     0.1  | 
                     99.9  | 
            
| 
                     6.00  | 
                     0.1  | 
                     99.9  | 
            
            
        WA64070, August 2009