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NEW BRUNSWICK SCIENTIFIC BioFlo 4500 Manual To Operations

Benchtop fermentor/bioreactor
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BioFlo 4500
Benchtop
Fermentor/Bioreactor
MANUAL NO: M1254-0050
Revision H
September 27, 2002
NEW BRUNSWICK SCIENTIFIC CO., INC.
BOX 4005 • 44 TALMADGE ROAD • EDISON, NJ 08818-4005
Telephone: 1-732-287-1200 • 1-800-631-5417
Fax: 732-287-4222 • Telex: 4753012 NBSCO
Internet: http://www.nbsc.com • E-mail: bioinfo@nbsc.com

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  • Page 1 Benchtop Fermentor/Bioreactor MANUAL NO: M1254-0050 Revision H September 27, 2002 NEW BRUNSWICK SCIENTIFIC CO., INC. BOX 4005 • 44 TALMADGE ROAD • EDISON, NJ 08818-4005 Telephone: 1-732-287-1200 • 1-800-631-5417 Fax: 732-287-4222 • Telex: 4753012 NBSCO Internet: http://www.nbsc.com • E-mail: bioinfo@nbsc.com...
  • Page 2 BioFlo 4500 M1254-0050 User’s Guide...
  • Page 3 Fax: 49 (0)7022 32486 E-mail: sales@nbssarl.fr E-mail: sales@nbsgmbh.de UNITED KINGDOM CHINA New Brunswick Scientific (UK) Ltd. New Brunswick Scientific Co., Inc. Edison House, 163 Dixons Hill Road 9A. Ai Bang Building North Mymms, Hatfield, Herts. AL9 7JE 585 Ling Ling Road United Kingdom Shanghai 200030, P.R.
  • Page 4 THIS PAGE INTENTIONALLY BLANK BioFlo 4500 M1254-0050 User’s Guide...
  • Page 5 Do not use this equipment in a hazardous atmosphere or with hazardous materials for which the equipment was not designed. New Brunswick Scientific Co., Inc. (NBS) is not responsible for any damage to this equipment that may result from the use of an accessory not manufactured by NBS.
  • Page 6 THIS PAGE INTENTIONALLY BLANK BioFlo 4500 M1254-0050 User’s Guide...
  • Page 7 Reproduction, adaptation, or translation without prior written permission from New Brunswick Scientific is prohibited. Disclaimer Notice New Brunswick Scientific Co., Inc. reserves the right to change information in this document without notice. Updates to information in this document reflect our commitment to continuing product development and improvement.
  • Page 8 THIS PAGE INTENTIONALLY BLANK BioFlo 4500 M1254-0050 User’s Guide...
  • Page 9 W A R R A N T Y Every Instrument manufactured by the New Brunswick Scientific Co., Inc. is warranted to be free from defects in material and work- manship. This apparatus with the exception of glassware, lamps and electrodes (where supplied), is warranted for 1 year against faulty com-...
  • Page 10 THIS PAGE INTENTIONALLY BLANK BioFlo 4500 M1254-0050 User’s Guide...
  • Page 11 Model Number: Voltage: Serial Number: Controller Type: The above information can be found on the electrical specification plate on the right side panel of the console. Purchased with the following installed options: New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 12 THIS PAGE INTENTIONALLY BLANK BioFlo 4500 M1254-0050 User’s Guide...

  • Page 13: Table Of Contents

    Piping and Instrumentation .................. 20 1.1.3 Utilities........................20 1.1.4 Optional ML-6100 Controller ................21 1.1.5 Other Options......................21 ......................22 PECIFICATIONS BIOFLO 4500 FEATURES....................27 .......................... 27 ESSEL 2.1.1 Viewing Window ....................28 2.1.2 Vessel Jacket ......................28 ................29 ESSEL AND...
  • Page 14 ................... 91 XHAUST REATMENT 12.2.1 Exhaust Air (Gas) Filters: ..................91 12.3 ..................92 PERATION OF PARGE YSTEM ML-6100 TWO-, THREE- & FOUR-GAS CONTROL..........93 13.1 -, T ........94 ETTING P THE HREE IXER PTION BioFlo 4500 M1254-0050 User’s Guide...
  • Page 15 Sparging with Nitrogen to Calibrate DO Zero Point ......... 134 ML-6100 LEVEL CONTROL ..................135 18.1 ........................135 ENSOR 18.2 ........................135 ONTROL 19 ML-6100 NUTRIENT CONTROL.................. 137 SAVING THE ML-6100 CONTROLLER CONFIGURATION ......139 New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 16 INOCULATION & ADDITION VALVE (OPTIONAL).......... 179 29.1 ......................179 TERILIZATION 29.2 & I ..................180 DDITION NOCULATION 29.2.1 Method A......................180 29.2.2 Method B......................181 SAMPLING VALVES (OPTIONAL)................. 183 30.1 ......................184 TERILIZATION 30.2 ........................184 AMPLING BioFlo 4500 M1254-0050 User’s Guide...
  • Page 17 APPENDIX A: CORROSION RESISTANCE TABLES......... 223 APPENDIX B: GENERAL CHARACTERISTICS OF EPR........231 APPENDIX C: STANDARD PID VALUES ............. 233 APPENDIX D: ADVANCED CONFIGURATION SETTINGS......235 APPENDIX E: FOUR-GAS MIX ALGORITHM ............ 237 INDEX..........................239 New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 18 BioFlo 4500 M1254-0050 User’s Guide...

  • Page 19: Overview

    The versatility of the BioFlo 4500 is derived from its state of the art ML-6100 instrumentation. This instrumentation package may be configured as an imbedded controller with touchscreen interface, a remote stand-alone system, or can be controlled by third-party SCADA systems.

  • Page 20: Features

    Minimum 350 mMoles/L /hr oxygen transfer rate (OTR) • Capable of minimum 1°C/minute temperature rise between 30° and 45°C 1.1.3 Utilities • Connections for water, air, gases, steam and drain are provided on either side of the base BioFlo 4500 M1254-0050 User’s Guide...

