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HPLC for GPC testing

Posted: Fri Sep 12, 2014 8:30 am
by cowdrey
Hi all

I have a low molecular heparin (nadroparin calcium) that I need to do molecular distribution testing on as per Ph. Eur monograph. I have no experience with this separation and I was hoping someone in the forum could assisst me. Based on the Ph Eur method it appears that the HPLC system is fairly standard with UV and RI detectors. I currently have a Waters Alliance 2695 with a 2487 UV detector. I was thinking of purchasing a 2414 RI detector and connecting it to my existing system. Would this be possible or are there any other modifications to the hardware that I need to do? We will need to upgrade our software (Empower) to include the GPC software.

Thanks in advance

Cowdrey

Re: HPLC for GPC testing

Posted: Fri Sep 12, 2014 9:36 am
by danko
You won’t be needing the UV detector in this context but you know that already – I presume.
To your question: The RI detector is what you need. The GPC option/add-on is nice to have in your case, but not absolutely “must-have” – all depending on how you’re going to handle your data.

Best regards

Re: HPLC for GPC testing

Posted: Fri Sep 12, 2014 9:40 am
by Gerhard Kratz
To connect an RI detector to your system should be not a problem and with GPC software on board you are well set up. Most important will be the right column. What column do you want to use for this GPC separation?
Good luck

Re: HPLC for GPC testing

Posted: Fri Sep 12, 2014 10:51 am
by kubowicz.tomasz
Hello

According to EP method you will need UV (see link below) - p.1717

http://lib.njutcm.edu.cn/yaodian/ep/EP5 ... /0828e.pdf

Regards

Tomasz Kubowicz

Re: HPLC for GPC testing

Posted: Fri Sep 12, 2014 12:46 pm
by cowdrey
Hi All

Thanks for the comments thusfar. I'll be using the following column:
TSK G2000 SW XL, 300mm x 7.8mm, 5um Silica, Porosity 125A, Proteine separation range 15000 to 100000 Da.
The data will be handled through Empower.

Thanks