Applied Separations Spe-ed SFE-Basic User manual

Spe-ed SFE-Basic
Operations Manual
Version 1.0
www.appliedseparations.com
Read Operations Manual for Instructions
BEFORE Setting Up or Running System
930 Hamilton St.
Allentown, PA 18101

Spe-ed™SFE Basic Manual
Disclaimer of Documentation Warranty
This publication is provided “as is” without warranty of any kind, express or implied.
Applied Separations does not warrant, guarantee, or make any representations regarding the
use, or the results of the use, of this publication in terms of correctness, accuracy, reliability,
correctness, or otherwise. Further, Applied Separations reserves the right to revise this
publication and to make changes from time to time in content herein without obligation of
Applied Separations to notify any person or organization of such revision or changes.

TABLE OF CONTENTS
INTRODUCTION
Section 1
Purpose of Spe-ed™SFE Basic
1.1
Specifications
1.2
Site Preparations
1.3
Pre-Installation
1.4
Overview of Features
1.5
REQUIRED UTILITIES
Section 2
Liquid Carbon Dioxide
2.1
Electricity
2.2
Compressed Air
2.3
Recirculating Coolant
2.4
ASSEMBLY AND SET-UP
Section 3
Front/Rear Views
3.1
Attaching the Liquid CO2 Tank
3.2
Power
3.3
Compressed Air
3.4
CO2 Safety Head Outlet
3.5
Connecting Recirculating Coolant
3.6
Connecting the Flow Meter
3.7
EXTRACTOR VESSEL PACKING
Section 4
Extractor Vessel Packing
4.1
In-Line Analyte Trapping
4.2
Vessel Packing Example
4.3
SYSTEM OPERATIONS
Section 5
Inlet, Outlet, Vent, Micro-metering,
and Fluid Delivery Valves 5.1
Operating the Spe-ed™SFE Basic 5.2
Pressurization of the Spe-ed™SFE Basic System 5.21
Depressurizing the Spe-ed™SFE via Vent Valves 5.22
Flushing the Transfer Lines 5.23

OPTIONS Section 6
Spe-ed™SFE Basic Modifier Pump Module 6.1
MAINTENANCE Section 7
Cleaning 7.1
Preventive Maintenance 7.2
TROUBLESHOOTING Section 8
Emergency Operating Procedures
ACCESSORIES Section 9
APPENDIX A
Example Applications

1
INTRODUCTION Section 1
Warranty
This manual provides basic information for the user to install and operate the Spe-ed™SFE
Basic, the maintenance required, and how to obtain necessary service or parts. WARNINGS,
CAUTIONS and NOTES are used throughout this manual to emphasize pertinent or critical
instructions to the user according to the following definitions.
Warning A procedure, that if not followed properly, can prove to be
extremely hazardous to the operator, the environment, or both.
Caution Emphasizes a procedure that must be followed to avoid possible
damage to the instrument.
Note Highlights important information about the instrument or procedures associated
with the operation of the instrument.
Several internationally accepted symbols are used to label certain parts of the system
and throughout this Operator Manual as required for CE Mark Safety Certification.
These symbols are defined below.
Caution Risk of Electric Shock- High voltage exists within enclosures. Allow
access to qualified personal only.
Easily Touched Higher Temperature Parts Surface is normally
hot during operation. Exercise caution when operating in this area.
Caution (refer to accompanying documents) Read Operators Manual
before applying power or operating system.