  • Page 21: Optional Ml-6100 Controller

    Automatic back pressure control • Choice of Ingold or Broadley James pH and DO sensors • Passivation package which includes all stainless steel valves and traps • Upgraded interior and exterior finishes • Electropolishing New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 22: Specifications

    Specifications BioFlo 4500 Fermentor/Bioreactor Vessel Design Code The vessel conforms to the latest issue of ASME, Sect. VIII Pressure Vessel Code. It is supplied with copies of the Manufacturer's Data Report (U-3). Vessel includes the ASME-Code Stamp. Total Volume Maximum Working Volume 15.0L...
  • Page 23 Optional Ingold gel or liquid- filled or Broadley James gel pH probe available Dissolved Oxygen (DO) Ingold Type Lower Side Wall Plugged as standard. Port 25 mm Optional Ingold or Broadley James polarographic O probe available …continued… New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 24 Air inlet filter Steam-sterilizable absolute 0.2µ filter that continuously removes viable organisms, cartridge- type 316 L, stainless steel housing, Domnick Hunter. NOTE: Relative humidity must be less than 95%. Other brands can be supplied upon request …continued… BioFlo 4500 M1254-0050 User’s Guide...
  • Page 25 Conductivity probe (Optional) Alarm Airflow and agitation are shut down in case of high foam alarm indication Sensor Choice of Ingold gel or liquid-filled, or Broadley James (Optional) gel pH probe available …continued… New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 26 Output type: PWM 0-100%, Mode: On/Off with Reverse action Display Current value, setpoint, controller output % displayed on virtual display gauge at touchscreen BioFlo 4500: General Specifications Weight 300 lbs. (136 kg) 420 lbs. (191 kg) Gross 425 lbs. (193 kg) 545 lbs.

  • Page 27: Bioflo 4500 Features

    The bottom harvest/drain port with a plunger-type valve is located on the centerline of the vessel. The port incorporates steam lines for continuous re- sterilization. Steam and/or condensate is then passed to a steam trap and drain connection. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 28: Viewing Window

    LAMP with MOTOR PUSHBUTTON RUPTURE DISK COMBINED FILLING (powder/liquid) / RUPTURE DISK LIGHT PORT PIPING BAFFLES IMPELLER SHAFT (4, removable) SET SCREW SIX-BLADED RUSHTON IMPELLERS (2) HEMISPHERICAL DISH BOTTOM BAFFLE RING SPARGER HARVEST/DRAIN/ SAMPLE PORT BioFlo 4500 M1254-0050 User’s Guide...

  • Page 29: Vessel And Jacket Penetrations

    • Two U-shaped handles Figure 3: Headplate Penetrations Top View RUPTURE DISK BEARING HOUSING EXHAUST PORT (plugged) HANDLE 19mm ID (28mm) FOAM & LEVEL PORTS PORTS with tube fittings Combined FILLING (power/liquid) & LIGHT PORT New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 30: Optional Headplate Equipment

    • Two movable six-bladed Rushton impellers Figure 4: Upper Side Wall Penetrations Top View Cross Section JACKET PRESSURE JACKET WATER OUT RELIEF VALVE OVERLAY PORT SPARGE VALVE (plugged) RING SPARGER SIX-BLADED RUSHTON IMPELLER 19mm PORTS 19mm PORTS VIEWING WINDOW BioFlo 4500 M1254-0050 User’s Guide...

  • Page 31: Optional Vessel And Vessel Upper Side Wall Equipment

    Optional Lower Side Wall Equipment • Re-sterilizable, spool-type sampling valve with ¼” quick-connect fittings • Ingold gel or liquid-filled or Broadley James gel pH probe • Ingold or Broadley James DO polarographic probe New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 32: Vessel Bottom

    • All process piping, valves and associated components are of type 316L stainless steel. All other piping is of type 316L or 304 stainless steel. • All gaskets are silicone. • All O-rings are Ethylene Propylene EPDM. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 33: Inspection, Verification & Unpacking Of Equipment

    Ensure that support structure and surface can bear the weight of the BioFlo 4500 (300 lbs/136 kg for the 20L vessel and 420 lbs/191 kg for the 30L vessel) plus vessel contents and any applicable auxiliary equipment.
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  • Page 35: Getting Started

    Set the console on a sturdy, level surface. Ensure that there is enough space around the back and the front of the BioFlo 4500 for proper operation and maintenance and that the surface can handle the weight of the unit, including vessel contents and any auxiliary equipment.

  • Page 36: External Computer Set-Up

    2. Place the computer on a sturdy, level surface in close proximity to the intended location of the fermentor. 3. Place the touchscreen monitor on the top of the computer. 4. Attach the power cord to the computer BioFlo 4500 M1254-0050 User’s Guide...
  • Page 37 The electrical connections to both the computer and the touchscreen should be made after the has been connected to the EXTERNAL COMPUTER CABLE fermentor/bioreactor. NOTE: Verify that the supply voltage matches the voltage of the computer. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 38: Computer Connections

    AUDIBLE ALARM EXTERNAL ALARM CONNECTOR Alarms An audible alarm is provided with the BioFlo 4500. It is mounted on the rear panel of the console (see drawing above). The alarm will sound when an alarm condition occurs. BioFlo 4500 M1254-0050...

  • Page 39: Biocommand Connections

    The BioFlo 4500 also has provision to allow the connection of an external alarm device. An external alarm device is connected to the console via the nine-pin on the rear panel (see drawing above). A cable must be ALARM CONNECTOR fabricated in order to use this option.

  • Page 40: Analog Inputs

    The local controller software used with the controller displays input and output connections as "J" numbers. The slot number, in this case, 2, refers to installed position of the I/O module in the ML-6100 controller. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 41: External Pump Connections

    External Pump Connections The BioFlo 4500 fermentation system is configured to operate a fourth, external pump. This fourth pump is controlled to turn on and off depending on system conditions. The supply voltage for this pump is supplied by a line cord connected to the .
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  • Page 43: Assembling The Fermentor/Bioreactor

    On initial set-up, prior to operation, check that all impellers are tightly fastened to the drive shaft. The BioFlo 4500 is equipped with two Rushton six-blade impellers as standard. The impellers’ position should be verified before securing the headplate to the vessel. The impellers are adjustable along the length of the shaft and are secured to the impeller by a locking collar.

