IKA® Works, Inc. 2635 North Chase Pkwy SE Wilmington, NC 28405 U.S.A Ph.: 1- 800-733-3037 Fax: 910-452-7693

IKA® BR 10

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CONTENTS WARRANTY .............................................................................................. 2 EXPLANATION OF SYMBOLS ................................................................... 3 SAFETY INSTRUCTIONS .......................................................................... 3 DESCRIPTION ........................................................................................... 4 UNPACKING ............................................................................................. 6 CONTENTS................................................................................................6 ASSEMBLY OF BR 10 ............................................................................... 7 START UP ............................................................................................... 13 PROGRAMMING ...................................................................................... 17 EXPERT MODE PROGRAMMING ............................................................. 27 ETHERNET AND EXTERNAL COMMUNICATION .................................... 32 SERVICING ............................................................................................. 34 ASSEMBLY DRAWINGS .......................................................................... 35 ORDERING SPARE AND REPLACEMENT PARTS ................................... 47 APPENDIX............................................................................................... 35 WARRANTY Congratulations on the purchase of an original IKA® laboratory machine that meets the highest engineering and quality standards. The IKA® warranty period is 24 months. For claims under the warranty please contact the local dealer. The instrument may also be sent directly to our factory, after prior return authorization from IKA, enclosing the delivery invoice, and giving the reason for the claim. The customer is liable for freight costs.

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EXPLANATION OF SYMBOLS

This symbol identifies information that is absolutely essential for safety and health. Failure to observe this information may result in a health hazards and injuries.

This symbol indicates information which is necessary to ensure the safe handling and operation of the bioreactor. Failure to observe this information could result in a test malfunction or damage to the instrument.

SAFETY INSTRUCTIONS

Use suitable personal protective equipment (PPE) when operating the laboratory reactor system. Make sure power cords are not in a position to be tripped over or kinked. The cords must have a drip loop lower than the instrument connection to make sure no fluids run back into the instrument. Implement the accident prevention regulations for this type of activity and workplace. Wear personal protective equipment at all times when working with the bioreactor. Do not use flammable or explosive gases, vapors or solvents in the vessel. The instrument is not designed for use in explosion proof areas.

The vessel is not designed to be pressurized. Always dedicate at least one port as a vent. Please follow the safe procedures specified for handling reactants and reaction products. Certain applications and materials may be hazardous. Take precautions to prevent contact with, or inhalation of toxic liquids, gases, fumes, vapors or powders. Risks may also be posed by biological or microbiological substances.

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Moving and rotating equipment parts also constitute a hazard. Take caution so clothing or hair does not get caught in the rotating coupling. Be aware that the drive can still be operated when the coupling is removed exposing a rotating shaft. The bioreactor vessel may only be heated, without pressure, through the exterior jacket. Do not under any circumstances use a heating mantel or a hot plate as the vessel could burst. The reactor system must always be ventilated when working under normal pressure in order to prevent any pressure build-up caused by highly volatile gases or unpredictable reaction pressure gradients. The maximum permitted temperature inside the reactor vessel is 200°C. Do not exceed this temperature. The heated vessel and cover can get very hot. Wear protective gloves when handling hot parts. Before the reactor vessel is filled, ensure that the reagents used do not corrode the Viton O-ring. Connect to power cord to 115 volt outlet only. Only use IKA approved accessories and original IKA® spare parts.

DESCRIPTION

The IKA® BR 10 system is a modular mini-plant reactor system. It has been designed to simulate and optimize chemical reaction processes and can also be used for mixing, dispersion and homogenization when modeling processes.

The BR 10 is a 10 liter bioreactor for use in the cultivation of cells under optimum conditions. The stirrer provides laminar agitation for oxygen transfer and mixing nutrients while keeping fermentation homogeneous. The photosynthesis configuration has lamp shields that provide efficient illumination to the process. A simple yet functional control system is conveniently located below the glass vessel for ease of operation and results in an exceptionally small foot print. The HMI (Human Machine Interface) for the control software is easy to read and operate. After the initial pH and temperature parameters are entered, the controller illuminates the fluorescent bulbs based on set temperature range and time. Luminosity can be reduced by

