3.0 NGC™ Chromatography Systems and ChromLab™ Software Instrument Guide Version 3.
NGC™ Chromatography Systems and ChromLab™ Software Instrument Guide Version 3.
Bio-Rad Technical Support Department The Bio-Rad Technical Support department in the U.S. is open Monday through Friday, 5:00 AM to 5:00 PM, Pacific Time. Worldwide technical support is available on the Web at www.consult.bio-rad.com. Phone: 1-800-424-6723, option 2 Fax: 1-510-741-5802 Email: LSG.TechServ.US@Bio-Rad.com (U.S.) LSG.TechServ.Intl@Bio-Rad.com (International) Web: www.consult.bio-rad.
Table of Contents Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11 The NGC Chromatography Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 Main Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 Site Requirements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14 Power Considerations. . . . . . . . . . . . . . . .
Table of Contents How the Sample Pump Works . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34 Valves . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36 Sample Inject Valve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 Detailed Attributes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 Ports on the Sample Inject Valve. . . .
Table of Contents Multi-Wavelength UV/Vis Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 Conductivity Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69 How They Work . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70 The Backpressure Regulator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71 pH Detector . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of Contents Connecting the Sample Injection Loops . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 95 Inserting Check Valves into the Sample Inject Valve . . . . . . . . . . . . . . . . . . . . . . 96 Preparing the Detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 UV Lamps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 97 Changing the Mixer Barrel. . . . . . . . . . . . . . . . . .
Table of Contents Appendix A Maintaining the Instrument . . . . . . . . . . . . . . . . . . . . 149 Cleaning the Outer Surfaces of the NGC Instrument . . . . . . . . . . . . . . . . . . . . . 150 Cleaning the NGC Fluidics System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 150 Storing the NGC Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 151 Recommended Maintenance Schedule . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of Contents Other Components . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220 Attaching an Expansion Tier to the NGC Instrument . . . . . . . . . . . . . . . . . . . 220 Replacing or Repositioning Modules on the NGC Instruments . . . . . . . . . . . 227 Converting Bays to Fit Modules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231 Cutting Replacement Tubing. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
Table of Contents Appendix D Connecting the C-96 Autosampler to NGC Systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 289 Using the C-96 Autosampler with NGC Systems . . . . . . . . . . . . . . . . . . . . . . . . 289 Connecting the C-96 Autosampler to an NGC System. . . . . . . . . . . . . . . . . . 289 Plumbing and Priming the C-96 Autosampler. . . . . . . . . . . . . . . . . . . . . . . . . 292 Setting Up the C-96 Autosampler for Use in ChromLab Methods . . . .
Table of Contents x | NGC Chromatography Systems and ChromLab Software
1 Introduction The NGC™ chromatography systems are preparative systems designed to rapidly automate the purification of biomolecules. The flexible, modular, and economical design makes NGC the instrument of choice for method development and scale-up. It is available in six preplumbed, factory-tested configurations at two different flow ranges. Each preconfigured system can be further customized and upgraded by adding valves, detectors, or pumps in order to meet specific application needs.
1 | Introduction The NGC Chromatography Systems The NGC chromatography systems are available in a series of combinations. Each system is available with either two 10 ml system pumps (the 10 series) or two 100 ml system pumps (the 100 series). Each system includes ChromLab™ software and the NGC touch screen.
Main Features NGC Discover Pro chromatography system includes All modules on the NGC Discover system Note: Only the multi-wavelength UV/Vis detector is available on the NGC Discover systems.
1 | Introduction Extend the preconfigured systems with additional valves for buffers, samples, and columns Organize the location of the modules to optimize separation performance based on method scale and complexity, and to minimize the system swept volume Minimize errors when connecting tubing using the Point-to-Plumb™ feature in ChromLab software Site Requirements Power Considerations Note: The power supply to the NGC system must be stable and within specifications at all times to ensure opti
Site Requirements Environment The NGC instruments are designed to work within temperatures of 4–40°C. The small footprint makes the NGC instruments ideal for placement into standard cold cabinets. The touch screen can be detached and placed outside the cold cabinet to control the instrument without opening the cabinet door, preventing temperature fluctuations. For specific temperature, humidity, and altitude ranges, see the NGC Chromatography Systems and ChromLab Software Installation Guide.
1 | Introduction Safety Requirements This section explains the different safety alerts in this document. This section also provides information about the safe use and setup of the instrument. Safety Alerts This document contains WARNINGS and Cautions pertaining to the installation and use of the NGC instrument. Alert Icon Definition Flammable/ Extreme Heat A hazardous situation that could result in serious personal injury or extreme damage to the instrument.
Finding Out More Finding Out More After you install NGC documentation from the NGC Chromatography Systems Documentation DVD, you can access installed NGC guides and tutorials on the Help menu in any ChromLab view. More information about the NGC chromatography systems and ChromLab software is available from the following sources. The NGC Chromatography Systems and ChromLab Software Installation Guide is available on your NGC Chromatography Systems Documentation DVD as a .pdf file.
