<!--
/* Font Definitions */
@font-face
{font-family:"Cambria Math";
panose-1:2 4 5 3 5 4 6 3 2 4;}
@font-face
{font-family:Calibri;
panose-1:2 15 5 2 2 2 4 3 2 4;}
@font-face
{font-family:Verdana;
panose-1:2 11 6 4 3 5 4 4 2 4;}
/* Style Definitions */
p.MsoNormal, li.MsoNormal, div.MsoNormal
{margin:0in;
font-size:12.0pt;
font-family:"Calibri",sans-serif;}
span.EmailStyle17
{mso-style-type:personal-compose;
font-family:"Calibri",sans-serif;
color:windowtext;}
.MsoChpDefault
{mso-style-type:export-only;
font-size:12.0pt;
font-family:"Calibri",sans-serif;}
@page WordSection1
{size:8.5in 11.0in;
margin:1.0in 1.0in 1.0in 1.0in;}
div.WordSection1
Rest of post
{page:WordSection1;}
-->
HI
I am considering running basic reverse phase C18 separation for peptides.
We are using an Easy Nano 1200 and a Orbitrap Fusion Lumos.
I am looking for suggestions on the basic reverse phase buffer system. I
most likely do not want to go above pH 10- and maybe pH 8-9 would even be
fine.
I also would be open to column suggestion- I am currently using Thermo’s
easy spray C18 Column. (EASY-SPRAY PEPMAP RSLC C18 2UM, 25CMX )
Any suggestions are appreciated.
Thanks
Robert R. Salzler Regeneron Pharmaceuticals, Inc. Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162 777 Old Saw Mill River Road Tarrytown, NY
10591-06707
Office: 914-847-3075
Cell: 716-864-6787
I do concat offline but check out this paper
https://pubmed.ncbi.nlm.nih.gov/28126900/.
It isn't clear what the buffer is (they give a product number) but other papers
that used the method have said it was pH 10 - sounds like ammonia. I don't
think you have to go to such a high pH - I think there was an RD Smith paper
that looked at lower pH's than 10 (better for media). Remember the main thing
you need to do is ionize the carboxyl groups which is pretty much done by pH 7.
Going up to 10 takes you into the mid point of the lysine pKa and that could
lead to greater variation from subtle effects.
D. Eric Anderson
Mass Spectrometry Facility, NIDDK
NIH
Building 8 Room B2A19, 9000 Rockville Pike, Bethesda MD, 20892 USA
(301)-496-7546
Rest of post
________________________________________
From: Robert.Salzler <email obscured>>
Sent: 07 October 2020 08:35
To: <email obscured>
Subject: [ABRF Discussion Forum] Basic Reverse Phase Separation of Peptides
― 1 file link ―
<!--
/* Font Definitions */
@font-face
{font-family:"Cambria Math";
panose-1:2 4 5 3 5 4 6 3 2 4;}
@font-face
{font-family:Calibri;
panose-1:2 15 5 2 2 2 4 3 2 4;}
@font-face
{font-family:Verdana;
panose-1:2 11 6 4 3 5 4 4 2 4;}
/* Style Definitions */
p.MsoNormal, li.MsoNormal, div.MsoNormal
{margin:0in;
font-size:12.0pt;
font-family:"Calibri",sans-serif;}
span.EmailStyle17
{mso-style-type:personal-compose;
font-family:"Calibri",sans-serif;
color:windowtext;}
.MsoChpDefault
{mso-style-type:export-only;
font-size:12.0pt;
font-family:"Calibri",sans-serif;}
@page WordSection1
{size:8.5in 11.0in;
margin:1.0in 1.0in 1.0in 1.0in;}
div.WordSection1
{page:WordSection1;}
-->
HI
I am considering running basic reverse phase C18 separation for peptides.
We are using an Easy Nano 1200 and a Orbitrap Fusion Lumos.
I am looking for suggestions on the basic reverse phase buffer system. I
most likely do not want to go above pH 10- and maybe pH 8-9 would even be
fine.
I also would be open to column suggestion- I am currently using Thermo’s
easy spray C18 Column. (EASY-SPRAY PEPMAP RSLC C18 2UM, 25CMX )
Any suggestions are appreciated.
Thanks
Robert R. Salzler Regeneron Pharmaceuticals, Inc. Therapeutic Proteins
Bioanalytical & Biomarker Technologies
Building 10, Room 162 777 Old Saw Mill River Road Tarrytown, NY
10591-06707
Office: 914-847-3075
Cell: 716-864-6787
― 1 file ―
□ image001.jpg (1.8kb)
http://list.abrf.org/r/file/nXcRkdb5l6zOyrvhS62HZcDJyur-ty-2OS9Zml/
――
View topic http://list.abrf.org/r/topic/4nXsou1La9R1NBdOqnoCC6
Leave group <email obscured>?Subject=Unsubscribe