Series 200 Vacuum Degasser

User’s Manual
Release History

Part Number Release Publication Date

09936206 D April 5, 2004

Any comments about the documentation for this product should be addressed to:

User Assistance
PerkinElmer
710 Bridgeport Avenue
Shelton, CT 06484-4794

Or emailed to: info@perkinelmer.com

Notices
The information contained in this document is subject to change without notice.
Except as specifically set forth in its terms and conditions of sale, PerkinElmer makes no
warranty of any kind with regard to this document, including, but not limited to, the
implied warranties of merchantability and fitness for a particular purpose.
PerkinElmer shall not be liable for errors contained herein for incidental consequential
damages in connection with furnishing, performance or use of this material.

Copyright Information
This document contains proprietary information that is protected by copyright.
All rights are reserved. No part of this publication may be reproduced in any form
whatsoever or translated into any language without the prior, written permission of
PerkinElmer, Inc.

Copyright © 2004 PerkinElmer, Inc.

Produced in the U.S.A.

Trademarks
Registered names, trademarks, etc. used in this document, even when not specifically
marked as such, are protected by law.

PerkinElmer is a registered trademark of PerkinElmer, Inc.

 Safety and Regulatory
Information S
Electromagnetic Compatibility (EMC)
United States
This product is classified as a digital device used exclusively as industrial, commercial, or
medical test equipment. It is exempt from the technical standards specified in Part 15 of
the FCC Rules and Regulations, based on Section 15.103 (c).

Europe
All information concerning EMC standards is in the Declaration of Conformity and these
statements will change as the European Union adds new requirements.
S-2 Safety and Regulatory Information Symbols and Conventions used in this Manual

Symbols and Conventions used in this Manual

In this manual the following graphic symbols and special text formats are used to set apart
important safety information.

A warning indicates an operation that can cause

personal injury if precautions are not followed.

A caution indicates an operation that can cause

instrument damage if precautions are not followed.

Notes emphasize significant information in a procedure or

description.

Symbols used on the Instrument

There are two different types of warning symbols that appear on the instrument.

This symbol indicates an operation (or condition on

the instrument) that could cause personal injury if
precautions are not followed. This can be any type of
hazard.
When you see this symbol, refer to the safety pages in
the manual for further information.
Consulter les documents d’accompagnement.

This symbol indicates the danger of electric shock, if

precautions are not followed, due to the presence of
high voltage.
Attention. Risque de choc électrique.
Warnings on the Instrument Safety and Regulatory Information S-3

Warnings on the Instrument

The following warning labels are affixed to the side panel of the instrument for all models
as shown in the figure below.

Figure S-1. Location of Warning Labels on the Series 200 Vacuum Degasser
S-4 Safety and Regulatory Information Warnings on the Instrument

Grounding circuit continuity is vital for safe operation of

equipment. Never operate equipment with grounding conductor
disconnected.

Débrancher le cordon d’alimentation avant d’ouvrir da

continuité des masses est essentielle. Pour un fonctionnement
sans danger. Ne jamais utiliser l’équipment si le fil de terre n’est
pas raccordé.

For protection against fire hazard, replace only with same type
and rating of fuse.

Afin d’assurer la protection contre les risques d’incendie,

remplacer uniquement par un fusible de même type et de même
courant nominal.

To reduce the chance of electric shock, do not remove this cover.

No user-serviceable parts are inside. Refer servicing to
PerkinElmer qualified service personnel.

Avertissement. Pour réduire le risque de chocs électriques, ne pas

ouvrir les couvercles si un outil est nécessaire. Ne contient aucune
pièce pouvant être réparée par l’utilisateur. Confier le
dépannage au personnel qualifié de PerkinElmer.

IMPORTANT Do not prime using pressure. Prime by suction only.

Internal membrane must not be pressurized above 10
psi.
Warnings on the Instrument Safety and Regulatory Information S-5

The following warning label covers the fuse receptacle located on the rear panel of the
instrument:

The Series 200 Vacuum Degasser is shipped from the

factory with:
- The main AC power switch set to the OFF
position.
- No fuses installed.
You need to install the proper fuse before installing
the line cord and applying AC power.

Additional graphic symbols used on the instrument are the following:

S-6 Safety and Regulatory Information Electrical Warnings

Electrical Warnings

Connect the instrument to an AC line power outlet

that has a protective ground connection. To ensure
satisfactory and safe operation of the instrument, it is
essential that the protective ground conductor (the
green/yellow lead) of the line power cord be connected
to true electrical ground. Any interruption of the
protective ground conductor, inside or outside the
instrument, or disconnection of the protective ground
terminal may impair the protection provided by the
instrument.

Do not operate the instrument with any covers or

parts removed.

Do not attempt to make adjustments, replacements or

repairs to this instrument except as described in the
accompanying User Documentation. Only a
PerkinElmer service representative or similarly
trained and authorized person should be permitted to
service the instrument.

Use only fuses with the required current rating and of

the specified type for replacement.
Quality Control/Good Laboratory Practices Safety and Regulatory Information S-7

Quality Control/Good Laboratory Practices

Quality Control
The user should develop appropriate quality control procedures for the LC Degasser (and
the entire LC system) to ensure suitability for its intended use. These procedures typically
consist of periodic performance verifications and routine inspections and suitability tests.

Certificate of System Control

Each Series 200 Vacuum Degasser is carefully built and tested in a controlled system in
accordance with the requirements specified in its applicable PerkinElmer Final Assembly
and Test Specification.

Each instrument is certified to meet its functional and performance specification upon
release to shipment. The integrity of this quality system is routinely audited and is
certified by a registrar.

