I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
Hello Adam
Do you see any ion spray current difference when spluttering versus spraying?
What ion spray voltage are you using? Are you using an EasynLC or something
else?
Cheers
Duncan
Sent from my iPhone
> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>
> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
Rest of post
>
> Best,
>
> Adam
> ――
> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
> Leave group <email obscured>?Subject=Unsubscribe
>
Hi Adam,
We usually have had a really good spray stability with EASY-Spray, however last
week we encountered the same issues as you described with our EASY-Spray .
After changing the column, we don't have sputtering anymore. We believe this is
related to the column/emitter production.
Hossein
To expand a bit, we are observing the same sort of spitting that you describe.
One column was within one day, another within a week. Normally they are much
more stable than this so that is why we are thinking a batch/production
problem.
Rest of post
Best,
Lynn
-----Original Message-----
From: ABRF Discussion Forum <email obscured>] On Behalf Of fazeliniah
Sent: Wednesday, September 12, 2018 1:59 PM
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] EASY-Spray C18 PepMap RSLC Emitter Spray
Sputter
Hi Adam,
We usually have had a really good spray stability with EASY-Spray, however last
week we encountered the same issues as you described with our EASY-Spray .
After changing the column, we don't have sputtering anymore. We believe this is
related to the column/emitter production.
Hossein
――
View topic http://list.abrf.org/r/topic/6TMquIdIG3uHAAZdTn8t6l
Leave group <email obscured>?Subject=Unsubscribe
Hi,
that was the reason we packed our columns ourselves, especially PepMap (many
years back) 7 - 8 out of 10 had been bad and we returned them. Column packing
is easy with some tricks, we used Reprosil and our longest living column was
retired, still ok, after nearly 10,000 injections - very short gradient. The
column got framed!
Manfred
Am 12.09.18, 20:03 schrieb "ABRF Discussion Forum im Auftrag von Spruce, Lynn"
<email obscured> im Auftrag von <email obscured>>:
To expand a bit, we are observing the same sort of spitting that you
describe. One column was within one day, another within a week. Normally they
are much more stable than this so that is why we are thinking a
batch/production problem.
Best,
Lynn
-----Original Message-----
From: ABRF Discussion Forum <email obscured>] On Behalf Of
fazeliniah
Sent: Wednesday, September 12, 2018 1:59 PM
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] EASY-Spray C18 PepMap RSLC Emitter
Spray Sputter
Hi Adam,
We usually have had a really good spray stability with EASY-Spray, however
last week we encountered the same issues as you described with our EASY-Spray .
After changing the column, we don't have sputtering anymore. We believe this is
related to the column/emitter production.
Hossein
――
View topic http://list.abrf.org/r/topic/6TMquIdIG3uHAAZdTn8t6l
Leave group <email obscured>?Subject=Unsubscribe
――
View topic http://list.abrf.org/r/topic/7HriWuVbyBWyPci21S46RU
Leave group <email obscured>?Subject=Unsubscribe
Hi Duncan,
I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA). I’ll
have to check what signal I have on a non-sputtering column. I’m running at 2.2
kV spray voltage. It’s a Waters NanoAcquity UPLC.
Thanks,
Adam
> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>>
wrote:
>
> Hello Adam
>
> Do you see any ion spray current difference when spluttering versus spraying?
What ion spray voltage are you using? Are you using an EasynLC or something
else?
Rest of post
>
>
>
> Cheers
>
> Duncan
>
> Sent from my iPhone
>
>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>
>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
>>
>> Best,
>>
>> Adam
>> ――
>> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>
> ――
> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
> Leave group <email obscured>?Subject=Unsubscribe
>
We have also noticed a significant deterioration in the quality of spray
and lifetime of the PepMap columns from Thermo. The proportion of duds that
die within a couple of weeks has been increasing alarmingly.
Bill Breuer
Rest of post
On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>> wrote:
> Hi Duncan,
>
> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA).
> I’ll have to check what signal I have on a non-sputtering column. I’m
> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>
> Thanks,
>
> Adam
>
> > On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>>
> wrote:
> >
> > Hello Adam
> >
> > Do you see any ion spray current difference when spluttering versus
> spraying? What ion spray voltage are you using? Are you using an EasynLC or
> something else?
