CombiFlash NextGen ®

Installation and Operation Guide

Part Number 69-5253-032

Copyright © 2020 Teledyne Technologies Incorporated. All rights reserved.
Rev. C, Dec 2020

Use and Disclosure of Data Information contained herein is classified as EAR99 under the U.S. Export Administration Regulations.
Export, reexport or diversion contrary to U.S. law is prohibited.
 CombiFlash NextGen Installation and Operation Guide
Safety Warnings

CombiFlash NextGen
Installation and Operation Guide
Safety Warnings

Safety Before installing, operating, or maintaining this equipment, it is

imperative that all hazards and preventive measures are fully
understood. While specific hazards may vary according to
location and application, take heed in the following general
warnings:

WARNING
Liquids associated with this instrument may be classified
as carcinogenic, biohazard, flammable, or radioactive.
Should these liquids be used, it is highly recommended that
this application be accomplished in an isolated
environment designed for these types of materials in
accordance with federal, state, and local regulatory laws,
and in compliance with your company’s chemical/hygiene
plan in the event of a spill.

WARNING
Avoid hazardous practices! If you use this instrument in
any way not specified in this manual, the protection
provided by the instrument may be impaired.

WARNING
If you are using flammable solvents or chemicals with this
system, vapor concentration levels may exceed the
maximum exposure levels as recommended by OSHA
Guide 1910.1000. To reduce those levels to a safe exposure,
Teledyne ISCO recommends that you place the system in
a laboratory hood designed for the purpose of ventilation.
This hood should be constructed and operated in
accordance with federal state and local regulations. In the
event of a solvent or chemical spill, your organization
should have a plan to deal with these mishaps. In all cases,
use good laboratory practices and standard safety
procedures.

Hazard Severity Levels This manual applies Hazard Severity Levels to the safety alerts.
These three levels are described in the sample alerts below.

CAUTION
Cautions identify a potential hazard, which if not avoided, may
result in minor or moderate injury. This category can also warn
you of unsafe practices, or conditions that may cause property
damage.

WARNING
Warnings identify a potentially hazardous condition, which
if not avoided, could result in death or serious injury.

3
CombiFlash NextGen Installation and Operation Guide
Safety Warnings

DANGER
DANGER – limited to the most extreme situations
to identify an imminent hazard, which if not
avoided, will result in death or serious injury.

Hazard Symbols The equipment and this manual use symbols used to warn of
hazards. The symbols are explained in the table below.

Hazard Symbols
Warnings and Cautions

The exclamation point within the triangle is a

warning sign alerting you of important instructions
in the instrument’s technical reference manual.

The lightning flash and arrowhead within the trian-

gle is a warning sign alerting you of “dangerous
voltage” inside the product.

Symboles de sécurité

Ce symbole signale l’existence d’instructions

importantes relatives au produit dans ce manuel.

Ce symbole signale la présence d’un danger

d’électocution.

Warnungen und Vorsichtshinweise

Das Ausrufezeichen in Dreieck ist ein Warn-

zeichen, das Sie darauf aufmerksam macht, daß
wichtige Anleitungen zu diesem Handbuch
gehören.

Der gepfeilte Blitz im Dreieck ist ein Warnzeichen,

das Sei vor “gefährlichen Spannungen” im
Inneren des Produkts warnt.

Advertencias y Precauciones

Esta señal le advierte sobre la importancia de las

instrucciones del manual que acompañan a este
producto.

Esta señal alerta sobre la presencia de alto

voltaje en el interior del producto.

4
 CombiFlash NextGen Installation and Operation Guide
Safety Warnings

For Additional Technical assistance for the Teledyne ISCO CombiFlash

Information NextGen can be obtained from:
Teledyne ISCO
4700 Superior St.
Lincoln NE 68504
Phone: (800) 775-2965 or (402) 853-5340
Fax: (402) 465-3001
E-mail: [email protected]

5
CombiFlash NextGen Installation and Operation Guide
Safety Warnings

6
 CombiFlash NextGen Systems
Table of Contents

Section 1 Introduction
1.1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
1.2 Product Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
1.3 Operating Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-6
1.3.1 System Control Possibilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-6
1.3.2 Data Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-6
1.4 Specification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-6
1.5 Controls, Indicators, and Features of the CombiFlash NextGen . . . . . . . . . . . . . . 1-9
1.6 Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-13
1.6.1 Hazard Severity Levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-14
1.6.2 Hazard Symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-14
1.6.3 Additional Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-15
1.6.4 System Pressure Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-15
1.6.5 Pressure Limits . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16
1.6.6 Overpressure Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-17

Section 2 Preparation
2.1 Unpacking the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
2.2 Instrument Location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
2.3 Connect Power . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
2.4 Plumbing Connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
2.5 External Gas . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-5
2.5.1 CombiFlash NextGen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-5
2.5.2 CombiFlash NextGen Equipped with Optional ELSD . . . . . . . . . . . . . . . . 2-6
2.6 Connect and Route Drain Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-6
2.7 Vapor Enclosure (Optional) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-9
2.8 Install Solid Load Cartridge Cap Ring Support and SLCC Storage Bracket (Optional)
2-10
2.9 Position the System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-11
2.10 Install Collection Tube Racks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-12
2.11 Powering on the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-14
2.12 Configure the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-14
2.12.1 Solvents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-15
2.12.2 General Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-16
2.13 Vapor Limit (if equipped) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-17
2.13.1 Set Default Tube Volumes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-18
2.13.2 User Preferences . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-18
2.13.3 User Management . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-20
2.13.4 Restart the System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-21
2.14 Prime the Solvent Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-21
2.15 System Verification . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-23
2.16 Installation Qualification Checklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-24

Section 3 Operation
3.1 Flash Sample Preparation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1
3.1.1 Liquid Sample Injection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

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CombiFlash NextGen Systems
Table of Contents

3.1.2 Solid Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

3.2 Loading a RediSep Rf Column . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2
3.3 Starting a Separation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-2
3.4 During the Separation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6
3.4.1 Run Control Buttons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-6
3.4.2 Bypassing the Solid Load Cartridge (Injection valve equipped systems only)
3-7
3.4.3 About Solvent Level Detection (systems equipped with this option only) . 3-7
3.5 Software Reference . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-8
3.5.1 Main Separation Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-8
3.5.2 Method Editor Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-10
3.5.3 Edit Gradient Options . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-10
3.5.4 Peak Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-12
3.5.5 Tools . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-16
3.5.6 Results Screen . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-17
3.5.7 PurIon Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-18

Section 4 Network Configuration

4.1 Network Configurations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1
4.1.1 Network PC Access . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2
4.1.2 Network Printing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2
4.1.3 Network File Save Configuration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2
4.2 Direct Connection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-2

Section 5 Maintenance
5.1 System Standby and Shut Down . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
5.2 Preventive Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-1
5.3 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-2
5.3.1 Collection Rack and Tray Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-2
5.4 Tubing Inspection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-3
5.5 Flow Cell Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-4
5.5.1 Post Separation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-4
5.5.2 Quick Cleaning when Recommended . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-5
5.5.3 Monthly Flow Cell Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-7
5.6 ELSD Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-7
5.6.1 Cleaning the ELSD Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-7
5.7 PurIon Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-10
5.7.1 Removing ESI Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-10
5.7.2 ESI Probe Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-11
5.7.3 APCI Probe Removal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-11
5.7.4 APCI Probe Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-12
5.7.5 Removal of Ion Source Housing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-12
5.7.6 ESI and APCI Removal from PurIon S and PurIon L . . . . . . . . . . . . . . . . 5-13
5.7.7 ESI and APCI Replacement from PurIon S and PurIon L . . . . . . . . . . . . 5-13
5.7.8 Cleaning the Ionization Source Capillary . . . . . . . . . . . . . . . . . . . . . . . . . 5-14
5.7.9 Replacement of Ion Source Housing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-14
5.7.10 Overpressure Error . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-14
5.7.11 Check Valve Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-15
5.7.12 Replacing Check Valves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-16
5.8 Electrical Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5-16

2
 CombiFlash NextGen Systems
Section 1 Introduction

1.1 Overview This Installation Guide provides:

• Safety information
• Unpacking instructions
• Installation instructions, including connections with
Windows and iOS operating systems
• Basic user operation
• Software overview
• Certification and warranty information

1.2 Product Overview The Teledyne ISCO CombiFlash NextGen flash chromatography
system gives you high-productivity automation, programmable
gradients, UV detection and peak separation, and automatic
detection of columns and collection tube racks (system
dependent). Its small size makes it a great “personal system” and
well-suited for operation within chemical hoods and other limited
indoor spaces. An optional fraction collection enclosure allows
usage on an open bench in some environments.

WARNING
Avoid hazardous practices! If you use this instrument in
any way not specified in this manual, the protection
provided by the instrument may be impaired; this may
increase your risk of injury.

1-1
CombiFlash NextGen Systems
Section 1 Introduction

The CombiFlash NextGen systems are available in different

configurations: NextGen, NextGen 300, and NextGen 300+.
CombiFlash NextGen – For general purpose Flash chromatog-
raphy needs, this configuration delivers flow rates from 1 to 100
mL/min with a maximum pressure limit of 150 psi. The system
delivers solvent gradients formed from the 2 solvent inlet lines.
This configuration has an easily accessible fluid path that sup-
ports liquid sample introduction, and a variable wavelength PDA
UV detector with optional UV-vis, Evaporative Light Scattering
Detection (ELSD) or PurIon Mass Spectrometer detection.

Figure 1-1 CombiFlash NextGen with 12 inch display, 1-100 mL/min, 150 psi (without solid load
injection valve)

1-2
 CombiFlash NextGen Systems
Section 1 Introduction

CombiFlash NextGen 300 – For more advanced needs, the

CombiFlash NextGen is a 1–300 mL/min, 300 psi pressure limit
(with injection valve option, otherwise 150 psi limit) system that
forms a binary gradient by selecting two solvents from four inlets
and also allows for the addition of a third solvent modifier. The
NextGen 300 offers the choice of UV (200–400 nm) or full
spectrum UV-vis (200–800 nm) detection along with optional
ELSD or PurIon Mass Spectrometer detection. This configu-
ration allows for the choice of optional features including an
automatic, self-cleaning injection valve for sample introduction,
RFID support for column and fraction collection rack identifi-
cation, and an internal air pump is used for post-run column air
purging, active solvent level sensing, and waste full bottle
detection. This model offers either a 12 inch or 15 inch touch-
screen display.

Figure 1-2 CombiFlash NextGen 300 system with 12-inch display, 1–300 mL/min, 300 psi (with
automatic injection valve)

1-3
CombiFlash NextGen Systems
Section 1 Introduction

CombiFlash NextGen 300+ – For more options, the Combi-

Flash NextGen is a 1–300 mL/min, 300 psi pressure limit system
that forms a binary gradient by selecting two solvents from four
inlets and also allows for the addition of a third solvent modifier.
This configuration has an automatic, self-cleaning injection valve
for sample introduction, RFID support for column and fraction
collection rack identification, an internal air pump for post-run
column air purging, active solvent level sensing, and waste full
bottle detection and the choice of either UV (200–400 nm) or full
spectrum UV-vis (200–800 nm) detection along with optional
ELSD or PurIon Mass Spectrometer detection. This model offers
either a 12 inch or 15 inch touchscreen display.

Figure 1-3 CombiFlash NextGen 300+ system with 15-inch display, 1–300 mL/min, 300 psi

1-4
 CombiFlash NextGen Systems
Section 1 Introduction

CombiFlash PurIon S or L – This optional mass spectrometer

detector has a detection range of 10 to 1200 Daltons (Da) (PurIon
S) or 10-2000 Da (PurIon L). During purification, this detector
can be combined with the UV or UV-vis detector to isolate UV
and visible light absorbing compounds, as well as compounds
with specific molecular weights or mass ranges.
All CombiFlash NextGen configuration are optimized for use
with Teledyne ISCO’s RediSep® Rf or RediSep Rf Gold® columns,
which are pre-packed with a variety of media. For example, the
CombiFlash NextGen can purify samples from 4 milligrams to
more than 33 grams using columns packed with 4 to 330 grams
of silica gel. Larger columns, up to 3 kg, can be used with an
optional external column mount (PN: 60-5394-551).
Applications include purification of organic compounds for drug
discovery, as well as research in agrochemicals, petrochemicals,
natural products, polymers, and catalysts.

Figure 1-4 CombiFlash NextGen 300+ with 15-inch display and PurIon

1-5
CombiFlash NextGen Systems
Section 1 Introduction

1.3 Operating Overview

1.3.1 System Control The CombiFlash NextGen system is equipped with a capacitive
Possibilities touch screen display for local control.
The system supports also TCP/IP communication to allow direct
or network control of the system by a computer using Internet
Explorer (IE10 or newer).

Note
Teledyne ISCO recommends that you obtain assistance from
your Information Technology department before attempting
direct or network connections

The system can be accessed from the built-in touch panel and up
to ten network devices. The touch panel shares control with all
connected devices. The system performs the most recent
command from any control input.

1.3.2 Data Storage To support operation from a variety of direct and network con-
nections, the software and all files are stored in the CombiFlash
NextGen on an internal hard drive. This ensures that your com-
pound purification methods and run history files can be viewed
from any connection. Optionally, run files may be saved to a USB
flash drive, a networked controlling computer, or a network
drive.

1.4 Specification
Table 1-1 CombiFlash NextGen System Specificationsa
NextGen Systems NextGen 300/300+ Systems
Overall Dimensions 66 x 36 x 43 cm (26 x 14.1 x 17")
(H WD)
Weight 27.7 kg (61 lbs) 27.7 kg (61 lbs)
33.6 kg (74 lbs) with Optional ELSD 33.6 kg (74 lbs) with Optional ELSD
Power Options Input voltage range from 100 to 240 VAC, 50/60 Hz, 2.0 Amps maximum.
Line cord is the disconnect device.
Line Frequency 50/60 Hz
Ambient Temperature 20 to 40 °C (maximum temperature must be at least 15 °C below the boiling point of the
solvent)
Humidity (when connected 95% relative humidity maximum at 20 to 40 °C
to power)
Flow Rate Range 1 to 100 mL/min 1 to 300 mL/min
Flow Rate Accuracy (tested ±10% ±5% (5-200 mL/min)
with water at 138 kPa or 20
psi)
Pressure Limit b c d e 1035 kPa (150 psi) Up to 2070 kPa (300 psi)
Up to 1035 kPa (150 psi) (if not equipped
with injection valve)

1-6
 CombiFlash NextGen Systems
Section 1 Introduction

Table 1-1 CombiFlash NextGen System Specificationsa

Pressure Display Accuracy 5% of full scale 5% of full scale
Gradient Formation Binary gradient from 1 and 2 solvent inlets. Binary gradient from 1, 2, and 3, 4 solvent
inlets. Solvent selection can be
automatically controlled by the method.
Gradient Accuracy ±2% of full scale, typical; ±2% of full scale
±5% maximum
Peak Detection Modes Slope or threshold
Flow Cell Pathlength 0.1 mm, ±25%
UV Detection Wavelength 200 to 400 nm, optional 200 to 800 nm UV-vis
Wavelength Accuracy ±1 nm
Fraction Accuracy ± [2mL + (flow rate  60)]
Optional ELSD
Gas Inlet Pressure 60 to 70 psig (414 to 483 kPa)
Gas Consumption <2.5 SLPM
Spray Chamber Setting range: 10 to 60 °C
Temperature Minimum guaranteed temperature is 5 °C below ambient
Drift Tube Temperature Setting range: 30 to 90 °C
Must be 5 °C above spray chamber temperature
Maximum temperature is 60 °C above ambient
Split Flow Rate 0.75 mL/min, ±10%
Flow Rate Range Minimum flow rate 5 ml/min. Lower flow rates will result in excessive detector signal
delay.
Electrical Safety per EN 61010-1
Pollution Degree 2
Installation Category II
Maximum Altitude 2000 meters

a. All specifications are subject to change.

b. For safety, maximum pressure limits are limited by each columns RFID-defined pressure limit. If the column is not RFID
recognized see section 1.6.5. Maximum pressure limit is also set at 200 psi for methods using a solid-load cartridge.
c. Solvent supply containers can be located up to 0.9 m below CombiFlash NextGen systems except when using
dichloromethane. Dichloromethane containers must be at the same level as the NextGen or above.
d. The syringe pumps on the NextGen 300 have a pressure limit of 300 PSI up to 200 ml/min. Above 200 ml/min the pressure
limit drops linearly to 250 PSI. Keep in mind that other factors such as the flow path and column size may cause the max-
imum pressure limit to be reduced.
e. NextGen systems may also exhibit excessive pressure at maximum flow rates with some viscous solvents due to the tub-
ing diameter used in the system.