Instrument Performance Verification (IPV)

To maintain functional performance, PerkinElmer recommends a yearly Instrument
Performance Verification (IPV) of the Series 200 Vacuum Degasser by a PerkinElmer
Service Engineer to ensure its operation within published specifications. These tests
consist of measuring the most important Degasser characteristics such as retention time
repeatability. Certification is available for regulatory compliance. Contact your local
PerkinElmer Sales and Service office.

Routine Inspection and Suitability Test

The Series 200 Vacuum Degasser housing should also be inspected weekly through the
ports for each Channel (A through D) and all 1/4-28 connectors for signs of liquid leaks.
Prior to any sample analysis, a system suitability test, which closely resembles the intended
assay, should be performed to ensure that the LC system is operating within established
criteria (e.g., peak resolution, peak asymmetry, precision, retention time, column plate
count, pressure limits, signal/noise ratio, etc.)

While the Universal Test Mix (UTM) Part No. 0089-0893, can be used for a system check,
we recommend that you develop a separate system suitability test and acceptance criteria
for each of your assays.
S-8 Safety and Regulatory Information Hazardous Chemicals

Hazardous Chemicals
Before using mobile phase solvents, you should be thoroughly familiar with all hazards
and safe handling practices. Observe the manufacturer’s recommendations for use, storage
and disposal. These recommendations are normally provided in the material safety data
sheets (MSDS) supplied with the solvents.

Some chemicals used with this instrument may be

hazardous or may become hazardous after completion
of an analysis. The responsible body (e.g., Lab
Manager) must take the necessary precautions to
ensure that the surrounding workplace and that the
instrument operators are not exposed to hazardous
levels of toxic substances (chemical or biological) as
defined in the applicable Material Safety Data Sheets
(MSDS) or OSHA, ACGIH, or COSHH documents.
Venting for fumes and disposal of waste must be in
accordance with all national, state and local health
and safety regulations and laws.

The degassing membrane in the Series 200 Vacuum

Degasser is manufactured from Teflon AF®. As with
older membranes manufactured from PTFE, Teflon
AF® is inert to all solvents normally used in HPLC.
However, Teflon AF® is soluble in perfluorinated
solvents such as Fluorinert® FC-75 and FC-40. Use of
such solvents in the Series 200 vacuum degasser will
result in the destruction of the membrane and
degasser.
Hazardous Chemicals Safety and Regulatory Information S-9

Definitions in Warning for Hazardous Chemicals

Responsible body. “Individual or group responsible for the use and maintenance of
equipment, and for ensuring that operators are adequately trained.” [per IEC 1010-1,
Amendment 2].

Operator. “Person operating equipment for its intended purpose.” [per IEC 1010-1,
Amendment 2].

OSHA: Occupational Safety and Health Administration (United States)

ACGIH: American Conference of Governmental Industrial Hygienists

COSHH: Control of Substances Hazardous to Health (United Kingdom)

S-10 Safety and Regulatory Information Temperature, Humidity and Environment

Temperature, Humidity and Environment

This instrument is designed for indoor use only.

Do not operate in a Cold Room or a refrigerated area.

The Series 200 Vacuum Degasser operates most
efficiently under the following conditions:
• Ambient temperature is 10 to 35 °C
• Ambient relative humidity is 20 to 80%
noncondensing
• Altitude is in the range of 0 to 2000 m.

This instrument is not designed for operation in an

explosive environment.

Installation Category

This instrument is able to withstand transient overvoltage according to Installation

Category II as defined in IEC 1010-1.

Pollution Degree

This equipment will operate safely in environments that contain nonconductive foreign
matter up to Pollution Degree 2 in IEC 1010-1.

Storage Conditions
The Series 200 Vacuum Degasser may be stored under the following conditions:

• Ambient temperature is -20 to +60 °C,

• Ambient relative humidity is 20 to 80% noncondensing,
• Altitude is in the range 0 to 12,000 m.
 Introduction 1
About the Series 200 Vacuum Degasser
The Vacuum Degasser is a high-efficiency in-line module that removes dissolved
gasses from HPLC solvents. Its unique design assures reliable continuous operation
and the highest level of continuous performance available without the need for
helium degassing. Up to five solvent lines may be degassed simultaneously by one
unit. The extremely low internal volume of each Teflon AF® channel offers very
quick equilibration and very short startup times compared with PTFE degassing
channels which have the same degassing efficiency.

Inside the unit, the solvent flows through a short length of Teflon AF® tubing which
is located in a vacuum chamber. Within this chamber a partial vacuum is maintained
by a constantly running, low RPM vacuum pump. Dissolved gasses migrate across
the tubing wall under a concentration gradient produced by the vacuum as the
solvent flows within the coil. Gasses removed are expelled, and the chamber is
maintained at a constant, preset vacuum level by varying the vacuum pump speed as
needed.

A special port in the vacuum pump continually flushes the pump head with a small
“bleed” of air to remove any solvent vapors which may enter the pump from the
vacuum chamber. This air bleed eliminates the need for any solenoid valves within
the system. This patented* design results in zero vacuum “hysteresis”. Previous
designs allowed the vacuum chamber pressure to fluctuate, with the pump cycling on
and then off in response to the vacuum level.

*Thisproduct is protected under U.S. patents 6,494,938; 6,248,157; 5,340,384 and 5,006,382. Other patents
pending.
ZHCR® is a registered trademark of Systec, Inc., New Brighton, Minnesota.
Teflon AF® is a registered trademark of E.I. du Pont de Nemours and Company.
1-2 Introduction

About the Controls and Indicators

Front Panel Controls and Indicators

A series of three LED’s is located on the front of the instrument above the solvent
inlets and outlets. These three LED’s are used by the microprocessor to indicate the
system status.

Immediately upon turning on the instrument, all 3 LED’s will momentarily flash on
then turn off. This confirms that the microprocessor has examined the vacuum sensor
and found that it is within the expected range of 0 to 810 mm Hg absolute pressure.

