Problem with rpc
Posted: Sat Apr 25, 2015 5:10 pm
Hello everyone,
I am using a 3ml resource RPC column from GE to fractionate resveratrol and any other impurities present. Since the powder is insoluble in water I have dissolved it in 30% ethanol. The concentration is 2 mg/ml and 1ml injected into the column. The buffers I used are 30% ethanol (buffer A) and 80% ethanol (buffer B).
So far I have completed three trials, but the peak heights and retention times keep changing, My sample volume and concentration is the same, so what could the cause be?
Also, RPC manuals say to use an a highly aqueous buffer with a small amount of organic solvent (~2%) for binding of the sample to the column matrix, how is that possible if the sample is insoluble in the said buffer?
Look forward to you responses
Regards,
Cedric
I am using a 3ml resource RPC column from GE to fractionate resveratrol and any other impurities present. Since the powder is insoluble in water I have dissolved it in 30% ethanol. The concentration is 2 mg/ml and 1ml injected into the column. The buffers I used are 30% ethanol (buffer A) and 80% ethanol (buffer B).
So far I have completed three trials, but the peak heights and retention times keep changing, My sample volume and concentration is the same, so what could the cause be?
Also, RPC manuals say to use an a highly aqueous buffer with a small amount of organic solvent (~2%) for binding of the sample to the column matrix, how is that possible if the sample is insoluble in the said buffer?
Look forward to you responses
Regards,
Cedric