Introduction to HPLC - Lecture 3: Reverse Phase HPLC

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A lecture series on HPLC covering everything from theory and background to practical trouble shooting. Lecture 3 provides an overview of reverse phase HPLC in which hydrophobic columns and polar mobile phases are used for separation and retention time analysis.
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[@shauryatiwari9400]

This is the most thorough HPLC series I found. Brilliant work. Thank you good person.

[@ChemComplete]

You're very welcome!

[@derekedington777]

thanks for the hplc vids. in biotech program in dayton and you do a great job explaining everything.

[@ChoocoCaramell]

Thanks for the effort in putting this material on youtube, I've been searching more of all of this topic and I don't find more clear material than this!! thanks a lot :-)

[@ChemComplete]

Glad it was helpful!

[@helenzhang173]

I love your video. You make things so clear. Thank you!

[@azarkhany7314]

thanks and can't wait for the next one!

[@thanukaratnayake2310]

Keep it coming , great stuff

[@akbarbahrami985]

Very impressive way of explaining materials. Loved that!

[@ChemComplete]

I am glad it was broken down in a way that was easy to understand. Reverse phase knowledge is very important for industry and academia.

[@EIsuruPriyaranga]

Thank you so much! You saved me and a great starter for my new PhD project!

[@lydiaamelia199]

I really love how you explain all of this, its so clear and i could understand it easily, thank you so much for making this lecture 🙏☺️

[@alexandermelchor9362]

I’ve been waiting for this haha

[@kuntarinirahsilawati920]

Thank you very much. Very helping to understand.

[@soheylasiavoshi7759]

Thank you Sir. It was so practical for me🌸

[@marcellagiovani3994]

Thank you so much for all these videos! it really helped me a lot in understanding and perhaps some troubleshooting on HPLC :)) maybe the next HPLC video could be somewhere around trouble shooting the result of the peaks? maybe on the shape of the peak, what is a good shape, what is a bad shape, what does it mean and what can we do to it. and maybe by then we can do some data analysis? :D thank you!!

[@mweilahmed9204]

😭❤️ thank you for the great job 💜💙💕

[@ChemComplete]

No problem 😊

[@RazaKhan-ds8by]

Simply excellent way too much clarity in his lecture. You are Star.

[@The_Amay1986]

Thank you so much it is very helpful

[@Swashbucklebuckle]

Up to this episode, this series has been extremely insightful and helpful! I think this knowledge will help me very much in my upcoming job interview(s).

[@ChemComplete]

Thank you so much! I hope the interviews go well.

[@Swashbucklebuckle]

@@ChemComplete Thank you! At least in my interview yesterday I nailed all the HPLC questions x) I will see...

[@pharmabot]

I love your lectures! - A small tip: PLEASE start using a wacom tablet instead of a mouse for writing on the screen

[@mhamado.abdullah6925]

Wow , thank you

[@noureddineraghdi4340]

thank you so much

[@winsontam3860]

Thank you

[@stijnstrotmann]

These lectures help me out so much!! Thanks a lot!

[@stijnstrotmann]

Heard the intro of lecture nr 4. The e-books go into far more details. Got it. ;)

[@MahmoodHashem]

thanks a lot i am waiting the coming videos.

[@ChemComplete]

Will upload soon!

[@azizabed7692]

Hi,
Do you have any lectures about different types of columns?
Analytical and preparative columns?
Thanks

[@yamawwad]

Kindly, where is the link to the video that talks about HPLC columns ? thank you.

[@godfredbiney9858]

Great

[@seyedmohammadseyedkhademi2454]

Thanks for this video, but I have one question at the time 43:13 of this video. you told that H2O/MeOH (in comparison to CH3CN/THF) has a higher Rt or longer spread. While in Revers Phase, the Stationary phase is Hydrophobic, and the more polar mobile phase should elute faster. Or I,m wrong? please explain a little more about this. Thanks a lot

[@AmineSamus]

Hi this was a great lecture. Im curious though, if the peaks give the same retention time in the chromatogram for multiple analytes, how do you separate them and determine which analyte is which?
Also does elution just mean how fast it will be detected by the detector? So a polar analyte will have a lower elution time?

[@ChemComplete]

If the peaks (two analytes) have the same retention time then you will need to do some experimentation to determine different solvent systems, flow rates, column typings etc. Confirmation of their identity can be accomplished through other spectroscopy such as MS and/or NMR which give more structural information.

[@roirat04]

And I paid over $1200 US dollars for a course over 27 years ago which didn't even touch HPLC , it was in the syllabus,, because the only Professor canceled the lab coursework..terrible to do that to eager mind at the time. I was sure to get my money back for the class.

[@pingu7075]

Okay so first, you'd play with the mobile phase. If that doesn't work change the colomn because of expenses?

[@pingu7075]

And stay strong with the shifting things in life

[@devanshshukla8857]

Dear Sir, this is a help request, I have a question :
is it true that more the elution power, less the retention time irrespective of the analyte?
for instance I have a sample of Lactic Acid.
I run HPLC using water as mobile phase and get retention time t1,
I do the same but with acetonitrile as the mobile phase and get retention time t2,
will t2 be < t1 ?
Will t2< t1 be true irrespective of the sample ( say i use bezoic acid instead of lactic acid)

[@ggstevo]

That should be true since you are now running a more nonpolar solvent than your starting solvent (water), your samples should elute quicker off the column.

[@BEH477]

hi

[@ChemComplete]

Hello my student!

[@q8051]

Hplc not easier before this.

[@KianaVision]

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