does method development actually matter or is it forum lore at this point
Asking properly rather than in a comment on somebody else’s thread: does method development actually matter or is it forum lore at this point. Retention time shifted 0.4 minutes and I assumed the worst. It was the column temperature. Ran a blank after a high-concentration injection and found the carryover I had been…
Mass spectrometry answers identity. UV purity answers relative quantity under the run conditions. A document with one and not the other is answering half the question, and the half it answers should be stated.
a blank injection between samples costs four minutes and settles most arguments
a blank injection between samples costs four minutes and settles most arguments
Adding one practical thing — run the blank. It answers this before anyone has to argue about it.
Small fix — 214nm, not 210. It matters for the comparison you are making with the other run.
UV response is not uniform across species. At 214nm you are looking at the peptide bond, which is why it is the working wavelength for this class; at 280nm you only see tryptophan, tyrosine and phenylalanine.
reproducibility beats resolution if you only get one of them
What did system suitability look like on that sequence?
baseline choice is a decision, not a measurement