Endress+Hauser KRFB User manual

Products Solutions Services
Operating Instructions
Raman flow assembly
KRFB, KRFC
BA02296C/66/EN/01.23
70209490

Operating Instructions Raman flow assembly
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Raman flow assembly Operating Instructions
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Table of Contents
1 About this document ........................... 4
1.1 Warnings ................................................................ 4
1.2 Symbols on the device ........................................... 4
1.3 U.S. export compliance .......................................... 4
1.4 Glossary ................................................................... 5
2 Basic safety instructions ..................... 6
2.1 Personnel requirements ........................................ 6
2.2 Designated use ....................................................... 6
2.3 Workplace safety ................................................... 7
2.4 Operational safety .................................................. 7
2.5 Service safety .......................................................... 7
2.6 Important safeguards ............................................ 7
2.7 Product safety ......................................................... 7
3 Product description ............................. 8
3.1 Micro flow bench .................................................... 8
3.2 Micro flow cell ........................................................ 9
4 Incoming product acceptance and
product identification ....................... 10
4.1 Incoming acceptance .......................................... 10
4.2 Product identification ......................................... 10
4.3 Scope of delivery.................................................. 10
5 Installation ........................................ 11
5.1 Installing the micro flow bench ......................... 11
5.2 Inspecting the micro flow cell............................. 12
5.3 Installing tubing sets to the micro flow cell ...... 12
5.4 Installing the micro flow cell .............................. 13
6 Commissioning .................................. 14
6.1 Receipt of product ................................................ 14
6.2 Calibration and verification ................................ 14
7 Operation ........................................... 15
7.1 Micro flow cell ...................................................... 15
7.2 Storing the micro flow cell .................................. 15
7.3 Storing the micro flow bench ............................. 15
8 Diagnostics and troubleshooting ..... 17
9 Maintenance ...................................... 19
9.1 Cleaning the micro flow cell window ................. 19
10 Repair ................................................. 20
10.1 Repairing the Raman flow assembly ................. 20
10.2 User serviceable parts ......................................... 20
11 Technical data ................................... 21
11.1 Micro flow bench ................................................. 21
11.2 Micro flow cell ...................................................... 22
12 Supplementary documentation ....... 23
13 Index ................................................... 24

Operating Instructions Raman flow assembly
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1 About this document
This manual provides information about the flow cell apparatus used in conjunction with the Endress+Hauser
Rxn-10 Raman spectroscopic probe. This includes the wetted flow cell and the flow bench optical interface.
Refer to the Rxn-10 Raman spectroscopic probe Operating Instructions for specific probe-related information.
1.1 Warnings
Structure of Information Meaning
Causes (/consequences)
Consequences of non-compliance (if
applicable)
‣ Corrective action
This symbol alerts you to a dangerous situation. Failure to avoid the
dangerous situation can result in a fatal or serious injury.
Causes (/consequences)
Consequences of non-compliance (if
applicable)
‣ Corrective action
This symbol alerts you to a dangerous situation. Failure to avoid this
situation can result in minor or more serious injuries.
Cause/situation
Consequences of non-compliance (if
applicable)
‣ Action/note
This symbol alerts you to situations which may result in damage to
property.
Table 1. Warnings
1.2 Symbols on the device
Symbol Description
The Laser Radiation symbol is used to alert the user to the danger of exposure to hazardous visible laser
radiation when using the system.
The High Voltage symbol that alerts people to the presence of electric potential large enough to cause
injury or damage. In certain industries, high voltage refers to voltage above a certain threshold.
Equipment and conductors that carry high voltage warrant special safety requirements and procedures.
The WEEE symbol indicates that the product should not be discarded as unsorted waste but must be sent
to separate collection facilities for recovery and recycling.
Table 2. Symbols
1.3 U.S. export compliance
The policy of Endress+Hauser is strict compliance with U.S. export control laws as detailed in the website of the
Bureau of Industry and Security at the U.S. Department of Commerce.