1-7
CombiFlash NextGen Systems
Section 1 Introduction

Table 1-2 CombiFlash NextGen with PurIon Systema b c

Dimensions (H x W x D) Mass Spectrometer 26 x 11 x 22 in (66 x 28 x 56 cm)
Roughing Pump 10 x 9 x 18 in (26 x 23 x 46 cm)
10 – 1200 Dalton, 1 Dalton Resolution
Mass Spectrometry Detection Electrospray Ionization (ESI) or Atmospheric Pressure Chemical
Ionization (APCI)
Positive or Negative Ionization Mode
Gas Consumption 4 SLPM
Split flow rates 1.47, 6, and 20 µl/min

a. All specifications are subject to change.

b. For columns undetected by RFID, the max pressure is limited to 50 psi.
c. NextGen 300 or 300+ systems with PurIon detector are not be capable of operation at flow rates greater than 200 ml/min
with some solvent systems due to excessive back pressure in the flow splitter valve.

Table 1-3 Component Materials List

Teledyne ISCO RediSep Rf Cartridge Virgin Polypropylene, silica-based media (alumina columns also available),
polyethylene (HDPE) frits
Process Tubing Fluoropolymer, Carbon impregnated fluoropolymer tubing and 316 stainless
steel tubing
Drain Tubing Vinyl with FEP liner
Process Valves PEEK, FFKM (NextGen 300, 300+) (NextGen includes PTFE)
Sample Loading Cap 316 stainless steel
Sample Loading Cap Seal PTFE, Elgiloy®a
Injection Valve PPS, 316 Stainless Steel
Flow Cell 303 SST, Type ES Quartz, FFKM (SIMRIZ SZ485)
Pump 316 stainless steel, PTFE, Elgiloy®

a. Elgiloy is a registered trademark of Combined Metals of Chicago, L.L.C.

1-8
 CombiFlash NextGen Systems
Section 1 Introduction

1.5 Controls, Indicators, Figures 1-5 through 1-8 illustrate the controls on the
and Features of the CombiFlash NextGen.
CombiFlash NextGen

11 2

1
8
5

7 12

3
6

10

Figure 1-5 CombiFlash NextGen features (front)

1. Touch Panel LCD display – Large 12 or 15 inch diagonal

High Definition display for system monitoring and control.
2. USB Port – Convenient, front panel port that accepts USB
Flash memory drive. A Flash drive may be inserted into
this port for transferring files, importing and exporting
methods, and performing system software updates.
3. Adjustable Flash Column Mount – The injection valve
assembly and upper column mount slide along this mount
so the system can accept a variety of column sizes.
4. On/Standby Switch – push button ON/STANDBY. Press
and hold for system shut down.
5. Fraction Collector Arm and Drop Former – The arm
and drop former move to deposit liquid in the collection
tubes.
6. Racks and Collection Tubes – Racks hold the fraction
collection tubes for all CombiFlash systems. Racks
include an RFID tag which the system uses to read the
rack type and collection tube size. The NextGen (basic)
cannot read the RFID tag.
7. Sample Injection Port – Luer-type fitting to accept the
sample though either a solid load sample cartridge (shown)
or a liquid injection using a syringe or similar device.
8. Solid Load Cartridge Cap (SLCC) Ring Support–
Supports the Solid Load Cartridge (optional).

1-9
CombiFlash NextGen Systems
Section 1 Introduction

9. SLCC Storage Bracket – Stores the solid load cartridge

cap (optional) when not in use.
10. Lower Flash Column Mount – Secures the column.
11. Top Shelf – Allows storage of accessories and solvent bot-
tles.
12. Vapor Enclosure – Encloses the fraction collection area,
allowing for hook up to external exhaust for use outside of
hood (optional).

Figure 1-6 CombiFlash NextGen features (sides)

1-10
 CombiFlash NextGen Systems
Section 1 Introduction

13
15
15
16
14 16

21

24
19

23

18

17
22 20 25

Figure 1-7 CombiFlash NextGen Features (Back)

13. Upper Drain Tube – Liquids spilled on the top shelf are
carried away through this tube to a user-supplied
container.
14. Ethernet Port – An 8P8C jack for a network connection
using a standard CAT5 cable, or for a direct connection to a
computer.
15. USB Port – For control of peripheral equipment such as
the PurIon mass spectrometer detector. This can also be
used for an external keyboard, or mouse.
16. Cooling Fan – Cools the internal electronic assemblies.

WARNING
Do not touch the fan.

17. Solvent and Waste Level Sensing Outlet Ports

(optional on some systems) – uses air or user-supplied
gas to measure the hydrostatic pressure of the solvent
above the weighted tubing outlet.
18. Solvent and Waste Level Ports (optional on some sys-
tems)- Supports use of sealed or almost sealed supply bot-
tle caps while maintaining solvent level sensing capability

1-11
CombiFlash NextGen Systems
Section 1 Introduction

19. Solvent Inlet Ports- A and B solvent supply. The

NextGen system allows only a single solvent each for A
and B (weak and strong solvent), while the remaining mod-
els support up to 4 solvents which can be used as either the
weak or strong solvent.
20. Optional External Gas Port – An inlet for an internal
pump which supplies pressurized air or user-supplied gas
for solvent level sensing, waste container full detection,
and system/column purges. The external gas should be
regulated to 2–5 psig before connecting the source to this
port. The port is present only on system equipped with sol-
vent level sensing.
21. Fraction Collection Exhaust Port – 2” port to connect
to vapor enclosure to an external exhaust.
22. Mains Power – Connects the system to AC line voltage.
23. Mains Circuit Breaker– This is a voltage independent
circuit breaker which also removes power from all active
components inside the instrument.
24. Waste Port – Fluid not directed to the fraction collector
exit to the waste container here.
25. Lower Drain Tube – This tube carries away liquids
spilled onto the rack area.
26. Cooling Fan – (ELSD equipped models only) Cools the
internal ELSD components.

31
28 26

29
30
27
31

Figure 1-8 NextGen ELSD (back) and P-Trap Drain Vent Assembly

27. ELSD Exhaust Port – (ELSD equipped models only)

Vents the carrier gas and vaporized solvents.
28. Nitrogen Inlet – (ELSD equipped models only) Connects
to the carrier gas supply (60 to 70 psi).
29. P-trap Drain – (ELSD equipped models only) During
operation, condensate from the Thermo-split™ process
drains from the system. An internal P-trap prevents aero-
sol particles from escaping through this drain.

1-12
 CombiFlash NextGen Systems
Section 1 Introduction

30. Pump Drain Outlet – (ELSD equipped models only) An

internal drip tray is located below the splitter for the
ELSD. Should the pump or its fittings leak, liquids will
drain from this port to protect internal components.
31. P-Trap Drain Vent Assembly – Fixture to allow P-trap
to drain. Vent portion should be aligned vertically.

CAUTION
Discontinue use of the CombiFlash NextGen if liquid is present
at the Pump Drain. Contact Teledyne ISCO technical service
for assistance with correcting the leak.

1.6 Safety Before installing, operating, or maintaining this equipment, it is

imperative that all hazards and preventive measures are fully
understood. While specific hazards may vary according to
location and application, take heed in the following general
warnings:

WARNING
Avoid hazardous practices! If you use this instrument in
any way not specified in this manual, the protection
provided by the instrument may be impaired.

WARNING
Liquids associated with this instrument may be classified
as carcinogenic, biohazard, flammable, or radioactive.
Should these liquids be used, it is highly recommended that
this application be accomplished in an isolated
environment designed for these types of materials in
accordance with federal, state, and local regulatory laws,
and in compliance with your company’s chemical/hygiene
plan in the event of a spill.

WARNING
If you are using flammable solvents or chemicals with this
system, vapor concentration levels may exceed the
maximum exposure levels as recommended by OSHA
Guide 1910.1000. To reduce those levels to a safe exposure,
Teledyne ISCO recommends that you place the system in
a laboratory hood designed for the purpose of ventilation.
This hood should be constructed and operated in
accordance with federal state and local regulations. In the
event of a solvent or chemical spill, your organization
should have a plan to deal with these mishaps. In all cases,
use good laboratory practices and standard safety
procedures.

1-13
CombiFlash NextGen Systems
Section 1 Introduction

WARNING
The CombiFlash NextGen has redundant safety devices to limit
pressure to less than 300 psi (2068 kPa), 150 psi (1034 kPa) for
systems without an automated injection valve. RediSep columns
smaller than 100 g are CE certified using standard IEC61010-1 for
use on the CombiFlash NextGen. RediSep columns 100 g or larger.
100 g meet Pressure Vessel Directive 97/23/EC.

1.6.1 Hazard Severity Levels This manual applies Hazard Severity Levels to the safety alerts.
These three levels are described in the sample alerts below.

CAUTION
Cautions identify a potential hazard, which if not avoided, may
result in minor or moderate injury. This category can also warn
you of unsafe practices, or conditions that may cause property
damage.

WARNING
Warnings identify a potentially hazardous condition, which
if not avoided, could result in death or serious injury.

DANGER
DANGER – limited to the most extreme situations
to identify an imminent hazard, which if not
avoided, will result in death or serious injury.

1.6.2 Hazard Symbols The equipment and this manual use symbols used to warn of
hazards. The symbols are explained in Table 1-4.

Table 1-4 Hazard Symbols

Warnings and Cautions

The exclamation point within the triangle is a

warning sign alerting you of important instructions
in the instrument’s technical reference manual.

The lightning flash and arrowhead within the trian-

gle is a warning sign alerting you of “dangerous
voltage” inside the product.

The pinch point symbol warns you that your fin-

gers or hands will be seriously injured if you place
them between the moving parts of the mechanism
near these symbols.

Symboles de sécurité

1-14
 CombiFlash NextGen Systems
Section 1 Introduction

Table 1-4 Hazard Symbols (Continued)

Ce symbole signale l’existence d’instructions
importantes relatives au produit dans ce manuel.

Ce symbole signale la présence d’un danger

d’électocution.

Risque de pincement. Ces symboles vous avertit

que les mains ou les doigts seront blessés séri-
eusement si vous les mettez entre les éléments
en mouvement du mécanisme près de ces sym-
boles.

Warnungen und Vorsichtshinweise

Das Ausrufezeichen in Dreieck ist ein Warn-

zeichen, das Sie darauf aufmerksam macht, daß
wichtige Anleitungen zu diesem Handbuch
gehören.

Der gepfeilte Blitz im Dreieck ist ein Warnzeichen,

das Sei vor “gefährlichen Spannungen” im
Inneren des Produkts warnt.

Vorsicht Quetschgefahr! Dieses Symbol warnt vor

einer unmittelbar drohenden Verletzungsgefahr für
Finger und Hände, wenn diese zwischen die
beweglichenTeile des gekennzeichneten Gerätes
geraten.

Advertencias y Precauciones

Esta señal le advierte sobre la importancia de las

instrucciones del manual que acompañan a este
producto.

Esta señal alerta sobre la presencia de alto

voltaje en el interior del producto.

Punto del machacamiento. Sus dedos o manos

seriusly serán dañados si usted los coloca entre
las piezas móviles cerca de estos símbolos.

1.6.3 Additional Technical assistance for the CombiFlash NextGen system can be
Information obtained from:
Teledyne ISCO
4700 Superior St.
Lincoln NE 68504
Phone: (800) 775-2965 or (402) 464-0231
Fax: (402) 465-3001
E-mail: [email protected]

1.6.4 System Pressure The NextGen series flash systems use a variety of techniques to
Management ensure user safety due to potential compound crashes and the
resulting pressure increase. The systems include redundant

1-15
CombiFlash NextGen Systems
Section 1 Introduction

pressure measuring devices to maintain safe operation in the

event of component failure. If the two pressure readings aren’t
within 15 psi of each other, it would indicate that one of the
transducers may have failed. An error message is displayed and
system operation is halted until the problem is resolved.

In addition, if both transducers readings match perfectly and

exhibit no change with time, it is an indication that the system
may have failed to update the pressure reading, which is also a
fault and will result in an error message.

1.6.5 Pressure Limits Teledyne Isco RediSep® columns have the pressure limit of the
column coded onto an RFID (Radio Frequency IDentification) tag
embedded in the column label. The encoded pressure limit is half
the average burst pressure for the column size in use. This infor-
mation is read by the NextGen system (if equipped with the
RFID option) and used to establish the maximum operating con-
ditions for the separation.

Note
4g silica columns are an exception; they have no RFID tag and
the system assumes a pressure limit of 200 psi which is less
than half their average burst pressure.

If a column without a functional RFID tag is used, the NextGen

systems assume that if a 4g column is being used, the maximum
acceptable pressure is 200 psi (150 psi for the NextGen only
system). All other column sizes default to a 100 psi pressure
limit. These limits are set to allow safe operation with most com-
mercially available flash columns.
If the NextGen system owner is confident of the quality of
columns procured, they can override the default (non-RFID
tagged column) pressure limits.
To access this override:
1. Select the HELP drop down and choose the SERVICE screen
option.
2. Entering the SERVICE screen requires using the adminis-
trative password. The default password on all new systems
is “combiflash”.
3. In the SERVICE screens, select PUMPS tab and the
NON-RFID COLUMN PRESSURE LIMITS option to program
the desired pressure limits for each column size.
4. Exit the SERVICE screen and allow the system to reboot.

Note
To allow a system administrator to preserve these settings, the
system administrator can change the password from the
default under the TOOLS drop down.

1-16
 CombiFlash NextGen Systems
Section 1 Introduction

Note
If the administrator password is changed, it is highly recom-
mended that the new password be saved in a secure location.

Note
The pressure limit is 200 psi, if using the solid-load car-
tridge loading method.

1.6.6 Overpressure If the pressure transducers are properly functioning and an over-
Conditions pressure condition is detected, the system will respond
depending on the severity of overpressure.
• If the blockage is severe and the pressure reading is 75
psi over the pressure limit, the system stops immedi-
ately.
• If the pressure exceeds the limit by less than 75 psi the
system drops the flow rate to 50% of the programmed
flow rate.
• If after 5 seconds the limit is still exceeded, the flow is
reduced by another 50%.
· In either of these conditions, the system will continue
the separation.
• If the blockage resolves due to increased gradient
strength and the pressure drops below 30% of the
pressure limit, the flow rate will increase stepwise to the
original flow rate.
• If the two flow rate reductions of 50% are insufficient to
resolve an overpressure condition, the system will stop
the separation to allow the user to take steps to resolve
the blockage.

1-17
CombiFlash NextGen Systems
Section 1 Introduction

1-18
 CombiFlash NextGen Systems
Section 2 Preparation

This section provides instructions for unpacking and installing

the CombiFlash NextGen system. The PurIon Mass Spec-
trometer has supplementary instructions included for instal-
lation of the PurIon in sections 2.1 through 2.16.

Note
CombiFlash PurIon system installation is not covered by this
manual. Instead, refer to instruction sheet 60-5233-638 for the
installation procedures.

Note
Section 2.16 contains an Installation Qualification checklist. If
required, sign off the checklist entries as you successfully
complete the following sections.

2.1 Unpacking the System The CombiFlash NextGen system is shipped in a single carton.
Carefully unpack the shipment and inspect the contents.

WARNING
The system is heavy. Use a two-person lift to prevent injury.