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activating only 2 or 4 of the 6 lamps. Constant air is supplied through an aeration tube until the pH falls out of set range. At this point, a control valve activates to mix carbon dioxide with the air which efficiently modifies the pH. The unit has an Ethernet connection which enables the monitoring of speed, temperature, time and pH. The monitoring is achieved by using existing IKA monitor software. No metal is exposed to the wetted surfaces inside the vessel making this system ideal for instances where bio-reactants are a problem such as with algae. All wetted parts are autoclavable. The glass vessel is fully jacketed on the sides and bottom enabling even heat distribution. Power for turning the stirrer is provided by an in-line, removable DC motor. The system comes with an excellent pH probe as well as a PT 100 Teflon coated temperature probe. The lid is made from glass filled Ultem and has additional ports venting and to add probes such as density and turbidity if so required. An aeration tube with a micro sparger and PTFE tubing that can be used for filling, dosing and extracting. Features of the BR 10

- 10 liter fully jacketed vessel - PTFE encapsulated stirrer with two 4-blade PTFE propellers providing minimal

turbulence - Wetted surfaces chemically inert and autoclavable - Overhead in-line drive motor with coupling for easy removal - Temperature probe – PTFE coated PT-100 RTD - pH probe – glass – gel filled - Automatic carbon dioxide (co2)/air control valve - Aeration tube with micro sparger and extractor tube - Integrated lighting controlled by timers and temperature - HMI Bio-controller integral with cabinet base - Ethernet connection for PC for monitoring temp, pH and stirrer speed - Real-time monitoring and data collection software - Two peristaltic pumps for removing and adding medium - Two rotameters with needle valve for controlling the flow of air and Co2

Technical Specifications

- Housing W x H x D : 510 x 920 x 550 mm - Required autoclave space: 270 diameter x 530 mm - Temperature Probe: -200 to 600°C, RTD PT 100, PTFE coated, 3 wire - pH Probe: 0-14 range, low maintenance, 0 to 100°C, 12mm glass/gel-

filled/Argenthal (Ag/AgCl)reference system - DC motor drive w/integral speed controller, 12VDC/40W, 15:1 gear reduction, 4-

255 rpm. - Power supply: 115VAC, 4 amp, 460 watts, 50/60 Hz

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- Vessel: 250 mm diameter glass, jacket including top and bottom fittings equipped with quick-disconnect fittings for 3/8” ID tubing.

- Working volume: 10 Liter - Cowie fittings: PTFE material, ¼” OD tubing, autoclaveable - Lighting: 6 compact double fluorescent lamps, 1800 lumen each, 24 watts - Illuminated area: 2000 cm² - 600 µmole/sec/m2 of photosynthetically active

radiation (PAR) - Rotameter for air w/needle valve – 299 mL/min - Rotameter for CO2 w/needle valve – 310 mL/min - Two variable speed peristaltic pumps – 450 ml/min

Electrical Specifications

- Motor: 12VDC/40W - Power supply: 115VAC, 4 amp, 460 watts, 50/60 Hz

UNPACKING

The BR 10 has many glass components. Take caution when unpacking not to break these delicate pieces.

Unpack the BR 10 bioreactor individual components and check for any damages. It is important that any damages which may have arisen during transport are ascertained when unpacking. If applicable a discrepancy report must be sent immediately to IKA®. Contact your local IKA® representative for assistance. Contents Verify that the following components are present: Vessel w/lid clamp Base with controls, DC motor,and Lamps Lid with fittings and coupling pH probe Temperature probe Vessel Outer-Jacket fittings w/quick-disconnect hose assemblies Paddle Stirrer Sparger glass tube 12mm od with frit Extractor PFA tube ¼” od x 18” long

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Auxiliary PFA tube ¼” od x 5” long (2) Power Cord Elbow fittings for ¼” od Extractor and Auxiliary tubes pH cable Customer supplied: 3/8” id tubing for outer-vessel jacket connections ¼” id tubing for air and CO2 supply lines to barbed input fittings on rear of unit. Also for peristaltic pump connections. ASSEMBLY OF BR 10 Unpack all the components and set the unit on a secure surface. DC In-Line Motor Instructions for connecting motor

(Note: Power cable motor connector is keyed to ensure proper connection.)

• Locate the motor power cord on the back panel of the unit.

• Securely insert the keyed connector of the power cord to the top of the

motor, and tighten the connector fitting finger tight.

• The motor is now ready to be mounted to the vessel lid.

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Figure 1.