2
Purpose of the Spe-ed ™SFE Basic 1.1
The Spe-ed™SFE Basic allows an analyst to perform supercritical fluid extractions simply
and efficiently. Developed in conjunction with the USDA, the Spe-ed™SFE Basic has
achieved simple sophistication in design and use. With its flexibility of both dynamic
(continuous flow) and static (where supercritical CO2 is pressurized in a sealed extraction
vessel for a specific period of time) modes, the Spe-ed™SFE Basic offers precise, repeatable
performances for a variety of laboratory procedures including, but not limited to:
• Extracting pesticides and PAHs from soil samples
• Extracting antibiotics and steroids from veterinary products
• Extracting natural products
• Extracting monomers from polymeric compounds
• Extracting fat from food products
Specifications 1.2
Physical Dimensions
Outside: 13” x 23 ¼” x 27 ½”
Electrical
P/N: 99550 P/N: 99551
220-240VAC, 50/60Hz @ 10A 110-120VAC, 50/60Hz @ 20A
Use 10A Time Lag 5x20mm Fuse Use 20A Time Lag 5x20mm Fuse
Warning To avoid fire hazard, replace fuses only with the same type and
rating as specified in Operator Manual and labeled on back of system.
Caution High Voltage exists within enclosure. Allow access to qualified
personal only.
Warning The Spe-ed™SFE Basic instrument may be pressurized to 10,000
psi (690 Bar) at temperatures up to 240°C. Check with the extractor vessel
manufacturer to determine whether vessels used can withstand the pressures and
temperatures that the SFE Basic is capable of delivering. Do not operate the Spe-
ed™SFE Basic beyond the MAWP (Maximum Allowable Working Pressure) of the
vessel. No warranties are given on the extraction vessels beyond those provided
by the manufacturers.

3
Caution The Spe-ed™SFE Basic was designed to use CO2 or CO2
containing organic solvent modifiers only. The use of any other fluid will render
all warranties void.
Caution This instrument is not intended for use with Nitrous Oxide, or with
corrosive compounds such as ammonia or sulfur hexafluoride, which can form an
explosive mixture with organic compounds.
Site Preparation 1.3
Location Bench Space Required
Width
13”
Depth*
23 1/2”
Height
27 1/2”
w/Cover up
43”
Weight
53lbs.
+ Vessel and Vessel
Heater
*Rear access occasionally required to drain air filter or replace broken rupture disc.
Compressed Air Line
The Spe-ed™SFE Basic requires a compressed air line (or cylinder) with a minimum
delivered pressure of 100 psig (6.9 BAR) and capacity of 6.2 actual cfm (cubic ft./min.) or
48 standard cfm. The compressor should be set to turn on at 95 psig (6.6 BAR) and turn off
at 125 psig (8.6 BAR)
Electricity
The Spe-ed™SFE Basic will need a dedicated electrical outlet of 120VAC, 50/60Hz,
20amps (P/N 99351) or 240VAC, 50/60Hz, 10amps (P/N 99350).
Vial Venting
Accessory connections and tubing are available for venting into collection vials and flow
meter hookup. Any extractions using hazardous materials should be done under a fume hood
or the flow meter outlet should have tubing venting into a fume hood.
Pressure Relief Connections
A 3/8” female NPT fitting can be connected to a CO2 safely head outlet. This connection
may be used to direct emergency venting to a remote location in the event of a rupture disc
failure.

4
Cylinder Safety Requirement
Provisions should be made for clamping the CO2 cylinder to either a permanent table edge or
wall by using the appropriate cylinder bracket.
Pre-Installation 1.4
The following items are included with your Spe-ed™SFE Basic. If any are missing,
please contact Applied Separations.
• Spe-ed™SFE Basic, P/N 9935
• Power cord (for oven), 120 VAC or 240 VAC
• Solvent collection rack, P/N 6366
• 6mL SPE discharge fitting (2), P/N 7712
• Discharge tube for collection of solvents (1), P/N 7713
• Samples of Applied Separations Spe-ed™SPE 1g/6mL cartridges (1 box), P/N 7965
• Sample of polypropylene wool, P/N 7952
• Polypropylene frits, P/N 7956
• SPE Vent Adaptor, P/N 7732
• Collection Tube Vent Assembly, P/N 7733
• Flow meter Kit, P/N 7928
• 60mL collection vials w/caps & septa, P/N 7735
Note Other necessary parts and accessories may have been on your order and are
included in the shipment. Check your packing slip and invoice.
Save the Spe-ed™SFE Basic boxes and packing materials for use if it becomes necessary to
return the Spe-ed™SFE Basic for service.
Tools needed for assembly:
Wrenches: 5/16”, 1/2”, 9/16” and 5/8” sizes
Extractor vessels are required for operation (See “Accessories” section of this manual).
If you have additional questions about the assembly or operation of the instrument, contact
Applied Separations Engineering Services Office at 610-770-0900.

