Antec ROXY User manual

ROXY
Installation Guide
210.7011, Edition 5, 2018
T [phone] | [e-mail] | www.AntecScientific.com
Antec Scientific
Industrieweg 12
2382 NV Zoeterwoude
The Netherlands

Copyright ©2018, Antec Scientific, The Netherlands. Contents of this publication may not be reproduced in any form or
by any means (including electronic storage and retrieval or translation into a foreign language) without prior agreement
and written consent from the copyright of the owner. Antec reserves the right to make changes to the design and speci-
fications of the instrument and this manual without prior notice.
ROXY, ALEXYS, DECADE, DECADE II, DECADE Elite, DECADE Lite, INTRO, Flexcell, ReactorCell, µ-PrepCell,
ISAAC, HyREF, SenCell and SynthesisCell are trademarks of Antec. Whatman™ (word and device) and Whatrnan™
(word only) are trademarks of Whatman lnternational Ltd. SOLVENT IFD™ and AQUEOUS IFD™ are trademarks of
Arbor Technologies, Inc. Clarity®, DataApex® are trademarks of DataApex Ltd. Microsoft® and Windows™ are trade-
marks of Microsoft Corporation. Excel is a registered trademark of the Microsoft Corporation. All other trademarks are
the property of their respective owners.
The software and the information provided herein is believed to be reliable. Antec shall not be liable for errors con-
tained herein or for incidental or consequential damages in connection with the furnishing, performance, or use of soft-
ware or this manual. All use of the software shall be entirely at the user’s own risk

Installation guide 3 (of 33)
Table of contents
Warning Symbols.................................................................................... 4
Intended use ........................................................................................... 6
WEEE directive....................................................................................... 7
Warranty, spare parts and service.......................................................... 8
Safety Instructions................................................................................... 8
Introduction.............................................................................................. 10
Preparing for installation........................................................................ 11
Pre-installation preparations ................................................................. 11
IQ and OQ............................................................................................. 11
PQ......................................................................................................... 11
Relevant documents for installation...................................................... 13
Software download................................................................................ 13
Contents of ROXY................................................................................. 14
Installation................................................................................................ 15
Electrical connections ........................................................................... 16
Software installation.............................................................................. 18
Installation of a flow cell........................................................................ 20
Installation of grounding kit ................................................................... 22
Performing an experiment..................................................................... 22
PQ Oxidative mode (metabolite synthesis) .......................................... 25
PQ procedure........................................................................................ 25
Results .................................................................................................. 26
PQ Reductive mode (disulfide bond reduction)................................... 27
PQ procedure TiBlue electrode (DC mode).......................................... 27
PQ procedure Ti electrode (Pulse mode) ............................................. 28
Results .................................................................................................. 29
Trouble shooting..................................................................................... 30
Dialogue Elite........................................................................................... 32
Hands-on training.................................................................................. 32

4 (of 33) ROXY installation guide, ed. 4
Warning Symbols
The following symbols are used in this guide:
This sign warns about the risk of electric shock. It calls
attention to a procedure or practice which, if not adhered
to, could result personal injury or even loss of life by elec-
trocution. Do not proceed beyond a danger sign until the
indicated conditions are fully understood and met.
The warning sign denotes a hazard. It calls attention to a
procedure or practice which, if not adhered to, could re-
sult in severe injury, loss of life or damage or destruction
of parts or all of the equipment. Do not proceed beyond a
warning sign until the indicated conditions are fully under-
stood and met.
The caution sign denotes a hazard. It calls attention to a
procedure or practice which, if not adhered to, could re-
sult in damage or destruction of parts or all of the equip-
ment and/or erratic results. Do not proceed beyond a
cautions sign until the indicated conditions are fully un-
derstood and met.
The biohazard sign draws attention to the fact that use
of biological materials, viral samples may carry a signifi-
cant health risk.
The toxic hazard sign draws attention to the fact that use
of toxic solvents or samples may carry a significant
health risk.

Installation guide 5 (of 33)
The attention sign signals relevant information. Read this
information.
The note sign signals additional information. It provides
advice or a suggestion that may support you in using the
equipment.

