WebIf the buffer pH is raised above a protein’s pI, it will carry a net negative charge. Because a protein’s pI is determined by its primary amino acid sequence and can thus be calculated, a buffer can then be chosen that … http://www.hplc.eu/Downloads/ACE_Guide_BufferSelection.pdf
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WebConsider an example of 90 % buffer solution and 10 % acetonitrile. (The buffer solution is 20 mM sodium dihydrogen phosphate adjusted to pH 2.5 by the addition of phosphoric acid.) A mixing ratio of 9:1 can be interpreted as a 9:1 volume ratio of buffer solution and acetonitrile. That is, the equivalent volumes are measured and mixed together. WebNov 20, 2024 · Since most compounds analysed by RP-HPLC contain one or more acidic or basic functional groups, most mobile phases require pH control. For this reason, buffers are widely used. Why Control pH? When an acid is more than 2 pH units above or below its pKa, it will be >99% ionised or non ionised, respectively. Bases are ionised below their pKa and ... team smandych
HPLC Tips & Tricks: Mobile Phase Preparation Part 2 - Buffers
WebJan 11, 2013 · Studies were performed to optimize the buffer and salt concentrations simultaneously to improve the retention of low pI mAbs and the resolution of high pI mAbs. The optimized salt-mediated pH-IEC method was not only applicable to mAbs over a broader pI range from 6.2 to 9.4, but also offered better resolution for mAbs with pI values … WebGenerally, organic solvents for HPLC, such as acetonitrile and methanol, are available in three qualities: Isocratic grade, gradient grade and hypergrade for LC-MS LiChrosolv ®. For MS analysis, hypergrade quality solvents should … WebHPLC teams manifest editor