Ionopticks Aurora series User manual

Recommendedguidelines for optimalsetupand

operation of Aurora Series columns:

IonOpticks accessories featured in these guidelines:

User Guide

Aurora Series™

UHPLCpacked emitter columns

Aurora Frontier Aurora Rapid150

AUR3-60075C18 AUR3-50150C18

AUR3-60075C18-CSI AUR3-50150C18-CSI

AUR3-60075C18-TS

Aurora Ultimate Aurora Rapid75

AUR3-25075C18 AUR3-5075C18

AUR3-25075C18-CSI AUR3-5075C18-CSI

AUR3-25075C18-TS

Aurora Elite

AUR3-15075C18

AUR3-15075C18-CSI

AUR3-15075C18-TS

IonOpticks 5cm Rapid Column Housing (5CMNFSH1)

IonOpticks TS Nanospray Flex Adapter (TSFLXAD01)

IonOpticks Column Heater (COLHTR01)

IonOpticks Heater Controller (IOHEATCON1)

IonOpticks Column Heater Extension Cable (EXTCABLE03)

Index.

Product Features 3

Compatibility 3

Video installationguides 4

Fitting Aurora Series columns to your UHPLC system

For Bruker CaptiveSpraysource 6

Using Bruker Column Toaster 7

Using Sonation Column Oven (PRSO-V1 and PRSO-V2) 8

For ThermoScientific Nanospray Flex source 9

with Sonation Column Oven (PRSO-V1 and PRSO-V2) 9

with an IonOpticks 5cm Rapid Column Housing (5CMNFSH1)

with an IonOpticks TS Nanospray Flex Adapter (TSFLXAD01) 11

For ThermoScientific EasySpray Source 13

with a TS column into an EasySpray source (ES081 and ES082) 13

Setting up the IonOpticks Heater and Heater Controller (COLHTR01 and IOHEATCON1) 5

Operationof Aurora Series columns 14

Initial operation 14

Standbyand idle conditions 14

Removal from a UHPLC 14

Recommended buffercompositions 14

Column volumes 14

Column temperature 14

Column equilibration 14

Sample loading 14

Example data 14

Recommended gradients 15

DISCLAIMER: The use of “we” “us” or “our” in this User Guide are references to IonOpticks Pty Ltd ACN 621 674 459. The information in this User

Guide including (without limitation) the recommendations, safety guidelines and product warnings with respect to the use of our products are to be read

strictly subject to our terms and conditions which can be located at www.ionopticks.com and the limitations and exclusions of our liability found within

those terms. Products should not be used if they appear damaged. We accept no liability for any loss or damage howsoever or wherever arising (including

death and/or personal injury)which results from or is connected with the failure by the customer to use our products strictlyin accordance with the

directions in this User Guide.

Spray voltage

Operating environmental conditions 14

Innovative Design.

Transforming Proteomics.

Our columns are differentiated by two key technological advances:

a unique packed emitter design that enables maximum mobile

phase velocity with no post-column dead volume; and our own

nanoZero® technology that provides user friendly ‘plug and play’

connections with true zero pre-column dead volume. Together,

these features combine to maximise chromatographic efficiency

and dramatically enhance performance, providing a best in class

solution for peptide and metabolite LC-MS separations.

Product Features

Integrated emitter with zero post-column dead volume.

Pre-fitted with nanoZero® to provide a zero-dead-volume

female union between a nanoViper™ Fingertight fitting (or

equivalent) and the Aurora series column.

nanoZero® fitting incorporates a 10-32 internal thread for

easy installation.

Designed to withstand ultra-high-performance LC

(UHPLC) backpressures of >1700 bar.

nanoZero® is electrically conductive to allow a voltage

connection to the entrance of the column.

Performance guarantee

Our columns are subjected to rigorous quality control procedures under

the direct eye of our Senior Scientists. All IonOpticks products are

covered by our 100% performance guarantee. Any item not meeting our

high-performance expectations due to manufacturing defects will be

replaced without charge to the customer.

