Introduction to HPLC - Lecture 1: HPLC Basics
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- เผยแพร่เมื่อ 22 พ.ค. 2024
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A lecture series on HPLC covering everything from theory and background to practical trouble shooting. Lecture 1 provides an overview of HPLC basics and how HPLC works including mobile phase, stationary phase and columns, as well as retention time and peak area.
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Just started working in a research lab and this is SUCH a helpful refresher. BIG thank you!
Thank You very much for sharing your lecture series! They are so simple and informative, and are fun to watch. Great resource and great reference I often use for work!
I'm technician of analytical instruments and working with dissolution apparatus and other things, my interviewer is not expecting me to have much knowledge of HPLC but I think you have helped a lot 💁💁💁
Have a great day!!
This is a great starter! In 1979 I worked as an intern in an analytical chemistry lab doing HPLC and I loved the technique, never forgot it. A few years ago I built a hobby lab, and bought a lot of Shimadzu LC-10 machines. Quite vintage 😊
Electronics all refurbished and OK, purchased LabSolutions PDA and now in the process of getting the pumps up and running. A bit costly with all the spare parts that are needed, but so nice getting it working!
I will check out all your YT's! Thank you for your dedication and sharing of knowledge, I really appreciate it! 🔆🔆🔆🔆🔆🤗👌🏻🙏🏻
Big thanks. I fully understood the basic concepts. The way of delivering the content is crystal clear and understandable.
Amazingly helpful! Especially for an RA working with HPLC who never took any HPLC classes.
Same! I majored in Biochemistry but only learned it briefly from the textbook.
I've been working only with a medium pressure chromatography system, because it is the one which is available in my lab. Despite this fact the exam questions require knowledge of running procedure for HPLC😑
YES! Thank you! You explain things so well. I'm going to watch this entire lecture series and share it with my supervisor and professor. This was so necessary and so easy to understand. Love it.
Fantastic! Your efforts are much appreciated.
Please keep creating such content. Bottom of my heart.. Thank you
This is great. Got a job doing downstream work and this helps clarify a lot of things.
This lecture was so enriching. Thank you so much.
Thank you for these lectures! Very helpful and thorough 🙂
Thank you for the lecture!! Very clear explanation!!
This is so easy to understand and at the same time detailed explanation for a complex topic of HPLC. Highly recommend it for anyone looking for an introduction to HPLC. Thank you so much for putting it out❤
Thank you! Your lectures are helping me study for my exams
This was extremely helpful prepping/reminding me how the instrument fully works for my interview. thank you very much
Best of luck!
Same reason that brought me here! I need that in-depth info because at my university I never got to use the hplc myself...
This is so helpful!! Super clear and informative, thank you
Subscribed to your channel to not miss out on any awesome videos! Keep up the good work :)
Very good introduction to HPLC!
Fantastic. Didn’t quite understand it well until now.
Amazing and a simplified informative lecture...Thanks
Amazing Lecture! Thank you
This is so helpful. Much appreciated.
Great video!
Using it to prepare for a new job. Thanks alot.
Great learning through this lecture
easy to understand, nice explanation!
I used HPLC in a pharma lab daily and I was curious to see based on my knowledge actually using it how it compares to your lecture, and I’ll say I learned a few extra things!
very good lecture!
Great lecture
Thank you. It really help me to understand.
This was very helpful, thank you!
Awesome video
Informative lecture. Thank you.
Explanation made hplc simple
very clear explanation, thank you very much
Amazing...just amazing
thank you for all of this!
subscribed! very informative video!
Great course. Thank you so much ❤
Great video!! Thank you!!
Great explanation!
Hmmo
Thanks so much. Very Educative
You are a great teacher, really enjoyed the class
Glad you enjoyed it!
Thanks for your time is very helpful
Top Notch!!!
thank you so much for the lectures great job it assist a lot 🥰
thank you for the lecture ^^
Good, it is really interesting...
Thank you for your video!!!!!
Thank you 😌🙏
Thank you, thank you, thank you🙏🏼
You are so welcome!
excellent video, VERY WELL EXPLAINED. Thank you so much
Glad it was helpful! Thanks for the feedback.
great vid
Clear explanation. Thanks
Glad you liked it
thank you it was useful
Great content!
Thank you! These take a while to put together so I am happy to hear they are appreciated!
superb
great..thank you
Thanks
Hvala.
thanks man!
thank you 😍
Was good❤
thanks!!