  • Page 44: Placing The Vessel On The Console

    The baffles may also be removed at this time. Mechanical assistance may be required to move the vessel and console. Position the console in its permanent location then proceed to reinstall the vessel, headplate and motor. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 45: Headplate & Motor Installation

    HEATER CONNECTOR Figure 14: Motor & Bearing Housing (shown with double mechanical seal option) MOTOR BEARING HOUSING BLOCK MOTOR MOUNTING BOLT SPEED ENCODER (1 of 4) HEADPLATE IMPELLER SHAFT New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 46: Service/Utility Connections

    Before making service/utility connections to the BioFlo 4500, become familiar with each service connection and its location. The following table describes each service connection. The BioFlo 4500 must be connected to regulated utilities as described in the order that they appear:...
  • Page 47 10 PSIG (0.67 bar) with unit in rapid cooling. NOTE: Utility steam to the fermentor/bioreactor should be free of particulates. If the utility steam is laden with particulates, a steam prefilter/regulator kit PN M1117-2020 is recommended. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 48 PN M1117-2020 is recommended. Figure 16: Service Connections For gas connections, see Figure 17 on the following page. DRAIN ½” FNPT WATER ¼” FNPT WATER RETURN ¼” FNPT STEAM ¼” FNPT POWER CORD PURE STEAM ¼” FNPT BioFlo 4500 M1254-0050 User’s Guide...
  • Page 49 Figure 17: Gas Connections CAUTION! Power fluctuations varying more than 10% from the nameplate rating can result in severe damage to the fermentor/bioreactor electronics. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 50: Steam Supplies

    Pure steam can be made available to the BioFlo 4500 either from a house supply or from a small, dedicated clean steam generator.

  • Page 51: Electrical

    5.5.5 Electrical The BioFlo 4500 is shipped with a line cord that does not include a plug. A suitable plug must be attached or the fermentor may be hardwired directly to a circuit breaker. Before making connections, verify that the...

  • Page 52: Water & Water Return

    Optional prefilter and regulator kits are available for installation on your BioFlo 4500 system: Optional Kit NBS Part Number Steam Prefilter/Regulator Kit M1117-2020 Water Prefilter/Regulator Kit M1117-2040 Water Regulator M1176-0060 Air Prefilter Kit M1117-2030 Air Prefilter/Regulator Kit M1230-3030 Air Regulator (Only) 0-125 PSI P0220-4080 BioFlo 4500 M1254-0050 User’s Guide...

  • Page 53: Preparation

    NOTE: Sections 6.1 through 6.3 refer to the use of the BioFlo 4500 with the ML-6100 controller only. If you are not using the ML-6100 controller, skip ahead to Section 6.4. Initial Preparation 1. Turn the power on by setting main power switch (on the side of the console) to position.

  • Page 54: Water Priming

    4. Set the operating setpoint of the temperature control loop to 10°C below the indicated temperature. 5. Observe the water return line. The vessel jacket is primed when a steady stream of water runs out of the line. If the line is not exposed, wait approximately two minutes. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 55: Calibrating The Touchscreen

    If the controller has been configured prior to or after initial start-up, will not appear blank after the start-up message GAUGE SCREEN disappears. The control loop display gauges from the last configuration entered will appear on the (see Figure GAUGE SCREEN 36). New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 56: Harvest/Drain Valve

    Harvest/Drain Valve The BioFlo 4500 is equipped with a manually-operated, re-sterilizable, plunger-type drain valve (see drawing below). The bottom drain port with valve is located on the centerline of the vessel, and is equipped with a ¼” quick-connect fitting for aseptic transfer of culture broth.

  • Page 57: Asceptic Harvest/Drain Method A

    1. Prepare an appropriate harvest vessel fitted with tubing (and a fitting that allows easy connection to the quick-connect). Sterilize if required. NOTE: Refer to your local NBS parts distributor or sales office for re- sterilizable quick-connect options. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 58: Asceptic Harvest/Drain Method B

    2. Autoclave harvest vessel and quick-connect fitting in the closed position (121ºC, 30 minutes minimum, depending on vessel). 3. Remove steam port cap from the harvest/drain port and connect the quick-connect fitting from the harvest vessel to the harvest/drain port. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 59 13. Close the valve on the quick-connect fitting. Detach the sterile quick- connect from the harvest/drain port. 14. Put the stainless steel port steam cap back on the port. Re-sterilize the port. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 61: Starting The System

    NOTE: The following paragraphs refer to the use of the BioFlo 4500 equipped with the ML-6100 controller only. Start-Up Do the following to start the BioFlo 4500 fermentation system: • Press the on the fermentor/bioreactor. Turn on the MAIN POWER SWITCH computer and touchscreen monitor if applicable to your system.
  • Page 62 This condition will not occur if the controller cannot communicate with the software, if the controller configuration has been deleted, or if there is no current controller configuration. All BioFlo 4500 fermentor/bioreactors are pre-configured at the factory for all standard process parameters and any factory-installed options (for example: automatic backpressure control).

  • Page 63: Display Gauge

    LOOP SETPOINT The setpoint value of the loop. MEASURED UNIT The unit of measurement of current output and setpoint. CONTROLLER The value of loop output as a percentage of full-scale controller output. OUTPUT % New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 64 BUTTON • Press to accept the loop setting changes. The new values you entered will be displayed in the gauge within a few seconds after returning to the GAUGE SCREEN BioFlo 4500 M1254-0050 User’s Guide...

  • Page 65: Ml-6100 Controller Configuration

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The ML-6100 controller of the BioFlo 4500 was configured prior to shipment. However, user adjustments may be required for certain processes. Configuring the controller includes specifying for each controlled parameter its display, input, output and control characteristics.

  • Page 66: System Information

    CONTROL TYPE BUTTON CONTROL TYPE DIALOG BOX Verify that the BioFlo 4500 is the selected controller type. Press A message will appear, asking if you want to change the control type. If you do, press the . If not, press the...

  • Page 67: Control Mode Selection And Node Number

    If you wish to change it, press the text box. INSTRUMENT NAME • A will appear. To clear the text out of the text box, RENAME MODE DIALOG BOX press . The can be blank. INSTRUMENT NAME CLEAR New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 68 AFS Mode must be selected and the must be set to an even NODE NUMBER number between 0 and 30. The first 10 loops in the BioFlo 4500 will appear at the selected . Any additional loops will appear at the next odd NODE NUMBER number (node number plus one).

