This is an Application Brief and does not contain a detailed Experimental section.
This application brief demonstrates the LC-MS gradient separation of buprenorphine and buprenorphine glucuronide on XBridge HILIC Columns.
The compounds used in this study are:
            
        | 
                     Column:  | 
                     XBridge HILIC, 3.5 μm, 2.1 x 100 mm  | 
            
| 
                     Part number:  | 
                     186004433  | 
            
| 
                     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:  | 
                     0.3 mL/min  | 
            
| 
                     Injection volume:  | 
                     5 μL  | 
            
| 
                     Sample concentration:  | 
                     50 ng/mL each  | 
            
| 
                     Sample diluent:  | 
                     75:25 ACN:MeOH with 0.2% HCOOH  | 
            
| 
                     Column temperature:  | 
                     30 °C  | 
            
| 
                     Instrument:  | 
                     Waters ACQUITY UPLC with TQD  | 
            
| 
                     MRM:  | 
                     Buprenorphine 468.3 > 54.7 Buprenorphine glucuronide 644.3 > 468.3  | 
            
| 
                     Dwell time:  | 
                     75 ms  | 
            
| 
                     ISD:  | 
                     10 ms  | 
            
| 
                     ICD:  | 
                     10 ms  | 
            
| 
                     Time(min)  | 
                     Profile  | 
            |
|---|---|---|
| 
                     %A  | 
                     %B  | 
            |
| 
                     0.00  | 
                     0.1  | 
                     99.9  | 
            
| 
                     1.05  | 
                     0.1  | 
                     99.9  | 
            
| 
                     4.35  | 
                     99.9  | 
                     0.1  | 
            
| 
                     5.00  | 
                     99.9  | 
                     0.1  | 
            
| 
                     5.01  | 
                     0.1  | 
                     99.9  | 
            
| 
                     6.00  | 
                     0.1  | 
                     99.9  | 
            
            
        WA64077, August 2009