BR 10 Back Base Plate

Position Qty IKA® Part

Number Description

16 1 2946700 Switch,Rocker,MF10 15 1 2279700 Receptacle, 6A, 250V 14 2 2946500 Screw, C’Sink, Socket HD, M3x10 13 1 CB393 Plug, Plastic, Rigid, Locking 12 1 CB133 Fitting, Conduit, Straight 11 2 CB262 Washer, Internal Tooth 10 2 CB261 Screw, Hex Soc Button Head 7 3 CB250 Fitting, Barb, 1/8 NPT x ¼” ID Hose 6 3 CB246 Fitting, Bulkhead, ¾-20 UNEF To 1/8-27 NPT 5 1 CB098 Connector, Female, Omega 4 1 CB225 Panel, Back, Cabinet 2 1 CB090 Connector, Bulkhead, BNC-F-f 1 1 CB066 Connector,CAT-6, Coupler Back Panel Connections The connections for the temperature probe, the pH cable, the air/CO2 outlet and inlet ports are located on the back panel. The temperature probe is a 3-blade connector that can only be plugged in one way. There is a pH probe cable, one end has a BNC connector which connects to the back panel pH connector then the other end will connect the pH probe itself. Connect ¼” ID tubing from the carbon dioxide and air supply source to the “IN” ports

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accordingly. Simply slide the tubing over the barbed fittings. Pressure regulators will be required to control the line pressures. Typically the CO2 and air pressure operate between 15 – 30 psi. There are needle valves on the unit that can regulate the flow. The tubing connected to the glass sparger tube is simply slid down into the sparger for an easy connection. The opposite end of this tube will connect to the air / CO2 “OUT” port by simply sliding the tubing over the barb fitting. It will best to do this once the vessel and lid are assembled. Install the power cord into the AC Inlet connector located on the back panel. (Refer to the Figure 1). Connect the other end to a wall power outlet and verify it matches the power requirements of the unit. 10L Vessel Set the 10L vessel on the white nylon support located in the center of the base unit. Connect the outer-vessel jacket hose assemblies to the glass jacket screw connections located at the top and bottom of the back of the vessel. The hose assemblies have disconnect coupler fittings that easily disconnect from the circulating bath. The coupler fittings also contain shut off valves so that no water leaks when the hoses are disconnected. Attach 3/8” ID tubing to the barbed connections of the coupler fittings and then connect to a recirculation bath for heating and cooling the vessel jacket. (Circulation bath is not included)

Do not apply hot water instantly to the jacket. Thermal shock will occur that could damage equipment and glass. Slowly heat up and cool down the vessel jacket to prevent thermal shock.

Vessel Lid Before placing the lid on the vessel, from the bottom side, install the dual 4-bladed stirrer into the center of the Stirrer Drive, (CB379), by inserting the rod through the large tightening nut and into the end of the driveshaft. Tighten the nut by hand, it may be necessary to insert a tool into the driveshaft coupling on the top side to hold the shaft from spinning while tightening the nut on the bottom side. Before placing the lid onto the vessel it is best to install the glass sparger. From the bottom insert the tubing that is attached to the top of the glass sparger through the largest compression fitting (12mm) and then pass the end of the glass tube through the top side of the fitting. It may be necessary to disassemble the compression fitting pass the tube through the fitting.

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Reassemble the fitting and slightly tighten the fitting to hold the sparger in place. Care must be taken not to break the glass sparger during this assembly. The sparger should be positioned slightly under the stirrer or to the side to avoid any interference with the stirring or bottom of the vessel.

Figure 2

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Installation of Temp and pH Probes

The Cowie fittings have ferrules in them that will fall out if the fitting nut is removed. Please refer to the diagram for reinstallation.

Use ¼” OD tubing with the Cowie fittings.

Remove the temperature probe from the box. Loosen the 5/16” Cowie fitting (item 3) but it is best not to remove the blue cap to maintain ferrule orientation. Insert the probe, from the top surface of the lid, into the correct fitting due to the fact it is the only one it will fit into. Tighten the blue nut to secure in place. Install the extractor tube and auxiliary tubes into the remaining ¼” compression fittings from the top of the lid by loosening the Cowie fittings. The tubes are ¼” OD PFA tubing that have 90° elbow Swagelok fittings. The other ends of the fittings have ¼” OD tube stubs. The operator may connect ¼” ID supply tubing by simply sliding the tubing on to the tube stub. Note that these 3 positions are interchangeable. Tighten the nuts to secure in place. Insertion depth is at operator’s discretion.

The sparger tube has a 90° bend that can interfere with the paddle of the stirrer upon installation. Take caution to make sure there is clearance between them.