> >
> >
> >
> > Cheers
> >
> > Duncan
> >
> > Sent from my iPhone
> >
> >> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
> >>
> >> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
> (ES802) column. We have an inline two-column system with trapping column.
> We've been diligent in using detergent clean up kits when needed and
> perform SPE on all samples. The set of samples where spray sputtering
> appeared were SCX fractions from a StageTip SCX fractionation approach
> (using ammonium acetate buffer). The column had 15 injections prior to this
> of which half were an E. coli peptide digest and the other half were
> acetonitrile ("blank") injections. Has anyone experienced spray sputtering
> issues with the Easy-Spray columns? In general what do you when you
> encounter spray sputtering ("solvent beading") at the emitter? Any
> suggestions for cleaning solutions other than high organic to flush the
> column?
> >>
> >> Best,
> >>
> >> Adam
> >> ――
> >> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
> >> Leave group <email obscured>?Subject=Unsubscribe
> >>
> >
> > ――
> > View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
> > Leave group <email obscured>?Subject=Unsubscribe
> >
>
> ――
> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
> Leave group <email obscured>?Subject=Unsubscribe
>
>
Try the µPAC columns from PharmaFluidics. These are great with very low
back pressure and low background and the separation of peptides is
superior as compared to any packed column.
Rest of post
Success
Goran
On 13-Sep-18 9:21 AM, William Breuer wrote:
> We have also noticed a significant deterioration in the quality of spray
> and lifetime of the PepMap columns from Thermo. The proportion of duds that
> die within a couple of weeks has been increasing alarmingly.
> Bill Breuer
>
> On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>> wrote:
>
>> Hi Duncan,
>>
>> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA).
>> I’ll have to check what signal I have on a non-sputtering column. I’m
>> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>>
>> Thanks,
>>
>> Adam
>>
>>> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>>
>> wrote:
>>> Hello Adam
>>>
>>> Do you see any ion spray current difference when spluttering versus
>> spraying? What ion spray voltage are you using? Are you using an EasynLC or
>> something else?
>>>
>>>
>>> Cheers
>>>
>>> Duncan
>>>
>>> Sent from my iPhone
>>>
>>>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>>>
>>>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
>> (ES802) column. We have an inline two-column system with trapping column.
>> We've been diligent in using detergent clean up kits when needed and
>> perform SPE on all samples. The set of samples where spray sputtering
>> appeared were SCX fractions from a StageTip SCX fractionation approach
>> (using ammonium acetate buffer). The column had 15 injections prior to this
>> of which half were an E. coli peptide digest and the other half were
>> acetonitrile ("blank") injections. Has anyone experienced spray sputtering
>> issues with the Easy-Spray columns? In general what do you when you
>> encounter spray sputtering ("solvent beading") at the emitter? Any
>> suggestions for cleaning solutions other than high organic to flush the
>> column?
>>>> Best,
>>>>
>>>> Adam
>>>> ――
>>>> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>
>>> ――
>>> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
>>> Leave group <email obscured>?Subject=Unsubscribe
>>>
>> ――
>> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>>
> ――
> View topic http://list.abrf.org/r/topic/2g4eUhZUTZWOtrz6ikV7UU
> Leave group <email obscured>?Subject=Unsubscribe
>
--
Goran Mitulović, Ph.D.
ORCID 0000-0003-1964-3965
Head Proteomics Core Facility
Medical University of Vienna
Clinical Department of Laboratory Medicine
Proteomics Core Facility, E5, LS5H
Waehringer Guertel 18-20
A-1090 Wien, Austria
Tel: +43 1 40400 53750
Mail: <email obscured>
Web: www.meduniwien.ac.at/proteomics-core
that's a very interesting suggestion - are the PharmaFluidics columns built
to fit into Thermo's Nano-spray source?
On Thu, Sep 13, 2018 at 11:08 AM, Goran Mitulović <
Rest of post
<email obscured>> wrote:
> Try the µPAC columns from PharmaFluidics. These are great with very low
> back pressure and low background and the separation of peptides is superior
> as compared to any packed column.
>
> Success
>
> Goran
>
>
>
> On 13-Sep-18 9:21 AM, William Breuer wrote:
>
>> We have also noticed a significant deterioration in the quality of spray
>> and lifetime of the PepMap columns from Thermo. The proportion of duds
>> that
>> die within a couple of weeks has been increasing alarmingly.