  • Page 69: Passwording And Locking

    BASIC OPERATIONS PASSWORD To change either password, press the in the PASSWORD BUTTON MAIN . The will appear, with the CONFIGURATION SCREEN PASSWORD DIALOG BOX default selection being the BASIC OPERATIONS PASSWORD New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 70 3. Touch the new password text area. 4. Touch the text area, then type in the new password CONFIRM PASSWORD utilizing the onscreen keyboard. 5. Press on the onscreen keyboard after completing all three fields. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 71 3. You will be prompted to enter the . Enter the BASIC OPERATIONS PASSWORD password and press . If the has been set BASIC OPERATIONS PASSWORD a blank value, you will not be prompted for it. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 72: Screensaver Option

    3. Select the time interval after which you wish the screensaver to activate. 4. Press to enable your selection CLOSE 5. To disable a screensaver, repeat the previous steps, pressing the ENABLE to remove the “ ” then pressing to enable your selection. CLOSE BioFlo 4500 M1254-0050 User’s Guide...

  • Page 73: Ml-6100 Control Loop Operations

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. Adding a New Loop to the Display Adding a new signal input (feedback signal) and control output (control signal) will require that the controller be “configured” for the new loop through the touchscreen interface.

  • Page 74: Configuring Control Loops

    To configure a new control loop or to modify a previously configured control loop: 1. Press the in the CONFIG BUTTON GAUGE SCREEN 2. The will appear. MAIN CONFIGURATION SCREEN 3. In the , touch the loop that you wish to configure. LOOPS WINDOW BioFlo 4500 M1254-0050 User’s Guide...
  • Page 75 ML-6100 controller to directly control the input signal of the control loop. will allow an analog 4-20 mA signal from an external PLC or ANALOG controller to be input to the controller, thus setting the controller in “slave” mode for the control loop. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 76 12. Select the to set the controller characteristics for this CONTROLLER ICON loop, i.e., how it will respond to differences between setpoint and current value. The will appear. CONTROLLER SETTINGS SCREEN BioFlo 4500 M1254-0050 User’s Guide...
  • Page 77 On and Off . The duration of the On state ranges between 0 and 100% of the period. The electrical characteristic of the PWM output signal depends on the Digital I/O Terminal Panel used. Enter a period between 1 and 600 seconds. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 78 • OUTPUT A varies linearly from minimum at 0% 2 CH DIRECT controller output to maximum at 100% controller output. • OUTPUT B varies linearly from maximum at -100% controller output to minimum at 0% controller output. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 79 The factory settings for the select calibration screen are: • Pointing Device • Pump 1 • Aux Pump • pH • Pump 2 • Hi Foam • DO • Pump 3 • Anti Foam New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 80 LOOP CONFIGURATION SCREEN. The Sterilization function will be enabled only if it was assigned an . Please refer to the Automatic OUTPUT TYPE CONNECTOR ID Sterilization section of this manual for detailed instructions. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 81: Deleting Loops

    4. Control will return to the . Press to return to CONFIGURATION SCREEN CLOSE . The display gauge for the deleted control loop will be GAUGE SCREEN removed. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 83: Ml-6100 Temperature Control System

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. There are four high watt density cartridge heaters at the bottom of the vessel. Water is heated by the cartridge heaters in the jacket, where heat is transferred through to the media in the inner vessel.
  • Page 84 PWM Period:10 Gains Sterilization P: 30 Output Device I: 1 Recorder D: 0 Norm Const:100 Input Device Enabled:No Conn:N/A Enabled:Yes Type:N/A Conn:S5-J3-1,16 Conn: S1-J3 Type:SSR-8 Type:RTD Scaling:100 Ω RTD Filter Gain:1.00 Filter Band:0 Meas. Limits:N/A BioFlo 4500 M1254-0050 User’s Guide...

  • Page 85: Ml-6100 Agitation Control System

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The top-entering agitation system incorporates a 1 HP direct drive on a 20- or 30- liter vessel, capable of agitation speeds from 50 to 1000 RPM. The speed is regulated electronically to compensate automatically for normal voltage fluctuations and changes in viscosity.

  • Page 86: Operation

    If the agitation control loop is cascaded from the DO control loop, the DO control loop may provide the setpoint for the Agitation control loop. The factory settings for the agitation control loop are as shown on the following page. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 87: Double Mechanical Seal (Optional)

    If the fermentor has the double mechanical seal option, the stream pressure between the seals must be set prior to operation of the agitation system. The pressure should be set 2-3 PSI greater than the vessel pressure. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 89: Ml-6100 Air Control System

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The Filter elements used must be changed periodically to maintain proper filter operation. Because many operating and environmental variations exist between fermentations, a periodic check of the filter condition is required to determine when replacement is necessary.

  • Page 90: Vessel Pressure

    The controller sends a 4- 20 mA signal that is converted to pressure by a current-to-pressure (I to P) device. Changes to vessel pressure are made at the touchscreen. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 91: Exhaust Gas Treatment

    The exiting air is filtered by a cartridge type 316L filter (supplied with the system). Each filter element is located in its own in-line stainless steel housing. The filter is sterilized during the last phase of vessel sterilization by steam passed through the filter. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 92: Operation Of Sparge System

    INLET FILTER CARTRIDGE • Adjust the airflow rate and the vessel pressure manually or at the touchscreen (if appropriate) in the airflow control loop. Make sure that another loop is not attempting to control air (gas) flow. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 93: Ml-6100 Two-, Three- & Four-Gas Control

    & & NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The optional gas mixing modules are activated when the GAS MIX CONTROLLER is selected on a specific loop. With the...

  • Page 94: Setting Up The Two-, Three- Or Four-Gas Mixer Option

    O Bi-directional DO Control 3-Gas Direct Air, O , and CO Oxygen enrichment w/ CO addition for pH control 4-Gas Center Off Air, N , and Bi-directional DO Control w/ addition for pH control BioFlo 4500 M1254-0050 User’s Guide...