6 (of 33) ROXY installation guide, ed. 4
Intended use
The ROXY EC(/LC) system or ROXY potentiostat for controlled REDOX
reactions up-front Mass Spectrometric detection. It can be used in a wide
range of application, for example:
•Fast synthesis of metabolites (µ-preparative)
•Rapid risk assessments of drug-protein binding
•Signal enhancement in MS
•Electrochemical cleavage of proteins/peptides
•Reduction of disulfide bonds in proteins/peptides
•Oxidative stress/damage of proteins, DNA, lipids, etc.
Operation of the ROXY EC(/LC) system can involve the use of hazardous
materials including corrosive fluids and flammable liquids. The system
should only be operated by users with the following expertise:
•Completed degree as chemical laboratory technician or
comparable vocational training.
•Fundamental knowledge of liquid chromatography & mass
spectrometry
•Participation in an installation of the system performed by the
manufacturer or a company authorized by the manufacturer
and suitable training on the system, flow cell and control soft-
ware.
•Knowledge and experience in the safe handling of toxic and
corrosive chemicals and knowledge of the application of fire
prevention measures prescribed for laboratories.
Information on safety practices is provided with your equipment operation
manuals. Before using your equipment or accessories, you must thor-
oughly read these safety practices. This manual is written for laboratory
technicians and scientists skilled in the art.
For research purposes only. While clinical applications may be shown,
this instrument is not tested by the manufacturer to comply with the In
Vitro Diagnostics Directive.

Installation guide 7 (of 33)
The ROXY EC system is ROHS compliant and in conformity with Directive
2011/65/EU Restricted use of Hazardous Substances in electrical and
electronic Equipment (ROHS).
Antec Leyden B.V. is an ISO 9001 certified company.
WEEE directive
All equipment of Antec Leyden which are subjected to the WEEE directive
shipped after August 13, 2005 are compliant with the WEEE marking re-
quirements. Such products are labelled with the “crossed out wheelie”, de-
picted on the left site.
The symbol on the product indicates that the product must not be
disposed as unsorted municipality waste. Please take care of proper
disposal and recycling.
If needed Antec offers the possibility for recycling. In that case
please ship the instrument back to the manufacturer together with a
decontamination form (Antec Leyden, the Netherlands). They will take
care of the proper disposal and recycling of the instrument.
Manufacturer shipping address for the end-of-life products:
Antec Leyden B.V.
Industrieweg 12
2382NV Zoeterwoude
The Netherlands
In case of questions, or if further information is required about the collec-
tion & recycling procedure, please contact your local distributor.
Unskilled, improper, or careless use of this equipment can create fire
hazards, or other hazards which can cause death, serious injury to
personnel, or severe damage to equipment and property. Observe all
relevant safety practices at all times. Only use the device for
applications that fall within the scope of the specified intended use.
Else the protective and safety equipment of the device could fail.

8 (of 33) ROXY installation guide, ed. 4
Warranty, spare parts and service
The warranty period of this ROXY EC system is 1 year on workmanship,
wear and tear parts are excluded. Manufacturer provides operational spare
parts of the instrument and current accessories for a period of five years
after shipment of the final production run of the flow cell. Spare parts will
be available after this five years period on an ‘as available’ basis.
Manufacturer provides a variety of services to support her customers after
warranty expiration. Repair service can be provided on a time and material
basis. Contact your local supplier for servicing. Technical support and
training can be provided by qualified chemists on both contractual and as-
needed basis.
Safety Instructions
Adhere to the following guidelines when using the ROXY EC system. The
safety practices are intended to ensure safe operation of the flow cell.
Working environment & safety
The intended use of the ROXY EC system is to perform controlled REDOX
reactions of target compounds (in a suitable liquid electrolyte medium) up-
front Mass Spectrometric detection. Operators using the system should
have the appropriate education an extensive understanding of GLP rules
and be skilled in the art. Use this system ONLY for the intended use. Use
of the system for any other purpose might cause unsafe situations.
Operation
To assure optimal performance it is recommend that the ROXY EC system
is checked regularly and maintenance procedures are carried out. Preven-
tive maintenance contracts are available for that Purpose. Please contact
your local dealer or the nearest sales office for more information.
Solvents
The solvents used may be flammable, toxic or corrosive. The room in
which the system is installed should be well ventilated to prevent that sol-
vent vapors cause poisoning or ignite and cause a fire. Use of open fire in
the vicinity of this system must be strictly prohibited. Do not install the sys-
tem in the same room with any other equipment that emits or could poten-
tially emit sparks. Provide protective equipment near the instrument, when
solvent gets into the eyes or on the skin, it must be flushed away immedi-
ately. Provide equipment, such eye wash stations and safety showers, as
close to system as possible. Use proper eye and skin protection when
working with solvents. Additional safety requirements or protection may be