Following the startup test, the microprocessor ramps the vacuum pump to high RPM,
to quickly exhaust atmosphere from the vacuum chamber. The left-hand LED
illuminates as the pump can be heard to run at high RPM. As vacuum is established
in the chamber, the middle yellow LED illuminates, indicating the vacuum within the
chamber has reached a value of less than 80 mm Hg absolute. As the vacuum pump
continues to remove air within the vacuum chamber, the right-hand green LED will
illuminate and the middle yellow LED extinguish, indicating that the vacuum
chamber has reached a vacuum of 54 mm Hg absolute, where the performance of the
degasser meets its specified level. When the vacuum level passes through 47 mm Hg
absolute, the microprocessor ramps the pump speed to the low RPM, continuous run
mode. At this low RPM, the vacuum will continue to fall to a constant value,
wherein the fluctuation in the vacuum is less than 1mm Hg. This “zero hysteresis
constant run” (ZHCR) mode is necessary, due to the extremely low mass, high
response, Teflon AF® degassing channel. The effect of vacuum changes within the
vacuum chamber can easily be seen in the UV absorbance of methanol (for example)
at 215 nM. Only a ZHCR design ensures a baseline which is unaffected by the
degasser.

An additional benefit of the constantly running pump is that a pressure rise in the
vacuum chamber can be observed. This “smart leak detection” is a benefit of the
patent-pending design of the degasser, and is best described as follows: Due to the
fixed rate at which the pump runs, the gasses crossing the membrane pass from the
chamber at a fixed rate as well. If a leak occurs within the chamber, the
microprocessor will see this as a rise in pressure related to the vapor pressure of any
leaking solvent. If the pressure within the chamber rises above 54 mm Hg absolute,
the yellow LED will once again illuminate, indicating a possible leak condition
within the chamber. If the pressure rises above 80 mm Hg, the system will shut down
and go into a “safe” mode.
 Introduction 1-3

Front Panel Connections

Depending upon the model, there are from 1 to 5 degassing channels. Pairs of
female ¼-28 connectors are located on the front of the Vacuum Degasser cabinet.
These are the input and output ports for running up to 5 solvent lines through the
Vacuum Degasser. Each channel has an input port and an output port on the same
level, labeled on the front panel as “A” through “E”. Flow direction is not critical.
Plugs are provided to seal the ports of unused channels.

Front Panel Indicators

Three LEDs are located on the front of the instrument above the solvent inlets and
outlets:

POWER (Green)

Indicates when power is applied to the Vacuum Degasser (plugged in and Power
switch ON).

STATUS (Yellow)

Indicates when vacuum level is outside acceptable operating range. Normally it

will come on at initial power-up and remain on during pump-down. It will go off
in a few minutes when the vacuum level goes below 100 mm of Hg absolute. If
an error condition occurs, this LED will flash in one of two modes:

• Flashing on and off in even 1-second intervals: pump was not able to reach
vacuum set point, indicating a possible leak in the system.

• Flashing on for 1 second and off for 2 seconds indicates a vacuum signal
error.

VACUUM (Green)

Indicates when vacuum level is within acceptable operating range. Normally it

will come on after the initial pump-down, and remain on as long as the Vacuum
Degasser is powered up and vacuum level is below 100 mm of Hg absolute.
1-4 Introduction

Figure 1. Front view of 5-Channel Vacuum Degasser

Rear Panel Connections and Indicators

Exhaust Port

The gas pumped out of the vacuum chamber leaves the unit through the
exhaust port.

Power Receptacle

The power receptacle accepts the DC plug of the supplied AC Adapter. See
Specifications section for further power requirement information.

Power Switch

This On/Off rocker type switch applies line power to the Vacuum Degasser. The
rocker indicators are as follows: “O” = Off and “|” = On.
Scope of the Manual Introduction 1-5

Validation Connector (Optional)

Depending on the model, there may be a 2-pin receptacle labeled

“Validation” located next to the power switch. This receptacle and its mating
screw-lock plug allow a validation signal from the Vacuum Degasser’s control
circuit to be sent to a computer or data system. This validation output indicates
vacuum level (see Specifications section for details).

Figure 2. Rear view of Vacuum Degasser

Scope of the Manual

This manual contains information about how to install, use, and troubleshoot the
vacuum degasser. This manual is written for a person who has developed a working
knowledge of liquid chromatography and understands:

• The function of each major component in a liquid chromatography system.

1-6 Introduction Warranty Exclusions and Limitations

• How to select a column based on the analysis you wish to perform.

• The principles of solvent selection and sample preparation.

If you require an introduction to these concepts, the following book is an excellent

reference: Practical Liquid Chromatography -- An Introduction by R. W. Yost, L. S.
Ettre, and R. D. Conlon, published by The Perkin-Elmer Corporation (1980), Part
No. 0993-9656.

Warranty Exclusions and Limitations

The Limited Warranty set forth on the PerkinElmer ORDERING INFORMATION
form applies to the Series 200 Vacuum Degasser. Among those items excluded from
the warranty under Part (B) "EXCLUSIONS AND LIMITATIONS" are:

l Fuses
l Nuts
l Ferrules
The items listed above are considered to be routinely replaced as part of the normal
wear of the instrument.

Conventions
The following conventions are used in this manual:

A warning indicates an operation that could cause

personal injury if precautions are not followed.

A caution indicates an operation that could cause

instrument damage if precautions are not followed.

Notes emphasize significant information in a procedure or

description.
Warnings and Precautions Introduction 1-7

Warnings and Precautions

Read and understand the following warnings and precautions.

Solvents

Give careful attention to the hazards associated with the

solvents you are using. Refer to the safety data sheets
provided by the manufacturer. For example, Material
Safety Data Sheets (MSDS) in the USA.