  • Page 95: Oxygen Enrichment

    Input Action Output Type Control Type None Direct 0-5 Volts None Agitation Gas Flow Oxygen Direct Gas Mix SSR4 13.2.2 Set-Up Procedure To set up the existing DO control loop for oxygen enrichment: New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 96 NONE the controller settings and return to the MAIN CONFIGURATION . If it is not, touch the . The select SCREEN OUTPUT DEVICE ICON will appear. Select as the OUTPUT DEVICE SCREEN NONE OUTPUT TYPE BioFlo 4500 M1254-0050 User’s Guide...
  • Page 97 13. Select the Oxygen control loop and press . The CONFIGURE LOOP will appear. CONFIGURATION SCREEN 14. Touch the on the CONTROLLER ICON LOOP CONFIGURATION SCREEN 15. The will appear. CONTROLLER SETTINGS SCREEN New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 98 2 Ch Comp 17. Enter the in seconds (the default value is 10 seconds) by PWM PERIOD pressing on the text box and entering in the time using the keyboard. 18. Press to accept these settings. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 99 21. Press to accept these settings, and return to the LOOP CONFIGURATION SCREEN 22. Verify that for the , and OUTPUT TYPE OUTPUT DEVICE B NONE Press to exit the LOOP CONFIGURATION SCREEN New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 100 They may be inappropriate for some fermentors. The control mode of the oxygen control loop display gauge in the GAUGE will now display SCREEN 2-GAS Following is a diagram of this cascade-driven oxygen enrichment configuration. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 101: Two-Gas

    In two gas mixing, air and oxygen mix for control of Dissolved Oxygen (DO) whenever demand exceeds the oxygen transfer rate obtainable with air alone, as indicated by the DO loop output. The controller maintains a constant and continuous gas flow. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 102: Control Procedure

    DO loop, to decline. 13. Exit the by pressing the MAIN CONFIGURATION SCREEN CLOSE . The control mode of the DO control loop display gauge in BUTTON will now display GAUGE SCREEN 2-GAS BioFlo 4500 M1254-0050 User’s Guide...

  • Page 103: Three-Gas

    (the group of four SSRs that the gas solenoid valves CONNECTOR ID will be connected to). 10. Press to accept these settings, and return to the LOOP CONFIGURATION SCREEN 11. Press to exit the LOOP CONFIGURATION SCREEN New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 104: Four-Gas

    PWM PERIOD the connected SSRs. 7. Press to accept these settings, and to return to the LOOP CONFIGURATION SCREEN BioFlo 4500 M1254-0050 User’s Guide...

  • Page 105: Viewing Gas Mix Status

    GAS MIX STATUS WINDOW • The current composition of the gas mixture is displayed in the percentage column. To override the gas mix controller, press the appropriate override button to force the selected gas to 100%. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 106 The gas mix cycle time is used to ensure proper operation of the gas mix impeller. The exact settings are process dependent and must be empirically established for each process. • To exit this window, press the CLOSE BUTTON BioFlo 4500 M1254-0050 User’s Guide...

  • Page 107: Ml-6100 Pump Control

    The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The BioFlo 4500 is designed to accommodate up to four optional peristaltic pumps for nutrient addition, pH control, foam control, or other processes. Three of these pumps are mounted on the piping front panel of the fermentor console.

  • Page 108: Pump Capacity

    26 in. Hg 26 in. Hg (660 mm Hg) (660 mm Hg) (660 mm Hg) Vacuum Suction Lift 29 ft. H 29 ft. H 29 ft. H (8.8 m H (8.8 m H (8.8 m H BioFlo 4500 M1254-0050 User’s Guide...

  • Page 109: Loading Tubing Into Pump Head

    TUBING RETAINERS the drawing on the following page, follow these steps to properly load tubing into PUMP HEAD • To open the , flip the black to the left. PUMP HEAD PUMP LEVER New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 110: Pump Calibration

    , press the black PUMP HEAD TUBING RETAINER inward and down until the pinches tubing. RETAINER 14.4 Pump Calibration NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 111 If the pump or actuator used has been calibrated, and its flow or transfer rate is known: • In the function window, select the . The RATE FUNCTION SET TOTAL will change to a BUTTON SET RATE BUTTON New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 112 • In the function window, select the . The RUN FUNCTION SET TOTAL BUTTON changes to a button. • Enter the amount of time, in seconds, you want the pump or actuator to run. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 113 . The controller will automatically calculate the rate. ENTER • Press the to exit this calibration screen. CLOSE BUTTON New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 115: Ml-6100 Foam Control

    A steam-sterilizable port or a septum port can be provided in the upper wall of the vessel for antifoam addition.) The BioFlo 4500 is preconfigured to include a base and antifoam control loop. The pump 3 loop is configured for the antifoam agent.

  • Page 116: Setup Of Foam Probe

    Teflon sleeve on the probe thus activating the system.) In order for the foam probe to function, the must FOAM PROBE GROUND CABLE be connected to the ground connection on the console and its clip must be clipped to the vessel. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 117: High Foam Safety System

    The same precautions for adjusting and care of the regular foam probe apply in this case. The ML-6100 may be configured to include a high foam control loop. The typical settings for this loop are as follows: New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 118 It is recommended that the high foam control loop be configured to trigger an alarm (both audible and visual) when the current value of control loop reaches the parameters below. The parameters for this alarm can be set on the ALARM SETUP SCREEN BioFlo 4500 M1254-0050 User’s Guide...
  • Page 119 To do so, press the boxes near the name of each loop to be shut down. A check will appear in the box to indicate that the loop was selected. For detailed instructions on the operations of alarms, refer to the Alarms section of this manual. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 120: Pump 3 - Antifoam Setup

    Input Device Input Device Enabled:No Enabled:No Conn: N/A Conn: N/A Enabled:No Enabled:No Type: N/A Type: N/A Conn: N/A Conn: N/A Conn: None Type: N/A Type: N/A Type: N/A Scaling:Linear Filter Gain:1 Filter Band:0 Meas. Limits:L BioFlo 4500 M1254-0050 User’s Guide...

  • Page 121: Conductance Probe Calibration

    • From the , press the . The GAUGE SCREEN CAL BUTTON CALIBRATION will appear. SELECTION WINDOW • Select the control loop you wish to calibrate by pressing it. A CALIBRATION will appear. SCREEN New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 122: Conductance Probe Grounding

    Conductance Probe Grounding In order for any of the conductance probes to function, the ground cable must be connected to the ground connection on the console and its clip must be clipped to the vessel. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 123: Ml-6100 Ph Control

    The following paragraphs only refer to the use of the BioFlo 4500 equipped with the ML-6100 controller. During the growth cycle, control of the pH may be required. The BioFlo 4500 is pre- configured at the factory to include a pH control loop. The pH control system is optional.