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1.4 Glossary
Term Description
ANSI American National Standards Institute
API active pharmaceutical ingredient
ATEX atmosphére explosible
°C Celsius
cm centimeter
°F Fahrenheit
CRS calibration reference standard
in. inches
IO immersion optic
kg kilogram
LED light emitting diode
m meter
μin microinches
μm micrometer
mm millimeter
mW milliwatt
NCO non-contact optic
nm nanometer
oct/min octaves per minute
PD process development
psi pounds per square inch
WEEE Waste Electrical and Electronic Equipment
Table 3. Glossary

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2 Basic safety instructions
The safety information in this section is specific to the flow cell that is compatible with the Raman Rxn-10
spectroscopic probe from Endress+Hauser. Refer to the Rxn-10 Raman spectroscopic probe Operating Instructions for
additional information related to probe and laser safety.
2.1 Personnel requirements
• Installation, commissioning, operation, and maintenance of the probe/optics may be carried out only by
specially trained technical personnel.
• Technical personnel must be authorized by the plant operator to carry out the specified activities.
• Technical personnel must have read and understood these Operating Instructions and must follow the
instructions contained herein.
• The facility must designate a laser safety officer who ensures staff are trained on all Class 3B laser operating
and safety procedures.
• Faults at the measuring point may only be rectified by authorized trained personnel. Repairs not described in
this document must be carried out only directly at the manufacturer's site or by the service organization.
2.2 Designated use
The Raman micro flow cell and flow bench are designed for flow rates which are most applicable to product and
process development and are optimized for biological samples. The micro flow cell is designed to connect to a flow
path creating an enclosed, wetted interface which is separate from the flow bench. The flow bench, which is
compatible with the Rxn-10, allows for the secure laser-sample interface as well as calibration and verification of
the apparatus. The flow bench is required for use of the micro flow cell.
Recommended applications for the optics include:
Apparatus Fields of application
Flow cell  Perfusion stream
 Material recirculation loop
 Tangential flow filtration
 Eluate monitoring
 Monomer-aggregate differentiation
Table 4. Fields of application
The above table lists common applications for the flow cell apparatus. There are other possible fields of application,
however, use of the device for any purpose outside of the fields of application described here poses a threat to the
safety of people and of the entire measuring system, and invalidates any warranty.

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2.3 Workplace safety
As the user, you are responsible for complying with the following safety conditions:
• Installation guidelines
• Local standards and regulations for electromagnetic compatibility
2.4 Operational safety
Before commissioning the entire measuring point:
• Verify that all connections are correct.
• Ensure that electro-optical cables are undamaged.
• Ensure fluid level and flow parameters are compatible with configured micro flow cell.
• Do not operate damaged products, and protect them against unintentional operation.
• Label damaged products as defective.
During operation:
• If faults cannot be rectified, products must be taken out of service and protected against unintentional
operation.
• When working with laser devices, always follow all local laser safety protocols which may include the use of
personal protective equipment and limiting device access to authorized users.
2.5 Service safety
Follow your company’s safety instructions when removing a process probe or optic from the process interface for
service. Always wear proper protective equipment when servicing the equipment.
2.6 Important safeguards
• Do not use the system for anything other than its intended use.
• Do not disconnect the Rxn-10 probe while the system is collecting Raman data.
• Do not remove the flow cell from the flow bench while the system is in use.
• Do not look directly into the laser beam.
• Do not point the laser at a mirrored or shiny surface, or a surface that can cause diffuse reflections. The
reflected beam is as harmful as the direct beam.
• When not in use, close the shutter on the Rxn-10 probe. If an optic cap is available, place it on the unused optic.
• Always use a laser beam block to avoid inadvertent scatter of laser radiation.
2.7 Product safety
The product is designed to meet all current safety requirements, and has been tested and shipped from the factory
in a safe operating condition. The relevant regulations and international standards have been observed. Devices
connected to an analyzer must comply with the applicable analyzer safety standards.