Do not use the Series 200 Vacuum Degasser to degas

perfluorinated solvents. Teflon AF® is soluble in
solvents such as the Fluorinert series from 3M company
and PF series from Ausimont. Where there is a question
about using a particular solvent that is fluorinated,
contact a PerkinElmer representative

• Wear appropriate eye protection at all times when handling chemicals. Use
safety glasses with side shields, goggles, or full-face shields, according to the
types of chemicals you will be handling.
• Wear suitable protective clothing, including gloves that are specifically resistant
to the chemicals being handled.
• Always use clean solvents. Solvents that have been distilled in glass (HPLC
Grade) are highly recommended.
• Filter the solvents through a 0.5-micron medium as an additional precaution.
• Carefully use and store flammable solvents, which may form hazardous by-
products when the instrument is shut down, by following the recommended
shutdown procedure.
• Check compatibility of solvent(s) with the type of column(s) being used.
• Know the relative polarity and miscibility of the solvents being used.
1-8 Introduction Warnings and Precautions

Solvents with Low Boiling Points

Do not use liquids that have a boiling point less than or equal to 30 °C (for example,
pentane and CH2CL2). When it is necessary to use these types of solvents, best
results can be obtained using pressurization in a solvent chamber, which reduces the
incidence of bubble formation in the pump inlet.

Buffers
Exercise care when using buffers in conjunction with organic solvents. NEVER
LEAVE BUFFERS IN THE SYSTEM OVERNIGHT. Buffers left in the system can
form salt crystals, which may cause premature pump seal failure, interfere with
proper check valve operation, and plug the connecting tubing or the detector
flowcell.

To remove buffers, flush the system first with water, then with methanol or
isopropanol. Change the mobile phase from methanol or isopropanol to water before
using buffers.

Corrosion
All parts that contact the mobile phase are made of PEEK, Kel-F®, Tefzel® or Teflon
AF®. PEEK is sensitive to Sulfuric acid and certain solvents. If you have questions
about your mobile phase, contact a PerkinElmer representative.

-1 . S o lv e n t s W h ic h M a y C o r r o d e t h e S e r ie s 2 0 0 V a c u u m D e g a s s e r

Aqua Regia (80% HCL, 20% HNO3 ) Chloride salt solutions*

Chlorinated solvents Halogenated solvents
Sulfuric Acid (Conc.)
* Titanium is more resistant to chloride salt solutions.
Warnings and Precautions Introduction 1-9

Air Bubbles
To prevent air from entering the system, and to ensure that pressure fluctuations do
not occur, observe the following precautions:

• Ensure that the fittings are tightened on the degasser input and output ports.
• Ensure that the pump's solvent inlet filter is below the solvent level in the solvent
reservoir.
• If the pump has not been used for an extended period of time, remove air bubbles
by connecting a priming syringe to the drain valve on the pump, opening the
drain valve, and fast-flushing the system to prime the pump. After priming the
pump, close the drain valve completely.

Pressure Buildup
Over time you may notice a gradual increase in the system operating pressure. If you
observe pressure readings greater than 3.45 MPa (500 psi) above the normal operating
pressure of your analysis, check the following items:

• If you are injecting “clean” samples and there is no other apparent cause for the
pressure buildup then remove, disassemble and clean the injector.
• If you are injecting “dirty” samples, the injector may be clogged or the column
packing material may have retained contaminated particulates. Remove and
clean the injector, and replace the column.
• The column end fitting or column frit may be plugged. Replace the end fitting or
frit to relieve excess pressure. (Refer to the instructions supplied with the
column.)
• In-line filters may be plugged. Replace the filter element.
• With the pump connected directly to the detector input, if you still experience
excessive pressure, the problem could be a plugged flowcell. You can flush, reverse-
flush, or rebuild the flowcell.
1-10 Introduction Warnings and Precautions

Specifications

T a b le 1 - 2 . S p e c if ic a t io n s o f t h e S e r ie s 2 0 0 V a c u u m D e g a s s e r

Specification Description
Dimensions: Height: 5.0 in.
Width: 2.87 in.
Depth: 9.81 in.
Weight 6 lb.
1-5 independent
Channels
Degassing Process
Gas permeation through a fluoropolymer membrane
1
Maximum Recommended Flow Rate 3.0 mL/min.
Pressure Drop2 1.37 mm Hg/mL/min.
~25% dissolved gases remaining in 60:40 MeOH/Water mixture at
Degassing Capacity 1 mL/min.
Dead Volume ~480 microliters per channel for standard channel
Materials contacting solvents PEEK, Glass-filled PTFE, Teflon AF®
Power:
Power requirement if using supplied AC 100 to 240 VAC (±10%), 1A, 50 to 60 Hz (±3 Hz)
Adapter
Power Requirement if not using supplied AC 15 to 24 VDC at 0.85 A maximum (0.5 A typical)
Adapter
Wall Sockets 4 supplied with AC Adapter, interchangeable: North
America/Japan, U.K., Continental Europe, Australia
Installation Over-Voltage Category II
Validation Output:
Signal 5 mVDC / 1 mm Hg absolute from 20 to 800 mm Hg
(0.100 VDC at 20 mm Hg; 4.000 VDC at 800 mm Hg)
Accuracy ±1.0% of reading ±0.010 VDC from 20 to 800 mm Hg
Operating Conditions:
Ambient Temperature 10 to 35 ºC
Ambient Relative Humidity (RH) 20 to 80 % RH (without condensation)
Altitude 0 to 2000 Meters
Indoor vs. Outdoor Use Indoor
Pollution Degree 2
Storage Conditions:
Ambient temperature -20 to +60 ºC
Ambient Relative Humidity 20 to 80% RH (without condensation)
Altitude 0 to 12000 M

1 Maximum recommended flow rate to prevent a 60:40 MeOH/Water mixture from outgassing. The estimate assumes low pressure
mixing and low flow restriction prior to the HPLC pump. MeOH/Water mixing represents the worst outgassing case and maximum
flow rate will likely increase with Acetonitrile/Water mixtures. High pressure mixing will also increase the maximum flow rate.
Degassing is still recommended.