  • Page 124: Calibration Of Ph Probe

    It should be about 1 cm below the filling orifices that are closed with rubber “T” stoppers. To add reference electrolyte, take the filling pipette (P0740-4820) and fill to 1cm below the orifices with Viscolyte B (P0860-0130) electrolyte. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 125: Gel-Filled Probes

    • Connect the pH cable with the probe attached to the coaxial connection on the console. • In the of the touchscreen, press the GAUGE SCREEN CAL. BUTTON will appear. CALIBRATION SELECTION WINDOW New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 126 Use the SPAN Clr, +/- to clear any unwanted information from the text box. Once BUTTON the reading has stabilized. Press the . The value SET SPAN BUTTON entered will then become the span value. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 127: Recalibration After Sterilization

    • Using an external pH meter, check the pH of the sample. • In the of the touchscreen, press the GAUGE SCREEN CAL. BUTTON will appear. CALIBRATION SELECTION WINDOW New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 128: Setting Up A Cascade From Ph To Pumps

    16.2 Setting Up a Cascade from pH to Pumps On the BioFlo 4500, the installed pumps are numbered 1 through 3. A fourth auxiliary pump may also be utilized. This is accomplished by connecting an auxiliary pump with the electrical receptacle provided on the rear panel of the console.
  • Page 129 , enter the minimum setpoint for pH. CASCADE MIN. SP Regardless of output of the cascade-from loop, the setpoint of this cascade to loop will not go below the value you enter here. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 130 PUMP 1 • is cascaded from pH in the same manner as described above by PUMP 2 entering in the data box. PUMP 2 • When data entry is complete, press BioFlo 4500 M1254-0050 User’s Guide...

  • Page 131: Ml-6100 Do Control

    ML-6100 controller. During the growth cycle, control of the dissolved oxygen (DO) level may be required. The BioFlo 4500 may be configured with an optional system to control DO. A typical control scheme for DO is as follows:...

  • Page 132: Calibration Of Do Probe

    A two-point calibration of the DO probe should be performed after each sterilization. Since the DO reading varies with temperature, calibration should be carried out at the growth temperature where DO control is required. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 133: Electronic Method To Calibrate Do Zero Point

    Use the ZERO STD Clr, +/- to clear any unwanted information from the text box. Once BUTTON the desired number is entered, press the number. SET ZERO New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 134: Sparging With Nitrogen To Calibrate Do Zero Point

    100%. NOTE: Monitoring DO during calibration can be done by viewing the DO gauge or by setting up a Trend Graph for DO and viewing the graph. (See section on Creating a Trend Graph). BioFlo 4500 M1254-0050 User’s Guide...

  • Page 135: Ml-6100 Level Control

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. 18.1 Sensor The optional conductivity (or Level) probe is located in the headplate and is adjustable in height. 18.2 Control Level is controlled by providing a signal to an assignable peristaltic pump located on the front panel of the console.
  • Page 136 Meas. Limits: N/A NOTE: When positioning the level probe, ensure that the probe does not contact any vessel parts. The probe should be placed as far as possible from any metal surfaces, otherwise false readings can occur. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 137: Ml-6100 Nutrient Control

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The addition of nutrient via an assignable peristaltic pump can be controlled up to 100%, set on a user-defined basis. The 0 to 100% setpoint adjustment will be directly proportional to the duty pump cycle.
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  • Page 139: Saving The Ml-6100 Controller Configuration

    The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. Once the controller on your BioFlo 4500 has been configured for use with your application, the controller settings (configuration) should be stored to disk for later use.

  • Page 140: Internal Floppy Drive

    5. Insert a diskette into the drive, and follow the procedures to save or to load a configuration file. 6. Remove the diskette when finished. 7. Replace the panel on the console and secure it in place with the two screws. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 141: Saving A Configuration

    . Enter the appropriate file name in DRIVE DIRECTORY the text area using the onscreen keyboard. NOTE If your BioFlo 4500 is equipped with an internal computer, the drive designations are as follows: • is the resident hard drive of the computer DRIVE C •...

  • Page 142: Loading A Controller Configuration

    When a configuration is loaded from disk, it will replace the current displayed configuration. NOTE If your BioFlo 4500 is equipped with an internal computer, the drive designations are as follows: • is the resident hard drive of the computer DRIVE C •...
  • Page 143 7. Press to load the file. 8. To return to the press in the GAUGE SCREEN, CLOSE MAIN CONFIGURATION SCREEN New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 145: Ml-6100 Cascade Control

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. A cascade is a control scheme in which the output of one control loop influences the setpoint of one or more other loops. In other words, it uses one parameter to influence others.
  • Page 146 5. In the first data entry box in the column, enter the minimum CASCADE MIN SP setpoint for this loop. Regardless of output of the cascade-from loop, the setpoint of this cascade-to loop will not go below the value you enter here. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 147: Controlling Dissolved Oxygen (Do) Using A Cascade

    DO will be maintained at its setpoint by automatic adjustments to the setpoints of agitation and airflow. Agitation in this example is limited to between 200 and 600 RPM. Airflow is limited to between 3 and 8 liters per minute. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 148 The following graph shows the influence of the DO controller output percentage on the agitation and airflow setpoints using the above-mentioned values: Agit and Airflow Setpoint vs. % Output of DISS O2 Controller Agit Airflow % Output of DISS O2 Controller BioFlo 4500 M1254-0050 User’s Guide...

  • Page 149: Ml-6100 Trend Button

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. allows you to display a graph of up to four hours of recent TREND BUTTON data. Up to four plots can be placed on the graph. The graph and data are only available when the fermentor is on.
  • Page 150 The default limits are 0.0 for Low and 100.0 for High. 5. After the limits have been set, press to display the graph or cancel to exit. 6. Press to continue, and the selected loop will appear on the trend graph. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 151 4 hours. Press the to scroll through increasing INC. SPAN BUTTON time spans, and press the to decrease the span. The DEC. SPAN BUTTON current time span will be displayed beneath the graph. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 153: Ml-6100 Gauge Button

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. The ML-6100 Controller has the capacity to control up to 16 control loops. In the , the operator can only view eight (8) loop...
  • Page 154 BioFlo 4500 M1254-0050 User’s Guide...

  • Page 155: Ml-6100 Alarms Button

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. 24.1 Setting Alarms To set alarms: 1. From the , press the . The GAUGE SCREEN ALARM BUTTON ALARMS SCREEN will appear.

  • Page 156: Alarm Mode

    The default value of the . To change the ALARM MODE ABSOLUTE , press the selection of choice. ALARM MODE BioFlo 4500 M1254-0050 User’s Guide...

