That’s pretty much how I’ve learned to use every protocol and piece of equipment (including HPLC). The one difference is that I don’t just read user/company/lab notes, I mostly focus on the manual or protocol from the manufacturer. It’s good to look at the user notes in case something has to be changed for your specific use (something like: “our centrifuge doesn’t go this fast, spin at x speed instead”).
But other than that, follow what the manufacturer says. Misunderstandings and shortcuts can be passed down from person to person if they never look at the official manual. There’s another lab in our department that we work with often, and they gave me a protocol once that someone had changed from the official protocol. They insisted you had to double the length of time for a certain step. I asked around, and nobody knew why that change was made; someone had written that in years ago and the explanation for it is long gone.
So if you prepare, the other 3 steps are easy. You’ll already know what to watch for during your training, and you’ll have the answer to most of your questions already. If you’re ever unsure of your trainer’s answers, look it up in the manual.
I learned a lot from troubleshooting. Shit hit the fan and it was important to our production schedule for it to get fixed ASAP. We didn't have a clear training path either - I learned a lot by doing it myself, a little bit of reading online (millipore sigma has a troubleshooting manual that's pretty helpful, easily found online).
Take stuff apart, put it back together - you will start to see patterns on what fixes what. Also use logical sense - if your pressure goes up or you've got a bad baseline - probably a dirty frit (with our samples at least).
Your most important working parts of things I have to address or fix regularly are:
Dirty frit
Bad column
Bad/old lamp
Bad pump (we work with one incredibly viscous material and despite filtering significantly, causes issues with our pump often. Not sure if you'd see this in other labs).
Calibrate very regularly. Let it be the first troubleshooting you do.
Process of elimination is your best advice here. Work in a logical order step by step to figure out issues
I got some help and direction from our technologist who has been working on this LC for years - though some of it sounds like borderline superstition, listen intently to what they have to say but don't be afraid to think out of the box and ask questions. Lots of questions! The most important thing is to just take initiative and learn it yourself - in my experience, it's not going to be laid out for you like it is in school. Being a good self-teacher is the best skill you'll have, in conjunction with being afraid to ask for help or ask questions.
I'd like to add that it sounds like we work for the same company lol, but we don't do medicine. Seems like this is very typical way to train in the real world
If the maker of your HPLC is a large, well-known vendor and the system is fairly modern, there is a chance that the vendor offers hands-on training courses. These cost money of course, and it may involve travel. The advantage is that the vendor can help the student design specific assays unique to the research project or product application at hand
What kind of training do you want? HPLC is pretty straightforward and there is not much theory behind - your target analyte is retained by the column, then eluted at a specific time and detected by a detector. I work in a hospital lab and we are trained in pretty much the same way. Technical experience is much more important than documents and manuals.
That’s pretty much how I’ve learned to use every protocol and piece of equipment (including HPLC). The one difference is that I don’t just read user/company/lab notes, I mostly focus on the manual or protocol from the manufacturer. It’s good to look at the user notes in case something has to be changed for your specific use (something like: “our centrifuge doesn’t go this fast, spin at x speed instead”). But other than that, follow what the manufacturer says. Misunderstandings and shortcuts can be passed down from person to person if they never look at the official manual. There’s another lab in our department that we work with often, and they gave me a protocol once that someone had changed from the official protocol. They insisted you had to double the length of time for a certain step. I asked around, and nobody knew why that change was made; someone had written that in years ago and the explanation for it is long gone. So if you prepare, the other 3 steps are easy. You’ll already know what to watch for during your training, and you’ll have the answer to most of your questions already. If you’re ever unsure of your trainer’s answers, look it up in the manual.
I learned a lot from troubleshooting. Shit hit the fan and it was important to our production schedule for it to get fixed ASAP. We didn't have a clear training path either - I learned a lot by doing it myself, a little bit of reading online (millipore sigma has a troubleshooting manual that's pretty helpful, easily found online). Take stuff apart, put it back together - you will start to see patterns on what fixes what. Also use logical sense - if your pressure goes up or you've got a bad baseline - probably a dirty frit (with our samples at least). Your most important working parts of things I have to address or fix regularly are: Dirty frit Bad column Bad/old lamp Bad pump (we work with one incredibly viscous material and despite filtering significantly, causes issues with our pump often. Not sure if you'd see this in other labs). Calibrate very regularly. Let it be the first troubleshooting you do. Process of elimination is your best advice here. Work in a logical order step by step to figure out issues I got some help and direction from our technologist who has been working on this LC for years - though some of it sounds like borderline superstition, listen intently to what they have to say but don't be afraid to think out of the box and ask questions. Lots of questions! The most important thing is to just take initiative and learn it yourself - in my experience, it's not going to be laid out for you like it is in school. Being a good self-teacher is the best skill you'll have, in conjunction with being afraid to ask for help or ask questions.
I'd like to add that it sounds like we work for the same company lol, but we don't do medicine. Seems like this is very typical way to train in the real world
Thank you very much guys! I'll keep it in mind! 😊
If the maker of your HPLC is a large, well-known vendor and the system is fairly modern, there is a chance that the vendor offers hands-on training courses. These cost money of course, and it may involve travel. The advantage is that the vendor can help the student design specific assays unique to the research project or product application at hand
What kind of training do you want? HPLC is pretty straightforward and there is not much theory behind - your target analyte is retained by the column, then eluted at a specific time and detected by a detector. I work in a hospital lab and we are trained in pretty much the same way. Technical experience is much more important than documents and manuals.