2 Calculated tubing pressure per unit change in flow assuming laminar flow with a viscosity of 1.0 cP. Inlet and outlet bulkheads may
contribute to the overall pressure, but are not included in the estimate.
 Installation 2
This chapter describes how to install your Series 200 Vacuum Degasser. The following
installation topics are covered in this chapter:

• Preparing your laboratory

• Unpacking your vacuum Degasser
• Electrical requirements and settings
• Connecting the tubing
• System connections
2-2 Installation Preparing Your Laboratory

Preparing Your Laboratory

Before installing your Series 200 Vacuum Degasser, prepare your laboratory according
to the following guidelines.

Required Air Quality

To minimize contamination problems in your laboratory, provide a relatively dust-free
environment. Make sure that the following gases or vapors are not present at levels that
exceed federal, state, and local ordinances for continuous human exposure:

• Flammable • Caustic
• Explosive • Corrosive
• Toxic

Make sure that your laboratory environment consists of the following temperature and
humidity levels:
• Ambient temperature between 10 and 35 °C
• Constant relative humidity between 20 and 80%, without condensation
Use care when working with hazardous solvents, or solvents that produce hazardous by-
products.

Solvent vapor levels that are high enough to interfere

with the detector performance should be considered
hazardous to someone who is continuously exposed to
the vapors.
Installation Preparing Your Laboratory 2-3

Solvents/Mobile Phase
Use only HPLC grade solvents in all analyses. HPLC grade water and methanol (1 liter
each) are required for performance verification.

The degassing membrane in the Series 200 Vacuum

Degasser is manufactured from Teflon AF®. As with older
membranes manufactured from PTFE, Teflon AF® is inert
to all solvents normally used in HPLC. However, Teflon
AF® is soluble in perfluorinated solvents such as
Fluorinert® FC-75 and FC-40 and Fomblin perfluoro
polyether solvents from Ausimont. Use of such solvents in
the Series 200 vacuum Degasser will result in the
dissolution and hence destruction of the membrane.

Corrosion
All parts that contact the mobile phase are made of PEEK, Glass-filled PTFE or Teflon
AF®. PEEK is sensitive to Sulfuric acid and certain solvents.

Space Requirements
The Series 200 Vacuum Degasser is designed to sit on a bench top, and is plumbed into
the LC system between the solvent supply and pump. A space 5 ¼-in high and 3-in wide
is sufficient. The case is 10-in deep (front to back), but additional space is required both
in front, to accommodate the tubing connected to the unit, and behind to accommodate
the power cord.
2-4 Installation Unpacking Your Series 200 Vacuum Degasser

Unpacking Your Series 200 Vacuum Degasser

Carefully unpack the Series 200 Vacuum Degasser and check for obvious signs of
damage that may have occurred during shipment. Immediately report any damaged or
missing items to the shipping carrier and PerkinElmer.

A Start-Up Kit is supplied with the Series 200 Vacuum Degasser. Use Table 2-1 to
inventory the Start-Up Kit parts.

Table 2-1. Start-Up Kit for the Series 200 Vacuum Degasser

Item Quantity
AC Adapter (including cord) 1
Interchangeable Wall Plugs 4 (North America/Japan, U.K., Continental Europe,
Australia)

Operator’s Manual 1
Plug, ¼-28, Rheodyne 6118 2

Electrical Requirements
The AC Adapter supplied with the Vacuum Degasser incorporates a universal AC input,
switching regulator. This allows the instrument to operate at any AC line voltage from
100 to 240 VAC (±10%) with a line frequency range of 47 to 63 Hz. The switching
regulator senses the incoming line voltage and automatically adjusts its operation
accordingly. The switching regulator assembly incorporates its own on-board line
voltage fuse. This fuse is not user serviceable. In the event that this fuse blows, it will
be necessary to replace the AC Adapter.

A set of four interchangeable wall plugs is included to allow the AC adapter to be

plugged into the standard electrical sockets in North America, Japan, the U.K., most
countries in continental Europe, and Australia.

It is recommended that the AC Adapter be used to supply power to the Vacuum

Degasser. If another means is used to supply power to the rear panel power jack, the
voltage must be within the range 15 to 24 VDC at 0.85 A maximum (0.5 A typical). The
rear panel power jack will accept a 2.1 mm female plug. Correct polarity must be
observed. The center connection of the plug must be positive, and the outside must be
negative. See figure below.
Installation Connecting the Tubing 2-5

Figure 3. Rear Panel Power Connector Polarity

Installing the AC Adapter with Power Cord

Locate the AC Adapter with power cord and the set of four interchangeable wall plugs.
Plug the round connector at the other end of the AC Adapter’s cord into the Power jack
on the Vacuum Degasser rear panel. From the set of four plugs, select the one
appropriate for the local electrical socket and install it onto the AC adapter. With the
power switch off, insert plug into the AC supply. The AC adapter should be positioned
for easy disconnection.

Connecting the Tubing

Solvent lines to be degassed are connected to the Vacuum Degasser’s front panel ports,
as detailed below. Unused ports must be plugged to enable the degasser to operate at its
peak level of performance.

To make a tubing connection:

1. Run a line of 1/8” O.D. x 1/16” I.D. Teflon chromatography tubing from the solvent
supply to the Vacuum Degasser.

2. Push the tubing through a PEEK 1/8” male ¼-28 fitting and slide a ferrule over the
tubing end (see figure below). Cut the Teflon tubing so the end is flat.