CAUTION
Do not lift the system by the fraction collector arm. Use the lift-
ing handles located on the instrument side panels. The left
side has a flush mount shipping handle, while the right side
can be lifted by gripping the side wall above the fraction collec-
tion area.

If there is any damage to the shipping carton or any components,

contact the shipping agent and Teledyne ISCO (or its authorized
representative) immediately.

WARNING
If there is any evidence that the system has been damaged
in shipping, do not plug it into AC power. Contact Teledyne
ISCO or its authorized representative for advice.

2-1
CombiFlash NextGen Systems
Section 2 Preparation

Compare the contents of the boxes with the enclosed packing

slips. If there are any shortages, contact Teledyne ISCO immedi-
ately.
The fraction collector arm is stowed to prevent damage during
shipping.
1. Loosen the screw retaining the arm to the right side of the
fraction collection area (Figure 2-1).

Figure 2-1 Remove screw holding the arm in place

2. Pull off the front cover of the NextGen by gripping the

edges of the cover and pulling forward off the instrument.
3. Store the screw by screwing it into the screw storage.
(Fig 2-2).

Figure 2-2 Location of screw storage

4. Replace the front cover by pressing back onto the instru-

ment.
5. The screw should be reinstalled if the system must be
shipped again.

2-2
 CombiFlash NextGen Systems
Section 2 Preparation

2.2 Instrument Location The CombiFlash NextGen systems have a relatively small foot-
print, requiring about 1550 square centimeters (240 in2) of level
bench space. Ensure that the system has at least 3 cm (1.25 in) of
air space behind it for ventilation. Additional space may be
required for solvent and waste containers.
Refer to Table 1-1 for environmental conditions and power
requirements.

WARNING
The system is heavy. Use a two-person lift to prevent injury.

Before making any connections to the NextGen system, place it

on the bench or in the fume hood where it will be operated. Tem-
porarily position the system so you can access the back panels to
complete the connections.

2.3 Connect Power Ensure that the mains breaker switch is in the OFF position.
Then, use the supplied IEC power cord to connect the system to
mains power.

WARNING
Mains power must meet the voltage, frequency, and
amperage requirements listed on the serial number label.

WARNING
As long as the AC mains power cord is connected, power
is inside the unit. The mains power cord is the disconnect
device.

Position the system so that the power cord can be

unplugged, or use a power strip where the plug can quickly
be removed from the outlet in the event of an emergency.

2.4 Plumbing
Connections
WARNING
Risk of fire or equipment damage. Failure to connect waste
port tubing may allow organic solvents to pool in unsafe
areas, possibly creating dangerous levels of flammable
vapors.

WARNING
Risk of fire ignited by electrostatic discharges. Never
substitute the black tubing on CombiFlash systems. The
black tubing (P/N 023-0503-06) is conductive. This tubing is
required to dissipate static electricity.

2-3
CombiFlash NextGen Systems
Section 2 Preparation

CAUTION
Elevated flammable vapor levels are possible. Ensure that the
waste container is adequately ventilated, preferably by placing it
in a fume hood.

The NextGen comes with the solvent inlet lines and waste lines
pre-assembled to the instrument. To install the NextGen:
1. Remove the 2 screws and shipping clamps identified in Fig-
ure 2-3. This releases the tubing lines for placement in the
solvent supply and waste reservoirs.

Figure 2-3 Location of the shipping clamps

Note
For convenience, the inlet filter and inlet line tubing assemblies
have been pre-assembled with a bottle cap that is typical for the
region (GL 38 in the USA and GL 45 in the rest of the world). If
these caps aren’t suitable for your containers, you will have to
make modifications as needed.

2. Place the solvent lines in the appropriate solvent supply

containers.

If the solvent lines are unable to reach the supply container (such
as when placed below the work surface), the third clamp that
retains the tubing above the inlet ports can be removed. In
addition, solvent supply extension lines are available and have

2-4
 CombiFlash NextGen Systems
Section 2 Preparation

been verified at flow rates up to 200 mL/min. If using dichloro-

methane (DCM) in a warm environment, flows may be limited to
lower values. The system (if equipped with solvent level-sensing
feature installed and enabled) will stop and notify the user that
the solvent bottle is empty when approx. xx cm (as set by the
user in the configuration menu) of liquid remains in the solvent
bottle, the level is solvent density dependent.
3. Place the waste line in a suitable waste container. The
waste line assembly has a level sense line installed into the
cap. The system (if equipped with solvent level-sensing fea-
ture) will consider the waste full when the end of the sense
line is submerged below the surface of the fluid (solvent
density dependent).
4. Place the waste cap on the desired waste bottle.
5. If you use a dedicated waste container instead of a bottle,
you may not be able to implement waste level sensing as
designed. In these cases, work with your Environmental
Health and Safety group to determine how to proceed. This
may require disassembly of the supplied waste line to meet
your needs.

Note
When using higher-density solvents such as dichloromethane
(DCM), place the solvent container level with or above the
NextGen system. Placing solvent containers below the level of
the system can contribute to decreased flow due to the high
vapor pressure of DCM. This problem becomes more pro-
nounced as the ambient temperature increases.

2.5 External Gas

2.5.1 CombiFlash NextGen The CombiFlash NextGen systems use an internal air pump to
provide compressed air for solvent and waste level sensing.
Optionally, the system can deliver a compressed gas. Inert or
other gases may be more suitable than air for some applications.
If ambient air is compatible with your solvents and waste, simply
leave the optional external gas port open.
If using a gas instead of air, connect the user-supplied 1/8 inch
I.D. tubing to the optional external gas port. Connect the other
end to the user-supplied compressed gas. The external com-
pressed gas should be dry, filtered, and regulated to 2–5 psi
(0.14–0.34 atm, 13.8–34.5 KPa).

2-5
CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-4 Location of external gas ports

2.5.2 CombiFlash NextGen ELSD operation requires a carrier gas. To connect the carrier gas
Equipped with to the NextGen:
Optional ELSD 1. Locate the 1/8 inch O.D. FEP tubing (part number
023-0503-02) from the accessory package.
2. Push one end of the tubing into the NITROGEN INLET port
on the back of the system. The tubing should be fully
seated in the port.
3. Cut the tubing to length and connect the other end to the
user-supplied carrier gas. An assortment of 1/8 inch adapt-
ers are supplied in the accessory kit to complete the con-
nection to your gas source.

Note
Teledyne ISCO recommends >99% pure nitrogen from a
source that can deliver 2.5 SLPM at 60 to 70 psi.

Figure 2-5 Location of ELSD gas port (Nitrogen input line circled)

2.6 Connect and Route

Drain Lines
WARNING
Risk of fire or equipment damage. Failure to connect drain
lines may allow organic solvents to pool in unsafe areas,
creating a potential for dangerous levels of flammable

2-6
 CombiFlash NextGen Systems
Section 2 Preparation

vapors. Improper draining may damage the instrument’s

internal components.

The CombiFlash NextGen system has drain tubes extending

from its back panel. The tubes drain away any liquid spilled on
the top shelf and the tray beneath the fraction collection racks.
1. Test the fraction collector drain by connecting a vacuum or
air supply source to the outlet end of the drain tube. Then,
verify the presence of such vacuum or air supply source on
the drain (Figure 2-6).
2. Test the top shelf drain by connecting a vacuum or air sup-
ply source to the outlet end of the drain tube. Then, verify
the presence of such vacuum or air supply source on the
drain (Figure 2-7).

Vacuum or pressurized air applied to the

outlet end of the drain tube must exist
at the collection tray drain hole.

Figure 2-6 Fraction collection tray drain hole

Vacuum or pressurized air applied to the

outlet end of the drain tube must exist
at the top shelf drain hole.

Figure 2-7 Top shelf drain

3. Route the end of both drain tubes to a suitable waste fluid

collection container.

Note
It may be necessary to extend the drain tube. If so, splice the
tubing with user-supplied tubing. The user-supplied tubing
should have an inside diameter no smaller than the existing
drain tubing, and must be compatible with the solvents used by
the system. Route this extension tubing to the waste collection
vessel.

2-7
CombiFlash NextGen Systems
Section 2 Preparation

CombiFlash with optional ELSD only) The CombiFlash

ELSD system has additional drains that must be connected
(Figure 2-8):
4. Locate the 1/4 inch I.D. silicone tubing and a nylon tubing
clamp from the accessory kit. Connect one end of the tub-
ing to the pump drain port and secure it with the clamp.
5. Install the ELSD P-trap vent assembly found in the acces-
sory package by placing the short tube into the port on the
rear panel, orienting the vent as shown in Figure 2-8 and
tightening the fitting nut finger tight plus a small amount
more with a wrench. Be sure the P-trap drain vent assem-
bly is oriented with the vent portion vertical.
6. The P-trap must be filled with fluid to prevent sample loss.
To accomplish this, raise the drain tubing attached to the
P-trap drain vent assembly above the instrument and
place 10 mL of liquid, such as isopropyl alcohol, into the
tube. Make sure the fluid level in the tubing doesn’t exceed
the level of the instrument case top. If the tubing is raised
too fast, fluid may flow out the top of the vent tube causing
a spill. Lift it high enough so the fluid enters the P-trap
drain (Figure 2-8).

P-trap drain vent assembly

drain line

Figure 2-8 Keep liquid level lower than the top of the system

7. Route the end of the P-trap drain tube to a suitable waste

fluid collection container. Please be aware that as the tub-
ing is lowered, several mL of isopropyl alcohol will run out
of the tubing. This is normal and means that the P-trap is
properly filled. Depending on your application, the P-trap
fluid may need periodic replenishment to ensure maximum
signal strength of the ELSD.

2-8
 CombiFlash NextGen Systems
Section 2 Preparation

Note
If accessible, the outlet end of the P-trap drain tube may be
used to refill the P-trap.

8. Route the attached tubing from the ELSD exhaust port

away from the system. This will prevent unnecessary sol-
vent vapor alarms.

Note
If using outside of the hood, ensure that the exhaust port tub-
ing is routed to an exhaust handling system that meets your
safety and environmental requirements. In normal operation,
the exhaust gas is dry and does not require a waste collection
container.

CAUTION
Discontinue use of the CombiFlash if liquid is present at the
Pump Drain. Contact Teledyne ISCO technical service for
assistance with correcting the leak.

2.7 Vapor Enclosure 1. 1. Secure the Vapor Enclosure with the three provided
(Optional) screws to the holes in the instrument (Figure 2-9). Two
along the back of the fraction collection area and one on the
side of the fraction collection area.

Figure 2-9 Location of holes for the vapor enclosure

2. The back of the NextGen has a 2 in. port for attachment to an

external exhaust (Figure 2-10). The external exhaust should
meet the specifications of your environmental, health, and safety
specifications.

2-9
CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-10 Location of the external exhaust port

2.8 Install Solid Load 1. Secure the SCLL storage bracket to the left side of the
Cartridge Cap Ring instrument with the screws provided (Figure 2-11).
Support and SLCC
Storage Bracket
(Optional)

Figure 2-11 Before (left) and after SLCC storage bracket is installed (right)

2. Place the SLCC Ring Support in the hole on the injection port
trolley (Figure 2-12).

2-10
 CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-12 SLCC ring support

3. Secure the support with the knobbed screw via the side hole
(Figure 2-13).

Figure 2-13 Location of the side hole to secure the SLCC ring support

2.9 Position the System After completing the various connections, the system can be
moved to its operating position. Turn the system so that the
operator can access all of the front view features and controls
(Figure 1-5). Use care not to damage the connections, tubing, and
cables while moving the system.

2-11
CombiFlash NextGen Systems
Section 2 Preparation

CAUTION
Ensure that the CombiFlash has at least 3 cm (1.25 inch) of air
space behind it for ventilation. Position the solvent and waste
containers as necessary.

Note
When using higher-density solvents such as dichloromethane
(DCM), place the solvent container level with or above the
CombiFlash NextGen system. Placing solvent containers
below the level of the NextGen can contribute to decreased
flow due to the high vapor pressure of DCM. This problem
becomes more pronounced as the ambient temperature
increases.

Note
When using higher-density solvents such as dichloromethane
(DCM), ensure that the waste container is no more than 3 feet
below the supply solvent levels. Placing solvent containers
above the system and waste containers on the floor may cause
the internal check valves to open and allow solvent to flow
through the system when in the standby state.

2.10 Install Collection Tube Before beginning a run, you must load collection racks with tubes
Racks onto the system’s fraction collector tray.
Your system was shipped with two collection tube racks. The
following tube rack sets are available:
• 60-5237-013 – Two racks for 13 x 100 mm test tubes
(8 mL). Total tubes: 216.
• 60-5237-061 – Two racks for 16 x 100 mm test tubes
(14 mL). Total tubes: 150.
• 60-5237-031 – Two racks for 16 x 125 mm test tubes
(15.5 mL). Total tubes: 150.
• 60-5237-032 – Two racks for 16 x 150/160 mm test tubes
(18 mL). Total tubes: 150.
• 60-5237-033 – Two racks for 18 x 150 mm test tubes
(25 mL). Total tubes: 140.
• 60-5237-034 – Two racks for 18 x 180 mm test tubes
(30 mL). Total tubes: 140.
• 60-5237-035 – Two racks for 25 x 150 mm vials (50 mL).
Total vials: 60.
• 60-5237-040 – One rack for twelve 480 mL French
square bottles.
• 60-5394-469 – Two racks for 20 mL (28 x 61 mm) scintil-
lation vials. Total vials: 54.
• 60-5394-468 – Two racks for 40 mL (28 x 95 mm) scintil-
lation vials. Total vials: 54.

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To load the racks:

1. Insert test tubes, vials, or bottles into the rack (Figure
2-14). The system assumes all tube positions have tubes. If
any tubes are missing, it is possible to have a fluid spill
when the fraction collection system advances to that posi-
tion

CAUTION
Risk of broken glass or equipment damage. Do not load test
tubes longer than the length listed on the tube size label.

Figure 2-14 Loading test tubes

2. While holding the rack with the tube size label visible (Fig-
ure 2-15), insert the racks into the system. Slide the rack
in until it stops. You can feel it drop into its seated position
when it is properly loaded.

Tube Size Label

Figure 2-15 Inserting the collection tube rack

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CombiFlash NextGen Systems
Section 2 Preparation

CAUTION
An incorrectly installed rack will cause the rack to be mis-
aligned under the fraction collector arm. Misaligned racks
might cause fractions to miss the tube opening or deposit in
the wrong tube. Always ensure the tube size label is visible (that
is, facing outward) and the rack is pushed in until it is seated.

When you turn the power switch to ON, the system will automat-
ically detect the type of rack and configure program settings
accordingly. Later, in section 2.13.1, you can configure the
maximum volume for each tube size if you want smaller fractions
than the tube capacity.

2.11 Powering on the System The system’s power switch is located on the lower right side
panel, near the front of the instrument.
1. Switch the rear panel circuit breaker to ON.
2. Press the switch to turn the system on. The system will
begin its startup routine which includes self- diagnostics.

Note
The momentary power button on the right side of the instru-
ment can be used to boot up the system or shut it down on a
daily basis. The rear panel power switch is only used when you
desire to completely remove power from all internal electron-
ics.

3. The system is ready for operation when the PeakTrak

screen is displayed.

2.12 Configure the System Before operating the system, it should be configured to best meet
your needs. To configure the system, open the CONFIGURATION
window by selecting TOOLS>CONFIGURATION from the PeakTrak
menu.
The configuration window has 3 tabs: I NSTRUMENT C ONFIGU-
R A T I O N , N E T W O R K C O N F I G U R A T I O N , U S E R P R E F E R E N C E S.
Network Configuration settings are discussed later in Section 4.
Adjust the selections on the I NSTRUMENT C ONFIGURATION or
USER PREFERENCES tabs for the desired operation and then click
OK to save the settings.

Note
Some configuration settings do not take effect until the system
is restarted. See Section 2.13.4.

