Thermo Scientific Dionex

Chromeleon
Operational Qualification/
Performance Qualification
for
Ultimate 3000 RSLCnano
Nano, Cap and MIC Systems

Annex to
Operating Instructions
Version 1.00

Revision: A
Date: May 2017

© 2017 Thermo Fisher Scientific Inc.

Doc.: Annex-A_RSLCnano_OQ_PQ_OI_V1_00.docx
Annex to OQ and PQ Operating Instructions

Contents
1 How to Use This Manual ................................................................................... 3

2 Process .............................................................................................................. 3
2.1 Required Materials .......................................................................................................................3
2.2 Performing the Checks ................................................................................................................3

3 Procedures ........................................................................................................ 4
3.1 Support of Thermo Scientific Dionex UltiMate 3000 RSLCnano ProFlow Flow Meter ..........4
3.2 Thermo Scientific Dionex UltiMate VWD-3x00 detector - Linearity with MIC flow cell .........4
3.2.1 Performing the Check .............................................................................................................4

Page 2 of 4 Annex to OQ and PQ Operating Instructions – Revision A of May 2017

 Annex to OQ and PQ Operating Instructions

1 How to Use This Manual

This annex is an addendum to RSLCnano OQ/PQ Operation Instruction, version 1.0. It describes the
changes on sequence templates with a version later than 4.4. The version number of the sequence
template is defined in the name of the report template.

2 Process
2.1 Required Materials
Part number 6720.0360 includes the RSLCnano OQ/PQ operating instructions, set of 8 caffeine
samples, 1 µL sample loop with nanoViper connections and connection tubing.
The parts (depending on the configuration) necessary to perform the OQ/PQ procedure, which are not
included in the OQ/PQ kit, are described in the RSLCnano OQ/PQ operating instructions.
The sample positions used in the sequence templates were changed to match with the ampoules in
the standards kit: The content of the 10 µg/mL standard ampoule has to be filled in only one vial that is
TM TM
placed in Thermo Scientific UltiMate WPS autosampler position RA4 (instead of RA4 and RB2).
Additionally, the content of the 40 µg/mL and the 60 µg/mL standard ampoules have to be filled in vials
and placed at the WPS autosampler positions RA6 and RA7 respectively. Thus, the following sample
positions are used with the new templates (differences to the operating instruction are marked bold):
Table 2-1 Sample Positions in the WPS Autosampler Rack

Position Substance Concentration Check(s)

RA1 Caffeine standard 1.0 µg/mL UV linearity
RA2 Caffeine standard 2.5 µg/mL UV linearity, UV linearity MIC
RA3 Caffeine standard 5.0 µg/mL UV linearity
RA4 Caffeine standard 10 µg/mL UV linearity, UV linearity
MIC, WPS carry over, VWD
wavelength check
RA5 Caffeine standard 20 µg/mL UV linearity, UV linearity
MIC, WPS reproducibility,
WPS linearity
RA6 Caffeine standard 40 µg/mL UV linearity MIC
RA7 Caffeine standard 60 µg/mL UV linearity MIC
RA8, RB1, and RB2 are empty!

RB3 Caffeine standard 1000 µg/mL WPS carry over

RB4 Mobile phase A: Mobile phase B 99:1 WPS carry over
RB5 Mobile phase A: Mobile phase B 99:1 WPS carry over
RB6 Mobile phase A: Mobile phase B 99:1 WPS carry over

2.2 Performing the Checks

The tests to be performed, the order in which they must be performed and the time required for each
individual test are listed in Table 2-1 (differences to operating instruction are marked bold). Performing
the OQ or PQ of an Ultimate 3000 RSLCnano system in a standard configuration will take
approximately 24 h.
Table 2-2 Test to be performed and time required for each test

Check Duration Comment

WARM_UP 4h Irrespective of the presence or
WARM_UP AND OVEN_TEST absence of a thermostated
column oven

Annex to OQ and PQ Operating Instructions – Revision A of May 2017 Page 3 of 4

Annex to OQ and PQ Operating Instructions

Check Duration Comment

FLOW CELL CHECK 5 min -
NOISE AND DRIFT 25 min
GRADIENT FORMATION TEST 6.0 h
WAVELENGTH CHECK 20 - 40 min Depending on the configuration
AUTOSAMPLER 1-2h (faster for MIC and slower for
REPRODUCIBILITY NAN)
UV LINEARITY
AUTOSAMPLER LINEARITY 45 min - 1.5 h
CARRY OVER TEST
Replumbing required
UV LINEARITY (MIC FLOW CELL) 30 min -
LOADING PUMP GRADIENT 5.0 h
FORMATION TEST

3 Procedures
3.1 Support of Thermo Scientific UltiMate 3000 RSLCnano ProFlow
Flow Meter
TM TM
Thermo Scientific Dionex Chromeleon RSLCnano Nan/Cap/Mic OQ/PQ supports the UltiMate
TM
RSLCnano ProFlow flow meter (since version 4.4 distributed with CM 6.80 SR15b).
The OQ/PQ is performed and evaluated against the specifications for the NAN (nano) configuration as
described in the RSLCnano OQ/PQ operating instructions, version 1.0.

3.2 Thermo Scientific UltiMate VWD-3x00 Detector - Linearity with

MIC Flow Cell
In order to qualify the loading pump of the UltiMate NCS-3500RS system, a MIC flow cell has to be
installed at the UltiMate VWD-3x00 detector. Therefore, the UV linearity is checked for the MIC flow
cell prior to the loading pump gradient formation test. The UV linearity check for the MIC flow cell is
automatically provided for all NCS-3500RS systems (since version 4.7 distributed with CM 6.80
SR16).

3.2.1 Performing the Check

At the beginning of the test, the flow of the NC pump is stopped and a message prompts the user to
change the configuration. The corresponding description in the operating instructions is obsolete.
The detector linearity is determined by injecting five different caffeine standards (2.5; 10; 20; 40;
60 μg/mL) and recording the resulting chromatogram at 272 nm. The regression coefficient is then
calculated based on peak heights and concentrations, and qualifies the detector linearity for the MIC
flow cell. The test results are displayed in the report under the “Det_Linearity” tab.

Page 4 of 4 Annex to OQ and PQ Operating Instructions – Revision A of May 2017