3. Screw the ¼-28 fitting into one port on the front of the Vacuum Degasser (Channel
A, for example). The direction of flow through the Vacuum Degasser is not critical.
Plastic connectors should be tightened by hand. Overtightening them will damage
the threads.

4. Repeat steps 1 through 3 to connect additional lines to be degassed.

5. Once all desired solvent lines have been connected to the Vacuum Degasser, any and
all unused ports should be plugged. Use the plugs supplied. Press in by hand.
2-6 Installation Connecting the Tubing

6. Prime each degassing membrane by pulling the solvent from the reservoir through
the degassing system. This can be done by connecting a syringe to the tubing or LC
pump priming port and drawing air and/or mobile phase into the syringe until no air
remains in the tubing, approximately 5 milliliters.

DO NOT prime the membranes by pushing solvent

through the degassing systems. This technique can
generate several hundred pounds of pressure which
might rupture the membrane, even though the Teflon
AF® membrane is quite rugged. The maximum
recommended pressure on the membrane is 1 mPa
(100 psig, 7 Bar).

Figure 4. Configuration of ¼-28 Nut, Ferrule and Tubing

Connecting the Vacuum Degasser in a Typical System

The following illustration shows the tubing connections that are typically made between
the Vacuum Degasser and other instruments in an LC system. The direction of flow
through the Vacuum Degasser is not critical.
Installation Connecting the Tubing 2-7

Figure 5. System Tubing Connections

Powering Up the Series 200 Vacuum Degasser

1. With the Vacuum Degasser plumbed into the system and the power cord installed, as
described in the above section, flip on the rear panel power switch. The green Power
LED should illuminate.

Immediately upon turning on the instrument, the microprocessor examines the

vacuum sensor signal to confirm that it is within an expected range. Following the
startup test, the microprocessor ramps the vacuum pump to high RPM, to quickly
exhaust atmosphere from the vacuum chamber. As the vacuum level approaches the
preset control value, the pump RPM will slowly ramp down to a low speed (typically
40 to 60 RPM) and will vary slightly as needed under the changing degassing load to
maintain a virtually constant vacuum level.

During initial pump-down, the yellow Status LED will be lit. Once the vacuum has
reached normal operating level, the yellow LED will extinguish and the green
Vacuum LED will illuminate. If you want to confirm that the pump is running,
beyond the front panel LEDs, the slight vibration caused by the microstepping of the
motor driving the vacuum pump may be felt by placing your hand on the instrument.

2. S ta r t s o lv e n t flo w th r o u g h th e s y s te m a n d c h e c k fo r le a k s a r o u n d th e ¼ -2 8
c o n n e c to r s .
If a leak occurs at the connection, tighten the fitting an additional 1/8 turn. If the
leak persists, disconnect the leaking fitting and inspect it. If the nut and ferrule
2-8 Installation Connecting the Tubing

appear to be in good condition, reconnect the fitting. If the leak persists, replace the
nut and ferrule and repeat the procedure until you achieve leak-free operation.

The Vacuum Degasser maintains a constant vacuum pressure of 50 mm Hg absolute

(nominal) by varying the speed of the vacuum pump as needed depending on the
degassing load in the system. The pump is designed for at least 5 years of constant
running and has integral in-pump venting, which eliminates the need for stop-start
running (U.S. Patent 6,248,157). The vacuum level and pump speed is constantly
monitored by the microprocessor for changes in operating conditions which might be
attributed to chamber internal leaks. If a potential leak is detected, the pump will be
shut down and the yellow Status LED will flash. The vacuum is maintained as long
as the Vacuum Degasser is powered on. Solvent flowing through the Vacuum
Degasser will continue to be degassed so long as the instrument is on and running.

3. Turn off the Vacuum Degasser when the LC to which it is connected is not in use.
The vacuum chamber(s) will slowly return to atmospheric pressure when the unit is
powered off. This is accomplished by a small, in-line vacuum bleed and reduces the
possibility of solvent vapors condensing in the vacuum tubing or pump head.

4. When flushing a line of solvent, the single lumen coil inside the chamber
contains a very small amount of solvent (approximately 480 microliters).
When changing from one solvent to another where the final solvent is
immiscible with the first, use an intermediate solvent miscible with both the
initial and final solvent. Carryover from solvent to solvent is much less than
previous PTFE designs. Once air bubbles have been cleared from the solvent
line, any further bubbles observed will be coming from the solvent reservoir or
from a leaking fitting.

Since there is virtually no solvent retained within the

Vacuum Degasser (~480 microliters per channel), priming
the system is relatively simple. Using the Prime mode on
the LC system pump, allow the pump to draw each
solvent to be used in the analysis at a flow rate of 2
mL/min. for 1-2 minutes. This ensures that the line from
the degasser channel being primed through the
proportioning valve on the pump has freshly degassed
solvent. This dynamic priming method will allow an
immediate startup of the analysis upon column
equilibration. Contrary to previous PTFE-based
degassers, the new Vacuum Degasser, which uses Teflon
AF® membranes, fully degasses solvents within the time it
takes for the volume to pass through the chamber, and yet
degasses the solvents as thoroughly as, or better than,
PTFE channels containing 40 times more solvent.
 Using the Series 200
Vacuum Degasser 3
This chapter describes how to use the Series 200 Vacuum Degasser.

Before beginning, make a final check of all tubing connections to and from the Series
200 Vacuum Degasser.

