 
            
        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