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 CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-16 Instrument configuration menu

2.12.1 Solvents When the system is installed, up to four solvents were connected
to the back of the system. This section of the INSTRUMENT CON-
FIGURATION tab allows you to name these solvents for conve-
nience. To do so:
1. Solvent names for drop down use are limited to a maxi-
mum of 16 including the prepopulated names, which can be
removed. Names are stored in the order created.
2. From the Solvent 1 drop-down list box, select the solvent
name connected to the 1 Solvent Inlet Port. Repeat for Sol-
vents 2, 3, and 4.
3. If your desired name is not listed by the system:
a. Click the ADD SOLVENT button.
b. Enter the solvent name and click the OK button.
c. Repeat steps 1 and 2 as needed.
4. Select the ENABLE SOLVENT LEVEL SENSING check box to
enable this feature (recommended). Clear this box to dis-
able this feature.

The solvent level sensing feature pauses the pump opera-

tion if the solvent level is below the configured level. In
addition, if the same supply container has been used for a
previous separation and has not been refilled, the system
will estimate the amount of solvent remaining. It compares
the volume required for a purification run with the esti-
mated volume in the solvent container and alerts you when
there is not enough before the separation begins.
5. If using the solvent level sensing feature, you can select
the Minimum Solvent Level (density dependent) as a range
in centimeters.

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CombiFlash NextGen Systems
Section 2 Preparation

Solvent level sensing works by sensing the fluid pressure

at the inlet line level. This is specified as a range to vari-
ance in the density of most common chromatographic sol-
vents. To understand this range, consider that
dichloromethane is ~2x more dense than hexane. If the
selected range is 2–4, the system triggers an alert when
the dichloromethane level falls to about 2 cm above the
weighted filter. At the same selected range, the system
would trigger an alert when the hexane falls to about 4 cm
above the weighted filter since the fluid pressure of hexane
is only half of dichloromethane of the same depth.

2.12.2 General Settings CombiFlash NextGen Name (optional) – Use this option to
name your system. The name will appear in operational displays
and separation reports. This feature is useful when your labo-
ratory has more than one CombiFlash system and you want to
determine which system performed a given separation.
Time Zone – Select your time zone from the drop-down list box.
This allows the factory set time to be updated to correspond to
your time zone. This also allows the system to adjust the system
time for seasonal time changes (such as daylight savings time in
the United States). If the date of this time change is changed by
the local government, the system may not be correct based on the
changed effective date.
Alert Sounds – Used along with red lights in the fraction area
(on some systems) to alert the user to items such as collection
rack fill, solvent low, and serious errors. The alert sounds also
occur to alert the user to when the run is complete or equili-
bration is complete for liquid or solid (pause) loading techniques.
End of Run Hold – When enabled the default system setting
will be for the run to wait for user input before undergoing the
automatic valve wash and air purge sequence at the end of the
run. This allows the user to extend and modify the run if they
find it necessary before purging the column.
Set Date/Time – Click this button to set the system date and
time.
To prevent unauthorized time changes, this feature is password
protected. The system is shipped with the password set to combi-
flash. Use the TOOLS>SET PASSWORD menu command to change
this password for greater security if desired
PurIon Delay – Configuration for the tubing included with the
PurIon. If your installation uses an additional length of tubing,
the volume of this tubing must be added.
Automatic Column Purchase – E n a b l e s a f l a s h c o l u m n
supply agreement that must be purchased separately from the
system. This allows us to supply columns as needed without need
for inventory management at the customer’s site.

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 CombiFlash NextGen Systems
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2.13 Vapor Limit (if The system has an optional internal vapor sensor that detects
equipped) vapors present inside the system. When the limit is exceeded, the
CombiFlash will shut down to avoid a hazardous condition.
Teledyne ISCO recommends using the default setting of low sen-
sitivity. This value represents an organic vapor level that is half
the explosive limit.
Medium and high sensitivity settings could cause random alarms
without significant vapors present. If PeakTrak displays a Vapor
Limit alarm perform the following checks on your laboratory and
the instrument:

Figure 2-17 Vapor Limit warning

• Ensure that no open containers or spills of organic

solvent are in close proximity to the system.
• Ensure that the system is located in a well-ventilated
area.
• Ensure that there is no visible solvent leakage from the
system.

Once you have remedied the cause for the vapor sensor
alarm, you may choose to change the default vapor
sensitivity level or ignore the vapor alarm and continue
operation without it for 10 minutes.

If PeakTrak continues to display the Vapor Limit alarm

after you have made these checks and corrected any
problems found, it is likely that organic vapors are
present in the ambient environment of your laboratory.
In this case, choose a less sensitive Vapor Limit setting.

Low sensitivity settings are appropriate for laboratory

environments with a somewhat elevated background
solvent vapor concentration, representing a safe setting,
but one with a reduced margin for error.

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CombiFlash NextGen Systems
Section 2 Preparation

2.13.1 Set Default Tube Click this button to set the default volume of the collection tubes.
Volumes There is some variation in the fraction size as a function of flow
rate. Be sure you don’t overflow the containers by setting the
default volume to close to the actual capacity.
The system will automatically advance to the next tube when
this default volume is met in each tube. Keep in mind that this
level can be set for each separation in the M ETHOD E DITOR.
During the separation, other factors such as automatic tube
advances for detected peaks will affect collected volume.

2.13.2 User Preferences View the USER PREFERENCES tab to configure system operation
for individual users. This is especially convenient if the system is
shared by several individuals with different preferences
(Figure 2-18).

Figure 2-18 User Preferences screen

Note
New systems will only have one user account named “com-
mon.

Use ELSD by Default – causes all runs after system boot up to

use the ELSD detector unless disabled in the run requirement
screen. After an initial separation, each subsequent separation
uses the same ELSD selection unless changed during the RUN
REQUIREMENTS screen.
Language – Select the user’s desired language from the
drop-down list box.
Automatically Print Report at End of Run – If a flash drive
is in the USB port at the end of the separation, the PDF file is
automatically saved on this drive in addition to the system’s
internal hard drive. The run (.txt) file must manually be saved to
the USB drive. If you want to automatically save to a network

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 CombiFlash NextGen Systems
Section 2 Preparation

location, see Section 4 Network Configuration.

Note
This option requires the system to be configured for network
operation and a connection to a printer on the network. Net-
work settings are discussed in Section 4. Do not select this
option for these initial installation steps.

Default Run Units – Run units are displayed along the X-axis
of the chromatogram. Select Time (in minutes) or Column
Volumes (CVs). A column volume unit is the time it takes to
pump enough solvent to exchange the volume held by the
column. The duration in minutes will vary according to the
column media and size, and the flow rate.
Enable Run Length Extension – When enabled, this option
automatically extends the run if a peak is eluting at the end of
the maximum %B gradient. This ensures that a late-eluting peak
fully comes off the column and is collected.
An automatic run extension is a five-minute isocratic hold added
to the end of the run’s maximum %B gradient profile. During the
extension, the system continues to pump the maximum %B
solvent mixture. Should the system still detect a peak after an
extension, the system will add another, up to a maximum of three
extensions.
Occasionally, compounds might come off the column once the %B
returns to the minimum value at the end of a run. If the RUN
LENGTH EXTENSION is enabled, it will automatically extend the
run one time to clear the column and plumbing of any remaining
material.

Figure 2-19 Automatic Run Extension Examples

Figure 2-19 illustrates the possible run extensions. The original

method was programmed to rise to the maximum %B over seven
minutes, hold for two minutes and return to the minimum %B for
a final minute. At nine minutes, a peak was eluting. The system
extended the run, holding the %B at the maximum level for
another five minutes. This also occurred at fourteen and
nineteen minutes, resulting in the second and third extensions.
At twenty-four minutes, the %B solvent strength returned to

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CombiFlash NextGen Systems
Section 2 Preparation

zero. Before the final minute elapsed, more compound was

detected, causing the system to extend the run for a final five
minutes at the minimum %B.
Enable rapid equilibration – select this option to equilibrate
the column at a higher flow rate.

Note
Due to the high pressure that is possible during rapid equilibra-
tion, this option may not be desirable for some column media
or purification methods.

Enable Automatic Peak Hold – inserts an isocratic hold for

the duration of each detected peak.

2.13.3 User Management User management allows creation of individual user accounts on
the system. Password access can be added if desired. When con-
figured, the user’s preferences are stored and their data is auto-
matically saved to the user’s personal folder on the system.
To implement control over system usage, a system administrator
can set varying levels of access for each user. The capability of
each user is summarized in the below chart.
User Management allows each user of the system to have some
customized settings.

Figure 2-20 User Management screen

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 CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-21 User Management guide

2.13.4 Restart the System Some configuration settings do not take effect until the system is
restarted. After completing the configuration, restart the system:
1. If the CONFIGURATION window is not closed already, click
the OK button to close it.
2. Momentarily press the power switch on the side of the
instrument and wait for the system to shut down.

CAUTION
Except during an emergency, never remove AC power immedi-
ately after placing the power switch in the OFF position. The
power down sequence includes file maintenance that takes up
to one minute to complete. Interrupting the file maintenance
may corrupt files causing abnormal operation or a complete
system failure.

3. After one minute or more, press the power switch.

4. The system is ready for operation when the PeakTrak
screen is displayed.

2.14 Prime the Solvent Before the first use, the system should be primed. Ensure that
Lines the solvent containers are filled, then (NextGen 300 or 300+):
1. Select the TOOLS>AUTO PRIME menu command. Automati-
cally primes each line with 50 mL of solvent in the order
directed by the user as seen in the Figure 2-21. You can
also rename your solvents by using this screen to switch
solvents. These changes are also reflected in the configura-
tion menu.

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CombiFlash NextGen Systems
Section 2 Preparation

Figure 2-22 Auto Prime screen

2. There are 4 solvent selection lists labeled from first solvent

through fourth solvent, describing the order in which sol-
vents will be pumped. The solvent choices for each list are
those defined by the CONFIGURATION window.
• If all solvents are of a similar polarity, it is recommended
that the solvent be primed in order of decreasing solvent
strength, with the weakest solvent primed last.
• If the 4 solvents are of mixed polarity (aqueous and
nonpolar), ensure that sequential solvents are
compatible, such as water, acetonitrile, ethyl acetate,
hexane.
• If you are performing an auto prime after redefining
some of the solvents, it may be beneficial to run 2
auto prime operations sequentially to get full flushing
of the lines.
3. Click the PLAY button to start priming the system.

Note
When priming the first time, inspect the solvent and waste con-
nections to the system. If any leaks are observed, click the
C ANCEL button to stop the Auto Prime. Correct the leak by
tightening the fitting an additional 14 turn, and then restart the
Auto Prime from step 1.

To terminate an auto prime step in process and advance to the

next Auto Prime step, you can click the FAST FORWARD button.
The Auto Prime window closes when finished.

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 CombiFlash NextGen Systems
Section 2 Preparation

After Auto Priming, the system is ready for operation. If you are
priming a new system for the first time, it may be beneficial to
flush all of the fluid lines with the desired solvent to remove any
isopropanol left from production testing.

Note
Use Auto Prime for quick solvent changes of similar polarity by
pumping a fixed amount of solvent through the waste fluid path
at 100 mL/min. For more advanced priming functions such as
varying the flow rates, fluid paths, etc., or to purge the system
with air before changing between normal and reversed phase
solvent systems, use the TOOLS>MANUAL CONTROL option.

Manual Prime (NextGen models only) – The pumping

system in the base NextGen model requires the pumps to be
manually primed. In order to do this there is a syringe port on
the front of the instrument under the screen. From TOOLS select
MANUAL PRIME and follow the directions on the screen. Pull the
syringe to remove air from each of the solvent lines as directed by
the on screen instructions.

2.15 System Verification It is recommended that the system operation be verified. To do

so, use the CombiFlash Universal verification kit Teledyne ISCO
part number 60-523-4317:
• The instructions for Universal Verification Mix can be
found in Instruction Sheet P/N 69-5233-870.
• For PurIon equipped systems the following test
mixtures are also needed:
· P/N 60-5234-315 Test Mixture for APCI probes, 5 mL.
· P/N 60-5234-627 Test Mixture for ESI probes, 10 mL.

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CombiFlash NextGen Systems
Section 2 Preparation

2.16 Installation Table 2-1 may be completed to verify and document the instal-
Qualification lation procedures contained in Section 2 of this guide. See fol-
Checklist lowing page:

Table 2-1 Installation Qualification Checklist

Installer Operator
Step Section Description Initials Initials
1 Unpacking the unit
2 Instrument location
3 Connect power
4 Connect solvent lines (if not pre-installed)
5 Connect waste lines (if not pre-installed)
6 External ELSD gas (optional)
7 Connect and route drain lines
8 Position the system
9 Installation of the collection tube racks
10 Install PurIon (optional)
11 Turn on the power
12 Configure the system
13 Prime the solvent lines
14 System verification
Certification of Section 2 Completion
Installer Name (print):

Installer Signature:
Date:

Operator Name (print):

Operator Signature:
Date:
Comments:

2-24
 CombiFlash NextGen Systems
Section 3 Operation

This section provides abbreviated operating instructions for the

CombiFlash NextGen systems. For complete instructions, refer
to HELP menu option from PeakTrak’s main menu.

3.1 Flash Sample Before starting a run, consider how the sample will be loaded
Preparation onto the column media. There are three different loading
methods: liquid injection, solid sample cartridges, and
pre-loading the sample on the column. Sample loading is covered
in the included CombiFlash NextGen Quick Start Guide.

3.1.1 Liquid Sample If the sample is soluble in the starting mobile phase or a solvent
Injection strength that doesn’t significantly impact the separation, it can
be prepared as a solution and injected onto the column when
prompted during the purification run.

3.1.2 Solid Samples Some compounds are not soluble in solvents that are compatible
with the chromatography. In addition, the compounds may have
very limited solubility in any solvent, resulting in sample disso-
lution volumes that are impractical for good chromatography.
The answer in this case is solid sample introduction.
Some samples of this type may be simply dissolved and placed
into a pre-filled solid load cartridge and allowed to absorb onto
the media in the cartridge. You can then use the cartridge imme-
diately or dry it before placing it on the system.
Other samples may need more care. Typically, you would dissolve
the sample using a suitable solvent, then create a mixture of
~20% sample load to media (w/w). Then dry the mixture under
conditions that will dry off the solvent without affecting the com-
pounds of interest. These compounds remain bound to the media.
Once dried, pour the media/sample mixture into an empty car-
tridge.

3-1
CombiFlash NextGen Systems
Section 3 Operation

3.2 Loading a RediSep Rf

Column
WARNING
The CombiFlash NextGen has redundant safety devices to
limit pressure to less than 300 psi (2068 kPa); 150 psi (1034
kPa) for systems without an automated injection valve.
RediSep columns smaller than 100 g are CE certified using
standard IEC61010-1 for use on the CombiFlash NextGen.
RediSep columns larger than 100 g meet Pressure Vessel
Directive 97/23/EC.

Note
For best results, always use RediSep Rf columns. The system
will not automatically detect other columns. The system also
limits the maximum operating pressure to 100 psi when the
column is not detected.

To load a RediSep Rf column:

1. Select the proper RediSep Rf column. The RediSep column
selection guide that can assist you in selecting a stationary
phase media and column size. In addition, the approximate
sample loading capacity is printed on the RediSep column
label.
2. Raise the column mount and insert the column into the top
column mount. Note that the column fittings are keyed to
ensure the correct flow direction.
3. Slowly lower the injection valve while aligning the bottom
column fitting. The spring-loaded column mount will hold
the column in place.
4. To seal the column fittings, give the column a slight twist
(1/4 turn).

Note
After loading a RediSep Rf column, systems configured with
RFID will use RFID technology to automatically detect the
media type and column size (if available on the column). Peak-
Trak displays the detected column size on the M AIN and
METHOD EDITOR windows. If the system does not detect the
column, manually select the column media and size.