>> Bill Breuer
>>
>> On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>>
>> wrote:
>>
>> Hi Duncan,
>>>
>>> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA).
>>> I’ll have to check what signal I have on a non-sputtering column. I’m
>>> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>>>
>>> Thanks,
>>>
>>> Adam
>>>
>>> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>
>>>> >
>>>>
>>> wrote:
>>>
>>>> Hello Adam
>>>>
>>>> Do you see any ion spray current difference when spluttering versus
>>>>
>>> spraying? What ion spray voltage are you using? Are you using an EasynLC
>>> or
>>> something else?
>>>
>>>>
>>>>
>>>> Cheers
>>>>
>>>> Duncan
>>>>
>>>> Sent from my iPhone
>>>>
>>>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>>>>
>>>>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
>>>>>
>>>> (ES802) column. We have an inline two-column system with trapping
>>> column.
>>> We've been diligent in using detergent clean up kits when needed and
>>> perform SPE on all samples. The set of samples where spray sputtering
>>> appeared were SCX fractions from a StageTip SCX fractionation approach
>>> (using ammonium acetate buffer). The column had 15 injections prior to
>>> this
>>> of which half were an E. coli peptide digest and the other half were
>>> acetonitrile ("blank") injections. Has anyone experienced spray
>>> sputtering
>>> issues with the Easy-Spray columns? In general what do you when you
>>> encounter spray sputtering ("solvent beading") at the emitter? Any
>>> suggestions for cleaning solutions other than high organic to flush the
>>> column?
>>>
>>>> Best,
>>>>>
>>>>> Adam
>>>>> ――
>>>>> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>
>>>>> ――
>>>> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>
>>>> ――
>>> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
>>> Leave group <email obscured>?Subject=Unsubscribe
>>>
>>>
>>> ――
>> View topic http://list.abrf.org/r/topic/2g4eUhZUTZWOtrz6ikV7UU
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>>
> --
>
>
> Goran Mitulović, Ph.D.
> ORCID 0000-0003-1964-3965
> Head Proteomics Core Facility
> Medical University of Vienna
> Clinical Department of Laboratory Medicine
> Proteomics Core Facility, E5, LS5H
> Waehringer Guertel 18-20
> A-1090 Wien, Austria
> Tel: +43 1 40400 53750
> Mail: <email obscured>
> Web: www.meduniwien.ac.at/proteomics-core
>
>
> ――
> View topic http://list.abrf.org/r/topic/khsuKotPcQmJIr9NzHy14
> Leave group <email obscured>?Subject=Unsubscribe
>
>
Actually, you can use them as a normal nano column. There are no
integrated emitters and you have to use either steel or glass emitters.
I am very happy with stainless steel emitters (ES542) from Thermo and
their classic nano-spray source, and, if you wish, I can send you some
pictures of the setup and chromatograms.
Rest of post
Goran
On 13-Sep-18 10:15 AM, William Breuer wrote:
> that's a very interesting suggestion - are the PharmaFluidics columns built
> to fit into Thermo's Nano-spray source?
>
> On Thu, Sep 13, 2018 at 11:08 AM, Goran Mitulović <
> <email obscured>> wrote:
>
>> Try the µPAC columns from PharmaFluidics. These are great with very low
>> back pressure and low background and the separation of peptides is superior
>> as compared to any packed column.
>>
>> Success
>>
>> Goran
>>
>>
>>
>> On 13-Sep-18 9:21 AM, William Breuer wrote:
>>
>>> We have also noticed a significant deterioration in the quality of spray
>>> and lifetime of the PepMap columns from Thermo. The proportion of duds
>>> that
>>> die within a couple of weeks has been increasing alarmingly.
>>> Bill Breuer
>>>
>>> On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>>
>>> wrote:
>>>
>>> Hi Duncan,
>>>> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA).
>>>> I’ll have to check what signal I have on a non-sputtering column. I’m
>>>> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>>>>
>>>> Thanks,
>>>>
>>>> Adam
>>>>
>>>> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>
>>>> wrote:
>>>>
>>>>> Hello Adam
>>>>>
>>>>> Do you see any ion spray current difference when spluttering versus
>>>>>
>>>> spraying? What ion spray voltage are you using? Are you using an EasynLC
>>>> or
>>>> something else?