5
OvervieworFeatures 1.5
The Spe-ed™SFE Basic is capable of pressures up to 10,000 psig and oven temperature up to
240° C. Within this range of pressures and temperatures the analyst can accomplish a wide
range of extractions.
Several unique features are incorporated into the Spe-ed™SFE Basic that overcome many of
the problems researchers currently encounter with other designs. For example, one of the
most troublesome problems encountered is the clogging of fixed flow restrictors.
The Spe-ed™SFE Basic system eliminates clogging by employing a patented, variable
micro- metering valve interface system. This non-clogging metering system has the capacity
to process even the most fat-laden samples as well as polymer, pharmaceutical and
environmental samples.
Another fixed flow restrictor problem is sample contamination carry over into the transfer
lines due to the inaccessibility of cleaning fixed restrictors.
Understanding the need to correct this frequent problem, the Spe-ed™SFE Basic’s valve and
transfer line systems were designed for simultaneous solvent flushing and with the provision
for an operator to collect any residue for further analysis. As a result, no contamination can
occur between extractions.
Additional features of the Spe-ed™SFE Basic include:
1. Control of flow rates between 0-5.5 LPM of expanded CO2 gas.
2. Increased liquid CO2 pressures of up to 10,000 psig (690 BAR), 10-100 times greater
than currently available.
3. Operation in either static and/or dynamic modes.

6
REQUIRED UTILITIES SECTION 2
LiquidCarbonDioxide(CO2) 2.1
Most analyses will require Instrument grade CO2 or CO2 passed through a CO2 clean-up
column. (See the “Accessories” section of this manual.) A dip/eductor tube will be required
in the CO2 cylinder.
Caution The CO2 cylinder must contain a dip/eductor tube to ensure that
liquid CO2 is delivered to the pump module.
Note The CO2 tank shut-off valve controlling the carbon dioxide must be opened before
the system can be pressurized.
Note Refer to section 3 for operating instructions with a refrigerated cooling bath
(RCB).
Note Prior to operating Spe-ed™RCB, it is important to pressurize the SFE Basic to a
minimum CO2 Pressure. Open the CO2 tank and then turn on RCB. When RCB
reaches the desired cooling temperature, operating pressure may be increased to
its optimum setting.
Electricity 2.2
The Spe-ed™SFE Basic will operate with a dedicated electrical circuit on either a 120VAC,
50/60Hz, 20amps (P/N 99351) or 240VAC, 50/60Hz, 10amps (P/N 99350) line.
For use outside the United States, the power cord will require an appropriate adaptor. Your
dealer will be able to assist you in the necessary adaptation for your specific power supply.
Caution High Voltage exists within enclosure. Allow access to qualified
personal only.
Compressed Air 2.3
Compressed air is necessary to drive the pump. Maximum air pressure should be 125 psig
(8.6 BAR) the minimum is 100 psig (6.9 BAR). The air supply may be obtained from an air
compressor or from air cylinders, although it is recommended that an air compressor be used.
If it is necessary to use gas cylinders, use a technical grade of air.
Recirculating Coolant 2.4
Recirculating Coolant is required by the pump for CO2 pressurization. The recirculating
coolant should be maintained at a controlled temperature of 0°C.