3.3 Starting a Separation After completing the system installation steps, preparing the
sample (See the CombiFlash NextGen Quick Start Guide for
sample loading procedures), and inserting the column, you are
ready to perform a run. The system will assume a default method
based on the column size placed in the system. Default methods
are optimized for the use of RediSep Rf columns with run param-
eters typically used by chemists.

3-2
 CombiFlash NextGen Systems
Section 3 Operation

The default settings include:

• Pre-configured equilibration settings based upon
column size and type.
• Set a flow rate and separation length suitable for the
column loaded.
• Set to collect all fluid in the fraction collector rack to
ensure peaks without UV absorbance are collected.
• Pre-programmed peak detection settings suitable for
typical peak sizes based on the column size.
• Automatically advance to the next tube when a peak is
detected and when a peak ends to maximize peak
concentration.
• Advance to the next tube if a partially resolved peak has
a shoulder or valley UV absorbing peaks based on slope
detection or level threshold.
To start a separation from the PEAKTRAK MAIN window:
1. Enter a sample name if desired. If no sample name is spec-
ified, PeakTrak will enter the date and time as the sample
name when you start the run.
2. Review the PEAKTRAK MAIN window settings. Some set-
tings can be edited from the main menu. For example:
• If the gradient conditions are not correct, the duration
can be adjusted using the RUN LENGTH input field.
• The gradient slope can be adjusted by grabbing an
inflection point and dragging to the desired position.
• Inflection points can be added or deleted if needed.
Select the ADD POINT button and touch the screen where
a point should be added, or DELETE POINT and touch the
point to be removed.
• For greater detail, the screen can be zoomed using a 2
finger pinch zoom, or panned using 2 fingers.
• Other parameters may be edited such as peak detection
parameters or numerical gradient point input using the
METHOD EDITOR selection at the top of the screen.
3. Click the PLAY button.
4. Select the sample LOADING TYPE from the list. This LOAD-
ING TYPE should support the sample preparation you
chose. Possible sample loading types are:
• Solid (requires the inject valve; optional on some
systems) – Select this option if you have prepared the
sample and placed it into a solid sample load cartridge
before starting the separation. (Column plan subscribers
must also specify the cartridge type and size).
· The system will proceed automatically until the end
of the programmed run length.

3-3
CombiFlash NextGen Systems
Section 3 Operation

• Solid (Pause) – Select this option if you will place the

sample into a solid sample load cartridge, but have not
yet prepared the sample. (Column plan subscribers
must also specify the cartridge type and size).

When you click the RUN button, the system will perform
a column equilibration and then wait while you prepare
the cartridge. After you have placed the cartridge on the
system, click OK to continue with the run.
• Liquid – Select this option if you have prepared a liquid
sample and plan to manually inject it into the injection
port or directly onto the column after column equili-
bration.
· If you inject using the luer fitting on top of the
column, inject the sample, then inject 3 mL of chase
fluid to wash the sample completely onto the column
· None (on column) – Select this option if you have
pre-loaded the sample on the column.
· The system will skip column equilibration so that the
sample will not be flushed from the column before the
run.
5. Review or update the START RACK and START TUBE.
6. Review the MINIMUM RUN REQUIREMENTS:
• The system will report the estimated solvent volumes,
expected waste, collection tube usage, etc. You can use
this information to verify that there will be enough
solvent to complete the run, the waste volume does not
exceed the collection container’s capacity, and whether or
not more tubes will be required during the run.
End of Run Hold- Select whether or not the run will
undergo automatic valve wash and air purging without
waiting for user input.

Note
The CombiFlash systems with solvent level sensing, estimate
the solvent volumes by monitoring the solvent level in the con-
tainer and the known usage rate. The system continues to
refine this.

Note
If equipped with solvent level sensing, the waste level sensing
tube is inserted correctly into the waste container, systems, will
automatically suspend operation before an overflow condition
might exist. To prevent the run from being suspended before
completing the run, ensure that the container will hold the
expected waste volume.

3-4
 CombiFlash NextGen Systems
Section 3 Operation

7. If equipped with an ELSD, ensure that the ELSD option is

selected if desired. The system defaults to ELSD detection
parameters that are compatible with fluids typically used
by the selected column. If performing a normal phase sepa-
ration using a high boiling point solvent such as heptane or
toluene, you may need to review the ELSD parameters in
the METHOD EDITOR; otherwise the ELSD may be disabled.
The system will automatically select or deselect use of the
ELSD based on the previous run.

Note
If you typically use normal phase solvents and the ELSD
P-trap drain fluid hasn’t been replenished recently, you may
need to ensure the P-trap is full for maximum ELSD signal
strength. This can be done by lifting the drain line above the
instrument and placing 10 mL of isopropyl alcohol in the drain
line. Lift the line sufficiently that the fluid flows into the P-trap
drain. Then place into a waste container so that the excess fluid
can drain out of the trap. If you normally use higher boiling point
solvents such as water, the P-trap will typically remain full and
will not require replenishment.

Note
If you are using a silica flash column and your solvent exceeds
7.5 pH, the ELSD should not be used. Basic conditions can
dissolve silica. When the solvent with dissolved silica is evapo-
rated in the ELSD, silica can build up over a period of time and
eventually clog the ESLD, resulting in an expensive repair.
Sample build up in the spray chamber is less of a concern
since it is sent to the chamber in smaller quantities only during
the peak and is more easily cleaned from the chamber during
regular cleaning (see Section 5.6 ELSD Maintenance).

Mass Spectrometer- (PurIon systems only) click this button to

enable mass spectrometer detection.
PurIon Loading- (PurIon systems only) sets the amount of
sample sent to the mass spectrometer. Default is set to LOW to
reduce saturation of the mass spectrometer detector. Select HIGH
for lightly loaded columns or samples with weak detection on the
mass spectrometer. For situations in-between, select MEDIUM.
Detection Ions- Enter up to 4 ions, or a range and up to 3 ions.
The ions MAY be a mix of positive and negative.

3-5
CombiFlash NextGen Systems
Section 3 Operation

3.4 During the Separation You may allow the system to proceed with the gradient while
monitoring the progress on the PeakTrak main window. If
desired, all method parameters may be modified during the sepa-
ration without stopping the separation. Changes to the gradient
may be made by dragging the solvent inflection points with your
finger without stopping the pump.
As the run progresses, the absorbance trace(s) is displayed while
the separation continues until the end of the run defined by the
run length setting.
During the run, data is automatically saved to the internal data
storage every few seconds. You can manually save a RUN FILE as
text (in XML format) or PDF to a flash drive while stopped or any
later time as long as the data file has not been deleted.
To automatically save a text or PDF file to a network location
after each run, configure the NETWORK FILE SAVE feature.

3.4.1 Run Control Buttons The following information will correspond with Figure 3-1.

Figure 3-1 MAIN SEPARATION screen

Play — The PLAY button starts the separation. During the sepa-
ration, this becomes the PAUSE button.
Pause — The PAUSE button holds the %B at the current value
while the system continues to operate (sometimes called an iso-
cratic hold). Note that pausing the run extends the run length.
While in the paused state, you can resume the gradient by
clicking the PLAY button, or stop the run by clicking the STOP
button. If you resume the run, the system continues with the gra-
dient curve at the %B when the system was paused.
Stop, Fast Forward, and Rewind — The S TOP button sus-
pends the entire run. Unlike the paused state, the pump, peak
detection, and fraction collection will stop. While in the stopped
condition, you can:
• Resume the run by clicking the PLAY button, skip the
rest of the separation by clicking the FAST FORWARD

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 CombiFlash NextGen Systems
Section 3 Operation

button which proceeds with the air purge (if the system
is equipped with this feature) or end the run by clicking
the REWIND button which clears the data from the
screen and displays the now truncated temporary
method.
The advantage of the FAST FORWARD vs. the REWIND is that the
fast forward advances to the air purge line flush, and detector
wash while REWIND performs none of these important mainte-
nance functions.
If REWIND is selected, it may be useful to manually flush the
lines so that strong solvent remaining in the lines doesn’t affect
subsequent separations.
Next Tube — This button advances the fraction collector to the
next tube position after the delay volume from the detector to the
fraction collector has elapsed. This ensures the fractionation
matches the detector signal at the time the button was pressed.

3.4.2 Bypassing the Solid Occasionally, impurities precipitate in the solid load cartridge as
Load Cartridge compounds are purified. This may cause high back pressures
(Injection valve resulting in reduced flow rates and long run times. Clicking this
equipped systems button changes the injection valve position, which removes the
only) cartridge from the solvent path and relieves the back pressure.
Any compound that hasn’t eluted from the solid load cartridge
will remain in the cartridge and not be purified.

3.4.3 About Solvent Level When enabled in the TOOLS >CONFIGURATION window, the system
Detection (systems will estimate the solvent volume usage by measuring the solvent
equipped with this level in the container and the monitoring the usage rate. The
option only) system continues to refine this estimate of available solvent
during operation.
At the beginning of a run, the system calculates the amount of
solvent required to complete the run. The system will alert you if
it detects that there is not enough solvent at the start of the run.
Likewise, the system will alert you during the run if the esti-
mated usage rate or the measured level indicates that there is
insufficient solvent. Note that pauses and automatic run exten-
sions during a run will increase the required solvent volume.
The options available on the alert message will vary depending
on when the condition occurs (before or during a run) and which
method (measured level or estimated usage rate) indicated the
condition.
To clear the alert message, the user must select one of the options
below.
• Continue – Select this option after you have added
more solvent to the container. After selecting this option
the system measures the solvent level again. If the level
is sufficient to complete the run, the run continues. If
not sufficient, the system displays the alert message
again.
• Ignore – This option is available when the estimated
usage rates indicates that the solvent levels are insuffi-

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CombiFlash NextGen Systems
Section 3 Operation

cient. Select IGNORE to clear the message and continue

the run. The system then ignores the estimated solvent
usage rate for the remainder of the run, but continues to
measure the solvent level. Should the level become
insufficient, the system will display another alert
message.
• Disable – Select this option to turn off solvent level
measurements and estimated solvent usage rates for the
remainder of the run.
• Cancel – This option appears before the run is started.
Select this option to cancel the run and return to the
main window which will display the current method.

3.5 Software Reference Many features of the software function like a typical PC or tablet
and therefore are not covered in detail. The remainder of this
section covers areas that may be less common or unique to
CombiFlash systems.

Figure 3-2 NextGen Login screen

The CombiFlash NextGen comes configured as an open access

system with all work done in a single common area. If so con-
figured, the above screen is not shown at startup. In addition, it
can be configured for multiple, individual users which can be
assigned individual passwords. In this case, the screen in Figure
Figure 3-2 is displayed after startup or individual logoff. In
either case, the SELECT USER drop down allows the user name to
be selected followed by a log in screen (if activated for the user).
Once logged in, the MAIN SEPARATION screen is shown (Figure
Figure 3-3).

3.5.1 Main Separation The MAIN SEPARATION screen (Figure 3-3) defaults to a 4g silica
Screen method until an RFID tagged column is sensed, or the user
selects a column via the column drop down (for support of non
RFID tagged columns).

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 CombiFlash NextGen Systems
Section 3 Operation

Figure 3-3 MAIN SEPARATION screen

• If the displayed method is acceptable, the START button

will start the separation.
• If a sample name is not entered, it defaults to the
current date and time. Also, there is no need to save the
method to perform a separation. The PeakTrak software
treats methods as a temporary method discarded after
each separation. If desired the user can save the method
at any time for future reuse. All method parameters are
stored with the data file and printed on the report so no
information about the separation is lost.
• If the displayed method doesn’t meet your needs, it can
be edited before use. For example if the suggested
gradient isn’t optimal, the gradient can be adjusted by
dragging the inflection points to the desired position. If
more gradient inflection points are desired, select the
INSERT POINT button followed by touching the point on
the gradient to insert the point. The METHOD EDITOR
button at the top left of the screen allows for further
method adjustment.
• ZOOM and PAN can be performed by using a 2 pinch
zoom (like many cell phones or tablets) or a 2 finger pan.
A single finger touch or drag is interpreted as a gradient
edit, not a pan function.
• Fraction collection defaults to collecting ALL to ensure
no peaks are lost due to lack of UV absorbing properties.
Even when selected, the peak detector operates with
default parameters based on the column size (unless
disabled by the user). This will cause the system to
advance to the next tube whenever a peak starts or ends
to ensure cleanly cut fractions with maximum concen-
tration. The tube changes and peaks will be marked and
the fraction collector will move after the delay volume
has passed.

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CombiFlash NextGen Systems
Section 3 Operation

· PEAKS collects fractions only when a peak is detected

and sends the non compound containing fluid to
waste.
· NONE is used conjunction with INITIAL WASTE AND
TIME windows (Figure 3-7) to minimize fractions or
when the user wants to manually select when to
collect while the remainder is sent to waste. This is
commonly used when a large single peak is desired
and is selected by the user.
• The MS button displays the current mass spectrum on
PurIon systems, and the current UV button displays the
UV (or UV-Vis) spectrum.

3.5.2 Method Editor Screen The METHOD EDITOR screen (Figure 3-4) allows control of less
commonly used parameters that aren’t available on the main
screen.

Figure 3-4 METHOD EDITOR screen

3.5.3 Edit Gradient Options The E DIT G RADIENT option is on the M ETHOD E DITOR screen.
This option:
• Allows you to more precisely define a gradient than
possible with a finger drag, by displaying the gradient in
a tabular format (Figure 3-5).
• Allows solvents to be changed with each segment of the
separation.
• Set a constant percentage of a third solvent as modifier
in this screen (in the lower right corner of the screen;
Figure 3-5).

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 CombiFlash NextGen Systems
Section 3 Operation

• The table can be collapsed using the EDIT GRADIENT

button again.

Figure 3-5 Edit Gradient screen

Other options on this screen include:

Equilibration Volume-Sets the amount of starting gradient
solvent to equilibrate the column. The volume can be adjusted
depending upon solvent used or column chosen (size and media).
The equilibration solvent mixture is the same as that defined at
the start of the run.
Air Purge-Sets the amount of time air is passed through the
column after completion of the separation to remove free solvent
from the column (only available on systems that have level
sensing capability)
Tube Volume-Sets the capacity for collected fluids. This volume
can be the default maximum volume (Max option) for the tube
size as defined in the configuration window, or a method-specific
volume less than the default maximum capacity.
· This is useful for collecting fractions smaller than the
maximum tube volume when the peak can’t be
detected and must be located after the run via other
detection techniques. Note that the actual fraction
size may be less if a newly detected peak causes a
tube change, or if you click the NEXT TUBE button.
· Fraction volume may also be less than the max
volume if the system is configured with an ELSD or
MS.

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CombiFlash NextGen Systems
Section 3 Operation

3.5.4 Peak Detection UV or UV-Vis – Selecting the λ button allows you to specify the
wavelength for detection
• Up to two different wavelengths can be monitored or
used to trigger fraction collection. This can be set up in
the details menu (described below).
Details-Allows customizing of the detection methods and
detection parameters.
• Signal Gain is a mathematical scaling of the collected
signal for display.
• Slope Based – Allows peaks to be cut based on the
slope of the signal.
· Select the average peak width for your chromatog-
raphy. Peak widths are measured at the baseline.
The slope detector will detect peak widths ranging
from about 0.2 to 2 times the peak width setting for a
moderately sized peak.
· For example, if you entered a peak width of 1 minute,
the range would be 12 seconds to 2 minutes.
· For best operation, the peak width should be set to
just over the average peak width being separated.
For instance, if the average peak width is 45 seconds,
you should enter a peak width of 1 minute. For most
flash chromatography, 1 minute is a good starting
point for a peak width.
• Threshold- Identifies a peak if the absorbance exceeds
the selected value.
· It will continue to identify a peak until the
absorbance drops 0.01 AU below the programmed
threshold to minimize false peaks due to a noisy
signal.
· If both slope based and threshold are selected, these
signals are logically “ORed” together; if either is
active a peak is collected.
• Monitor- Displays the detector signal for reference, but
won’t trigger fraction collection based on the signal.
• Baseline Correction- Allows subtraction of absor-
bance due to solvent UV absorbance.
· Selecting this option enables a pre-run gradient to
provide a ‘blank’ run that can be used to adjust for
absorbance changes due to changing concentration of
UV-absorbing solvents throughout the run.
· Baseline correction may impact peak intensity.
Additionally, this setting may effect peak shape as
the software distinguishes between sample and
solvent.