>>>>
>>>>>
>>>>> Cheers
>>>>>
>>>>> Duncan
>>>>>
>>>>> Sent from my iPhone
>>>>>
>>>>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>>>>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
>>>>>>
>>>>> (ES802) column. We have an inline two-column system with trapping
>>>> column.
>>>> We've been diligent in using detergent clean up kits when needed and
>>>> perform SPE on all samples. The set of samples where spray sputtering
>>>> appeared were SCX fractions from a StageTip SCX fractionation approach
>>>> (using ammonium acetate buffer). The column had 15 injections prior to
>>>> this
>>>> of which half were an E. coli peptide digest and the other half were
>>>> acetonitrile ("blank") injections. Has anyone experienced spray
>>>> sputtering
>>>> issues with the Easy-Spray columns? In general what do you when you
>>>> encounter spray sputtering ("solvent beading") at the emitter? Any
>>>> suggestions for cleaning solutions other than high organic to flush the
>>>> column?
>>>>
>>>>> Best,
>>>>>> Adam
>>>>>> ――
>>>>>> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
>>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>>
>>>>>> ――
>>>>> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>
>>>>> ――
>>>> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>
>>>>
>>>> ――
>>> View topic http://list.abrf.org/r/topic/2g4eUhZUTZWOtrz6ikV7UU
>>> Leave group <email obscured>?Subject=Unsubscribe
>>>
>>>
>> --
>>
>>
>> Goran Mitulović, Ph.D.
>> ORCID 0000-0003-1964-3965
>> Head Proteomics Core Facility
>> Medical University of Vienna
>> Clinical Department of Laboratory Medicine
>> Proteomics Core Facility, E5, LS5H
>> Waehringer Guertel 18-20
>> A-1090 Wien, Austria
>> Tel: +43 1 40400 53750
>> Mail: <email obscured>
>> Web: www.meduniwien.ac.at/proteomics-core
>>
>>
>> ――
>> View topic http://list.abrf.org/r/topic/khsuKotPcQmJIr9NzHy14
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>>
> ――
> View topic http://list.abrf.org/r/topic/1a779oyxaNBVPYP25A9xdM
> Leave group <email obscured>?Subject=Unsubscribe
>
--
Goran Mitulović, Ph.D.
ORCID 0000-0003-1964-3965
Head Proteomics Core Facility
Medical University of Vienna
Clinical Department of Laboratory Medicine
Proteomics Core Facility, E5, LS5H
Waehringer Guertel 18-20
A-1090 Wien, Austria
Tel: +43 1 40400 53750
Mail: <email obscured>
Web: www.meduniwien.ac.at/proteomics-core
Thank you very much Goran, I'd love to see pictures of your setup.
Bill
On Thu, Sep 13, 2018 at 11:27 AM, Goran Mitulović <
Rest of post
<email obscured>> wrote:
> Actually, you can use them as a normal nano column. There are no
> integrated emitters and you have to use either steel or glass emitters. I
> am very happy with stainless steel emitters (ES542) from Thermo and their
> classic nano-spray source, and, if you wish, I can send you some pictures
> of the setup and chromatograms.
>
> Goran
>
>
>
>
>
> On 13-Sep-18 10:15 AM, William Breuer wrote:
>
>> that's a very interesting suggestion - are the PharmaFluidics columns
>> built
>> to fit into Thermo's Nano-spray source?
>>
>> On Thu, Sep 13, 2018 at 11:08 AM, Goran Mitulović <
>> <email obscured>> wrote:
>>
>> Try the µPAC columns from PharmaFluidics. These are great with very low
>>> back pressure and low background and the separation of peptides is
>>> superior
>>> as compared to any packed column.
>>>
>>> Success
>>>
>>> Goran
>>>
>>>
>>>
>>> On 13-Sep-18 9:21 AM, William Breuer wrote:
>>>
>>> We have also noticed a significant deterioration in the quality of spray
>>>> and lifetime of the PepMap columns from Thermo. The proportion of duds
>>>> that
>>>> die within a couple of weeks has been increasing alarmingly.