Remove the pH probe from its box. Take great caution as it is fragile. Note that the ends of pH probes must never dry out. For brand new probes, use the knife provided in the box to remove the protective tip. Refer to the appendix for more detailed instructions on activation and storage. The probe sent with the unit has been activated and calibrated with the unit, therefore the activation step has already been completed. Gently place the pH probe into the pH adapter fitting in the lid from the top side. The probe screws in to the adapter.

Ph probe Activation Instructions for new pH probes.

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Completed lid with all components

Figure 3

The completed installation should look like the figure above. The cables are not shown for clarity.

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START UP Gently install the lid onto the vessel, center it, and clamp the lid in place. Make sure while installing the lid, that the glass sparger tube does not interfere with the side and bottom of vessel to avoid breaking the glass frit off the end. Adjust the vessel relative to the base so that the outer vessel jacket fittings face the rear and the IKA logo is facing forward. The lid should be oriented so the pH probe is located back left side about the 11:00 o’clock position when looking from the top and the temp probe will be in the 1:00 o’clock. Notice the slots in the lid, these slots need to align with the 4 handles of the vessel clamping ring. Once lid is positioned correctly flip the handles up into the slots and tighten the knobs slightly to secure the lid. Connect the pH cable to top of probe by pushing on then tightening the nut. Connect the tubing from the sparger to the port labeled Air/CO2 OUT on the back panel. Once the lid is secured, insert the motor into the driveshaft coupler. The shaft of the motor has a drive pin perpendicular to the shaft that will align with the slot in the coupler. As the motor is inserted into the coupler the drive pin will self align into the slot. As the motor is being installed notice there is a slot on the side of the housing that needs to align with the locking knob. A locking knob (Position 6 in Figure 2) secures the motor.

Turn on the power switch located on the back panel. The HMI will initialize and then display Time, Date, Temperature, pH, and RPM. The stirrer motor on/off is controlled by the fourth (bottom) button on the keypad, notice the word Start on the display next to the bottom button. The rotation speed is controlled by adjusting the parameters, which will be explained later in the manual. The lights are also controlled through the HMI only.

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Rotameters Rotameters are located on the left side of the BR 10. These meters are designed to monitor the flow of CO2 and Air into the vessel. The top rotameter needle valve regulates the flow of CO2 and the bottom measures and regulates the flow of air into the vessel. The flow of gas or air raises a float in the tapered tube. As the flow of CO2 or air is increased into the vessel, the higher the float will rise. A needle valve knob is located at the bottom of each measuring window that is used to regulate the flow.. This will assist in the regulation process of the gasses depending on the goals of the user. Move the knob to the right (clockwise), to decresae the flow, and to the left or counter-clockwise to increase the flow. The tubes have graduation marks of 0-65 that are for reference. All gasses measure differently due to their density, so to determine actual flow, correlation charts are provide below for determining the actual flow of carbon dioxide and air through the rotometers.

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CO2 Metering Temperature: 70.0º F Metering Pressure: 14.70 psia Metering Density: 0.001835 g/m Metering Viscosity: 0.01470 cp Density at STD. Cond: 0.001835 g/m Accuracy: ñ2% F.S. Barometric Pressure: 14.70 psia Note: The following calculations are based on the flow of carbon dioxide under the above conditions. Air Flow Metering Temperature: 70.0º F Metering Pressure: 14.70 psia Metering Density: 0.001200 g/m Metering Viscosity: 0.01812 cp Density at STD. Cond: 0.001200 g/m Accuracy: ñ2% F.S. Barometric Pressure: 14.70 psia Note: The following calculations are based on the flow of air under the above conditions.

Carbon Dioxide Flowmeter Scale

Reading (mm)

Flow(mL/min)

65 310 60 284 55 256 50 229 45 204 40 177 35 149 30 122 25 100 20 82.0 15 64.0 10 48.0 5 34.0

Air Flowmeter Scale

Reading (mm)

Flow(mL/min)

65 299 60 267 55 238 50 207 45 181 40 160 35 138 30 118 25 97.4 20 80.5 15 63.2 10 48.4 5 36.2

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Peristaltic Pump There are two peristaltic pumps, or roller pumps, located on the base of the BR 10. These positive displacement pumps are designed to assist in the addition and regulation of fluids into the vessel. This flexible option allows the operator to add a variety of fluids, regulate or circulate the flow according to the goals desired. Each pump has an input and output connection. Both pumps provide an efficient and consistent method for adding or extracting liquid from the vessel according to the desired goals. The black power button is located next to each pump (Pictured in Figure 4) Located beside the power button is the potentiometer which regulates the voltage to the motor, and directly affects the flow rate. Turning the potentiometer clockwise, (right) increases the flow, and counterclockwise, (left), decreases the flow.