7
ASSEMBLY AND SET-UP SECTION 3
Front View of the Spe-ed ™ SFE Basic 3.1
Figure 1

8
Rear View of the Spe-ed ™ SFE Basic
Figure 2

9
ConnectingtheCO
2
Cylindertothe Spe-ed ™ SFE Basic 3.2
Attach the flex hose with a CGA fitting
(P/N 7118) from the CO2 cylinder to the
CO2 inlet port on rear panel (Figure 3).
Tighten the nut on the rear panel
connection with a 9/16” wrench. (See the
“Accessories” section of this manual for
tubing and wrenches.)
Note Always use latex gloves during the
hook-up or change-over of a CO2 cylinder
to avoid contamination of the CO2. Skin
oils may cause interferences during
subsequent analysis.
Figure 3
Power 3.3
Connect the AC power cord (provided) from the back of unit to the dedicated circuit

10
Compressed Air 3.4
Connect 1/4”OD nylon tubing
(P/N 7115) from the compressed
air source (compressed air
cylinder or house air supply) to
the air filter on the back of the
unit (Figure 4). Press the tubing
into the fitting port firmly until
secure. (See the “Accessories”
section of this manual.)
To remove the tubing from the
air inlet port, press the connector
nozzle firmly while removing the
tubing.
Figure 4
CO2 SafetyHead Outlet 3.5
The safety head (shown in Figure 5) on the rear panel
of the pump module, contains a rupture disc that will
burst if the CO2 outlet pressure exceeds 12,500 psi
(862 BAR). In the event of a ruptured disc, the CO2
will vent from the safety head outlet until the pressure
in the system is relieved.
Note This outlet has a 3/8” female NPT
pipe thread fitting. This fitting
allows the user to vent, if desired,
the discharging gas to a housing
ventilation system.
Note In the event of a burst rupture
disc, a new disc must be installed
to seal the port. The system cannot
be pressurized unless a new
rupture disc has been installed.
(Please see the “Accessories”
section of this manual.)
Figure 5

11
Connecting Recirculating Coolant 3.6
Set-up and Use
1. Attach the flexible tubing fluid lines to the
inlet and outlet adaptors on the rear of the SFE
Basic (Figure 6) to the Spe-ed™RCB
Refrigerated Circulating Bath (P/N 7025,
120VAC or P/N 7027, 240VAC).
2. Set cooling temperature in accordance with
CO2 consumption and CO2 pressure (usually
0°C).
Figure 6
Note Prior to operating Spe-ed™RCB, it is important to pressurize the SFE pump to a
minimum CO2 Pressure. Open the CO2 tank and then turn on RCB. When RCB
reaches the desired cooling temperature, operating pressure may be increased to
its optimum setting.

12
Connecting the Flow meter 3.7
The flow meter is attached to the right side of the unit. It is calibrated for gaseous CO2.
Instructions for using the flow meter for SPE collection and for solvent collection follow.
For SPE Collection
Slide the 6mL SPE cartridge onto the
cartridge adaptor located on the Flow
Control Module and connect the tubing
adaptor to the 6mL cartridge. Affix a length
of Tygon tubing to the ends of both the
tubing adaptor and to the flow meter. The
micro-adjustment valve at the outlet of the
flow meter should be full open. If the balls
in the flow meter oscillate during use,
slightly close the valve until the oscillations
stop.
For Solvent Collection
Perforate the septum on the collection vial
with the collector vessel vent. Affix the
tubing adaptor to the flow meter (Figure 7).
Figure 7
Note If a solvent is used in the collection vial, the vial must be positioned so that the
solvent level is below the tip of the vent needle.