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3 Product description
The Raman flow cell and associated flow bench, powered by Kaiser Raman technology, enable enhanced signal
generation while monitoring material in a flow path. The system is an optimized non-contact apparatus that
leverages the internal design characteristics of the flow cell to enhance the total signal-to-noise ratio for faster
sampling times and lower limits of detection. Refer to the sections below for descriptions of the parts and their
functions.
3.1 Micro flow bench
The micro flow bench is configured and factory-tuned for the unique flow cell and application space needs, where
the fluid-path thickness and the refractive index of the sample are known or well approximated. For optimal
performance, it is recommended that the sample refractive index at the laser-wavelength be characterized so that
proper factory-setting can be achieved. The fluid-path thickness is defined by the flow cell configuration. It is
recommended that the micro flow bench and micro flow cell be configured together to ensure optimal performance
in the field. The micro flow bench has been optimized for use in the near infrared (NIR) region (785 nm).
The micro flow bench comes standard with a light shield to safely acquire the Raman spectra while eliminating
noise from the environment. In addition, a beam dump is available which safely blocks the amplifier to collect
spectra in back-scatter mode only. This is useful for running experiments that do not require the signal
enhancement.
The micro flow bench comes standard with an amplifier to enhance the Raman signal per unit-time.
3.1.1 Micro flow bench options
The micro flow bench provides application flexibility, and can be configured to the unique and demanding needs of a
variety of flowed-sample conditions. The micro flow bench is currently available in an Rxn-10 compatible
configuration which is factory tuned for the specific application needs. In most cases aqueous environments are an
acceptable approximation; however, extension beyond these spaces is available.
If a unique mounting solution is required, the micro flow bench can be configured with OEM-specific hardware to
directly integrate into the unique environment.

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3.2 Micro flow cell
The micro flow cell has been designed to produce laminar flow within a wide range of volumetric flowrates,
pressures, tubing diameters, and fitting options. Micro flow cells are ASME BPE SF4 Bio-Compatible and can be
cleaned and sterilized for re-use, or can be discarded after each use similar to sleeved immersion optics from
Endress+Hauser. Additionally, micro flow cells are configurable with a variety of fluid path thicknesses if the
standard 2.5 mm option is not sufficient.
3.2.1 Micro flow cell options
The micro flow bench and micro flow cell system has been designed to ensure maximum transferability between
cells and benches, allowing micro flow cells to be replaced on the flow bench with minimal impact to the resulting
spectrum.
3.2.2 Micro flow bench and flow cell data collection zone
The micro flow bench and flow cell are configured together to maximize the performance of the system. The
amplifier enhances the signal by collecting both forward and backward propagating scatter. The data collection zone
has been designed for optimal use in clear samples. If collection in opaque samples is required, contact Service
(Repairing the Raman flow assembly) → .

Operating Instructions Raman flow assembly
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4 Incoming product acceptance and product identification
4.1 Incoming acceptance
• Verify that the packaging is undamaged. Notify the supplier of any damage to the packaging. Keep the damaged
packaging until the issue has been resolved.
• Verify that the contents are undamaged. Notify the supplier of any damage to the delivery contents. Keep the
damaged goods until the issue has been resolved.
• Check that the delivery is complete and nothing is missing. Compare the shipping documents with your order.
• Pack the product for storage and transportation in such a way that it is protected against impact and moisture.
The original packaging offers the best protection. Make sure to comply with the permitted ambient conditions.
• If you have any questions, please contact your supplier or your local sales center.
Incorrect transportation can damage the micro flow cell and micro flow bench.
4.2 Product identification
4.2.1 Label
At a minimum, the optics are labeled with the following information:
• Manufacturer information
• Part number
• Serial number
• Wavelength
• KMAT (product configuration code)
Compare the information on the label/tag with the order.
4.2.2 Manufacturer address
Endress+Hauser
371 Parkland Plaza
Ann Arbor, MI 48103 USA
4.3 Scope of delivery
The scope of delivery comprises:
• Raman micro flow bench
• Raman micro flow cell
• Raman flow assembly Operating Instructions
• Local declarations of conformity, if applicable
• Material certificates, if applicable
If you have any questions, please contact your supplier or local sales center.

