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Compact HPLC

November 4, 2019

High performance liquid chromatography (HPLC) is a nearly universal analytical tool in organic synthesis and biochemical development laboratories. With the advent of commercially available automated sample extraction and dilution systems for heterogeneous systems (e.g. Mettler EasySampler, GlobalFIA FloPro, etc.), process development laboratories are increasingly moving towards automated analyses during synthetic route development and scale-up. Automated measurement enables unattended operation with consistent sampling at regularly scheduled intervals, increasing data density and leading to more efficient process understanding. A current limitation of the technology is related to analysis using conventional liquid chromatography systems. The chromatography system must be located proximal to the extraction and analysis or transferred over large distances. The former requirement imposes restrictions in confined spaces near the face of the laminar flow hoods where these studies are typically undertaken, and the latter approach imposes additional transfer time while increasing the risk of hold-up or carryover between successive samples. The ETC desires an integrated system (sample preparation and analysis) in a compact form factor that would enable mounting inside a lab hood. Depending on configuration, a compact system might enable “on demand” operation, due to minimal dead volume and small volume components that could quickly equilibrate prior to performing measurement. 


Download the Request for Information and submit your response.





What is the method of injection? Automated valve or hand turn valve?  

An automated valve with the ability to accept a sample and trigger from an external sampling module would be desired, and/or the ability to interface an external autosampler (ie PAL, HTA).

When asking for gradient capability do you want high pressure mixing ,or low pressure mixing with a quaternary valve and built in degassing or both options?

High pressure mixing is preferable, but both options would be desirable.

What size columns and how many need to fit the temperature controlled column compartment?

1-column, up to 150mm. Multiple column capacity is optional.

Under software requirements, can you provide guidance on exactly what is required to achieve an intuitive interface?

Graphic and tabular interface with limited number of tabs, using conventional terminology, such that routine operations can be performed following a 10-15 minute tutorial or using a quick start guide.

Would a touch panel control interfaced to a standard chromatography package be acceptable? What sequence of screens and options would you want on the touch panel?

Touch screen would be acceptable, run control with single and multi-injection capability, instrument status and real time chromatogram display with the potential for reaction kinetic plot. 

Would there be only one configuration or would you prefer a modular rack system that would enable a choice of pumping options as well as detector options to be configured for each application?

Modular would be desirable.

I was reviewing the RFI and realized that one of the application areas might be monitoring proteins. If this is correct, I believe that adding a pH/ Conductivity monitor as one of the modules would be useful.

In order to accommodate biological measurements, optional features (e.g. passivation/injection port flush) could be included, as well as  pH/conductivity sensors. Engineering of those features is left to the vendor’s discretion, but would be considered favorably in the review process.

What would be the preferred flow rate?  It is currently not specified in the proposal.  Should we assume analytical flow rate ranges? 

Yes, 0.1-2 mL/min, as provides appropriate linear velocity for the system.


What would be the preferred method of sample introduction?  Is the system required to pull the sample outside of the instrument?  Does it also need to incorporate vial introduction and if so, how many?   

As described in the proposal, valved injection from an externally prepared sample is the anticipated routine operation, with optional coupling to vendor or third party autosamplers.


You specified the single column compartment capable of 20°C – 65°C temperature control.  Can this be expressed as “ambient to 65°C”?  Some labs may operate above 20°C as ambient (depending on season and location).  Does the 20°C requirement indicate an expectation of some level of compartment cooling capability? 

Ambient is sufficient for initial implementation, later versions sub-ambient would be desirable, but not necessary.


Can we assume the wetted parts compatibility requirement is for typical LC solvents (acetonitrile, methyl alcohol)?

Yes, the basic configuration is envisioned for “typical” organic synthesis support. If a bio-inert system can be defined, that is a desirable option.


Is there an expectation of sample throughput capability?

This question is somewhat unclear. The typical method cycle times are viewed to be < 8 minutes, and the system should be able to support 24 hours of unattended operation (mobile phase capacity, etc.).


With respect to the output via OPC-UA, are there a specific set of commands that should be enabled?

Status signals, such as system ready/busy, and report files would ideally be rendered through OPC-UA, but if there are other standard file formats for export, please identify. It is also desired that the raw detector signal can be passed out to an external CDA system (Empower, OpenLab, etc.)


It is mentioned that an integrated system in a compact form factor with dimensions in the order of
16” x 14” x 18” (W x H x D) is desired. Would it be acceptable if the dimensions are not met exactly but the system volume is in a similar range (i.e. if the footprint is smaller whilst the height is > 14”)? 