Please contact us at [email protected] if you have any concerns

relating to your column. Our terms and conditions are provided with

every quote, but we want to make sure you are receiving a quality

product every time so please do not hesitate to get in touch with any

feedback or concerns.

Compatibility.

Aurora columns are compatible with a wide range of LC-MS

systems. Compatibility is not limited to equipment presented in

this guide. For further information about instrument

compatibility, please visit helpcentre.ionopticks.com

Product Specifications.

Column format:Analytical column

Pore size120Å

Particle size 1.7μm

Column type:Reversed-phase

Max pressure >1700 bar

pH stability 1-8

For use with:UHPLC

Temp limits60oC (lowpH)

Stationary phaseC18

Mass Spectrometer Ion Sources

•Thermo Scientific EasySpray

Thermo Scientific Nanospray Flex

Bruker CaptiveSpray

•

UHPLC

•Bruker nanoElute

•Bruker nanoElute 2

•Evosep One (Whisper methods only)

◦Aurora Elite; 20SPD, 40SPD

◦Aurora Rapid75; 80SPD

•Thermo Scientific Dionex UHPLC systems

•Thermo Scientific Easy-nLC 1000 / 1200

•Thermo Scientific Vanquish Neo

•Waters M-Class

•Waters nanoAcquity

Fittings

•IDEX MarvelX

•Thermo Scientific nanoViper

•Waters ZenFit

Column Ovens

•IonOpticks Column Heater

•Bruker Column Toaster

•Sonation Column Oven

Spray voltage

Video installation guides.

Access to online video content

We’ve created a series of online tutorials for easy setup, information

on compatibility and useful tips.

To watch installation videos specific to your laboratory setup, visit

helpcentre.ionopticks.com and search for “Installation Guides”.

For all other online content, find us on social media @IonOpticks,

or see our videos on YouTube at www.youtube.com/@ionopticks

Regulatory Compliance

IonOpticks evaluates its products to ensure full compliance with applicable North American and European regulations. The IonOpticks columns, used in conjunction

with the column heater, heater controller and other accessories, are intended for use in the basic electromagnetic environment as defined in IEC 61326-1. For detailed

information on emission level and permissible performance under the electromagnetic immunity conditions, please contact IonOpticks.

IonOpticks products are compliant with the Restriction of Hazardous Substances (RoHS) directive. This equipment has been tested and found to comply with the limits

for a Class A digital device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the

equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in

accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause

harmful interference in which case the user will be required to correct the interference at their own expense.

Changes or modifications to these products not expressly approved by the party responsible for compliance could void the user's authority to operate the equipment.

Setting up the IonOpticks heater

andheater controller. (COLHTR01 and IOHEATCON1)

Plug heater cable into channel 1 of Heater Controller unit.

2Ensure settings on the heater controller unit

match these found here:

3Set heater to desired temperature. We recommend using a

temperature between 40-60°C. The maximum recommended

temperature is 60°C.

4The heater is now installed and ready for use.

1.1

If using an IonOpticks Heater Extension Cable (EXTCABLE03):

Plug extension cable into channel 1 of IonOpticks Heater Controller

unit. Plug cable from IonOpticks heater into the receiving plug on

the Heater Extension Cable

Note: Only a single extension cable can be used with the IonOpticks

Heater. The use of multiple extension cables may cause a failure in the

operation of the heater and could damage the IonOpticks Heater and

IonOpticks Heater Controller.

1.2

This device complies with part 15 of the FCC Rules.

Operation is subject to the following two conditions:

(1) This device may not cause harmful interference,

and (2) this device must accept any interference

received, including interference that may caused

undesired operation.

24 V, 0.8A, 20 W

Heater Rating

For Bruker CaptiveSpraysource.

Disassemble rear metal housing

of CSI probe insert (2 screws)

Carefully remove Aurora CSI protective

cap.

Insert Aurora CSI fitting into CSI probe.

Insert using a twisting motion to prevent

pinching of rubber seal.

Remove the protective plug from the

nanoZero® union.

