This is an Application Brief and does not contain a detailed Experimental section.
This application brief demonstrates the gradient separation of histidine dipeptides on ACQUITY UPLC BEH HILIC Columns.
The compounds used in this study are:
            
        | 
                     Column:  | 
                     ACQUITY UPLC BEH HILIC, 2.1 x 50 mm, 1.7 μm  | 
            
| 
                     Part Number:  | 
                     186003460  | 
            
| 
                     Mobile Phase A:  | 
                     50/50 ACN/10 mM ammonium formate, w/ 0.125% HCOOH, pH 3.0  | 
            
| 
                     Mobile Phase B:  | 
                     95/5 ACN/10 mM ammonium formate, w/ 0.125% HCOOH, pH 3.0  | 
            
| 
                     Flow Rate:  | 
                     0.5 mL/min  | 
            
| 
                     Injection Volume:  | 
                     5.0 μL  | 
            
| 
                     Sample Diluent:  | 
                     75:25 ACN:MeOH  | 
            
| 
                     Sample Concentration:  | 
                     Creatinine 1μg/mL; Carnosine 5 μg/mL; Anserine 5 μg/mL; Creatine 5 μg/mL  | 
            
| 
                     Column Temperature:  | 
                     30 °C  | 
            
| 
                     Weak Needle Wash:  | 
                     ACN/H2O 95/5  | 
            
| 
                     Instrument:  | 
                     Waters ACQUITY UPLC with ACQUITY SQD  | 
            
| 
                     Time (min)  | 
                     Profile  | 
            |
|---|---|---|
| 
                     %A  | 
                     %B  | 
            |
| 
                     0.00  | 
                     0.1  | 
                     99.9  | 
            
| 
                     5.00  | 
                     99.9  | 
                     0.1  | 
            
| 
                     5.01  | 
                     0.1  | 
                     99.9  | 
            
| 
                     6.00  | 
                     0.1  | 
                     99.9  | 
            
| 
                     Ionization Mode:  | 
                     ES+  | 
            
| 
                     Capillary:  | 
                     2.5 kV  | 
            
| 
                     Cone:  | 
                     20 V (Carnosine; Creatinine, Anserine); 25 V (Creatine)  | 
            
| 
                     Source Temperature:  | 
                     120 °C  | 
            
| 
                     Desolvation Temperature:  | 
                     400 °C  | 
            
| 
                     Desolvation Gas Flow:  | 
                     800 L/Hr  | 
            
| 
                     Cone gas Flow:  | 
                     5 L/Hr  | 
            
| 
                     SIR m/z:  | 
                     227.1 m/z (Carnosine); 132.1 m/z (Creatine); 114.05 m/z (Creatinine); 241.1 m/z (Anserine)  | 
            
| 
                     Dwell Time:  | 
                     0.1 s  | 
            
            
        WA60139, August 2009