>>>> Bill Breuer
>>>>
>>>> On Wed, Sep 12, 2018 at 9:52 PM, Adam Rabalski <email obscured>>
>>>> wrote:
>>>>
>>>> Hi Duncan,
>>>>
>>>>> I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1%
>>>>> FA).
>>>>> I’ll have to check what signal I have on a non-sputtering column. I’m
>>>>> running at 2.2 kV spray voltage. It’s a Waters NanoAcquity UPLC.
>>>>>
>>>>> Thanks,
>>>>>
>>>>> Adam
>>>>>
>>>>> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>.
>>>>> uk
>>>>> wrote:
>>>>>
>>>>> Hello Adam
>>>>>>
>>>>>> Do you see any ion spray current difference when spluttering versus
>>>>>>
>>>>>> spraying? What ion spray voltage are you using? Are you using an
>>>>> EasynLC
>>>>> or
>>>>> something else?
>>>>>
>>>>>
>>>>>> Cheers
>>>>>>
>>>>>> Duncan
>>>>>>
>>>>>> Sent from my iPhone
>>>>>>
>>>>>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>>>>>
>>>>>>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC
>>>>>>>
>>>>>>> (ES802) column. We have an inline two-column system with trapping
>>>>>>
>>>>> column.
>>>>> We've been diligent in using detergent clean up kits when needed and
>>>>> perform SPE on all samples. The set of samples where spray sputtering
>>>>> appeared were SCX fractions from a StageTip SCX fractionation approach
>>>>> (using ammonium acetate buffer). The column had 15 injections prior to
>>>>> this
>>>>> of which half were an E. coli peptide digest and the other half were
>>>>> acetonitrile ("blank") injections. Has anyone experienced spray
>>>>> sputtering
>>>>> issues with the Easy-Spray columns? In general what do you when you
>>>>> encounter spray sputtering ("solvent beading") at the emitter? Any
>>>>> suggestions for cleaning solutions other than high organic to flush the
>>>>> column?
>>>>>
>>>>> Best,
>>>>>>
>>>>>>> Adam
>>>>>>> ――
>>>>>>> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
>>>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>>>
>>>>>>> ――
>>>>>>>
>>>>>> View topic http://list.abrf.org/r/topic/5niTIEbK4QBs3xViQV8Pxd
>>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>>
>>>>>> ――
>>>>>>
>>>>> View topic http://list.abrf.org/r/topic/1sNclGDvclhHtvSY5NYa01
>>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>>
>>>>>
>>>>> ――
>>>>>
>>>> View topic http://list.abrf.org/r/topic/2g4eUhZUTZWOtrz6ikV7UU
>>>> Leave group <email obscured>?Subject=Unsubscribe
>>>>
>>>>
>>>> --
>>>
>>>
>>> Goran Mitulović, Ph.D.
>>> ORCID 0000-0003-1964-3965
>>> Head Proteomics Core Facility
>>> Medical University of Vienna
>>> Clinical Department of Laboratory Medicine
>>> Proteomics Core Facility, E5, LS5H
>>> Waehringer Guertel 18-20
>>> A-1090 Wien, Austria
>>> Tel: +43 1 40400 53750
>>> Mail: <email obscured>
>>> Web: www.meduniwien.ac.at/proteomics-core
>>>
>>>
>>> ――
>>> View topic http://list.abrf.org/r/topic/khsuKotPcQmJIr9NzHy14
>>> Leave group <email obscured>?Subject=Unsubscribe
>>>
>>>
>>> ――
>> View topic http://list.abrf.org/r/topic/1a779oyxaNBVPYP25A9xdM
>> Leave group <email obscured>?Subject=Unsubscribe
>>
>>
> --
>
>
> Goran Mitulović, Ph.D.
> ORCID 0000-0003-1964-3965
> Head Proteomics Core Facility
> Medical University of Vienna
> Clinical Department of Laboratory Medicine
> Proteomics Core Facility, E5, LS5H
> Waehringer Guertel 18-20
> A-1090 Wien, Austria
> Tel: +43 1 40400 53750
> Mail: <email obscured>
> Web: www.meduniwien.ac.at/proteomics-core
>
>
> ――
> View topic http://list.abrf.org/r/topic/2ErXbw0TzhalSlRkYaauiy
> Leave group <email obscured>?Subject=Unsubscribe
>
>
Hello Adam
I was referring to the ion spray current read back not the TIC. The current
read back will allow you to see if there is an issue that correlates with the
spitting.