Principles of Operation

The Series 200 Vacuum Degasser consists of a vacuum chamber, degassing tube,
variable speed vacuum pump, microprocessor controller, sensor, and check valves. The
solvent (mobile phase) flows into a degassing tube, which is inside a vacuum chamber.
Decreased pressure in the chamber causes the outward movement of gas dissolved in the
mobile phase across the tube wall, in accordance to Henry’s Law, thus degassing the
mobile phase. The pressure in the vacuum chamber is established by the vacuum pump
and monitored by the microprocessor through an integrated absolute pressure sensor.
Degassed mobile phase exits the vacuum degasser and enters the pump.
3-2 Using the Series 200 Vacuum Degasser Principles of Operation

Figure 3-1. Schematic of the Series 200 Vacuum Degasser

Principles of Operation Using the Series 200 Vacuum Degasser 3-3

How the Series 200 Vacuum Degasser Operates

Immediately upon turning on the instrument, the microprocessor examines the vacuum
sensor signal to confirm that it is within an expected range. Following the startup test,
the microprocessor ramps the vacuum pump to high RPM, to quickly exhaust atmosphere
from the vacuum chamber. As the vacuum level approaches the preset control value, the
pump RPM will slowly ramp down to a low speed (typically 40 to 60 RPM).
Afterwards, the pump RPM will vary slightly, as needed under the changing degassing
load, to maintain a virtually constant vacuum level (50 ±0.5 mm Hg). This "zero
hysteresis, constant run" (ZHCR®) mode is necessary, due to the extremely low mass,
high response, Teflon AF® degassing tubing. Only a ZHCR® design ensures a baseline
which is unaffected by the degasser.

Smart Leak Detection

An additional benefit of maintaining a constant vacuum level is that a potential leak in

the vacuum degassing system can be observed by monitoring the RPM of the pump.
This “smart leak detection” is a benefit of the patented design of the degasser. If a leak
occurs within the chamber, the microprocessor will increase the pump RPM in an
attempt to maintain the vacuum level. If the pump cannot maintain the vacuum level (if
it runs at an elevated RPM for more than 2 minutes), the yellow LED will flash,
indicating a possible leak condition, and the system will shut down and go into a “safe”
mode.

Principles of degassing using Teflon AF® membranes

This relatively recent addition to the field of degassing has properties not found in other
fluoropolymers. The fully amorphous nature of this fluoropolymer and its molecular
structure creates a molecular level porosity unlike the mechanically induced porosity in
PTFE extruded tubing. In addition, unlike the process used in extruding PTFE, no
extrusion agents are needed (like kerosene, etc.) which contaminate mobile phases until
they are extracted by the mobile phase over time. Likewise, this molecular structure,
combined with the very small surface areas required to degas the mobile phase, reduces
the possibility of carryover from one solvent or mobile phase to another to virtually zero.

Teflon AF® is so non-polar that it is both solvophobic and hydrophobic. This feature of
Teflon AF® reduces the possibility of cross-channel contamination from one channel to
another, and when combined with the ultra-low internal volumes of Teflon AF® channels
needed for HPLC flow rates, all but eliminates this cross contamination concern by the
chromatographer. Teflon AF® has been used in certain optical systems associated with
HPLC for a few years without concern for normal HPLC solvents. However, Teflon
3-4 Using the Series 200 Vacuum Degasser Principles of Operation

AF® is soluble in certain solvents (see cautionary statements) and must not be used to
degas these types of solvents.

Teflon AF® is permeable to some degree to water vapor whereas PTFE is not. While the
vacuum pump in the Vacuum Degasser contains internal provisions for sweeping water
or solvent vapor from the pump continuously, it is possible that over time, high
concentration buffers may form crystals within the channel due to the loss of water
within the channel. The same precautions should be taken to prevent crystallization
within these channels as are taken for the HPLC pump. See the “Short-term Shutdown”
procedures.
Operating Summary Using the Series 200 Vacuum Degasser 3-5

Operating Summary

The following steps summarize how to use the vacuum degasser:

1. Select and fill each solvent reservoir with the mobile phase for your analysis.

2. Verify that the Series 200 Vacuum Degasser is properly installed as described in
Chapter 2, “Installation.”

Never connect the Series 200 Vacuum Degasser to the

output side of the pump. The high pressure may cause
permanent damage to the degassing membrane.

3. Verify that the tubing to your injector, column, and detector is properly connected.
Also verify that plugs are installed in the unused ports.

4. Disconnect and remove the tubing that is connected to the output port, connect the
priming syringe to this port, and pull the solvent through the degasser until bubbles
no longer appear. Then reconnect the tubing to the output port.

5. Switch on the degasser and start the pump at 1.0 mL/min. Allow the system to
equilibrate for 5-10 minutes. The small volumes contained in the Series 200 Vacuum
Degasser should only be considered in chromatograph equilibration time when flow
rates less than 1 mL/min are used.

Note Use of this product outside the scope of this

manual may present a hazard.
3-6 Using the Series 200 Vacuum Degasser Operating Summary

Extending the degassing flow rate range

Certain organic solvents used in reversed phase chromatography outgas upon

mixing with water, if not properly degassed. These solvents are generally
alcohols (e.g. methanol), acetonitrile and tetrahydrofuran. Passing water and
methanol through a single channel is generally sufficient to degas these solvents
so outgassing does not occur upon mixing when a 60:40 methanol/water mixture
is generated by your Vacuum Degasser or pump at a flow rate of 3 mL/min. If
outgassing does occur, or if a flow rate higher than 3 mL/min. is required, it is a
general rule that only the organic portion of the mobile phase needs to be
passed through a second degassing channel to ensure outgassing does not
occur. This is due to the ability of all organic solvents (e.g. methanol) to hold at
least 10 times more dissolved atmosphere than water can.

To more thoroughly degas a mobile phase, connect the outlet of the organic
channel to the inlet a second channel and the outlet of the second channel to the
pump. This places the two channels in series and doubles the degassing
capacity for the organic portion of the mobile phase.
Shutdown Using the Series 200 Vacuum Degasser 3-7

Shutdown

There are two types of shutdown procedures: long-term and short-term.