Installation guide 9 (of 33)
necessary depending on the chemicals used in combination with this
equipment. Make sure that you understand the hazards associated with
the chemicals used and take appropriate measures with regards to safety
and protection. Sample containers (vials) should be sealed to minimize any
risks related to solvent vapor.
Biological Hazard
When you analyze biological fluids you need to take the necessary precau-
tions and treat all specimens as potentially infectious. Always wear protec-
tive and gloves when handling toxic or biologically infectious samples to
prevent biohazards or hazards while working with the system. When the
system or parts of it needs to be shipped to Antec for service/repair or dis-
posal, it has to be accompanied with a decontamination form which should
be completely filled in and signed by the customer. Without this decontami-
nation form the system or parts of it will not be processed by Antec (either
repaired or disposed).
Waste disposal
Perform periodic leak checks on LC tubing and connections. Do not close
or block the drain in the oven compartment. Do not allow flammable and/or
toxic solvents to accumulate. Follow a regulated, approved waste disposal
program. Never dispose of flammable and/ toxic solvents through the mu-
nicipal sewage system.
Using the ROXY EC system in other ways than indicated in the manual
might result in erratic or unsafe operation.

10 (of 33) ROXY installation guide, ed. 4
C H A P T E R 1
Introduction
To assure an efficient and successful onsite installation of the ROXY EC
system, we have written this guide. This documents describes all the steps
that should be taken. It contains references to relevant user manuals and
qualification documents.
There are a few different configurations applied for electrochemistry cou-
pled to MS (EC/MS). The EC reactor can be used for continuous flow ex-
periment, or flow injection analysis both are not using chromatography.
Sometimes a secondary solvent is added to study post EC reaction conju-
gation with glutathione for example. In combination with HPLC the reactor
can be placed prior to the HPLC injector (EC/HPLC/MS) or post column
(HPLC/EC/MS).
This document primarily outlines the ROXY EC/MS installation, for the
other configurations most of the described procedure is the same. Except
that for installation of a autosampler or pump the corresponding user man-
uals should be used.
Fig. 1. ROXY EC/MS with syringe pump for continuous flow experiments.
This document is intended as a general check list to guide you through
the installation of a ROXY EC system and by no means a replacement
of the installation sections in the individual user manuals supplied with
the system.

Installation guide 11 (of 33)
C H A P T E R 2
Preparing for installation
Pre-installation preparations
Installation requirements document
To assure an efficient and successful onsite installation of the ROXY EC
system by engineers of Antec or its distributors, we have a guideline with
pre-installation requirements available (document 210.7050). This docu-
ment contains hardware and site requirements as well as information
about chemicals required for the installation and performance qualification
of the ROXY EC system. The ‘site preparation reply form’ in the document
confirming that all requirements are met, has to be signed and returned by
the user prior to installation.
It is the customer’s responsibility that the installation site require-
ments are met and all chemicals are available onsite at the date of in-
stallation. It is the installation engineer’s responsibility to send this
pre-installation requirement document (well in advance of the instal-
lation date) and get a signed copy in return.
IQ and OQ
A printed copy of an Installation Qualification (IQ) document is provided
with each ROXY. The IQ/OQ document is also available for download from
Antec’s website. It is a document for installation and training/instruction of
the user that should be signed-off. An Operational Qualification (OQ) of a
ROXY is a dummy cell test and output measurement. This test is always
part of the installation procedure.
A completed and signed IQ/OQ document will serve as an objective proof
that the installation was completed successful and was carried out to the
satisfaction of both parties. Familiarize yourself with the procedure, per-
form the installation according to the guidelines given herein, and fill in the
IQ/OQ document during the installation.
PQ
A performance Qualification (PQ) for the ROXY demonstrates the perfor-
mance of the system using a solution with a standard (substrate). In a con-
tinuous flow experiment, the substrate is converted in the electrochemical
reactor and the reaction products are measured using an MS. For the