Provide the current dimensions you think are reasonable for your system. The dimensions provided in the RFI are suggested dimensions.


Is an integrated system mandatory? Would it be acceptable to have a modular system which matches the desired form factor or comes close?

A modular system is acceptable, as long as simplicity of operation and robustness are retained.


Should the specified dimensions include the solvent reservoirs and is there flexibility on the size and location of the solvent reservoirs? 

Ideally that would include the solvent reservoirs, but there is flexibility on the size and location. The team will select the unit that most closely matches the desired attributes, but is not anticipating any unit as a precise match.


There is no information about the operating temperatures. Could you please specify the desired temperature range as this might have impact on the temperature range of the column thermostat?

The first generation will be in use in laboratory environments, so 60-80°F. Subsequent iterations may have more expansive requirements for ambient temperature.


It is mentioned that a migration path to GMP must be available. Does availability of compliance services (IQ/OQ), compliant software (i.e. 21CFR part 11 support) cover this or are there further requirements (i.e. ATEX certifications, specific material requirements)?

GMP operation is  currently out of scope, but the team desires a path to GMP use (e.g. coupling with an existing 21CFR Part 11 compliant CDS or path to certification). Other considerations can be discussed with the group during the presentation.


What are the boundary conditions to execute a project together with ETC? Is there financial funding available from the possible project sponsors for the development? What is the typical timeframe for an ETC funded project? 

To execute a project with ETC there is a standard project agreement and confidentiality agreement that are executed between ETC and the 3rd party collaborator.  In addition a statement of work (SOW) needs to be agreed upon by all parties.

While there are examples where ETC members have fully funded a project, it is expected that the collaborator also invest in the project.  Historically ETC members participating in a project have funded 0-100% of the project costs and also provided subject matter expertise. 

Regarding the timeline, once a collaborator is selected and the SOW agreed it can take 1-4 months before the project can begin. During this time period ETC members complete their internal project Charter process and whenever possible work in parallel with the collaborator to finalize the agreements. 

The software environment – for our compact HPLC prototype we have developed a simple GUI in visual basic (Visual Studio 2019 MS) to control the system. However, our firmware has been developed in the Arduino Integrated Development Environment (IDE). Does the consortium consider this an issue – the Arduino IDE open source environment?

Arduino is seen as a development prototype only at this stage and would not be considered robust moving forward. The proposal should discuss how this would be managed.

What is the consortium’s expectation/preference for the system control software executable file – is it to be embedded in the system or loaded on an external IT asset in a windows environment communicating wired or wireless?  Describe your current system. Robustness and simplicity are the key performance attributes.

How are software upgrades expected to be installed – through the internet; flash drive; or other mechanism? 

Web delivery is preferred, thumb drives and other portable media are not consistently supported throughout the vendor companies.

Does the consortium have a cost:performance expectation?

Is there scope for a minimally viable solution to achieve the essential separation requirement? 

Comparable to a 600 bar benchtop system with respect to theoretical plates.

Specifically, do you need a 400 bar pressure minimum or can we meet the separation objective by design?

Describe what your system is capable of, and direct comparison of performance to a benchtop is desirable.

Does the detector need to be 60 Hz or is there a minimum number of data points per peak that is acceptable (for e.g. 20 data points)?

The digitization should be appropriate to the system separation speed (minimum 10 points/peak).

How important is the minimum UV wavelength of 210 nm?  Would 235 nm be acceptable?

Comparison to benchtop units is a significant consideration, so a path to obtain 210 nm should at least be under consideration. This topic can be discussed during the presentation if your proposal is selected for the next step.

In addition, would the consortium value a modular conductivity detector – a capacitively contactless coupled conductivity detector (C4D)? 

Other detectors are fine, with the caveat the comparison to existing benchtop systems is the standard.

Have the consortium considered other external detectors beyond MS? For example RI or ELSD detector which is supported by an external trigger.

See above. Correlation to data from existing systems will be necessary to drive broad deployment of the technology.

Would interfacing with other sample preparation modules be valued by the consortium? 

Yes, some of these features are called out in the existing RFI.

Controlling the workflow through our software which integrates with many 3rd party CDS. 

Describe in your proposal.

Commercial Autosampler integration with tools such as micro-Liquid Liquid Extraction and powder dispensing. 

These attributes are desired by sponsor companies. Please describe in your proposal.

Is there a target timeline for project completion in Section 5.3? 

The detailed timelines will be negotiated in the proposal webex and subsequent communications.

Is there a target cost range for the project in Section 5.4? 

Your proposal should inquire your estimated development costs, which will be divided among the organizations that decide to participate in the development program.

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