Unscrew CSI probe from CSI housing

Feed Aurora column through the rear metal housing and reassemble CSI probe (2 screws),

ensuring rubber seal is in place between metal housing and Aurora CSI fitting.

Feed column through CSI housing and screw CSI probe insert into housing.

Feed Aurora column with nanoZero®

fitting through CSI probe housing.

The CSI housing is now ready to connect

to your LC-MS system.

3 4 5

66.1 7

88.1 9

For Bruker CaptiveSpraysource

Retract the column heater from

the CaptiveSpray source

Insert the CSI Housing into the

source, press with a small

amount of pressure and tighten

the screw until finger tight.

Using a gloved finger, block the

air inlet and monitor the Fore

pressure. The Fore vacuum

needs to drop to below 3e-01

mbar within 10 seconds to be

considered usable.

If not sealed, try refitting before replacing

rubber seals (Refer Bruker CSI

troubleshooting guide).

Move the column heater in front

of the CaptiveSpray source.

10 11 12 13

16 17

15.2

Place column directly into the heater recess. Hold in position by

placing the holding arm onto the nanoZero®. This requires a

replacement metal holding arm (supplied by IonOpticks) to allow an

earth connection with the nanoZero®.

For Aurora Rapid75 and Aurora Rapid150 columns

Curl the column inside the

heating plate and hold in

position by placing under

the holding arm.

15.1

Tighten the earth screw onto the

nanoZero® to ensure that the

nanoZero® is grounded and held

in position.

For Aurora Frontier, Aurora Ultimate and Aurora Elite columns

Place lid on heater

Holding the nanoZero® with a

spanner, tighten the nanoViper

fitting finger tight until you reach

a firm stop.

Install filter into the CSI housing

air inlet.

if usingBruker Column Toaster

The column is now

installed in the heater

and ready for

operation.

15.2

Place the earth cable clip over the thin section of the nanoZero® and

slide over the hexagonal section to make a firm connection. If the clip

feels loose, remove the clip, pinch in the clip arms and repeat the

process.

Earthing

For Bruker CaptiveSpraysource

if using Sonation Column Oven (PRSO-V1and PRSO-V2)

Retract the column heater from

the CaptiveSpray source

Using a gloved finger, block the

air inlet and monitor the Fore

pressure. The Fore vacuum

needs to drop to below 3e-01

mbar within 10 seconds to be

considered usable.

If not sealed, try refitting before replacing

rubber seals (Refer Bruker CSI

troubleshooting guide).

Move the column heater in front

of the CaptiveSpray source.

10 11 12 13

Closing the heater

Holding the nanoZero® with a

spanner, tighten the nanoViper

fitting finger tight until you reach

a firm stop.

Curl the column inside the

heating plate and place the

nanoZero® and earth clip into

the plastic holder.

Press the CSI Housing with a

small amount of pressure and

tighten the screw until finger

tight.

Place top plate on heater and secure in position using the press pins.

Be careful to avoid pinching the column during this process.

Install filter into the CSI housing

air inlet.

The column is now

installed in the heater

and ready for

operation.

For Thermo Scientific NanosprayFlex

Curl the column inside the

heating plate.

1 3

Carefully slide protective sheath backwards towards the nanoZero®

fitting to expose the emitter tip. Press the holding clamp on the heater

open and place the column into the heater. The column emitter should

extend 15-20mm beyond the heating plate.

Preparing the column

Holding the nanoZero® with a

spanner, tighten the nanoViper

fitting finger tight until you reach

a firm stop.

with Sonation Column Oven (PRSO-V1and PRSO-V2)

You will need:

1 x HVCABLE01

for connection of the

source high-voltage to

the nanoZero® fitting.

Connect the HVCABLE01 to the

nanoZero as shown.

Place top plate on heater and secure in position using the press pins.

Be careful to avoid pinching the column during this process.

Ensure that the stage is fully

retracted in the z-axis before

moving the source towards

the mass spectrometer.

Slide the source into position

and adjust the x, y and z axis

on the stage to ensure a good

emitter position. The emitter

should be 3-5mm from the

ion transfer capillary.

7The column is now