Raman flow assembly Operating Instructions
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5 Installation
5.1 Installing the micro flow bench
The Endress+Hauser micro flow bench slips onto the Rxn-10 probe and is secured by a torque limiting thumb screw
clamp. The thumb screw on the Rxn-10 probe should never be fully removed.
When installing or removing optics, ensure the laser and emission shutter are in the closed position.
To install the Rxn-10 probe onto the micro flow bench:
1. Loosen the metal screw on the Rxn-10 probe by turning the screw counter-clockwise approximately one
turn (do not remove).
2. Insert the end optic clamp of the probe onto the Rxn-10 adapter of the micro flow bench.
Figure 1. Inserting the Rxn-10 probe onto the Rxn-10 adapter of the micro flow bench
3. Slide the probe over the Rxn-10 adapter of the micro flow bench until it stops.
Figure 2. Final position of the Rxn-10 probe with the micro flow bench
4. Tighten the thumb screw by gently turning it clockwise until there is an audible “click” sound. This indicates
the thumb screw has reached the desired torque. Failure to tighten the screw will result in the optic coming
loose, potentially damaging the optic.
5. After installing the micro flow bench, use the micro flow bench calibration kit to perform an intensity
calibration for the probe with the new optic.
To remove the Rxn-10 probe from the micro flow bench:
1. Loosen the torque limiting thumb screw by turning it counter-clockwise approximately one turn so that the
Rxn-10 adapter is released from the clamp. Do not remove the screw.
2. Slide the probe off of the adapter.
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Operating Instructions Raman flow assembly
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5.2 Inspecting the micro flow cell
It is important to inspect the exterior window surfaces of the micro flow cell before installation of any tubes. See
inspection and cleaning procedures in Maintenance → .
Figure 3. Normal (left) and off-axis (right) inspection of a cell under a microscope
5.3 Installing tubing sets to the micro flow cell
The micro flow cell comes standard with 5/16-24 flat-bottom threaded ports for adapting to the flexible tubing
ferrules commonly used in bioprocessing and HPLC streams. However, the micro flow cell can be configured with
OEM-specific ports to enable unique applications. Tubing must be installed by trained and qualified personnel
according to standard operating procedures (SOP).
Endress+Hauser recommends that all tubing ports be inspected before installing tubing or sealing ferrules. All tubes
should be cut square with approved cutting tools and techniques to ensure leak-free connections. When installing
tubing and ferrules into the ports, ensure the tubing and ferrules are flush for the best performance.
Wear appropriate gloves when attaching tubing to avoid possible contamination of micro flow cell window
surfaces.
Figure 4. Placement of tubing sets to the micro flow cell
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5.4 Installing the micro flow cell
Before installing the micro flow bench, ensure the Rxn-10 laser shutter is closed and confirm that no data collection
is occurring. Ensure all flow connections are correctly affixed and are functioning as intended.
To install the micro flow cell:
1. Pull back on the light shield. The light shield stroke path is embossed on the top of the light shield.
2. Install the flow cell with the probe-indicating arrows pointing toward the Rxn-10 probe.
3. Adjust the light shield forward until the plunger contacts and fixes the micro flow cell in place.
4. Confirm the beam dump is open or closed, according to the application requirements.
5. Close the light shield completely.
Before collecting data, open the Rxn-10 shutter.

Operating Instructions Raman flow assembly
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6 Commissioning
The micro flow bench is delivered ready to connect to the Rxn-10 probe. Follow the instructions below to
commission the micro flow bench for use in conjunction with the probe.
No additional alignment or adjustment to the system is required and we strongly recommend that customers
not attempt adjustments.
6.1 Receipt of product
Perform the steps for incoming product acceptance described in Incoming acceptance → .
6.2 Calibration and verification
The probe and the analyzer must be calibrated before use.
6.2.1 Raman flow assembly calibration and verification kit
After installing the Rxn-10 probe onto the micro flow bench, use the Raman flow assembly calibration and
verification kit to perform an intensity calibration and verification for the probe head with the new micro flow
bench.
Refer to the Raman flow assembly calibration and verification kit Operating Instructions for additional information
about the Raman flow assembly calibration and verification kit.
6.2.2 Performing calibration and verification of the probe
Refer to the Raman flow assembly calibration and verification kit Operating Instructions for steps to:
• Perform internal analyzer calibration which may include alignment calibration, full wavelength calibration
and/or full laser wavelength calibration depending on status of analyzer
• Perform probe calibration: requires Raman flow assembly calibration and verification kit
• Perform probe verification: verifies the calibration results using a standard reference sample; verification cell
• View calibration and verification reports
Do NOT immerse any components of the micro flow bench or Rxn-10 directly into a sample.
 The verification cell accessory should be used for verification of the micro flow bench.
The Raman RunTime software will not allow spectra to be collected without passing internal and probe calibrations.
Passing the probe verification step is not required but is highly recommended.
Raman Rxn analyzer operating instructions are available by searching the Downloads area of the Endress+Hauser
website: https://endress.com/downloads.