If you are using a nAcquity with a trap I'm guessing you are using a vented
configuration. Is this the case? If so, this involves having no flow through
the analytical column, liquid junction and emitter whilst you load the sample.
If you are applying HV during this phase of the method, thats not good for the
EasySpray liquid junction or emitter. When I used a vented n Acquity and easy
spray setup I turned off the HV during the loading phase and only introduced it
a couple of minutes after the valve switched and flow was returned to the
analytical column. this means you only have HV supply whilst the liquid
junction is fully flowing. I found this massively improved EasySpray life span
and eliminated spitting all together.
Hope this helps
Rest of post
Duncan
________________________________________
From: ABRF Discussion Forum <email obscured>] on behalf of Adam Rabalski
<email obscured>]
Sent: 12 September 2018 19:52
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] EASY-Spray C18 PepMap RSLC Emitter Spray
Sputter
Hi Duncan,
I observe 1E5 signal for the 371 ion in the Orbitrap at 98% A (0.1% FA). I’ll
have to check what signal I have on a non-sputtering column. I’m running at 2.2
kV spray voltage. It’s a Waters NanoAcquity UPLC.
Thanks,
Adam
> On Sep 12, 2018, at 12:33, Duncan Smith <email obscured>>
wrote:
>
> Hello Adam
>
> Do you see any ion spray current difference when spluttering versus spraying?
What ion spray voltage are you using? Are you using an EasynLC or something
else?
>
>
>
> Cheers
>
> Duncan
>
> Sent from my iPhone
>
>> On 12 Sep 2018, at 17:44, Adam Rabalski <email obscured>> wrote:
>>
>> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
>>
>> Best,
>>
>> Adam
>> ――
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>>
>
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Hi Duncan,
Yes that's correct I'm using a vented trap configuration. The spray current is
about 0.05-0.10 uA when I observe sputtering. The voltage is off during the
trapping phase as the instrument is in standby. I'll add in an additional
segment at the beginning of the method to turn the spray voltage off for a few
minutes after valve switching as you suggest.
Thanks,
Adam
PS
Goran, I'm interested in seeing your setup as well! What length of the uPAC do
you use and what are your ID rates like for a complex sample such as mammalian
proteome?
Dear all,
as I can see there is some interest in µPAC. I have a presentation on
that and can load it on some place for download. Please give me a day or
two to figure out how to do that since I am at
https://reduse2018.meduniwien.ac.at/ and have no direct access to my PC.
After that, we can discuss all you are interested about. There are many
results I have generated with this column, 2 m flow rates of 800 nl/min
and 600 nl/min, respectively. All samples analyzed, so far are of human
origin - serum, urine, tissue. The column enables better sequence
coverage as compared to the PepMap 50 cm and, simultaneously, higher ID
rates. As I wrote, I have a lot of material but I cannot discuss all of
it due to fact that I do not "own" the data and I need to ask for
permission on showing the most complex ones. You might want to contact
me privately for detailed description of the methods used.
Many greetings
Goran
On 13-Sep-18 4:40 PM, Adam Rabalski wrote:
> Hi Duncan,
>
> Yes that's correct I'm using a vented trap configuration. The spray current
is about 0.05-0.10 uA when I observe sputtering. The voltage is off during the
trapping phase as the instrument is in standby. I'll add in an additional
segment at the beginning of the method to turn the spray voltage off for a few
minutes after valve switching as you suggest.
>
> Thanks,
>
> Adam
>
> PS
>
> Goran, I'm interested in seeing your setup as well! What length of the uPAC
do you use and what are your ID rates like for a complex sample such as
mammalian proteome?
Rest of post
> ――
> View topic http://list.abrf.org/r/topic/1WobORw8tlQAinZpt7ST0F
> Leave group <email obscured>?Subject=Unsubscribe
>
--
Goran Mitulović, Ph.D.
