Dear scientific community. In order to analyze the amino acid profil of proteic matrices we need a purification after the hydrolysis of our sample. We intend to use the Dowex 50WX8 hydrogen form as the solid phase for column chromatography. We need a protocol for column conditioning, and buffer solution in order to have the total amino acid profil as a result. Many thanks in advance.
Strong Cation Exchange SPE cartridge must be sufficient and easy to do. Check Phenomenex Strata SCX standard protocol.
Why do you think that you need a clean-up step? We analyzed quite complex samples without problems. A centrifugation and/or filtration step should suffice in many cases.
Cation exchange resins are very convenient for purification. Matrix components may interfere with HPLC analysis and clean-up may be beneficial at this point. In order to decrease the amount of work at sample preparation and for more accurate results you may select GC-MS or LC-MS technique after derivatization. In this case, you can exclude the SPE process. If you have to use HPLC any format (cartridge SPE, in tip SPE, or column chromatography) of cation exchange resin will do the job.
Thank you all of you for your precious responses.indeed, we intend to quantify the amino acids by HPLC DAD and we don't have a protocole for the cean up as there will be many impurities like sugars lipids polyphenols...we work on pulses and sure they are proteic rich matrices this fact didn't exclude impurities. We want to use column chromatography with the dowex 50wx8 as a solid phase, but we need a tested protocole if you have it it will be very useful for us.many thanks in advanve
Use the link below for BioRad resin which has similar properties to Dowex cation exchanger. You may find protocols at the end of the document related to amino acid concentrations relatively old but they could be useful for giving a try or to compose your own experimental design...
https://www.bio-rad.com/webroot/web/pdf/lsr/literature/LIT203.pdf
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