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 CombiFlash NextGen Systems
Section 3 Operation

Figure 3-6 DETECTION OPTIONS screen

All Wavelength Detection- Enable this option to detect peaks

within a user-selected range of wavelengths.
· This algorithm compares the current total spectrum
of absorbance and compares it to the solvent
spectrum. The more that the spectrum deviates from
the solvent spectrum, the higher the signal. It is
useful to limit the range of wavelengths used in this
algorithm to better differentiate the compound
spectrum from the solvent background spectrum.
ELS Detector- Enable this option to use the optional evapo-
rative light scattering (ELS) detector.
When enabled, click the DETAILS button to configure additional
settings:
• Sensitivity; Low or High sets the preamp gain in the
detector.
· Most flash work should be done at low sensitivity.
High sensitivity is preferred when using higher
boiling point solvents such as heptane, water,
methanol, or acetonitrile.
Signal Gain – You may modify the gain to scale the detector
signal.
Slope Based and Threshold – Refer to these features
described.
Spray Chamber Temperature – The default settings are 30 °C
(normal phase default methods) and 15 °C (reverse phase).
· This setting may be adjusted from 10 to 60 °C so that
detection can be optimized for the solvent system in
use. However, the lower limit is 5 °C below the
ambient temperature.
Drift Tube Temperature – The default settings are 60 °C
(normal phase default methods) and 60 °C (reverse phase).

3-13
CombiFlash NextGen Systems
Section 3 Operation

· This setting may be adjusted from 30 to 90 °C, but is

limited to a range of 5 °C below and 60°C above the
ambient temperature.
Mass Spectrometer – Enable this option to monitor or detect
compounds with a PurIon mass spectrometer system (PurIon
systems only).
• Threshold – Signal level used to detect a peak. This is
based on the baseline noise. The baseline noise is
measured during the first column volume; this is multi-
plied by the value entered in the THRESHOLD control to
generate a trigger value. A peak is collected if the signal
is greater than this trigger value.
· For the PurIon mass detector, only threshold peak
detection is allowed. In this case the threshold is set
as a factor of the measured noise signal. The noise for
the first few seconds of the chromatogram is used to
determine average noise level. Any mass signal that
exceeds this level by the selected factor will be
collected as a peak.
• Monitor – Clicking on this option prevents fraction-
ation based on the mass spectrometer signal.
• Terminate On Target – Stops the run after all mass
spectrometer detection ions have been detected.
• Detection Ions – Used to set ions for detection or to be
monitored. Up to 4 single ions may be chosen, or a range
of ions and up to 3 single ions may be chosen. Detected
ions may be a mixture of positive and negative ions.
Purity Measure – When using two wavelength detection
options, a ratio of the two wavelengths can be displayed which
can provide an indication of compound purity.
• For example, if a pure compound is eluting, the absor-
bance is linearly related to the concentration of the
compound in the solvent regardless of wavelength. If the
compound absorbs differently at different wavelengths,
the absorbance at each wavelength may be different, but
is still linearly related to the concentration. If the
concentration doubles, the absorbance at each
wavelength doubles (at least within the limitations for
Beer’s Law). Since the relationship of absorbance to
concentration is not variable, the ratio remains steady
while the concentration changes from the beginning to
the end of the peak.
• During the duration of the peak, this constant value is
displayed as a horizontal line.
• Now assume a case where there is a second compound
eluting, only slightly shifted in time from the original
compound. It is possible that the detection absorbance
trace alone would indicate a single, valid chromato-
graphic peak. In reality, it is a combination of two peaks.
By monitoring a second wavelength, it may be possible
to reveal the second compound. Because of the slight

3-14
 CombiFlash NextGen Systems
Section 3 Operation

shift in time and the different absorbance properties of

the two compounds, the changing ratio during the
detected peak would reveal the impurity. Therefore, one
can therefore assume that if the ratio is not constant for
the entire duration of the peak, the compound eluting
may not be pure.
• The SHOW SPECTRAL PURITY selection measures purity
by using a comparison of UV-spectra measured at
differing time; it is not limited to 2 wavelengths. The
algorithm used is the "similarity index".
· The SPECTRAL PURITY DETECTION button allows
fractionation based on spectral purity. If the spectra
within the peak are similar, it is considered 1 peak. If
the composition of the peak changes during the peak,
the spectra will not be similar and the purity index
will decrease from the default value of “1” indicating
a change in purity from the early portion of the peak.
This allows the system to advance to the next
fraction as the purity within a detected peak
changes.
Initial Waste & Time Windows – The number of fractions col-
lected can be minimized by setting PEAK COLLECTION to NONE on
the MAIN screen and selecting only portions of the separation for
fraction collection.
• Initial waste can be used to send fluid to waste before
the first peak of interest. Time windows can be used to
identify areas of interest that are collected within the
separation. For example, if you know your peak of
interest will elute between 3 and 4 minutes, you can
create a time window to collect all during this period, or
to collect peaks only during this period.

Figure 3-7 Time Windows screen

Time to CV – Allows the user to change the units used for the
horizontal axis during the separation from the configured units.

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CombiFlash NextGen Systems
Section 3 Operation

Column Information – Displays column usage history. This is

useful for reusable columns such as C18. It can display the age of
the column, number of separations, and fluids left in column in
the previous use.
Gradient Optimizer – Allows a gradient prediction based on
TLC values and is targeted at improving the results for difficult
separations.

3.5.5 Tools

Figure 3-8 Manual Control screen

Manual Control – Capability is also included to allow the user

to wash individual flow paths.
• This is useful for troubleshooting or cleaning flow paths
of potential obstructions or changing from normal to
reverse phase solvent systems.
• If changing solvent systems from reverse to normal
phase or vice versa, the entire flow path should be
flushed with an intermediary solvent that is miscible
with both reverse and normal phase solvents such as
isopropanol, ethanol, or acetone.
• Able to set the flow rate, solvent mixture ratio, and
volume pumped in this menu. Also able to choose to
collect into a certain tube or rack in order to flush
fraction collection arm tubing.
• Contains a lamp energy window to indicate if the flow
cell windows are dirty. An indicator graphically shows
the current level along with a numerical value within
the graph area. Any value in the mid-range of the graph
is normal.
• Offers the ability to manually air purge the cartridge or
column manually (if the system is configured with
solvent level sensing capability).

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 CombiFlash NextGen Systems
Section 3 Operation

3.5.6 Results Screen

Figure 3-9 Results screen

At the end of a separation, the final chromatogram report is

shown. At this point, the user can: print the report; customize the
displayed report by adding multiple UV absorbance or mass
spectra to the report; format the display, reuse the final (poten-
tially modified) method for a subsequent separation; scale up the
method for a separation on a different column size; or add time to
the current separation to continue if the desired compound
hasn’t eluted yet.
If SCALE METHOD is selected, simply chose the size of column for
scaling from the displayed drop down (Figure 3-10). The method
will be scaled for immediate use, or can be saved for later use
(Figure 3-11).

Figure 3-10 Scale Method screen

3-17
CombiFlash NextGen Systems
Section 3 Operation

Figure 3-11 Files screen

3.5.7 PurIon Operation The PurIon functions like another detector in the system. In
addition, operation of the Mass Detector has been integrated into
the PeakTrak control software. To use the PurIon, turn the
PurIon power ON (if not already on). When turned on, the PurIon
requires 30 minutes to achieve proper vacuum levels. A
countdown window will monitor progress. Separations can be
completed without the PurIon during this period.
Operate/Standby/Shutdown: When ready to use the PurIon,
select the MS drop down and select OPERATE. This will turn on
the gas flow, heaters, and detector voltage. When not in use,
select STANDBY to turn of the detector voltage and heaters. The
standby mode will prolong the life of the detector when not in
use. The SHUTDOWN command is used to power down the turbo
pump in the PurIon in preparation for repair or repositioning of
the system. Moving the system without shutting down can cause
major damage to the PurIon. Turbo speed is monitored during
the shutdown process.
Method Development (for PurIon): Choose this command to
open the MS METHOD DEVELOPMENT window to verify ionization
conditions for the compounds to be purified. This command is not
available from a remotely connected computer via a web browser.
• To use the METHOD DEVELOPMENT screen, dissolve a
minute amount of crude sample in a minimal amount of
suitable solvent. Dilute this by placing 1 drop in
~ 20 mL of solvent. Place the front panel inject valve
into the load injection and inject >20 μl using a 22 gauge
blunt tipped needle. Within a few seconds the mass
spectra will be displayed.
• If your ion of interest isn’t visible, it could be due to the
formation of adducts or fragmentation. If you select the

3-18
 CombiFlash NextGen Systems
Section 3 Operation

ION FINDER and enter your desired mass, it will compare

the 10 most intense ions with a list of common adducts
or fragments to determine if your desired mass was
detected as one of the peaks. If so, you can select the
peak for detection in your separation.
• The displayed spectrum will continue to collect mass
information until the window is closed or the display
cleared operation of the injection valve will clear the
current display.
• If you compound isn’t detected, you can adjust the
ionization settings by selecting from the drop down
menu (Figure 3-12). The factory selections include:
· ROBUST for compounds that do not easily ionize.
· TYPICAL works well for most compounds.
· FRAGILE for those compounds that are delicate or
easily fragment.

Figure 3-12 Selecting ion settings

When one of the ion settings is selected, you have the option of
using the suggested setting, or editing the parameters to
optimize the ionization settings. The following are setting
examples:
• Factory Settings- Restore the ROBUST, TYPICAL, and
FRAGILE ion settings to the factory default values.
• Probe - The probe nebulizes and ionizes the sample.
There is a choice of ESI (electrospray interface) or APCI
(atmospheric pressure chemical ionization) probes. The
software will change the labels on the control to reflect
the probe installed in the mass spectrometer.
• Gas Temp-The temperature of the nebulization gas for
the probe. Lower temperatures are used for more
delicate, heat labile compounds. The temperature is set
to quickly evaporate the carrier solvent (note that

3-19
CombiFlash NextGen Systems
Section 3 Operation

PurIon S and PurIon L systems will display a single

temperature for both positive and negative ionization).
• Voltage (Current) - This displays a voltage setting (ESI
probes) or current value (APCI probes). Lower values
are used for more delicate compounds.
· The capillary is heated to complete the evaporation of
solvent. It also carries a voltage; lower voltages are
used for more delicate compounds.
• Source Voltage - The source voltage settings have the
greatest effect on fragmentation. Higher values induce
more fragmentation but also reduce adduct formation.
• Offset- OFFSET is the voltage applied to all masses.
Large values tend to increase fragmentation but reduce
adduct formation.
• Span - SPAN voltage defines an increased voltage
applied as the mass increases. As with the offset, larger
values increase fragmentation.
• Close - Closes the window.

Figure 3-13 Ionization Settings

Once the Mass Spectrum is displayed and represents the infor-

mation you want to see, it can be printed or saved for future ref-
erence. To view more Mass labels, use the FORMAT GRAPH button.
The fluid pressure indicator is used during troubleshooting if
there is a blockage in the fluid interface. The TIC indicator is a
relative measure of background Total Ion Count and therefore a
measure of PurIon cleanliness. It will never read completely 0,
but rather indicates progress of any cleaning activity performed.

3-20
 CombiFlash NextGen Systems
Section 4 Network Configuration

Note
The procedures described in this section will require assis-
tance from your network administrator. Contact your Informa-
tion Technology department before proceeding.

4.1 Network The system has a factory-assigned IP address—192.168.1.51. In

Configurations many cases, this address must be reconfigured for use on a cor-
porate network. To reconfigure the IP address:
1. From the touch screen panel, select the TOOLS>CONFIGU-
RATION menu command.

2. Select the NETWORK CONFIGURATION tab.

3. Select Static IP as the NETWORK TYPE.

Note
The CombiFlash NextGen system also provides limited sup-
port for DCHP connections. If DHCP is desired, select DCHP
as the network type and skip steps 4 through 7. The system
will display instructions required to complete a DHCP connec-
tion.

4. Enter the IP Address, Netmask, and Gateway information

provided by your network administrator.

Note
Should your network administrator request the MAC address, it
can be found in the HELP>ABOUT PEAKTRAK
window.

5. Click the OK button to save the settings.

6. Confirm that your network administrator has completed
any necessary network changes to support the system.
7. Locate the CAT5 connection cable (P/N 480-6545-01) in the
accessory package. Insert one end into the Ethernet port
on the back panel. Connect the other end of the cable to
your network access port.

You should be able to connect to the CombiFlash NextGen from a

PC on the network (Section 4.1.1) and configure other network
features such as network printing (Section 4.1.2) and network
file saving (Section 4.1.3). Networks with Wi-Fi access points
may also support remote control with mobile digital devices.

4-1
CombiFlash NextGen Systems
Section 4 Network Configuration

4.1.1 Network PC Access A successfully networked CombiFlash NextGen system can be accessed
by a PC that meets the recommendations listed in Table 4-1.

Table 4-1 Personal Computer Recommendations a

Operating Systems: Microsoft Windows 7, Windows 8, Windows 10
Hardware: The computer hardware must meet the minimum
required specifications of the selected operating
system.
Network Connection: Ethernet,100 Mbit/s or faster
Display: 1280 x 800 pixels, minimum
Internet Browser: Microsoft Internet Explorer IE 10 or newer

a. This table shows supported PC configurations. Other configurations or

AJAX-compatible browsers may be possible but are unsupported by
Teledyne ISCO.
To access the system, simply open an Internet Browser and enter
the IP address as “http:// ...”, where the ‘...’ are replaced by the
selected address. The browser window will load PeakTrak after
you enter the address.

4.1.2 Network Printing The CombiFlash NextGen system can print to a network printer
and supports both JetDirect and line printer (LPR) queues.
Consult with your network administrator to determine the IP
address of selected printer. If the printer uses an LPR print
queue, you must also find out the queue name. If the printer uses
a JetDirect print queue, also ask for the port number.
When this information is known, you can proceed with config-
uring system for network printing.
1. Select the TOOLS>CONFIGURATION menu command.
2. Select the NETWORK CONFIGURATION tab.
3. Follow the on-screen instructions for entering the address
and queue information.
4. Click OK to save the settings. The CombiFlash NextGen
system will send a test page to the printer.
After successfully printing a test page, the network printer will
be available for printing using the FILE>PRINT menu command
or through the Automatically Print Report at END OF RUN OPTION
ON THE TOOLS>CONFIGURATION USER PREFERENCES tab.

4.1.3 Network File Save To save all run histories automatically as a text or PDF file, con-
Configuration figure the NETWORK FILE SAVE option. This allows the Combi-
Flash NextGen system to access the corporate network and save
the file in a selected Network Share folder. Consult with your
network administrator to configure these fields.

4.2 Direct Connection A direct connection supports communication between the Combi-
Flash NextGen system and a single PC that meets the recom-
mendations in Table 4-1.

4-2
 CombiFlash NextGen Systems
Section 5 Maintenance

5.1 System Standby and During extended periods of inactivity, you can place the system in
Shut Down STANDBY to conserve power. To do so, log off the system (FILE >
LOG OUT) and place the On/Standby switch in STANDBY.
When in the S TANDBY state, normal system operation is no
longer available from the touch screen or remotely. However,
some internal components are still powered.

WARNING
As long as the AC mains power cord is connected, power
is inside the unit. The mains power cord is the disconnect
device. Position the CombiFlash NextGen system so that
the power cord can be unplugged, or use a power strip
where the plug can quickly be removed from the outlet in
the event of an emergency.

When you first place the system in STANDBY, internal compo-

nents continue to operate for almost one minute while per-
forming file maintenance and preparing the system for possible
power removal.