Figure 4

Additional insertion points are provided on the vessel’s lid for direct ports into the container, (Refer to Figure 2).

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UNLOCKED HOLD ON TO

EXPERT

PROGRAMMING

HMI Main Display

In order to program the parameters you must bypass the safety lock. The safety lock is there to avoid unwanted modification to the parameters. To bypass the safety lock, it is necessary to press one of the top three buttons for more that 5 seconds to access setup menus. Proceed as described below. 1. Press and hold one of the top three buttons to enter the setup menu. The prompt is as follows:

2. Keep the button pressed until the screen changes and the controller prompts as shown below. (If the button is continually pressed, the screen goes into the EXPERT mode, which will be described at a later time.)

LOCKED

HOLD TO UNLOCK

12:00 MON 24/01/11

T1° = 24.0 pH1 = 7.40

Start

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3. Release the button and the controller prompts as follows and proceed as desired:

The HMI is a simple design with easy to use menus. All the menus have four lines and each line corresponds to the corresponding button as shown in the above figure. In the figure shown the first button is used to set the clock, the second button is used to set the timer, the third button is used to set the temperature and the fourth button is used to activate the CONTINUE command and to accept a prompted value.

HMI menu operation instructions

The software has been designed for several applications so there will be prompts that do not pertain to the Bioreactor configuration. Note: The Appendix contains a flow diagram showing the nesting of the HMI menus. Initial Setup There are five main parameters that need to be programmed to run your Bioreactor, the clock, timers, temperature settings, pH and stirrer speed. The first time power is connected the initial setup menu appears on the HMI and requires time and date input. Time is measured in military time. Pressing the + button increases the value and the – button decreases the value. Pressing the CONTINUE button accepts the input and moves to the next screen.

<---> Clock <---> Timers <---> Temp <---> Continue

<---> Clock <---> Timers <---> Temp <---> Continue

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Clock setup Clicking the “Clock” label will allow the following input: Hour of the day (00-24) (Military Time) Minute (01-59)

Day of the week (Mon – Sun) Day of the month (01-31) Month MM (01-12) Year (2004-2999) Note: the date is displayed in the European format of DAY/MONTH/YEAR. Loading Parameters

The default menu while under power shows time, day, date, Temperature, pH and RPM.

To enter parameters pick one of the top three buttons and hold it while the “UNLOCKED” prompt is showing. The following menu will appear:

The appendix has a flow diagram of the various menus that will come up on the screen.

Time Day Date T1° = XX.X pH1 = XX.XX Stop rpm = XXX

<---> Clock <---> Timers <---> Temp <---> Continue

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Press continue for more of the parameters menu:

Setting TIMERS The timers control the time (military time) of day the lights come on and off. There are six timers that are available, but currently only Timer 1 and Timer 2 need programming. Timer 1 controls two lights and Timer 2 controls four lights. There are six lights total. From the menu, select Timers for this menu:

Press Set Points for this menu:

<---> Set points <---> Info <---> Expert <---> Exit TIMERS

<---> pH <---> Multi Timer <---> Dosometric Pump <---> Continue

<---> Stirrer Speed <---> Info Multi Plug <---> Dosometric Pump <---> Continue

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To turn Timer 1 ON, enter time by clicking the + for more time and – for less (gives 10 minute intervals). Example: To turn lights on at 6:00 AM adjust time to 06:00. Press Continue for this menu:

To turn Timer 1 OFF, enter time off by clicking the + for more time and – for less. Example: To turn lights off at 8:00 PM adjust time to 20:00. Repeat steps for Timer 2. Setting TEMPERATURE The lights will generate heat and could cause the vessel temp to increase if not be controlled with a circulating bath. The temperature setting will cut the lights off and on when the temperature goes out of range. The user can set this range with two temperature inputs,Temp 1 and Temp 2 setpoints that will turn the lights off to reduce the heat being generated. This action supersedes the original light timer temporarily. The timer continues as normal after interruption. Go back to main menu and press Temp for this menu:

<---> Timer 1 ON = XX.XX <---> + <---> - <---> Continue

<---> Timer 1 OFF = XX.XX <---> On: + <---> Off: - <---> Continue

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Press Set Temp 1 for this menu:

Enter temperature by clicking the + for more time and – for less. This is for max temperature. Temp 1 works in conjunction with Timer 1, so when the temperature exceeds the setpoint, Timer 1 will be temporarily disabled as long as the temperature is above the setpoint, therefore the two lights will turn off. Repeat for Temp 2 and set the temperature. Temp 2 works in conjunction with Timer 2, so when the temperature exceeds the setpoint, Timer 2 will be temporarily disabled as long as the temperature is above the setpoint, therefore the four lights will turn off. Setting pH The desired pH is entered and a control valve switches from constant air supply to carbon dioxide when the pH value is out of range. Once in range, the air supply is returned. The desired pH level is entered first then the allowable pH tolerance range is entered through the expert mode. An alarm tolerance range is also entered in the expert mode. Go back to main menu and press pH for this menu:

<---> Set Temp 1 <---> Set Temp 2 <---> Expert <---> Exit TEMP

<---> Opt. Set T = XX.XX <---> + <---> - <---> Continue

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Press Set pH 1 for this menu:

Enter desired pH by clicking the + for more pH and – for less. Press Continue for this menu:

Press Expert (sets alarm and the allowable pH tolerance range) for this menu:

<---> Set pH 1 <---> . . . . . . . . <---> Expert <---> Exit

<---> Opt. set pH 1: XX.XX <---> + <---> - <---> Continue

<---> Set pH 1 <---> . . . . . . . . <---> Expert <---> Exit

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Press Continue for this menu:

Enter pH tolerance range to activate alarm by clicking the + to increase and – to decrease. Press Continue for this menu:

Enter desired pH tolerance range by clicking the + to increase and – to decrease. Press Continue for this menu:

<---> Acoustic alarm <---> YES <---> NO <---> Continue

<---> Alarm Start: +/- X.XX <---> + <---> - <---> Continue

<---> pH control: +/- X.XX <---> + <---> - <---> Continue

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This menu is used to assign plug values to the inputs. Do not enter information to modify the plug assignments, as the plugs are factory set by IKA. Keep pressing Continue to return to main menu. Setting STIRRER SPEED The Stirrer Speed is adjusted by clicking the + to increase and – to decrease the rpm. The Stirrer is started and stopped by pushing the fourth (bottom) button on the Main menu. Go to the set-up menu and press Stirrer Speed for this menu:

Setting MULTI TIMER The multi timer is designed to allow the light timer to operate more than one time cycle in a day. This allows the lights to come on and off for a maximum of four different times a day. Go back to main menu and press Multi Timer for this menu:

<---> Set points <---> Info <---> Expert <---> Exit

<---> Task: decrease P1 (1-) <---> Assigned plug: A3 <---> Modify <---> Continue

RPM = 030 (0-255) <---> + <---> - <---> Continue

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Press Set points for this menu:

Press Continue for this menu:

Enter time on for the first cycle by clicking the + for more time and – for less. Press Continue for this menu:

Enter time off for the first cycle by clicking the + for more time and – for less. Press Continue to enter up to three additional cycles.

<---> Clear Points? : Yes <---> Yes? <---> No? <---> Continue

<---> T4 1 on = XX.XX <---> + <---> - <---> Continue

<---> T4 1 off = XX.XX <---> + <---> - <---> Continue

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UNLOCKED HOLD ON TO

EXPERT

EXPERT MODE PROGRAMMING Expert Mode Programming is used to do pH and temp calibrations, plug output assignments and data collection settings. Enter the expert mode by pressing and holding one of the top three buttons to enter the setup menu. The prompt is as follows:

Keep the button pressed until the screen changes and the controller prompts as shown below. Continue pressing the button. Release the button and the controller prompts as follows:

LOCKED

HOLD TO UNLOCK

EXPERT

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Press Continue for the following menu:

Calibration

Calibration is used to calibrate the pH and temperature probe. Make sure both probes are plugged into the back panel otherwise the menu will read empty. Temperature Probe The temperature probe calibration requires the use of a second temperature measuring device. Calibrate at 0°C first then calibrate against a known temperature source. Press Calibrations for the following menu:

Do not dry pH probe with a towel as this will damage it. Remove moisture tip before usage and replace during storage. pH probes should never dry out.