12 (of 33) ROXY installation guide, ed. 4
oxidative mode and reductive mode a separate performance test is availa-
ble based on the conversion of Amodiaquine and Insulin, respectively.
Which test is required is depending on the specific ROXY system installed
and/or customers application (OX/RED).
A PQ should be performed as last step of the installation a to assure the
proper operation of the full ROXY EC system (Syringe pump, ROXY poten-
tiostat, flow cell) in combination with MS. See chapter 4 and 5 of this instal-
lation guide and familiarize yourself with the procedure.
For the PQ specific chemicals and solutions should be available at the cus-
tomer site at the day of installation
Test solutions for oxidative mode
To perform a performance qualification in oxidative mode the following so-
lution should be available:
1. Mobile phase: about 100mL of 20mM Ammonium formate (pH 7.4
adjusted with ammonium hydroxide) in 50% acetonitrile.
2. Sample: 20mL of 10µM Amodiaquine in the solution described in
point 1.
Test solution for reductive mode
To perform performance qualification in reductive mode the following solu-
tion should be available:
1. Mobile phase: about 100mL of 1% formic acid in 50% acetonitrile.
2. Sample: 20mL of 5μg/mL insulin in the mobile phase described in
point 1.
The customer should prepare the solution(s) preferable a day before
the installation day. The sample solution of both Amodiaquine and
Insulin should be stored at 4°C in the dark (light sensitive).

Installation guide 13 (of 33)
Relevant documents for installation
The detailed description of ROXY EC system installation is part of the fol-
lowing user manuals:
210.7050 - ROXY Installation Requirements (prior to installation!)
210.7011 - ROXY EC Installation guide (this document)
210.7010 - ROXY Potentiostat user manual
210.7014 - ReactorCell user manual
204.0010A-µ-PrepCell2.0 user manual
206.0010 - SynthesisCell user manual
175.0015 - Dialogue Elite user manual
For the latest versions of the Antec’s manuals and application notes visit
www.antecscientific.com.
Software download
To assure that the system will be installed with the most recent software
version available we recommend to download the most recent copy of the
Dialogue Elite software and drivers (i.e. USB/COM port driver) before the
installation.
http://www.antecscientific.com/support/documents-and-downloads/soft-
ware
When preparing for an installation, we advise to print out any
document/manual that needs to be signed off.

14 (of 33) ROXY installation guide, ed. 4
Contents of ROXY
Although there are a few exceptions, most ROXY EC systems are deliv-
ered as the following parts:
Part no.
Description
Qty
210.0010A
ROXY Potentiostat, High Current
1
210.0200D
ROXY Potentiostat accessory kit
1
188.0035
Dual syringe infusion pump*
1
188.0300
Syringe 1mL, TLL
2
188.0302
Syringe 100µL, TLL
1
250.0123C
Dual port USB-RS232 converter
1
180.0161A
ROXY LC conn. kit, EC*
1
250.0139L
ROXY cell cable incl 0.5uF, 2m
1
250.0035
ROXY grounding kit
1
171.9015
Dialogue Elite OQ, PQ, ROXY (standard)
1
204.43101
µ-PrepCell 2.0 GC/BDD
1
204.43122
µ-PrepCell 2.0 TiBlue
1
204.43023
µ-PrepCell High Pressure Ti
1
210.00404
ReactorCell kit (incl. BDD, GC, Au Pt)
1
1 ROXY EC system with µ-PrepCell 2.0 (210.0074A)
2 ROXY EC system for S-S reduction (210.0072A)
3ROXY EC system for HDX (210.0073A)
4 ROXY EC system with ReactorCell (210.0070A)
* A ROXY for HDX has no syringe pump.
The details are in the packing list that comes with the delivery.

