3 - UPLC_Work_Practical_Considerations

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Running U-HPLC: Practical Considerations Strong Wash Solvent • Choose based on the chemistry application • 100% organic solvent is acceptable – Except THF – Do not add acid or base in 100% organic solvent • Prime using the needle wash function • Default value is 200 µL – 200 µL is a typical value for this function Weak Wash Solvent • Purges needle and syringe fluid path • Must be compatible with sample solvent • For best results, weak wash solvent should be equivalent to the following (excluding buffers): – mobile phase composition (for isocratic separations) – initial gradient condition (for gradient separations) – If you dilute the samples, match the weak wash solvent to the sample diluent • Degassed for good hydraulic properties Shula Levin, Waters Israel 26
Running U-HPLC: Practical Considerations Weak Wash Solvent • Compatible with initial gradient conditions and sample solubility • Avoid buffers – increases the risks of precipitation at re- equilibration • Five prime cycles fully replace wash solvents • Default value is 200 µL ACQUITY UPLC Sample Manager Injection Methods THREE injection methods available: 1. Full Loop Injection Method/Mode as the injection technique using “Pressure Assist” • Injects 100% of actual loop volume 2. Partial Loop Injection Method/Mode using “Needle Overfill” as the injection technique • Selectable: Recommended from 10% to 75% of total loop volume 3. Partial Loop Injection Method/Mode using “Pressure Assist” as the injection technique • Selectable: Recommended from 10% to 50% of total loop volume Shula Levin, Waters Israel 27
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3 - UPLC_Work_Practical_Considerations

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