ORCID 0000-0003-1964-3965
Head Proteomics Core Facility
Medical University of Vienna
Clinical Department of Laboratory Medicine
Proteomics Core Facility, E5, LS5H
Waehringer Guertel 18-20
A-1090 Wien, Austria
Tel: +43 1 40400 53750
Mail: <email obscured>
Web: www.meduniwien.ac.at/proteomics-core
Hi Adam,
We also had this issue time and again with the EasySpray and eventually stopped
using them. It was quite recently. We made a huge effort to troubleshoot and
Thermo helped us a lot but at the end of the day they were just not performing.
Best,
Yishai
> On 12 Sep 2018, at 19:44, Adam Rabalski <email obscured>> wrote:
>
> I'm running into sputtering issues on an EASY-Spray C18 PepMap RSLC (ES802)
column. We have an inline two-column system with trapping column. We've been
diligent in using detergent clean up kits when needed and perform SPE on all
samples. The set of samples where spray sputtering appeared were SCX fractions
from a StageTip SCX fractionation approach (using ammonium acetate buffer). The
column had 15 injections prior to this of which half were an E. coli peptide
digest and the other half were acetonitrile ("blank") injections. Has anyone
experienced spray sputtering issues with the Easy-Spray columns? In general
what do you when you encounter spray sputtering ("solvent beading") at the
emitter? Any suggestions for cleaning solutions other than high organic to
flush the column?
Rest of post
>
> Best,
>
> Adam
> ――
> View topic http://list.abrf.org/r/topic/6AYYvJVGzRqCESWv4mJVdK
> Leave group <email obscured>?Subject=Unsubscribe
>
Hello,
As a long time member of the ABRF (sPRG through three studies),
I regret that your experiences with Thermo Fisher Scientific products have not
met the expectations that we have set in our product specifications for
EasySpray columns. I was hesitant to post since I don't want to compromise the
instrument manufacturer / ABRF member relationship that I value very highly. We
would like the opportunity to correct this through our link in blue on the
right side of the page https://www.separatedbyexperience.com/chromexpert, the
button is labeled “Contact Technical Support >>”. This starts a discussion with
us here in Chromatography Columns and Consumables support group. We can
diagnose your problems and help you achieve success.
Craig P. Dufresne, Ph.D.
Staff Scientist, Field Applications
Chromatography Columns and Consumables
Thermo Fisher Scientific
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Cell: 561-951-5582
<email obscured>
Hi Craig,
I use the EasySpray Columns exclusively (ES802, ES803, ES804, and ES805) and
the nanoTrap columns (PN 164535). Like others, I too found the learning curve
modestly steep and not inexpensive, but worthwhile in the end.
I have both the Easy nLC 1000 and two 3000 RSLC nano UHPLC systems. Sputtering
was only problematic for the nLC 1000. After several column exchanges from
Thermo tech support, I finally figured out the sputtering was due to voltage
leaking back through the column. For me, the fix was PN 6041.5114 - Grounding
Adapter, that looks just like a nanoViper fitting but is metal and mounts
between the column and valve. No more sputtering.
With the 3000 RSLC nano systems, sputtering is only observed at the end of the
column's lifetime.
Just my $0.02
Rest of post
Regards,
Jeff
-----Original Message-----
From: craig.dufresne <email obscured>>
Sent: Tuesday, October 02, 2018 11:54 AM
To: <email obscured>
Subject: Re: [ABRF Discussion Forum] EASY-Spray C18 PepMap RSLC Emitter Spray
Sputter
Hello,
As a long time member of the ABRF (sPRG through three studies),
I regret that your experiences with Thermo Fisher Scientific products have not
met the expectations that we have set in our product specifications for
EasySpray columns. I was hesitant to post since I don't want to compromise the
instrument manufacturer / ABRF member relationship that I value very highly. We
would like the opportunity to correct this through our link in blue on the
right side of the page https://www.separatedbyexperience.com/chromexpert, the
button is labeled “Contact Technical Support >>”. This starts a discussion with
us here in Chromatography Columns and Consumables support group. We can
diagnose your problems and help you achieve success.
Craig P. Dufresne, Ph.D.
Staff Scientist, Field Applications
Chromatography Columns and Consumables
Thermo Fisher Scientific
1400 Northpoint Parkway, Suite 10
West Palm Beach, FL 33407
Cell: 561-951-5582
<email obscured>
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