depending on the model of jasco HPLC, the parameter separation may vary. you can use the following link for this purpose.

https://jascoinc.com/products/chromatography/hplc/hplc-software/

https://www.brechbuehler.ch/HPLC-SFC-SFE.1796.0.html?&no_cache=1&cid=54920&did=4404&sechash=5b3d8ec4

Also videos will available on you tube. this can be more useful than the pdf version.

Achieving conditions automatically (and preferably w/o work :-) ) is a dream many of us chromatographers have. However, the reality is against it.

There are some tools that might be of help. Software tools like DryLab and Chromsword (and there will be other software tools available doing the same job, I only know those two) help with the design of separation conditions. This - at least the last time I looked into it, and used it in one of my previous laboratories - involves some needed isocratic test runs (I think to recall something like four to six runs) with the analytes of interest, and then perform the calculation.

Realistically speaking: The modulation returns OK approximations for standard solutions, but when matrix kicks in...

In short, such tools are excellent helpers to design promising staring conditions, but the proof lies in the pudding. Run the experiments, and follow the rules of method development. No cutting corners, but sometimes such tools help to save some time.

Does this help?

Cheers,

Detlef.

Hi,

I am using Jasco as well. Which model of Jascoare you using? With what software? In our lab, we have Galaxie as the software. What separation parameters are you interested in? These details will me or others to help more.

You can also contact the product manager of Jasco.

With best regards,

JPM

Nasser wrote: "I am looking for how to automatically obtain the parameters of separation?"

The request is unreasonable as no such automated system exists (However in Television and in the Movies, such science fiction can be seen). Over the past decades, I have personally designed and sold many types of commercial automated HPLC, LC-MS and SFC screening systems which are designed to assist chromatographers in identifying and developing methods for known and unknown samples in the most efficient manner. However, the success of these system relies mostly on the practical experience and skill of the end-user. These sophisticated automated "tools" simply help in speeding-up the entire process. Finding the proper conditions which result in satisfactory LC separation requires many years of hands-on practical knowledge and skill. Instrumentation and software can help in this regard, but at this time, NO SOFTWARE or SYSTEM can determine these parameters without the aid of a skilled human at the helm.

BTW: The HPLC brand is not relevant here as we are discussing a proposed strategy, not a vendor specific aspect.

Of greatest importance in finding the optimum parameters to successfully retain and resolve apart four separate peaks by HPLC requires a solid understanding of the fundamentals of liquid chromatography. I strongly recommend that you begin by reading and studying some of the better books on the topic such as "Introduction to Liquid Chromatography" (Dolan, Synder, Kirkland) as well as working with an experienced liquid chromatographer.

Chromatography simulators (e.g. DryLab) can be useful teaching/learning tools to help you better understand the roles of the key parameters such as flow rate, temperature, % composition, dwell volume, particle size and so on. However, these tool require that YOU first provide the system with actual data obtained by initially running very specific methods to collect data for the system to interpolate or extrapolate from. The data the system provides is only as good as the data you input. The data provided can be useful to speed up the fine - tuning process for novice and intermediate users, but any experienced chromatographer would be able to optimize methods without the need for such simplistic software. That said, theses simulators are excellent teaching aids and I do recommend them for novice users such as yourself.

More advanced simulators designed to predict separation conditions also exist (e.g. ChromSword), but just like the simpler simulators, they rely on first coming up with and running your own trial methods and then inputting the data to the software for use in analyzing the data to make suggestions. Just as before, the data input by you directly effects the quality of suggested methods provided by the system (bad examples = poor results). Because they rely heavily on your input data and methods (which are a result of your experience level in chromatography) it is my opinion that this type of predictive system is not suitable for most users (because they do not have the advanced skills and experience to use the HPLC system).

There are also method prediction software based on structural information. For known pure compounds, these can provide a good starting place to investigate. For mixtures of compounds, their value decreases as the mixture complexity increases.

To initially develop an HPLC method for four known compounds, I would suggest you:

(1) Work with an experienced liquid chromatographer and

(2) review the literature for published examples of separations for each of the compounds to guide you in developing a new method for all four compounds.

If you have unknown compounds then you will need the assistance of an experienced liquid chromatographer to develop a new method. If this is the case, then please refer to your school's analytical chemistry dept for personal assistance in this matter as this is not something that we can assist you via the web.

Your molecular targets determine the amount of baseline separation and the chromatography conditions (column, mobile phases...). Thus, you will have to experiment and perform an online search.