<---> Calibrations <---> Ass. Task to Plug <---> Reset <---> Continue EXPERT

<---> Memory Modes <---> Logout Modes <---> History View <---> Exit

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Press Cal. T1 for the following menu:

Press Yes for the following menu:

Press Continue for the following menu:

Place Temp probe in the same place of a digital thermometer <---> Continue

<---> Cal. T 1 <---> Cal. pH 1 <---> . . . . . . . . <---> Continue Cal.

Are you sure? <---> YES <---> Exit

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Press Continue for the following menu:

Correct the temperature value with the + and – control based on the reading from the second temperature device. pH Probe Note: Test requires clean water, pH 4, and pH 7 solutions. The pH probe requires periodic calibration to maintain accuracy. The probe is calibrated through a simple process of placing it into a standard solution of known water quality and following the prompts on the display. Typical standard solutions are pH 4 and pH 7. Set up two cups, one with each solution. Rinse the probe with water but do not dry off with a towel. This could damage the probe. Place probe in the pH 4 solution. Return to the calibration menu:

Press Cal pH 1 for the following menu:

Continue when stabilized **XXXX** <---> Continue

<---> T = XX.XX° <---> + <---> - <---> Continue

<---> Cal. T 1 <---> Cal. pH 1 <---> . . . . . . . . <---> Continue Cal.

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Press Yes for the following menu:

Press any key for the following menu:

Press Continue for the following menu:

Are you sure? <---> YES <---> Exit

Place pH probe in pH 4 Any key to continue

Continue when stabilized pH4 **XXXX** <---> Continue

Place pH probe in pH 7 Any key to continue

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Press any key for the following menu:

Press Continue to finish the process. Remove the probe from the pH7 solution and rinse with water. If the probe is not within the tolerance limits, the reading will not get stable or show erratic values. Should this occur, clean the probe and try to recalibrate it. If the probe is out of specification the controller will prompt that there is an error.

If the probe has been cleaned twice and there are still problems, it is time to replace the probe. They should be replaced every 16 to 18 months or as needed for accuracy and reliability. The EXPERT mode has the following other options: Assign Task to Plug Outputs Reset Memory Modes Logout Modes History View Expert modes per Timers, Temp, pH and Multi Timer also exist for the non expert menus. Refer to the Appendix for a flow diagram showing the nesting of these menus. ETHERNET AND EXTERNAL COMMUNICATION The instrument can be connected to an external computer to enable monitoring of pH, temperature and RPM by the Ethernet connection. The software can be configured to monitor several instruments at once. Software needed: MOXA CD IKA Monitor CD

Continue when stabilized pH7 **XXXX** <---> Continue

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Minimum computer: Windows XP 256 meg RAM 50 meg hard drive Ethernet connection Installing software To install MOXA software on external computer: Insert the MOXA CD into a computer drive and browse the CD. Select Folder: DE-311 Select Folder: Software Select correct Operating System of PC Select: magsuite_setup_3.4 Select: Next (multiple times) Select: Install Do not check either box when prompted as follows: Launch configurator to configure N Port parameters……… Select: Close Plug Ethernet cable into the back of the IKA Bioreactor Wait 30 seconds Select on computer desktop: Start Select: All programs Select: Import management suite Select: Configurator Select: Locate Select: Broadcast Search The computer is now searching for the drive in the Bioreactor and finding the IP address of the drive. When it finds it a line will appear on the MOXA interface panel with the IP address. Select by double clicking: the unit found It is possible to assign a static (new) IP address at this point. If changing IP address, check box and proceed. We have found it is best to reassign the IP address, it defaults to 192.169.127.254, change it to something different for instance 192.168.127.240. If you have multiple units then make sure when you reassign their IP addresses they are different from each other. Select: OK Select: Close Select: Back to software Select: Close

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Return to Import Management suite Select: COM port mapping Select: real COM installer PC will search network for N port Select: N port Select: Add server wizard Select: Next Select: manually enter IP address Select: Next Choose the COM port that you wish to assign Select: Next (multiple times) Select by checking: Yes, please enable COM ports now Select: Finish Select: Yes to warning The MOXA software is now installed To install IKA Monitor software on external computer: Inset IKA Monitor CD in computer drive Select: Browse Select Folder: Monitor Select Folder: Setup Select: Next (multiple times) Select: Close On PC desktop Select: Programs Select Folder: IKA Select: IKA Monitor Select COM # that was chosen in previous installation Select: Connect The software will begin collecting data and present it in graph form for each parameter being monitored. Each COM port can support a different Bioreactor which enables the user to monitor many units at once through a new IKA Monitor window. SERVICING Periodically check the following for flaws, wear or fractures:

Glass vessel

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Glass fittings

O-ring in lid

Lip seal in drive coupling

Coupling metal interfaces

Monitor hose connections for leaks APPENDIX Appendix I Engineering Drawings BR 10

Appendix II pH Probe Installation Instructions

Appendix III pH Probe Data Sheet

ASSEMBLY DRAWINGS AND PARTS LIST The following drawings give a breakdown of the parts used in the Bioreactor. Appendix I. provides engineering drawings of the Bioreactor to give a breakdown of the parts used. Engineering Drawings: CB372 Assembly BR-10 Bioreactor

CB198 Assembly Stirrer Drive

CB371 Assembly Cabinet Cover

CB189 Assembly Base

CB361 Assembly Right-Wing Lamp

CB360 Assembly Left-Wing Lamp

CB191 Assembly Cover-Flange

CB379 Assembly Drive

CB273 Assembly BR 10 Vessel

CB403 BR 10 Stirrer

CB226 Assembly Back Pannel

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Assembly Drawing CB372 BR 10 Bioreactor

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Assembly Drawing CB198 Stirrer Drive

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Assembly Drawing CB371 Cabinet Cover

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Assembly Drawing CB189 Base

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Assembly Drawing CB361 Right-Wing Lamp

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Assembly Drawing CB360 Left-Wing Lamp

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Assembly Drawing CB191 Driver

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Assembly Drawing CB379 Stirrer Drive

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Assembly Drawing CB273 BR 10 Vessel

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Assembly Drawing CB403 Stirrer

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Assembly Drawing CB226 Back Panel

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ORDERING SPARE AND REPLACEMENT PARTS To order repair parts please use the form below. This allows us to process your order quickly. Provide your complete address and, if different, the desired delivery addresses. Enter the part information in the table below. The exact name and part number can be found in the parts list in the previous section. Always give the position, name and the identification number.

Pos. IKA Part Number

Item Description Assembly Drawing

Material

Send to: IKA® Works, Inc. 2635 North Chase Pkwy SE Wilmington, NC 28405 U.S.A Ph.: 1- 800-733-3037 Fax: 910-452-7693 www.ikaprocess.com

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Appendix II pH Probe Installation instructions 1. When unpacking, check electrode for possible mechanical damage to the membrane, the diaphragm or the body. Electrodes subject to complaints must be returned to us or to our agent in the original box indicating the invoice number. 2. Take off watering cap and flush electrode with distilled water. After rinsing with distilled water, wipe the electrode with a tissue. Do not rub as an increase of the response time might result. 3. Remove rubber cap which covers the electrolyte filling aperture or cut off nipple to ensure pressure equalization with the atmosphere. 4. Eliminate any air bubbles inside the membrane space of the measuring electrode by smoothly swinging the unit in the vertical plane. 5. To renew a dried out glass membrane, soak it for several hours in the appropriate storage solution. It is recommended to store glass electrodes in distilled water, combined and reference electrodes in the reference electrolyte solution. 6. After a long and dry storage period, the glass membrane can be reactivated with hydrofluoric acid (code no. 51 340 073): the electrode membrane is dipped for 1 minute in this solution followed by rinsing with distilled water and soaking for 12 hours in the storage solution. 7. Electrodes with ground sleeve diaphragm: slightly lift the ground sleeve so that some reference electrolyte drops out. 8. Check level of electrolyte which should be approximately 10 mm below refilling aperture. Refill, if necessary with: 3M KCI+AbCI (order no. 51 340 045) for electrodes with Ag/AgCI wire 9. Electrodes with gellified electrolyte without refilling port are treated as per items 1,2, 4, 5 and 6 above. For storage we recommend a solution 3 mol/I KCI. 10. Before calibrating or taking measurements, attention should be paid to the instructions of the relative pH instrument manufacturer. Additionally, please refer to the METTLER TOLEDO brochure “Practice and Theory of pH Measurement”.

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Appendix III pH Probe Data Sheet

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IKA® WORKS, INC. LABORATORY- TECHNOLOGY

ANALYZING TECHNOLOGY PROCESSING EQUIPMENT

IKA® WORKS, INC.

2635 NORTH CHASE PKWY, SE WILMINGTON, NC 28405-7419

TEL.: 800/733-3037 TEL.: 910/452-7059 FAX: 910/452-7693

E-mail: process@ikausa.com Website: http://www.ikausa.com