CAUTION
Removing the AC mains power cord before the file mainte-
nance is complete might corrupt files on the internal hard drive.
These corrupted files can cause abnormal operation or a com-
plete system failure that requires service. Unless power must
be removed due to an emergency, always wait at least one
minute after placing the system in STANDBY before removing
the AC mains power cord.

5.2 Preventive The system requires preventive maintenance for safe and
Maintenance reliable operation. Refer to the schedule below for the minimum
periodic maintenance requirements.
As Needed – Perform these tasks as conditions require:
• Cleaning (Section 5.3).
• Quick flow cell cleaning when recommended by a system
alert message (Section 5.5).
• Wipe cone on PurIon system with wipe soaked with
methanol or water to remove visible residue near cone
inlet. (PurIon systems only).

5-1
CombiFlash NextGen Systems
Section 5 Maintenance

Every Run – Perform these tasks at the end of each run:

• Allow the separation run to finish with a high
percentage of solvent B to flush residual compounds
from the column, internal tubing, and flow cell. Refer to
section 5.5.
• Allow Cone wash to run to completion (PurIon systems
only) to wash residual compounds from the fluid
interface, probe, and to clean the cone area.
• Allow Valve Wash sequence to run to completion to wash
residual compounds from the injection valve and ELSD
(if installed) flow path.
Monthly – Perform these tasks at least monthly, more fre-
quently if conditions warrant:
• Tubing Inspection (Section 5.4.
• Collection rack and tray cleaning (Section 5.3.1).
• Monthly flow cell cleaning (Section 5.5.3).
Annually – Perform these tasks at least annually, more fre-
quently if conditions warrant:
• Change roughing pump oil (PurIon systems only).

5.3 Cleaning To clean the exterior surfaces, use a cleaning cloth dampened
with a mixture of distilled water and a mild detergent. Use iso-
propyl alcohol for tougher stains.
On printed areas such as labels, avoid rubbing vigorously or
using aggressive solvents like acetone. Each will ruin the printed
text.

CAUTION
Do not immerse the instrument in a water bath or subject it to a
liquid spray. The instrument is not watertight and these actions
could damage the internal electronics.

5.3.1 Collection Rack and

Tray Cleaning
WARNING
Risk of fire or equipment damage. Unclean collection racks
and tray might inhibit their conductive properties. The
racks and tray must be kept clean to dissipate static
electricity.

The collection tube racks and tray are made of conductive plastic.
Dirt, film, or coatings might prevent their ability to dissipate
static electricity. To avoid problems that possibly result from an
electrostatic discharge, clean the racks and tray monthly. Use
distilled water with a mild detergent. For tougher stains, use iso-
propyl alcohol.

5-2
 CombiFlash NextGen Systems
Section 5 Maintenance

5.4 Tubing Inspection

WARNING
Risk of fire or equipment damage. Faulty tubing, fittings,
and drains may allow organic solvents to pool in unsafe
areas, creating a potential for dangerous levels of
flammable vapors. Improper draining may damage the
instrument’s internal components.

Perform a tubing inspection monthly:

1. Visually inspect the solvent, waste, and drain tubing. The
tubing must be free of any damage, kinks, or deterioration.
Fittings should show no signs of leaks.
2. Test the collection tray drain and top shelf drain by con-
necting a vacuum or air supply source to the outlet end of
the drain tubes. Then, verify the presence of such vacuum
or air supply source on the drain hole (Figures 5-1and 5-2).
Correct any deficiencies before returning the instrument to oper-
ation.

Vacuum or pressurized
air applied to the outlet
end of the drain tube
must exist at the
collection tray drain hole.

Figure 5-1 Fraction collection tray drain hole

Vacuum or pressurized air applied

to the outlet end of the drain
tube must exist at the top self
drain hole.

Figure 5-2 Top shelf drain hole

5-3
CombiFlash NextGen Systems
Section 5 Maintenance

5.5 Flow Cell Cleaning

5.5.1 Post Separation As a preventive measure, all default column methods finish the
separation run with a high percentage of solvent B (Figure 5-3).
This brief time (one to six column volumes) of strong solvent
flushes residual compounds from the column, flow cell, and
internal tubing.

Figure 5-3 Default post-separation column and flow cell

flush

Skipping the post-separation flush may cause residual com-

pounds to build up and crystallize, which might result in:
• Cross contaminating later separation runs.
• Higher operating pressures.
• Reduced flow cell lamp energy.
• A noisy baseline on the absorbance trace.
• Frequent messages recommending flow cell cleaning
(Figure 5-4).
Typically, chemists STOP and then FAST FORWARD the run after
the last compound elutes. This action skips the post-separation
flush. If any of the above conditions appear, consider allowing
some of the runs to continue through the flush, or run a high per-
centage of %B solvent through the system for a few minutes at
the end of each day.
If the separation runs always continue through the flush and the
conditions still occur frequently, edit the D EFAULT COLUMN
METHODs to extend the flush duration.

CAUTION
Do not use polar, basic solvent systems with silica column
media. These solvent systems may break down the silica
structure, possibly causing obstructions in the flow path.
Examples of such solvent systems include, but are not limited
to, those containing more than 20% methanol with ammonia.

5-4
 CombiFlash NextGen Systems
Section 5 Maintenance

5.5.2 Quick Cleaning when When the lamp energy is lower than normal, the system will rec-
Recommended ommend flow cell cleaning (Figure 5-4) before starting a sepa-
ration run.

Figure 5-4 Flow cell cleaning message window

When the system displays this message you can:

• Cancel Run (recommended) – Click the CANCEL RUN
button so you can perform a quick cleaning described in
the following steps.
• Continue Collect All – Click this button to ignore the
message. Because the peak detection operation might be
impaired, the system automatically collects all fluids to
avoid diverting compounds of interest to waste.
• Help – Click this button to display the flow cell cleaning
on-line help topic.
To perform a quick cleaning:
1. After clicking the CANCEL RUN button, select the
TOOLS>MANUAL CONTROL menu option.
2. From the MANUAL CONTROL window, note the Raw Lamp
Energy level at 254 nm. The Raw Lamp Energy gauge has
two ranges: red and green. (Figure 5-5),

Figure 5-5 Raw lamp energy gauge

• Red – Lamp energy is obstructed to a degree that the

system might not reliably detect peaks. If you attempt to
operate the system, peak collection will be forced to
collect all. This prevents diverting desired compounds to
waste.
• Green – Lamp energy is sufficient to detect peaks
within typical system limits.

5-5
CombiFlash NextGen Systems
Section 5 Maintenance

3. Remove the column and insert a bypass tube between the

upper and lower column mounts. A bypass tube (P/N
209-0165-46) can be found in the accessory kit shipped
with your system.
4. Set the Flow Rate to 40 mL/min (Figure 5-6).

Figure 5-6 Manual Control Settings - Quick Cleaning

5. Select THROUGH COLUMN for the VALVE POSITION option.

6. Select NEXT for the Pump into Tube # option. By pumping
into a collection tube, the diverter valve is also cleaned of
any residue during this operation.
7. Click the PRIME B button to pump 100% Solvent B through
the bypass tube and into the collection tubes.

Note
Pumping solvent B at a moderate flow rate (25 to 45 mL/min)
overtime will usually solubilize obstructions. Generally, the rec-
ommended solvent is the highest polarity solvent you have
recently used (solvent B).

8. Monitor the Raw Lamp Energy gauge. As the system

pumps solvent, the raw lamp energy should gradually
improve. Pump solvent for two to five minutes or until the
indicator reaches the far-right of the green range.
If after five minutes the indicator is still in the red range, repeat
the cleaning steps using an alternative solvent. Or, complete the
steps in the Monthly Flow Cell Cleaning procedure (section
5.5.3).

Note
If the numerical values of the Raw Lamp Energy (Figure 5-5)
do not change, or if the first number remains at zero, contact
Teledyne ISCO’s Technical Service department.

5-6
 CombiFlash NextGen Systems
Section 5 Maintenance

5.5.3 Monthly Flow Cell Perform this procedure as part of your scheduled preventive
Cleaning maintenance, or when QUICK CLEANING AS REQUIRED (section
5.5.2) does not improve the lamp energy.
1. Remove the column and insert a bypass tube between the
upper and lower column mounts. A bypass tube
(P/N 209-0165-46) can be found in the accessory kit
shipped with your system.
2. From the menu, select TOOLS>MANUAL CONTROL. This
opens the MANUAL CONTROL window.
3. Set the Flow Rate to 40 mL/min.
4. Select THROUGH COLUMN FOR THE VALVE POSITION option.
5. Select NEXT for the Pump into Tube # option.
6. Place the B1 Solvent inlet line into a reservoir of methanol,
acetone, or a strong solvent that readily dissolves residual
sample material.
7. Click the PRIME B button to pump 100% Solvent B through
the bypass tube and into the collection tubes.
8. After three minutes, click the STOP button. Allow the sys-
tem to stand for at least six hours. Overnight is recom-
mended.
9. Return the B1 solvent line to the original solvent container.
10. Perform the QUICK CLEANING AS REQUIRED (Section 5.5.2
and monitor the Raw Lamp Energy. (Figure 5-5)

If the lamp energy is in the green range return the system

to operation. If the lamp energy is red, contact Teledyne
ISCO’s Technical Service department for assistance.

5.6 ELSD Maintenance Periodic cleaning of the spray chamber will keep the NextGen
operating at maximum performance. Use the following steps if
spray chamber cleaning is recommended.

5.6.1 Cleaning the ELSD 1. From the run screen select the METHOD EDITOR.
Detector 2. In the Peak detection window, select the ELS DETECTOR
radio button then select DETAILS.
3. Set the spray chamber temperature to 40°C.
4. Exit the Edit Method screen and accept the changes, then
allow approximately 5 minutes for the chamber to reach
the set temperature.

5-7
CombiFlash NextGen Systems
Section 5 Maintenance

Figure 5-7 Detection Options screens

5. Lift the p-trap drain tubing up to approximately case top

level. Using a wash bottle, syringe, or suitable measuring
device, slowly fill the drain line with up to 40 mL of ace-
tone. Lift the end of the tubing as needed to transfer most
of the liquid into the spray chamber. Make sure the fluid
level in the tubing doesn’t exceed the level of the instru-
ment case top. If the tubing is raised too fast, fluid may
flow out the top of the vent tube causing a spill. Hold the
tubing up for at least 1 minute after the fluid has been
transferred to the spray chamber.

Figure 5-8 Line filled with 40 ml of acetone

5-8
 CombiFlash NextGen Systems
Section 5 Maintenance

6. Place the open end of the p-trap drain line in a 100 mL (or
greater) beaker.

Figure 5-9 Draining the acetone into a 100 mL

beaker

7. Allow the acetone to drain out of the unit by dropping the

end of the p-trap drain line, while it is in the 100 mL bea-
ker, below the p-trap drain line level.
8. Fill the p-trap pump drain line with up to 40 mL of Isopro-
pyl alcohol using the same procedure as step 5.
9. Place the open end of the p-trap drain line in a 100 mL (or
greater) beaker.
10. Allow the Isopropyl alcohol to drain out of the unit by drop-
ping the end of the p-trap drain line, while it is in the
100 mL beaker, below the p-trap drain line level.
11. Use the DETAILs menu to set the spray chamber to 60°C.
12. Fill the p-trap with up to 40 mL of Isopropyl Alcohol as
before and drape the p-trap drain line of the top of the unit
and let it set for at least 20 minutes.

Figure 5-10 P-trap pump drain line draped over the unit

5-9
CombiFlash NextGen Systems
Section 5 Maintenance

13. Allow the Isopropyl alcohol to drain out of the unit by drop-
ping the end of the p-trap drain line, while it is in the
100 mL beaker, below the p-trap drain line level.
14. To ensure that there is no fluid remaining in the drift tube,
set the drift tube to 90°C. Go to the MANUAL CONTROL win-
dow to turn the gas on and let run 10 minutes. Turn off the
gas.
15. Press FILE and then NEW to reset the method tempera-
tures. Use the instrument normally.

Note
During the rinse steps, it is normal to have flakes or particles in
the wash liquid.

5.7 PurIon Maintenance

5.7.1 Removing ESI Probe Refer to Figure 5-11 for procedure illustration.
1. Use menu item MS -> STANDBY to place the PurIon into
standby mode.
2. Disconnect the high voltage connector by pulling straight
up (do not twist).
3. Unscrew the 10-32 PEEK fitting from the probe.
4. Disconnect the 1/8 inch PTFE nebulization gas tubing from
the push-to-connect fitting.
5. Loosen the thumb screw on the ion source block.
6. Pull out the ESI Probe directly up and out of the ion source
chamber.

Figure 5-11 Removing the ESI probe

5-10
 CombiFlash NextGen Systems
Section 5 Maintenance

5.7.2 ESI Probe 1. Insert the ESI probe into the source chamber (ensure the
Replacement corona discharge pin is removed from the source chamber.
Refer to Removing the Ion Source Chamber and Reinstall-
ing the Ion Source Chamber to remove the corona dis-
charge pin- (APCI equipped systems only) (Section 5.7.5).
2. Push down on the ESI probe and tighten the thumb screw
on the ion source chamber to ensure the ESI probe is fixed
in position.

Note
Failure to tighten the thumbscrew will result in Error 317, indi-
cating the probe is not fully seated in the ion source housing.

3. Connect the 1/8 inch PTFE tubing into the push-to-connect

fitting labeled as 'Neb Gas'.
4. Finger-tighten the 10-32 PEEK fitting.
5. Connect the high voltage connector.

5.7.3 APCI Probe Removal Refer to Figure 5-12 for procedure illustration.
1. Disconnect the high voltage connector by pulling straight
up (do not twist).
2. Unscrew the 10-32 PEEK fitting from the probe inlet.
3. Disconnect the 1/8 inch PTFE tubing from the push-to-con-
nect fitting labeled as 'Neb Gas'.
4. Disconnect the 1/8 inch PTFE tubing from the push-to-con-
nect fitting labeled as 'APCI heated gas'.
5. Loosen the thumbscrew on the front of the ion source hous-
ing.
6. Carefully lift and pull out the APCI probe from the source
chamber.
7. If changing to ESI, remove the corona discharge pin (see
removing the ionization source chamber Section 5.7.5).

5-11
CombiFlash NextGen Systems
Section 5 Maintenance

Figure 5-12 Removing the APCI probe

5.7.4 APCI Probe 1. Remove the ion source housing and install the corona dis-
Replacement charge pin. The pin should not be placed over the cone inlet
hole, but rather at the "9:00" position.
2. Insert the APCI probe into the source chamber.
3. Push down on the probe and tighten the thumbscrew on
the front of the ion source housing.
4. Connect the 1/8 inch PTFE tubing labeled "Neb Gas" into
the push-to-connect fitting labeled as 'Neb Gas'.
5. Connect the 1/8 inch PTFE tubing labeled "APCI" to the
push-to-connect fitting labeled as 'APCI heated gas'.
6. Finger-tighten the 10-32 PEEK fitting.
7. Connect the high voltage connector.

5.7.5 Removal of Ion Source 1. Follow the steps in "Removing the ESI Probe" (5.7.2) or
Housing "Removing the APCI Probe" (5.7.4) as appropriate to dis-
connect the lines.
2. Loosen the two thumb screws and unscrew the ¼-28 PTFE
tube fitting labeled as 'heated desolvation' from the side of
the chamber.

5-12
 CombiFlash NextGen Systems
Section 5 Maintenance

Figure 5-13 Ion source chamber

3. Gently lift and pull out the ion source chamber.

5.7.6 ESI and APCI Removal Refer to Figure 5-14 or procedure illustration.
from PurIon S and 1. Place the mass spectrometer STANDBY mode.
PurIon L
2. Unscrew the ¼-28 PEEK fitting at the top of the ion source
housing.
3. Loosen the two clamps at both sides.
4. Gently lift and pull out the source housing.

