HPLC Data Analysis

[IshaanG29]

Hello,

I am a student researcher working on the simultaneous estimation of two drugs via the HPLC. I was able to run the samples in the HPLC and downloaded the raw data files. This is the first time I have been introduced to this machine so I don't exactly know how to get the chromatograms from the raw files. Also, I am not sure which software to use that will help me with my data.

I would really appreciate some help regarding this.

Thank you!

[Consumer Products Guy]

Get some hands-on instruction.
Get lots of hands-on instruction.

[Andy Alpert]

I agree with Consumer Products Guy. Having said that, here's what you can do with the raw data you describe. I presume that it's something like an ASCII file: A series of (X,Y) data points. The X numbers would be the time while the Y numbers would be the response of the detector in whatever units the detector uses (absorbance units, etc.). You need a program that will make a graph out of such data. One fairly cheap program that does this handily is Psi-Plot ($99 educators' price). Opening the program, you would copy and paste the values for X into one column of a spreadsheet. Paste the values for Y in another column. Then, again within the program, select an (X,Y) data graph, specify the values in the X column as the data points for the X-axis and the values in the Y column as the Y-axis, and click on the button to graph these. You will then get a graph consisting of the chromatogram. You can export this graph in Windows Metafile (WMF) format. Insert that into a Powerpoint slide and you will get a slide in which the chromatogram is fully editable: You can change the color, the line thickness, even edit individual data points (although of course we scientists never edit the data). Other scientific graphics programs will allow you to do the same thing. Having said this: I'm surprised that your HPLC control program doesn't have a feature permitting the export of a chromatogram in which this hasn't been done already. Start reading about the features of the program that controls your machine.

[lmh]

If you can find anyone who knows anything about the system, get them to do a training session with you. If you are stuck in a remote place with no hope of any proper training, then do a lot of google-searches. You will find general information on how to do chromatography - for example if you have an academic e-mail account you should be able to get access to ChromAcademy which does general training. Then you should also be able to google for the name of the software that controls your hplc. The software will probably contain all the tools you need for examining chromatograms, measuring the areas of peaks, creating calibration curves, and reading off concentrations of analytes. Most manufacturers have good documentation and even tutorials on how to do it (though admittedly a few will try very hard to get you to go on their training courses instead of reading! Training courses are usually very good, but often very expensive).
Good luck!

[James_Ball]

Hello,

I am a student researcher working on the simultaneous estimation of two drugs via the HPLC. I was able to run the samples in the HPLC and downloaded the raw data files. This is the first time I have been introduced to this machine so I don't exactly know how to get the chromatograms from the raw files. Also, I am not sure which software to use that will help me with my data.

I would really appreciate some help regarding this.

Thank you!

The type or raw data file will depend on what software acquired the data. Some systems will save data into a proprietary format which may require the original system to be able to display the information.

Can you tell us what the manufacturer of the instrument and software is?