This video was great! The more detailed picture of the HPLC system was great and clears things up a lot. I have a question about the chromatogram: Does one substance, I mean like one chemical compound, always only have one single peak in the chromatogram, and the other peaks must be something else?
In general, yes each unique compound will show up as a single peak on a chromatogram. That being said many times suboptimal conditions can lead to "peak tailing" which spreads out or gives the illusions of multiple peaks. The only time a compound might give 2 signals is if somehow it was breaking down into two separate sub-compounds when exposed to the HPLC conditions.
@@ChemComplete Thanks so much for the reply! :)
Great Class, please what software did you use to draw during the lecture ?
thank you very much . you are amazing
Happy to help
I really like your accent. Thanks for this explication!
Great
So cool
thanks duuude
You need a high temprature to push the compounds in. So does that mean the GC works with (air) pressure too? 2:34
I would like to know why and how we use standarts of these samples?
May God bless u
Do i need to have certification to operat this machine
Thank you
Keep this up.
Gostei!
Thanks a lot for the explanation. How many types of calibration curves are there that can be used to calculate the concentration of a certain constituent in a sample ?
There is technically an infinite number because you can use any variety of known concentrations. There are two major points to keep in mind when designing an unknown calibration curve:
1. You want to use multiple known concentrations with a good spread of values. I always teach a minimum of 5 known concentration values for a solid correlation.
2. The range of known concentrations must capture the unknown value. This is to say that the unknown concentration must fall between your lowest and highest known concentration.
This is a great video will be waiting for more. I am using a reagent to convert dimethyl sulfide to dimethylsulfoxide, can use HPLC (as you have mentioned using calibration curve) to get catalytic turnover numbers?
you would probably be better served trying a GC method for this.
Extremely helpful, thank you!
I have a question: is retention time calculated at the moment of the peak or where the curve of that substance begins (and therefore where it starts to come out of the column)?
Hiiii where are you from
@@AKMirzaganj_2023 cringe
Peak
Im currently doing an Internship at a company and my boss suggested to me I should write my bachelor thesis about hlpc analysis for a special kind of oligosaccharifes. Im a bit scared about the fact that nobody at the company including me worked with hlpc before. Can anyone give me advice If it is a good Idea??
Sir i have a question.. Does a compound that is to detect have same retention time while using different solvent system?
Great lecture! Thank you. One think that is unclear to me is how the HPLC system collects the different fractions after separating a mixture into pure compounds. Does it do this automatically?
A standard HPLC System will often not collect the compounds after they pass the detector, only analyze them. With some advanced detectors such as MS it is not even possible, because the analyte is destroyed in the detector.
However, with many detectors (such as standard UV/VIS-detectors for example) you can isolate a compund or fraction by simply collecting the eluent (mobile phase) after it passes the detector. For that you just take the capillary exit and collect everything that comes out during a certain time frame (normally, all eluent exiting the detector will just go into your solvent waste container).
You will see the peak of the compund(s) that interest you on your software screen (so you know when it is in the detector) and then immediately collect the eluent in a vial or similar container. When exactly your compound gets from the detector into the vial depends on the flow rate and length of the capillary after the detector.
Separating two compounds with very similar retention times this way is pretty difficult. Also, whatever fractions you collect this way will usually go through some more steps, such as evaporation of the solvents and further purification steps.
So called preparative HPLC setups are you used to collect bigger amounts of a chemical (or a fraction) separated by chromatography. They use higher injection volumes/amounts, bigger columns and higher flow rates (as well as pressures) than standard setups, so that the amount collected at the end is worth the effort.
@@CompletedChaos Thank you for the detailed explanation!
@@CompletedChaos v well explained..I hv done this in impurity profile lab for some time 👍
❤️
👏👏👏
Im an Engineer who is working as a service engineer in hplc agilent modules. Being from the mechanical background I am weak in chemistry like what is polar, non polar, ionic bonds, chains, silica, different phase, etc so can amyone fuide me and explain it in easy words
sir i need the software of hplc.. of ( Hitachi ) company
khan academy who? tysm!!!
What a compliment, thank you!
В институт привезли и установили этот аппарат, очень дорогой и грузом лежит безхозный. Дело в том что нету специалистов умеющих работать на этом приборе, с английским плоховатои нет руководства по эксплуотации. Если есть что то то на английском языке. Можно русифицировать прогграммное обеспечение этого прибора??????????
Great presentation but pleeease stop making that lip slapping sound
Great presentation and thank you for taking the time to share your knowledge and make this video but the noise of your lips or tongue or mouth is distracting and annoying.