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7 Operation
7.1 Micro flow cell
Hydrocarbon solvents can degrade hardware performance and invalidate the warranty.
 Do not use the micro flow cell with hydrocarbon solvents (including ketones and aromatics).
 The micro flow bench and micro flow cell are NOT intended to be submerged into any liquid.
7.2 Storing the micro flow cell
Store the micro flow cell in the original factory packaging until its intended use and installation into a flow-system.
If re-use is intended after flowing sample through the micro flow cell, it is recommended that the flow cell be purged
by standard operating procedures (SOP) and dried with clean and filtered air. Refer to flow cell inspection and
cleaning section on air cleanliness in Cleaning the micro flow cell window → .
Remove all residual moisture from the cell through a clean drying process such as bake-out. Endress+Hauser does
not recommend a specific cleaning procedure as application SOPs may necessitate different parameters. Any bake-
out procedure should be done within the qualified storage temperature limits found in Micro flow cell → .
Exceeding these can damage the flow cell.
7.3 Storing the micro flow bench
When storing the micro flow bench, keep the system closed to minimize contamination. To do this, adjust the beam
dump to closed position, close the light shield fully, and place the cap over the Rxn-10 adapter as shown in figures 5
and 6 below. Store the micro flow bench in the original shipping box and materials. If these are not available,
storing the device in a clean bag within the specified environmental conditions will provide sufficient protection.
Figure 5. Shutter closure open, close beam dump
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Figure 6. Shutter closure closed
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8 Diagnostics and troubleshooting
Endress+Hauser Raman flow assemblies provide consistent and reliable long-term and repeated operation. Refer to
the Rxn-10 Raman spectroscopic probe Operating Instructions (BA02160C) to troubleshoot issues with the Rxn-10
probe and accessory optics.
Symptom Possible cause Action
1 Substantial
reduction in signal
External surfaces of windows are
dirty
1. Carefully remove the cell from the flow
bench and inspect both optical windows
for residue, fingerprints, etc.
2. If necessary, clean the windows with
isopropyl alcohol and a lint free wipe or
swab before returning to the flow bench.
Beam dump is in the closed
position
Verify the position of the beam dump using
the control wheel and correct if necessary.
Cell is not correctly installed into
bench
Open light shield and verify that the flow cell
is properly installed in the bench, and that the
plunger is fully contacting the cell without any
gaps. Cell should rotate slightly relative to the
probe axis but should not wiggle free or move
around easily.
2 Complete loss of
signal while laser is
powered and laser
emission indicator is
lit
Material is not flowing through
the cell or beam path is
obstructed by bubble
Ensure the cell is charged with liquid and free
of bubbles. If bubbles are a consistent problem,
consider mounting the bench such that the
flow path is vertical with material moving
from the bottom up.
Rxn-10 is not properly connected
or Rxn-10 shutter is closed
Ensure the Rxn-10 is properly attached, and
the laser beam shutter is in the open (I)
position.
3 Flow bench not
calibrating
Beam dump is not in the “open”
position
Verify the position of the beam dump using
the control wheel and correct if necessary.
Calibration cell is not installed
correctly
Ensure that the calibration cell is installed in
the correct direction with probe-indication
arrows pointing correctly towards the Rxn-10
Probe.
Light shield is still in the
retracted position
Fully close the light shield to ensure the optic
is properly engaged with the cell.
Rxn-10 Probe is not properly
connected or Rxn-10 shutter is
closed
Ensure the Rxn-10 is properly attached, and
the laser beam shutter is in the open (I)
position.
4 Flow bench not
passing verification
Bubbles in the verification cell Carefully open the cell and use the supplied
blunt tip syringe to fill the cell with 70 % IPA.
70 % IPA is contaminated Ensure clean and un-contaminated 70 % IPA is
used in the cell. Flush the cell and replace the
volume.
Calibration cell or verification
cell have contamination in the
clear aperture.
Remove the cell and inspect visually for
contamination. Follow proper cleaning
procedures to clean any contamination from
the optical surfaces.