  • Page 157: Visual & Audible Alarms

    To enable the shutdown feature, press the . The SHUTDOWN BUTTON ALARM will appear (see sample screen below). SHUTDOWN SELECTIONS SCREEN New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 158: Responding To Alarms

    When an alarm is triggered, a thick red flashing line appears around the affected and, if was selected in the alarm set-up, a repeating DISPLAY GAUGE AUDIBLE beep is sounded from the console. TEMP loop in Alarm condition BioFlo 4500 M1254-0050 User’s Guide...

  • Page 159: Acknowledging The Alarm

    ACKNOWLEDGE Press to return to the . The red outline will remain in GAUGE SCREEN continuous (non-flashing) mode around the control loop DISPLAY GAUGE until the condition that triggered the alarm is resolved. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 161: Ml-6100 Deadband Feature

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. To apply a deadband to a control loop: 1. Touch the from the CONFIG BUTTON NBS LOCAL CONTROLLER INTERFACE . The BioCommand...
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  • Page 163: Ml-6100 Use With Biocommand

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. BioCommand is New Brunswick Scientific’s process control supervisory software system. It enables supervisory control of up to eight controllers from a single computer.
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  • Page 165: Preparing For Ml-6100-Controlled Fermentation

    ML-6100 controller. After becoming familiar with the components and operation of each of the systems, subsystems, options and ML-6100 controller that are included on your BioFlo 4500, you are ready to set up the fermentor/bioreactor for a run.

  • Page 166: Adding Media To The Vessel

    During operation, the lamp housing gets hot. Allow it to cool thoroughly before attempting to open the powder or liquid charging (and light) port. To add media through the powder or liquid charging/light port, remove the light by opening the Tri-clamp connector. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 167: Sterilization Of Vessel Using Heat-Labile Media

    (Refer to the Section 28, Automatic Sterilization.) New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 169: Ml-6100-Controlled Automatic Sterilization

    NOTE: The following procedures refer only to the use of the BioFlo 4500 equipped with the ML-6100 controller. Automatic Sterilization is activated at the touchscreen interface. The controller is preconfigured for this feature. 28.1 Sterilization (with Liquid In Vessel) NOTE: Recommended sterilization parameters are 30 minutes at 121.5°...
  • Page 170 NBS AUTOMATIC STERILIZATION CONTROL DIALOG appear. 13. Select the Temperature control loop by pressing on its name. The will appear. The selected AUTOMATIC STERILIZE PARAMETERS WINDOW control loop name will appear at the top of the window. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 171: Resetting Sterilization Parameters

    °C. The valve must be less than or equal to (≤) the STER TEMP • DRAIN TIME Drain Time, range: 0-500 minutes. At the factory it is set to 0. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 172: Valve Sequencing

    Enter the password on the CONFIGURATION PASSWORD keyboard and press . If the is set to SYSTEM CONFIGURATION PASSWORD blanks, the prompt will not appear. The software will then return to the AUTOMATIC STERILIZE PARAMETERS WINDOW BioFlo 4500 M1254-0050 User’s Guide...
  • Page 173 Cooling with temperature more 10°C above normal operation temperature setpoint. • COOL B Cooling with temperature within 10°C above normal operation temperature setpoint. • GROWTH Cooling with temperature within than 1°C above normal operation temperature setpoint. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 174 The factory settings for the sterilization valve sequence with media in the vessel are the following: The factory settings for the sterilization valve sequence with the vessel empty are the following: NOTE * only first valve shown on screen BioFlo 4500 M1254-0050 User’s Guide...

  • Page 175: Shutdown During Sterilization

    Shutdown of control loops is optional, depending on whether or not they are connected to the vessel. If liquid is in the vessel, the agitation loop should not be shut down. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 176: Enabling Sterilization

    The sterilization process may be stopped or aborted at any time after the process has been started. To abort the sterilization process: 1. From the , press the . The GAUGE SCREEN STERIL. BUTTON NBS AUTOMATIC will appear: STERILIZATION CONTROL DIALOG BioFlo 4500 M1254-0050 User’s Guide...
  • Page 177 GAUGE SCREEN 4. The word will be displayed in the control mode of the control loop COOL display gauge until the current temperature of the loop is within its setpoint limits. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 178: Sterilization (Vessel Empty)

    The sterilization valve sequence must be changed to reflect the valve sequence diagram with the vessel empty. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 179: Inoculation & Addition Valve (Optional)

    19 mm port. STEAM INLET VALVE 3. Allow steam to pass through the valve for about 15 minutes. 4. Close the STEAM INLET VALVE 5. Allow the valve to cool thoroughly before use. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 180: Addition & Inoculation

    5. Add the liquid inoculum by gravity feed or via a peristaltic pump. 6. After inoculation, close the inoculation valve: press the black polymer handles in and turn counterclockwise, then release. 7. Break the addition vessel connection by removing the connecting tubing from the quick-connect fitting. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 181: Method B

    10. Add the liquid inoculum by gravity feed or by a peristaltic pump. 11. After inoculation, close the inoculation valve: press the handles in and turn counterclockwise, then release. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 182 12. Close the valve on the quick-connect. 13. Detach the sterile quick-connect from the inoculation/addition valve. 14. Replace the stainless steel cap. 15. Re-sterilize the port. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 183: Sampling Valves (Optional)

    Information section of this manual or contact your local NBS sales representative or parts distributor for details. Figure 38: Sampling Valve Detail STEAM INLET VALVE SAMPLING VALVE CONTROL RING PURE STEAM CONTAMINATED CONDENSATE DRAIN New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 184: Sterilization

    6. Remove the sampler vessel connecting tubing from the quick-connect fitting. 7. Replace the stainless steel cap, and sterilize the valve. 30.2.2 Method B 1. Prepare an appropriate presterilized sample vessel fitted with tubing and a quick-connect. BioFlo 4500 M1254-0050 User’s Guide...
  • Page 185 7. To close the valve, turn the counterclockwise, close CONTROL RING the valve on the quick-connect, and detach the quick-connect from the sampling valve. 8. Replace the stainless steel cap, and sterilize the valve. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 187: Septum Ports (Optional)

    NOTE Several other types of Septum Ports are available for use on the BioFlo 4500. Contact your local NBS representative for the information on these options. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 188: Syringe Port Assembly

    The syringe port assembly kit allows sterile access to the culture vessel. This kit requires a 19 mm headplate port equipped with a septum port kit. The syringe port assembly is available with 3 mm bore (PN M1214-5011) or 6 mm bore syringe (PN M1214-5013) BioFlo 4500 M1254-0050 User’s Guide...