13
EXTRACTOR VESSEL PACKING SECTION 4
PackingtheExtractorVessel 4.1
1. Choose an appropriate sized vessel rated at the pressure
required for your application.
Note The following instructions describe the use of the
analytical scale extraction vessels available from
Applied Separations. Vessels obtained from other
suppliers may vary slightly in design.
2. Remove one of the end-fittings and push a plug of glass
or polypropylene wool or a polypropylene frit (P/N 7956) to
the closed end of the vessel with a tamping rod.
Caution In the end-fittings of the extractor
vessels are metal frits that hold the sample in
place and prevent particulate matter from
entering the fluid transfer lines. Since the frits
can be easily clogged by the sample matrix, it is
important to place some kind of coarse
filtration material such as polypropylene wool
in the vessel to protect the frits from fine
particulate matter.
3. Fill the vessel with the sample and tamp the bed. See the
“Accessories” section of this manual for tamping rods.)
Be sure to pack the sample bed firmly to ensure that the
supercritical fluid diffuses uniformly through the sample
matrix.
4. Add a second plug of polypropylene wool or a frit to the top of the sample bed (Figure 9).
If a large void remains, fill the additional space in the vessel with Spe-ed™Matrix (P/N
7950).
5. Add a final plug of polypropylene wool or a frit and tighten the end-fitting.
6. Set the vessel aside or refrigerate until ready to perform the extraction.

14
Warning Make sure that the Seals or O-Rings/Backup Rings are properly
installed and the end caps are properly tightened.
Note For many samples, packing the tube in this manner would result in a sample bed
that would not permit even diffusion of the supercritical fluid. In such instances,
some materials must be used to disperse the original sample. Among the materials
which have been used are glass and metal beads, sand, silica gel, sodium sulfate,
and Spe-ed™Matrix (P/N 7950).
Note Extraction vessels usually have recessed seals or o-rings/backup rings at each
end. During cleaning of the vessels after the extraction, the seal may become
dislodged. The vessel will not fully pressurize unless the seal is replaced in the
vessel.

15
Packing the Extraction Vessel for
In-lineAnalyteTrapping 4.2
Below is a procedure for packing an extractor vessel with SPE columns (in a Teflon sleeve)
to perform in-line SFE applications.
1. Attach the top end cap to the vessel and
tighten. Engrave this end of the vessel as
“TOP” (Figure 10).
Warning Make
sure that the end
caps of the
extractor vessel are
secured tightly to
prevent CO2
leakage and
subsequent analyte
loss.
2. Carefully
remove the
sealing ring
from the
other column
end (Figure
11).
3. Insert the
Teflon
sleeve/SPE
column and
tamp it into
place (Figure
12).
4. Add a plug of
polypropylene wool
or a frit and tamp
(Figure 13).
5. Add the sample
matrix and tamp it
into place (Figure
13).
6. Fill the remaining portion of the vessel
with Spe-ed™Matrix and polypropylene
wool or a frit and tamp them into place
(Figure 13).
7. Clean the extractor vessel to prevent any
CO2 leakage and replace the seal.

16
8. Attach the bottom end cap to the extractor vessel and
tighten (Figure 14).
9. Set the vessel aside or refrigerate until ready to perform
the extraction.
Caution At the ends of the extractor vessels
are metal frits that hold the sample in place
and prevent particulate matter from entering
the fluid transfer lines. Since the frits can be
easily clogged by the sample matrix, it is
important to place filtration material (ex. wool,
Spe-ed™Matrix) in the vessel to protect the
frits from fine particulate matter.
VesselPacking Example 4.3
It is beyond the scope of this manual to provide examples of all CO2 extraction sample
preparation techniques. One example has been chosen to serve as a guide for other
applications.
Preparation of a Potato Chip Sample for SFE
1. Grind a sample of potato chips with a mortar and pestle. (Note: Ground potato chips
will ensure a higher fat extraction output.)
2. Pour the dry mixture into a high pressure extraction vessel fitted with a plug of
polypropylene wool or a polypropylene frit (P/N 7956) in the TOP end cap.
3. Add a second smaller plug of polypropylene wool or a frit. Tamp to tightly compact
the dry mixture in the vessel.
Note This step may be critical to the success of the extraction. Loosely packed material
may allow the supercritical fluid to flow through the vessel without adequately
penetrating the sample matrix.
4. Fill the void volume in the extractor vessel with Spe-ed™Matrix.
5. Push a third polypropylene wool plug or a frit into the vessel and press it firmly
against the Spe-ed™Matrix.
6. Screw the bottom vessel end cap onto the vessel.