Operating Instructions Raman flow assembly
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Symptom Possible cause Action
Beam dump is not in the “open”
position
Verify the position of the beam dump using
the control wheel and correct if necessary.
Light shield is still in the
retracted position
Fully close the light shield to ensure the optic
is properly engaged with the cell.
The incorrect source spectral file
(SSF) was used during calibration
of the instrument
Confirm the correct source spectral file (SSF)
and calibration date of the Calibration Cell.
Rxn-10 probe is not properly
connected or Rxn-10 shutter is
closed
Ensure the Rxn-10 is properly attached, and
the laser beam shutter is in the open (I)
position.
Calibration cell is not installed
correctly
Ensure that the calibration cell is installed in
the correct direction with probe-indication
arrows pointing towards the Rxn-10 probe.
Calibration temperature was
incorrectly input into RunTime
software
Ensure that the proper units (°F or °C)
correspond on the temperature meter and in
RunTime.
Calibration cell temperature was
not stable during calibration
Ensure that the calibration cell has reached
thermal stability* before calibrating the
instrument. Ensure that the system is not
subject to excessive heat sources such as
exhaust fans, hot plates, or handling.
* Refer to the Raman flow assembly calibration and
verification kit Operating Instructions (BA02294C) for
instructions on measuring the temperature of the micro
flow bench calibration cell.
5 Flow cell tubing
ports are leaking
Tubing was not cut square or was
crushed during cutting
Use only approved tubing cutting tools
designed for the tubing material. Use only
tubing sizes intended for the flow cell. Re-cut
ends if necessary.
Fittings incorrectly installed Confirm that fittings are for the correct tubing
size and are compatible with the flow cell
threads. Replace if necessary.
Table 5. Diagnostics and troubleshooting

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9 Maintenance
If the micro flow cell windows have come in contact with a sample, dust or fingerprints, etc., they may need to be
cleaned.
For all other maintenance, it is recommended that the optics are serviced at the manufacturer’s site.
9.1 Cleaning the micro flow cell window
The optical surfaces of the micro flow cell must be clean before use. If cleaning is required, Endress+Hauser
recommends the process below. Clean gloves are recommended whenever handling the micro flow bench and micro
flow cell to minimize the transmission of oils or other film-type contamination to the exterior optical window
surfaces.
Contaminant Cleaning process
Large particles and dust
 Blow off the surface with clean, dry air.
Oil contamination in the air lines can result in film on the optical
surface.
 Ensure only clean air is used.
 Inspect the surface. Repeat the cleaning process if large particles or dust
remain.
Small particles
 Gently clean with the lens wipes provided with the Raman micro flow cell.
o Do not damage the optical surface with excessive or aggressive
cleaning.
o Use a fresh cleaning wipe to avoid scratching the surface with trapped
debris.
o Minimize liquid buildup on the window surface to avoid leaving film
contamination on the surface.
 Inspect the surface. Repeat the cleaning process if contaminants remain.
Film-type contamination
 Gently clean with the lens wipes provided with the Raman micro flow cell.
o Do not damage the optical surface with excessive or aggressive
cleaning.
o Use a fresh cleaning wipe to avoid scratching the surface with trapped
debris.
o Minimize liquid buildup on the window surface to avoid leaving film
contamination on the surface.
 Inspect the surface. Repeat the cleaning process if contaminants remain.
Table 6. Cleaning the micro flow cell window
Figure 7. Cleaning the optical surface with a lens wipe
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10 Repair
10.1 Repairing the Raman flow assembly
Repairs not described in this document must be carried out only at the manufacturer's site or by the service
organization. For Technical Service, refer to our website (https://endress.com/contact) for the list of local sales
channels.
If a product must be returned for repair or replacement, follow all decontamination procedures indicated by your
service provider.
Failure to properly decontaminate wetted parts before return can result in a fatal or serious injury.
To ensure swift, safe and professional product returns, please contact your service organization.
For additional product return information, refer to the following site and select the applicable market/region:
https://www.endress.com/en/instrumentation-services/instrumentation-repair.
10.2 User serviceable parts
There are no user serviceable parts for the Raman flow assembly.