Installation guide 15 (of 33)
C H A P T E R 3
Installation
Take the following steps to unpack and install the devices:
1. Check all carton boxes for damage. We advise to store the
transport boxes in case of further transport.
2. To unpack the detector, lift it from its box by both hands. Never lift
the detector at its front door, but at its sides.
3. Unpack the syringe pump.
4. Align the system components as indicated in the photo below
5. Check the part and serial numbers of the individual system parts
and write it down in the IQ document.
6. Check the contents of accessory kits using the packing list, to be
sure it is complete and undamaged.
7. Follow the installation instructions in the next section of this docu-
ment.
Fig. 2. ROXY EC with syringe pump.
A tutorial movie how to install a ROXY with an µ-PrepCell
is available on the Antec website:
https://www.antecscientific.com/support/tutorials/roxy-installation

16 (of 33) ROXY installation guide, ed. 4
Electrical connections
1. Connect the power cable, trigger cable and serial communication ca-
ble to the ROXY Potentiostat as it is shown in the Fig. 3. Use cables
provided by Antec or ensure that the used cables meet all relevant
safety and EMC and local requirements.
The manufacturer is not liable if power cords are used which do not
meet the relevant safety and EMC requirements (CE, CSA and UL).
Fig. 3. Back panel of a ROXY Potentiostat (left) and VScom RS232-USB
converter. Converter is connected to computer using an USB port.
2. Install the driver of the VScom USB-to-RS232 converter.
3. Connect the VScom port “Serial 1” to the ROXY with the serial cable.
4. Connect the VScom port “Serial 2” to the syringe pump “RS232 IN”
port (Harvard 11 pump), or connect the PC directly to the pump using
the USB cable supplied with the pump (Legato 101 pump).
5. Connect the cell cable to the Cell 1 connector inside the potentiostat.
6. Connect the USB cable(s) to computer and wait for Windows to finish
installation, in case needed. Right bottom of the Windows screen
shows the progress, in case a Windows driver is loaded.

Installation guide 17 (of 33)
7. Check in the Windows Device manager the numbers assigned to USB
serial ports. Look for “USB Serial Port”. The highest number is “Serial
2” the other is “Serial 1”. Also the Legato port is recognizable as such.
In case of doubt unplug the cable and identify the port that disappears
in the device manager.

18 (of 33) ROXY installation guide, ed. 4
Software installation
1. Install Dialogue Elite and connect the license key (dongle). Without a
license, a message appears and the program starts with limited func-
tionality (demo mode).
2. After installing Dialogue software, switch on potentiostat and syringe
pump.
3. Run the Dialogue software from Windows start menu under 'Antec
software'. The program starts with the 'device selector', where active
devices and ports can be selected. Choose the correct port numbers
to connect Potentiostat and syringe pump, respectively.
4. In case an update is available a message appears. It is advisable to al-
ways use the latest updates.

Installation guide 19 (of 33)
5. The green LEDs in the left bottom corner of the Dialogue window show
that the devices are successfully connected.
6. Use the 'Send to device' button to apply changes in settings as the
green arrow (right bottom) indicates. 'Read from device' refreshes the
current settings.
7. The monitor tab shows the actual status of most relevant parameters.
The S pump tab is for control of the syringe pump.
8. For more details about the Dialogue Elite software, see the user man-
ual. Our website contains a number of screen recordings of most com-
mon actions.

20 (of 33) ROXY installation guide, ed. 4
Installation of a flow cell
1. Follow the specific procedure to assemble the flow cell as de-
scribed in the flow cell user manual.
2. The schematic drawing how to connect the inlet and outlet tubing
with the cell and to place the grounded union is delivered with the
ROXY EC system (see Fig. 4).
1. Place the Luer Lock connector on the syringe. Fill the syringe with sol-
vent containing the substrate and connect it with the inlet tubing. En-
sure that the tubing is filled with liquid and does not contain any bub-
bles.
2. Connect the tubing with the cell. The outlet tubing and REF electrode
should not be connected yet. Hold the cell tilted (45° angle) and fill it
until the REF chamber of cell is filled.
3. Mount the REF electrode and remove excess of the liquid around it.
Push the solution firmly through the outlet port. You may see some air
escaping from the outlet.
4. Place the syringe in the infusion pump and start the flow.
For successful installation it is of utmost importance to install the
reactor cell bubble free.