Automation can help enhance the throughput and improve the sample-to-sample consistency of experiments. With HPLC CONNECT™, DYNAMICS® can be used to simply control your Agilent HPLC autosampler for sample injection into the Mobius™ flow cell, enabling automated size and zeta potential measurements with your Mobius. Automated measurements can also be performed using a native HPLC program separately from DYNAMICS. Setting up automated measurements with your Mobius is simple and straightforward.
Setting up multiple injections in DYNAMICS
DYNAMICS makes running multiple injections with your autosampler effortless by providing a simple way to set up automated experiments with presets. If you are working with an Agilent autosampler using HPLC CONNECT, connecting and controlling your autosampler is straightforward with DYNAMICS.
In DYNAMICS, select the designated Mobius and Autosampler. Once an autosampler is selected, the Auto-sampler control panel button will enable. Within this control panel, a test injection can be performed and information regarding the last injection is displayed.
Using the Event Schedule for automated Mobius measurements
DYNAMICS has multiple presets that can quickly set up automated Mobius measurements, either operating separately from a HPLC injection program or integrated with HPLC CONNECT using an Agilent autosampler.
The perfect workflow can be tailored for your experiments using the event schedule. In addition to flow and injection presets, parameters specific to your sample for DLS and PALS can be set within the event schedule to match your experimental needs. The event schedule can then loop through wash-sample cycles to ensure the flow cell is properly flushed between injections and accurate, high throughput data is collected.
To create an experiment with an event schedule from a preset, navigate to File -> New from Preset… and select the appropriate preset experiment. This will open an experiment with a predefined event schedule that can be modified for your specific needs.
In this example, the Do-Loop will proceed with a wash injection from the first vial position. The Wash-Sample cycle will loop three times, where each loop consists of 5 wash injections followed by 5 sample injections. Additionally, a Sample definition can be applied to sample measurements to streamline analysis if the sample is the same for the three cycles. The preset event schedule then concludes with 5 additional final wash injections.
Best practices for automated Mobius experiments
The following recommendations are great practices to ensure success with your automated DLS and PALS experiments with your Mobius. It is very important the sample is fully cleared from the flow cell prior to injecting the next sample. In addition to the numerous wash injections included in the event schedule, we recommend using a Wash Injection Volume that is ~100-200 μL greater than the Sample Injection Volume. We also recommend using a Wash Flow Rate of 2 mL/min and Wash Pump Run Time of 5 min or equivalent to ensure that 5-10 mL of solvent is flushed between injections.
During sample injection, a Sample Flow Rate of 0.5-1 mL/min is recommended with a Sample Injection Volume of at least 500 μL. Note that the Sample Pump Run Time is the time the pump flows to deliver the sample to the cell prior to stopping the flow for measurement.
To minimize dilution effects, a back pressure regulator added after the Mobius is recommended to ensure flow is properly stopped during measurement.
Mobius Agilent autosampler support via HPLC CONNECT is only available in DYNAMICS 8.1.2 and higher with HPLC CONNECT 184.108.40.206 and higher. Note that only one software can control the autosampler at a time. For more details see the DYNAMICS 8 User’s Guide or the Mobius Hardware User’s Guide.
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