Figure 5-14 ESI and APCI removal from PurIon S and PurIon L

5.7.7 ESI and APCI 1. Carefully place the ion source housing on top of the base
Replacement from plate and line up with the rear electrical connection. Push
PurIon S and PurIon L down until source chamber is seated evenly across the base
plate.
2. Lock down two housing clamps at both sides.
3. Finger-tighten the ¼-28 PEEK fitting at the top of the
source housing.

5-13
CombiFlash NextGen Systems
Section 5 Maintenance

5.7.8 Cleaning the Plugging of the capillary (either in the ESI or APCI probe) causes
Ionization Source the pressure of the carrier fluid from the fluid interface to exceed
Capillary the maximum operating pressure indicated by Error 310 or Error
316. This error can be avoided by filtering directly injected
samples through a 0.45 µ syringe filter. To unplug the capillary,
complete the following:
1. Complete the appropriate procedure to remove the probe
from your system.
2. Using a 1 mL syringe, with Luer-Loc and adaptors, con-
nected to ESI or APCI capillary and push fluid through to
remove plug. If this procedure does not resolve the issue,
an HPLC pump can be connected to the inlet fitting.

Figure 5-15 ESI probe with syringe

3. If this procedure fails to unplug the capillary, refer the

instructions for the rebuild kit provided with your PurIon
system.
4. Reinstall the probe following the appropriate procedures
for your system.

5.7.9 Replacement of Ion 1. Place the ion source housing on top of the base plate and
Source Housing line up with the rear electrical connection. Push down until
source chamber is seated evenly across the base plate.
2. Finger tighten the two thumb screws and finger tighten
the ¼-28 PTFE tube fitting labeled as 'heated desolvation'
on the side of the housing.
3. Follow the steps in "Replacing the ESI Probe" (5.7.2) or
"Replacing the APCI Probe" (5.7.4)as appropriate to recon-
nect the lines.

5.7.10 Overpressure Error The fluid interface has a pressure transducer to monitor
pressure of the carrier fluid. Since the sample is introduced at
the splitter valve, plugs usually occur between the valve and the
PurIon source sprayer. The most common location for a plug is
within the probe capillary. The occurrence of plugs can be
reduced by using a 0.45 µ syringe filter when injecting samples
for Method Development and Ionization Settings. To trouble-
shoot an overpressure error, complete the following:

5-14
 CombiFlash NextGen Systems
Section 5 Maintenance

• If the error occurs during a run, select CONTINUE

WITHOUT PurIon, and the purification can continue
without the PurIon signal or peak detection, but will
rely on any other detector selected such as UV or ELSD.
This allows the run to be completed before trouble-
shooting the plug.
• Use the menu item MS-> Manual control.
• Press the START CARRIER PUMP button.
• Watch the pressure on the ribbon gauge.
• Loosen the fitting at the source inlet. If the pressure
drops, then the source capillary is plugged.
• If there is still an error or the pressure remains high,
then the plug is between that point and the splitter
valve (or the valve itself). Continue to loosen fittings
going back the to the splitter valve until the error is
corrected.

5.7.11 Check Valve Cleaning If the check valves are allowed to dry out after using volatile
salts (e.g., ammonium acetate or ammonium formate), they may
stick and fail to function. Complete the following to clean the
check valves:
1. Remove the inlet and outlet lines from the check valve
holders (Figure 5-16), and then remove the check valve
holders. Pliers may be needed to remove the holders.

Figure 5-16 Check valve holder

2. Remove check valves from pump head and place in a bea-

ker of methanol. Sonicate check valves for 15 to 20 min-
utes.
3. Reinstall check valves into pump head making sure that
the ends of the check valves (with multiple holes) are fac-
ing up towards the outlet of the pump.
4. Reinstall check valve holders and tighten finger tight.
Then using pliers tighten an additional ¼ turn.
5. Check the flow rate delivery. If the flow rate delivery is still
incorrect, replace the check valves. The correct flow rate

5-15
CombiFlash NextGen Systems
Section 5 Maintenance

depends on the back pressure of the carrier solvent. The

flow will be 0.5 mL/minute until the fluid interface reaches
operating pressure, then reduces the flow rate to 0.2
mL/minute under typical operating conditions for the fluid
interface. The operating mode can be determined by listen-
ing to the pump operate. While running at 50 mL/min, the
pump motor speed is continuous. Once operating pressure
is reached, the pump has a rapid refill stroke approxi-
mately that occurs ~ 8 seconds.

5.7.12 Replacing Check Valves 1. Install new check valve cartridges (P/N: 250-0001-17) into
pump head housing making sure the ends of the cartridges,
with three small holes, are facing upwards towards the
outlet (Figure 5-17).

Figure 5-17 Correct orientation of cartridge

2. Tighten check valve holders finger tight, then with pliers

tighten an additional ¼ turn.

5.8 Electrical If your instrument stops working and the touch panel display is
Troubleshooting off, check the line cord connection.
If the line cord is connected properly, check circuit breaker on the
system’s rear panel to ensure it is switched to the ON position.

Table 5-1 Common PurIon

Error Codes and Resolutions
PurIon temperatures are stabilizing. The PurIon has several areas with heaters. The software
has a set timer to allow temperatures to come up to oper-
[r] seconds remain. (where [r] is a number)
ating conditions. After that time, a separate error is thrown
if the heaters are not within an acceptable band. The
default time is 300 seconds after entering the operate
condition.
During the standby condition most heaters are set to OFF
except the inlet capillary with is set to 50 °C during
standby.
The PurIon vacuum level is too low to operate. Please The Pirani pressure must be below 5.5E-3 mbar before
verify that the roughing pump is on and operating cor- the PurIon turbo pump will operate. If trying to place the
rectly. PurIon in operate without turning on the roughing pump,
this message will appear.
Pirani Pressure: [s1] mbar. (where s1 is the vacuum read-
ing). This error generally occurs if the user forgets to turn the
roughing pump back on after cleaning the capillary or
changing the pump oil.

5-16
 CombiFlash NextGen Systems
Section 5 Maintenance

Table 5-1 Common PurIon

Error Codes and Resolutions
The PurIon has been shut down. It will be unavailable for This message is displayed after the PurIon has been suc-
use until the NextGen has been rebooted. Please ensure cessfully shut down through the SHUTDOWN command. It
the PurIon and fluid interface are both turned on before serves as a reminder that an NextGen reboot will be nec-
rebooting the NextGen. essary for the PurIon to be enabled again by the Next-
Gen.
The splitter valve seals have exceeded their recom- The splitter valve supplier has stated that typical valve life
mended life. The PurIon will continue to operate, but exceeds 1,000,000 actuations. After that time, there is no
there is an increased possibility of leakage at the splitter method of determining when it will leak. The valve can be
valve and loss of PurIon detection during a separation. rebuilt using the valve rebuild kit (P/N:60-5234-629) or
continue to use and monitor for leakage. Leakage should
not drip out the bottom of the fluid interface front cover.
No ion source is detected on the PurIon. Please ensure The PurIon has reported that the ion source high voltage
that the ion source is properly installed and connected. If cable is not properly plugged into its socket. This is the
the source is still not detected, contact a qualified service cable with the round connector and is to the right and
technician. Error 309 behind the ion source.
This generally occurs upon changing or cleaning probes.
A plug has occurred in the PurIon fluid lines. The separa- This error message is displayed if a plug is detected dur-
tion can be continued without PurIon detection or contin- ing a separation.
ued if the plug is corrected. Error 310
The PurIon inlet cone may be plugged which could pre- During normal operation, the Pirani pressure should be
vent detection. Continued operation will not cause dam- >1.5E-3 mbar. Anything less is an indication of either par-
age. Please contact a qualified service technician to tial or complete plugging of the capillary cone entrance to
clean the cone; Pirani Pressure: [s1] mbar. Error 315 the vacuum area. See cleaning the cone capillary (5.7.8).
A plug has occurred in the PurIon fluid lines. The plug This error occurs if the tubing is plugged while the
must be corrected to allow continued operation. Error method development screen is in use. See cleaning the
316. probe capillary (5.7.8).
The ionization probe (ESI or APCI) isn’t fully seated into On PurIon systems (not PurIon S or PurIon L), the probe
the ion source housing. Please ensure the probe is fully isn’t seated properly. Loosen the thumbscrew on the front
seated, then press OK to continue. Error 317. of the ion source housing, push down on the probe, then
tighten the thumbscrew.
The fluid interface pressure is too low. Error 325 This could be due to lack of carrier fluid, loss of pump
prime, or leakage.

Table 5-2 General Troubleshooting

Error/Symptom Solution
No signal on any mass, or very weak • Verify nitrogen pressure. Nitrogen isrequired to nebulize the
signal sample.
• Check carrier solvent level, refill carrier solvent and re-prime the
carrier solvent pump. Carrier solvent is needed to bring sample to
the mass spectrometer.
• Check that the priming valve is tight. Aloose priming valve
prevents sample from getting to the mass spectrometer.
• Verify the splitting valve is not leaking.This causes loss of carrier
solvent preventing sample from getting to the mass spectrometer.
• Check for leaks around other fittings on the solvent interface.

5-17
CombiFlash NextGen Systems
Section 5 Maintenance

Table 5-2 General Troubleshooting

Delayed signal during run. • Verify nitrogen pressure. Sample may drip through the cone and
still be detected later.
• Check priming valve tightness. A slight leak will delay sample.
• Verify the splitting valve is not leaking.
• Check for leaks around other fittings on the solvent interface.
Sample masses observed but weak, or • Check polarity to see if the sample ionizes under the opposite
masses other than the analyte polarity.
observed.
• Try another ionization settings.
• Compound fragments- use fragment mass or try another
ionization setting.
• Compound forms adducts.
• Compound fails to ionize using the selected probe. Some
compounds ionize better using APCI rather than ESI, for example.

5-18
 CE ISMI-A

EU DECLARATION OF CONFORMITY
Application of Council Directive: 2014/30/EU - T h e EMC Directive
2014/35/EU - T h e Low Voltage Directive
2014/53/EU - Radio Equipment Directive {RF ID reader)
2011/65/EU - T h e RoHS Directive

Manufacturer's Name: Teledyne lSCO

Manufacturer's Address: 4700 Superior
Lincoln, Nebraska 68504 USA
Mailing Address: P.0. Box 82531, Lincoln, NE 68501

Equipment Type/Environment: Laboratory Equipment for Light Industrial/Commercial Environments

Trade Name/Model No: CombiFlash NextGen100, NextGen300, and NextGen300+ all including optional
ELSD and Purl on

Year of Issue: 2018

EN 55011:2009/2010
EN 61326-1:2013
Harmonized Standards: EN 61000-3-2:2014
EN 61000-3-3:2013
EN 61010-1:2010
EN 61010-2-081:2015
EN 61010-2-010:2014

RF ID reader:
EN 300 330-1 V2.l.1
EN 301489-1 V2.l.1
EN 301489-3 V2.l.1
EN 50364:2010

I, the undersigned under sole responsibility, hereby declare that the design of the equipment specified
above conforms to the above Directive(s) and Standards as of JUNE 18, 2018.

USA Representative

Signature:

Printed Name:
Title:

605252081
Rev - B

END-04-005 11/30/2020
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O: Represent the concentration of the hazardous substance in this component’s any homogeneous pieces is lower than the
ST/ standard limitation.

X烉堐䣢宍㚱㭺㚱⭛䈑峐军⮹⛐宍悐ẞ䘬㝸ᶨ⛯峐㛸㕁ᷕ䘬⏓慷崭↢ST/ 㞯Ⅾ奬⭂䘬旸慷天㯪ˤ
(ẩ᷂⎗⛐㬌⢬炻㟡㌖⭆旭ね⅝⮡ᶲ堐ᷕㇻ“X” 䘬㈨㛗⍇⚈徃埴徃ᶨ㬍宜㖶ˤ)
X: Represent the concentration of the hazardous substance in this component’s at least one homogeneous piece is higher
than the ST/ standard limitation.
(Manufacturer may give technical reasons to the “X”marks)

䍗ᾅἧ䓐㛇䓙乷樴䠖⭂ˤ
The Environmentally Friendly Use Period (EFUP) was determined through experience.

䓇ṏ㖍㛇塓亾䞩⛐䲣↿⎟䞩ᷕˤ⇵ᶱỵ㔘⫿ᷢ䓇ṏ⸜(207 ẋ堐2007⸜) ˤ昷⎶䘬ᶨ᷒⫿㭵ẋ堐㚰ấ烉A ᷢᶨ㚰炻

BᷢḴ㚰炻䫱䫱ˤ
The date of Manufacture is in code within the serial number. The first three numbers are the year of manufacture with the
second digit removed (218 is year 2018) followed by a letter for the month. "A" is January, "B" is February and so on (I is
not used).

CombiFlash and ACCQPrep Hazmat Table 60-5233-406 Rev. C

 Teledyne I6&2 One Year Limited Factory Service Warranty*
This warranty exclusively covers Teledyne ,6&2 The warranty for any instrument is the one in effect on date
instruments, providing a one-year limited warranty of shipment. The warranty period begins on the shipping
covering parts and labor. date, unless Teledyne ,6&2 agrees in writing to a
Any instrument that fails during the warranty period due to different date.
faulty parts or workmanship will be repaired at the factory Excluded from this warranty are normal wear; expendable
at no charge to the customer. Teledyne I6&2’s items such as desiccant, pH sensors, charts, ribbon, lamps,
exclusive liability is limited to repair or replacement of tubing, and glassware; fittings and wetted parts of valves;
defective instruments. Teledyne ,6&2 is not liable for check valves, pistons, piston seals, wash seals, cylinders,
consequential damages. pulse damper diaphragms, inlet lines and filter elements;
Teledyne ,6&2 will pay surface transportation charges both and damage due to corrosion, misuse, accident, or lack
ways within the 48 contiguous United States if the of proper LQVWDOODWLRQRUmaintenance. This warranty does
instrument proves to be defective within 30 days of not cover products not sold under the Teledyne
shipment. Throughout the remainder of the warranty period, ,6&2 trademark or for which any other warranty is
the customer will pay to return the instrument to Teledyne specifically stated.
,6&2 and Teledyne ,6&2 will pay surface transportation No item may be returned for warranty service without a
to return the repaired instrument to the customer. return authorization number (RMA) issued by Teledyne
Teledyne ,6&2 will not pay air freight or customer’s ,6&2.
packing and crating charges. This warranty does not cover This warranty is expressly in lieu of all other warranties
loss, damage, or defects resulting from transportation and obligations and Teledyne Isco specifically disclaims
between the customer’s facility and the repair facility. any warranty of merchantability or fitness for a
particular purpose.
The warrantor is Teledyne ,6&2, 4700 Superior, Lincoln,
NE 68504, U.S.A.
*This warranty applies to the USA and countries where Teledyne ,6&2 does not have an authorized
dealer.Customers in countries outside the USA, where Teledyne Isco has an authorized dealer, should
contacttheir Teledyne ,6&2 dealer for warranty service.

Problems can o f t e n be diagnosed and corrected without returning the instrument to the
factory. Before returning any instrument for repair, please contact the Teledyne Isco Service
Department for instructions and to obtain a return material authorization number (RMA).
Instruments needing factory repair should be packed carefully and shipped to the attention of
the service department. Small, non-fragile items can be sent by insured parcel post. PLEASE
WRITE THE RMA NUMBER ON THE OUTSIDE OF THE SHIPPING CONTAINER and
enclose a note explaining the problem.

Shipping Address: Teledyne ,6&2 - Attention Repair Service

4700 Superior Street
Lincoln, NE 68504 USA
Mailing Address: Teledyne ,6&2PO Box
82531 Lincoln, NE 68501
USA
Phone: Repair service: (800) 775-2965 (lab instruments)
(866) 298-6174 (samplers & flow meters)
Sales & General Information: (800) 228-4373 (USA & Canada)
Fax: (402) 465-3001
Email: [email protected]

March 2, 2016 P/N 60-1002-040 Rev J

b