Instructions for tuning Sciex API for MSMS data collection
in RAD:
1)
Sign up, also sign log book
2)
Hardware --- checklist
3)
Software
open tune 2.5
Open
files:
Q3 Calibration
Q3 State
These
files can be found on HD#2, daily data files folders
If
starting in the morning, Electronics 2 should be on.
Username
: MassSpec, PW: KFF
Calibration
Calibrant:
PPG
Set up syringe in the pump and tighten the block against the
plunger.
Turn on power on back.
Check flow. Set flow
to change. Set at 10 µL/min.
Check diameter and set at 14mm.
To start push Run.
Turn
on electronics 1
Also
open settings file, which can be found in the following folder.
[HD]--> [api ... ] -->
[settings]
select file PPG Q3 with color
To
start scanning:
click
both buttons apple/2 ---------- or you can click SCAN menu and
select
Q3 M1
Make
sure [MCA] is ON (MCA affects
if scans are additive or not).
Collect
~10 scans
You
want to get a V shape for the middle bottom 2 panels.
Under
the UTILITIES menu, select CALIBRATE and delta mass should be less than 0.05. If the delta mass is too high, repeat the
scan.
Replace
Calibration
Repeat
When
happy with calibration you save calibration file and save state file both in the daily data file folder. Create a new folder for the current day.
Need
to redo with wide calibrations, then with Q1.
8 files should
be saved:
Q1
state file
Q1
cal
Q1
state file wide
Q1
cal wide
Q3
state file
Q3
cal
Q3
state file wide
Q3
cal wide
When
done, also turn off electronics 1 ,,, so not to shock yourself when
touching
the cover.
Settings
for proteins:
Orifice (OR) 60
CC 1
These
values for PPG are OR 35, and CC 10
TO SET UP FOR
MSMS:
1.
Open
Q1 state file wide and record: RE1 and DM1
Open
Q3 state file wide and change these parameters:
RE1=RE1(from
Q1)-20
DM1=DM1
from Q1
Other
parameters from sticker on the machine:
R2=0,
L7=5, Rx=-10, R3=-5
2.
Foe
negative MSMS
change
polarity under utilities and
change
ISV=3500
CC=1
CAD
gas on (under CC)
Save
as MSMSnegative state file in Tune
3.
To
save in RAD:
Save
as: Daily data files/RADdata
à
create daily data file and 3 subfolders: data, state, cal.
NO
SPACES
Save
Q1cal and Q3cal (wide)
Save
MSMSnegativestate
4.
Quit
tune and open RAD
Open
MSMSsatatefile
Open
Q1cal and Q3cal
On
1090 HPLC:
Open
method with correct masses.
Batch
– data – this folder
Number
of samples, highlight #1 and start
