I need to work at pH 3.70 with high regulatory capacity and low Abs. at 240 nm. In this time I use buffer acetate 500 mM but I have problems with it. Could anybodie help me?
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By H W Mueller on Wednesday, November 22, 2000 - 06:23 am:
What problem do you have? Is the absorbance too high? How about a phosphate-acetate buffer, you can reduce the acetate to about half your present concentration. Another possibility: Formate buffer. If MP absobance is a problem you should forget citrate.
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By Uwe Neue on Wednesday, November 22, 2000 - 03:00 pm:
500 mM is fairly high. Can you reduce the concentration? Alternatively, you may still get away with a phosphate buffer. While this is far away from the pKa, you are working at a concentration that is 10 to 50 times higher than common RPLC buffer concentrations. This may still give you a reasonable buffer capacity. Check how reproducibly the pH can be adjusted.
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By John Tarr on Wednesday, December 6, 2000 - 04:19 pm:
Acetate pKa = 4.76 and formate pKa = 3.75. Therefore, at pH 3.7 a 50 mM formate buffer would have about the same buffering capacity as 500 mM acetate.
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