32 August / September 2019


and total method cycle time of 5.7 minutes with PMAX


of 510 bar. This provides a >9


times increase in sample throughput and a >89% reduction in runtime / cycle time. The modifi ed rapid HPLC format data in Figure 2C, has a new run time of 13.4 minutes and a total cycle time of 21.3 minutes (including equilibration) with PMAX


of 193 bar. This


represents a >2.5 times increase in sample throughput and a >60% reduction in runtime / cycle time. Solvent consumption for the total cycle times for each format (but excluding initial equilibration and shutdown methods) for 100 samples can be calculated as 5,400 mL, 718 mL and 1,515 mL for the original, UHPLC and modifi ed rapid HPLC methods respectively.


Concluding remarks


Using simple fi rst principles or free software tools, it is possible to signifi cantly increase sample throughput and reduce solvent consumption for many legacy HPLC methods providing a boost to lab capacity


and speed of data output. The magnitude of the improvement will depend upon the laboratory instrumentation available; UHPLC offers impressive numbers but modifi ed rapid HPLC options are still worthy of consideration. In this article, isocratic and gradient examples have been used to demonstrate the approach with calculations of sample throughput, runtime savings and solvent savings included. When exploring these exciting potential improvements, it is important to remember that certain system characteristics (dispersion, dwell time, detector settings, etc.) should be determined and factored into experiments to ensure method translations are as accurate as possible [4, 5]. These concepts and how they can affect isocratic and gradient method translations are also discussed elsewhere [6]. Although the method translation approach is often discussed in the context of migrating existing methods to UHPLC, this article demonstrates how signifi cant improvements can also be made to maximise the utilisation of existing HPLC instrumentation.


References


1. ACE Knowledge Note AKN0023: Gradient Method Translations Using the ACE Translation Tool (accessed at https:// uk.vwr.com/cms/ace_knowledge_notes)


2. ACE LC Translator (download at https:// uk.vwr.com/cms/ace_knowledge_zone)


3. ACE Knowledge Note AKN0015: How to determine column volume and particle porosity (accessed at https://uk.vwr.com/ cms/ace_knowledge_notes)


4. ACE Knowledge Note AKN0001: How to Determine System Dwell Volume (accessed at https://uk.vwr.com/cms/ace_knowledge_notes)


5. ACE Knowledge Note AKN0017: How to Determine Extra Column Dispersion and Extra Column Volume (accessed at https:// uk.vwr.com/cms/ace_knowledge_notes)


6. Webinar by Alan P McKeown, “Practical UHPLC: Selectivity and Rapid Method Development, Method Translations and Instrument Transfers” (accessed at https:// uk.vwr.com/cms/uhplc_webinar_7)


Runge Mikron Miniature Pocket Sized Detectors


Small and modular. Robust and intelligent. The new range of Runge Mikron detectors recently introduced by Biotech AB, unites many attractive features in a tiny package. Different modules can be combined to measure in a variety of fluidic systems, including liquid chromatography.


Thanks to several available LED-based fixed-wavelength light sources with a lifetime of more than 5000 hours, power consumption below 2.5 watts, and start-up within seconds, the Runge Mikron detectors are ideal for incorporation into portable field instruments and online monitoring devices. This also allows the detectors to be operated in refrigerated laboratory environments down to 4°C (39°F), without virtually any heat production. Flexibility in sampling frequencies, detector cell sizes, port connections and choice of wetted materials, make the Runge Mikron detectors suitable for many flow rate conditions and eluent compositions.


The Runge Mikron detectors are easy to connect as they communicate through and draw power from one single USB-C port. Drivers are provided for several chromatography software, alternatively, an open protocol can be used for customised implementation. The detectors conform to international standards and are CE marked.


More information online: ilmt.co/PL/RJqg


Robust (U)HPLC Columns for Proteins/mAbs


YMC-Triart Bio C4 is a new wide-pore phase for (U)HPLC based on the established hybrid-silica particle, YMC-Triart. Due to its 300 Å pore size, it is designed for peptide, protein or monoclonal antibody separations. High temperature (up to 90°C) and pH (1-10) stability is provided! Excellent column-to-column as well as lot-to-lot reproducibility is guaranteed.


More information online: https://ymc.de/rp-bioseparation.html


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