Re: HPLC Ghost Peaks

James Farmar (jfarmar@nybc.org)
Mon, 28 Sep 1998 15:46:09 -0400

At 9/28/98, you wrote:
>
>We are having problems with 'ghost' -peaks that are appearing in are RPC
>runs. We are using a spheri-5 rp-18 column with a precolumn ( rp-8)
>The buffers are:
>A: 0.1M Triethylamine (TEA, >99%) in HPLC-grade water brought to pH7 with
>acetic acid (>99.8%)
>B: 80% acetonitrile in water

Dear Ben,
You don't say in your note, but are you filtering your buffers (with
dedicated glassware)?
I encountered a triplet or quartet of peaks when I used 0.1% TFA buffers as
well as the TEAc buffers. I spent almost a month eliminating sources.
I tracked the ghost source to the disposable filter I was using for the
buffers. When I switched from nylon to polypropylene, I lost the peaks.
(One test is to send Buffer A through the same filter several times to see
whether an increase in the size of the ghost peaks results.)
You can also try filtering the ACN/H2O buffer first which might reduce the
amount of material extracted by the subsequent filtration of the no/low
organic buffer.
Good hunting.

Jim Farmar
MicroChemistry/NYBC