Short-Term Shutdown (Overnight and Weekends)

Observe all precautions pertaining to hazardous solvents and/or those solvents that form
harmful deposits or by-products.

1. Remove harmful mobile phases from the vacuum degasser and other instruments in
the system.

2. Flush the column according to the instructions supplied with the column. Flush
buffer salts from the system with water. Evaporation leaves salt crystals that may
form harmful deposits. Remove chloroform or solvents that can decompose to form
hydrochloric acid from the system.

Damage caused by precipitating buffer salts in

capillary tubing, or damage resulting from this
condition, is specifically excluded from warranty.

3. After removing harmful mobile phases, prepare the detector for most mobile phases
by flushing it with isopropanol. To avoid contaminating the system, refilter or
discard solvents (including water) that were exposed to the environment for
more than 24 hours before use.

4. For weekend storage we recommend flushing 60/40% MeOH/Water through the

vacuum degasser, pump, column, and flowcell (provided your column is compatible
with MeOH/Water). Then turn off the vacuum degasser, pump, and detector.
3-8 Using the Series 200 Vacuum Degasser Shutdown

Long-Term Shutdown

1. Follow Short-Term Shutdown procedure Steps 1 and 2.

2. Remove the column and direct the pump output tubing to a beaker. Flush the
vacuum degasser first with water and then with isopropanol.

3. Turn off the vacuum degasser. Then disconnect the tubing between the vacuum
degasser and solvent reservoirs and the vacuum degasser and pump. Plug all of the
ports on the vacuum degasser.

4. Store the vacuum degasser in a clean, dry location.

5. Before using the vacuum degasser, completely purge it with the correct solvent for
the column before reconnecting the column and restarting the system.
 Maintenance 4
You can experience long and trouble-free performance from your Series 200 Vacuum
Degasser by performing both routine and preventive maintenance procedures.

Preventive Maintenance

Perform preventive maintenance to ensure that your Series 200 Vacuum Degasser will
perform consistently at an optimal level. To maintain the Degasser in the best condition,
the following measures are recommended:

1. Adhere to standard laboratory cleanliness practices.

• Use only high-purity solvents (preferably HPLC Grade) for mobile phases. (Water
should be bottled HPLC grade, or filtered and deionized tap water.)
• Filter all solvents to prevent particulate contamination and tubing blockages.

2. Use only high-purity gases when drying contact areas.

• Ensure that all new tubing (stainless steel) is passivated and thoroughly flushed
before making pump connections. (The tubing available from PerkinElmer is
passivated.)
• Follow the short- and long-term shutdown procedures that are described in Chapter
3 of this manual.
4-2 Maintenance Routine Maintenance

Routine Maintenance

Routine maintenance is defined as replacing the normal wear items when you notice
degradation in performance.

If you have a problem, you can save time and money by referring to the Troubleshooting
Guide (on the next page) before calling your local PerkinElmer Service Engineer. Your
problem may be minor and you may be able to correct it yourself by using the
Troubleshooting Guide to pinpoint the cause.

Never remove the Series 200 Vacuum Degasser cover.

There is nothing inside the detector that requires
customer maintenance. If you encounter a problem, call
your local PerkinElmer Service Engineer for assistance.
Troubleshooting Guide Maintenance 4-3

Troubleshooting Guide

Problem Probable Cause Solution

1) Power switch is ON but all 3 (a) Series 200 Vacuum Degasser (a) Plug the AC adapter into the AC
LED’s are off, indicating no power to AC adapter is not plugged into outlet.
the degasser. the AC outlet.
(b) Blown fuse.
(b) Replace the fuse.
2) Yellow Status LED is on steadily, 2) Pump is in initial pull-down phase 2) Typically normal operation,
pump is running and RPM seems or system’s degassing demand has although if pump speed continues to
high. increased. rise for an extended period of time
(as heard by the pitch of the stepper
motor) it could indicate a potential
fault condition.
3) Yellow Status LED is flashing 3) Possible system leak. 3) Contact your Service
approximately 1 second off, 1 second Representative.
on. Vacuum pump is not running.
4) Yellow Status LED is flashing 4) Possible sensor or Control Board 4) Contact your Service
approximately 2 seconds off, 1 fault. Representative.
second on. Vacuum pump is not
running.
5) Is there a way to check whether the 5) Due to the design of the pump and 5a) Place a hand on the top of the
system is operating correctly when degasser, the pump is virtually silent unit. A slight vibration can be felt
Power and Vacuum green LEDs are at low RPM, even though vacuum is indicating the pump is operating at
illuminated, but pump can’t be heard good and degassing is normal. low RPM.
running?
5b) Monitor the UV absorbance of
non-degassed methanol at 215 nM
versus degassed methanol coming
through the degasser. Proper
performance of the degasser should
decrease the UV absorbance of the
methanol significantly.
6) Bubbles appear through the output 6) Loose fitting(s). 6) Tighten the input and output
tubing. fittings.
7) No solvent flow. 7a) Air in the HPLC pump head. 7a) Prime/purge the pump head.
7b) If a buffer solvent was left in the 7b) Use a different channel, or
degasser for some time after use, it connect the channel to a beaker of the
may plug the degasser elements. solvent without the buffer. Draw the
solvent through the channel to
dissolve the buffer. Do not push the
solvent through the channel. If this
flushing action does not work,
contact your Service Representative.
4-4 Maintenance Series 200 Vacuum Degasser Performance

Series 200 Vacuum Degasser Performance

The following graph illustrates the performance of the Series 200 Vacuum Degasser.

Figure 4-1. Graphical representation of the performance of the Series 200 Vacuum
Degasser
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