  • Page 189: Luer Lok Syringe Adapter

    1. Remove the from the on the headplate port. PLUG SEPTUM COVER 2. Remove the from the syringe assembly. PROTECTIVE SLEEVE 3. Place the syringe assembly into the port and puncture the septum membrane. New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 190 6. To remove the assembly, unscrew the from the SYRINGE GUIDE SEPTUM and remove. COVER 7. Replace the on the PLUG SEPTUM COVER Figure 42: Syringe Assembly Connection SYRINGE GUIDE HEADPLATE SEPTUM COVER SEPTUM MEMBRANE SYRINGE NEEDLE BioFlo 4500 M1254-0050 User’s Guide...

  • Page 191: Optional Hose Barb Ports

    31.3 Optional Hose Barb Ports There are a variety of hose barb port options that can be used on the BioFlo 4500 headplate. Single, double, and triple ports are available. These ports are steam- sterilized with the vessel and are not re-sterilizable.

  • Page 192: Hose Barb Addition Port Options

    Addition Port Assembly, 1/16" (1.5mm) M1254-5042 Addition Port Assembly, 1/8" (3.17mm) M1254-5043 Addition Port Assembly, 1/4" (6.35mm) M1254-5044 Double Port Assembly, 1/16" (1.5mm) M1254-5045 Double Port Assembly, 1/8" (3.17mm) M1254-5046 Triple Port Assembly, 1/16" (6.35mm) BioFlo 4500 M1254-0050 User’s Guide...

  • Page 193: Shutdown Procedure

    • Connect an appropriate hose to the quick-connect fitting on the port, to transfer vessel contents. 9. If the fermentor is to remain off for an extensive period of time, close all service supply valves. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 195: Post-Run Vessel Cleaning Procedure

    7. Fill the vessel with tap water again, up to or near the headplate, and leave the Agitation on at 200 rpm for an hour or overnight. 8. Drain the water before the next run. New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 197: Preventive Maintenance

    34.3 Three-Month Intervals Perform the following checks and inspections every three months: New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 198: One-Year Intervals

    Note any vessel pressure decrease. If there is more than a 2 PSI (0.13 bar) drop within one hour, check for leakage. 34.4 One-Year Intervals Once every year, perform all the three-month checks and inspections plus the following: BioFlo 4500 M1254-0050 User’s Guide...

  • Page 199: Air Filter, Inlet & Exhaust

    CAUTION! Ensure that the Controller configuration is saved to disk before attempting to replace the batteries inside the Controller. New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 200: Battery Replacement

    5. Observing the polarity markings on the , replace the old BATTERY HOLDER batteries with size AA 1.5V alkaline batteries. 6. Reattach the fan harness. 7. Reinstall the back panel with the panel screws and secure. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 201: Motor Removal

    2. Unscrew the four black T-handled bolts that secure the motor. Remove them and set them aside for reuse. 3. Lift the motor directly upward. Figure 45: Motor Removal MOTOR BEARING HOUSING BLOCK MOTOR MOUNTING BOLT SPEED ENCODER (1 of 4) COUPLING HEADPLATE IMPELLER SHAFT New Brunswick Scientific Co., Inc. User’s Guide...

  • Page 202: Motor Installation

    1. Align the coupling on the driveshaft with the coupling on the motor, and ease the motor into place. 2. Reinsert and screw in the four black T-handled bolts to secure the motor in place. 3. Plug in the speed sensor cable and the motor cable. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 203: Troubleshooting

    If any problems occur with your BioFlo 4500, do not attempt to perform any service on the equipment. Unauthorized servicing may void the warranty. Please contact your local NBS Representative. In any correspondence with NBS, please refer to the Model Number and Serial Number of your BioFlo 4500.
  • Page 204 Poor loop control Sensors and probes miscalibrated. Recalibrate probes and sensors. Agitation loop See Below Inaccurate agitation Cable connections may be Check cable connections. If all loop readings loose. connections are firmly seated, replace speed sensor. BioFlo 4500 M1254-0050 User’s Guide...

  • Page 205: Replacement Parts & Accessory Information

    & & When ordering replacement or accessory parts, or requesting service information, please provide the Model Number, and Serial Number of your BioFlo 4500. This information is on the on the side panel of the console. ELECTRICAL SPECIFICATION PLATE 36.1 Descriptions &...
  • Page 206 Replacement Parts, 20 L - Customer M1254-6010 Replacement Parts, 20 L Double Seal- Customer M1254-6012 Replacement Parts, 30 L Double Seal- Customer M1254-6014 Replacement Parts, 30 L - Customer M1254-6016 Top Port Syringe Assembly - 3 mm bore syringe M1214-5012 BioFlo 4500 M1254-0050 User’s Guide...
  • Page 207 External Computer Cable (20 ft.) M1218-8039 Replacement Vessel Lamp Bulb with pushbutton P0300-0209 Vessel Light with pushbutton P0300-0201 Metaclamp 2” Tri-Clamp P0300-0207 Flow Controller P0460-7040 Solid State Relay Board M1218-7008 Motor Coupling Insert P0160-7270 New Brunswick Scientific Co., Inc. User’s Guide...
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  • Page 209: Descriptions & Part Numbers Glossary

    Oxygen Transfer Rate Proportional/Integral/Derivative Pounds per Square Inch PSIG Pounds per Square Inch, Gauge Pulse Width Modulation Revolutions Per Minute Resistance Temperature Device SCADA Supervisory Control And Data Acquisition SLPM Standard Liters Per Minute New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 210 Solid State Relay Uninterruptible Power Source Vacuum Connector O-ring Vessel Volume per Minute W x D x H Width x Depth x Height BioFlo 4500 M1254-0050 User’s Guide...

  • Page 211: Drawings

    38.1 Control Schematics Figure 46: Basic Control Schematics—Detail A SEE DETAIL B New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 212 Figure 47: Basic Control Schematics—Detail B SEE DETAIL A BioFlo 4500 M1254-0050 User’s Guide...
  • Page 213 Figure 48: Power Schematics DETAIL A (Figure 50) DETAIL B (Figure 51) New Brunswick Scientific Co., Inc. User’s Guide...
  • Page 214 Figure 49: Power Schematics, Detail A BioFlo 4500 M1254-0050 User’s Guide...

Table of Contents