1 | 18 Introduction | NGC Chromatography Systems and ChromLab Software
2 The NGC Instrument The NGC™ instrument ships preassembled with the components necessary to perform gradient separations. The modular components slide into slots in the system known as bays. Some modules fit into single-wide bays while others require double-wide bays. Bays can be converted from one size to the other by adding or removing a center divider. Each module has a unique electronic ID that enables the system to recognize its function when the module is placed into a bay.
2 | The NGC Instrument The NGC Instrument Illustrated The illustrations in this section display the main components of the NGC Discover™ Pro chromatography system: Front View Back View Right Side View Left Side View Note: Your system might not include all the modules shown in these illustrations. For a list of modules on the NGC Quest™, NGC Scout™, and NGC Discover chromatography systems, see Table 9 on page 86.
The NGC Instrument Illustrated This page is intentionally blank.
2 | The NGC Instrument Front View 22 | NGC Chromatography Systems and ChromLab Software
The NGC Instrument Illustrated LEGEND 1 Touch screen monitor 2 Sample pump 3 Sample inlet valve 4 Buffer tray 5 Empty bay, covered 6 LED display 7 Multi-wavelength UV/Vis detector 8 Conductivity monitor 9 Outlet valve 10 pH detector 11 Column and peripheral mount 12 Columns 13 Column grip 14 Column-switching valve 15 Buffer inlet B 16 Peripheral ports 17 USB ports and soft power switch 18 System pump B 19 Drip tray 20 Mixer 21 Buffer blending valve 22 System pump
2 | The NGC Instrument Back View LEGEND 24 1 Cable connector port/hand grip 2 Cable connector port 3 Touch screen arm 4 Touch screen connector cable 5 Cable connector port/hand grip 6 Instrument’s serial number label 7 Touch screen connector port 8 Ethernet connector port 9 Power outlet ports 10 Power inlet port 11 Fuse cover 12 Power on/off switch | NGC Chromatography Systems and ChromLab Software
The NGC Instrument Illustrated Right Side View LEGEND 1 Drain hole 2 Peripheral port 3 Peripheral ports 4 USB ports 5 Soft power switch 6 Drip tray 7 Touch screen cable cover 8 Air sensor module 9 Column and peripheral mount Instrument Guide | 25
2 | The NGC Instrument Left Side View LEGEND 26 1 Buffer tray 2 Drain hole 3 Touch screen cable cover 4 USB ports 5 Peripheral port 6 Peripheral ports 7 Vents | NGC Chromatography Systems and ChromLab Software
The NGC Instrument Illustrated Liquid Flow Path of the NGC Discover Pro System At startup, ChromLab displays a default fluidic scheme in the System Control tab (as well as in the Method Editor tab and on the touch screen). The fluidic scheme depicts the plumbing flow for the modules. The image that follows displays the plumbing flow of the modules in the NGC Discover Pro system during a sample run. The modules in the fluidic scheme are described in detail in the sections that follow.
2 | The NGC Instrument Pumps The NGC systems can have up to three high-precision pumps: two system (or gradient) pumps (pump A and pump B) and one sample pump. The system pumps can create isocratic or linear gradients at a range of precisely controlled flow rates and pressures. The sample pump can load large sample volumes onto a column or fill large sample loops. Flow rates on all pumps can be controlled to avoid overpressure.
Pumps System Pumps The system pumps perform isocratic or gradient elution at the specified flow rates.
2 | The NGC Instrument Detailed Attributes The NGC systems include two system pumps: pump A (the left pump) and pump B (the right pump). The system pumps are available in two flow rates: 10 ml/min (the F10 pumps) and 100 ml/min (the F100 pumps). F10 Pump Flow rate delivery: 0.001–10 ml/min specified Operating pressure range: 0–3,650 psi (0–25.2 MPa) Operating viscosity range: 0.35–10 cP F100 Pump Flow rate delivery: 0.
Pumps Each system pump module has an emergency Pause button and a Purge button. Pressing the emergency Pause button on either system pump stops both pumps. If a run is in progress when you press Pause on either system pump, both system pumps stop and the run pauses. You can resume or cancel the run through ChromLab. If you choose to cancel the run, you can save the run data up to that point. Pressing Purge on a system pump runs that pump at full speed, replacing any air in the lines with buffer.
2 | The NGC Instrument Piston Wash System The pumps on the NGC system automatically rinse behind the piston with a constant low flow of 20% ethanol. This prolongs the working lifetime of the seal by keeping the seal wetted and preventing salt crystals from depositing onto the pistons. When the pumps are running, the pistons and the check valve located in each chamber automatically pump the rinse fluid, which circulates through the system.
Pumps Sample Pump The sample pump is used to automatically load large sample volumes onto a column or to automatically fill large sample loops. When sample inlet valve modules are installed along with the sample pump, the sample pump performs as an autosampler and enables the system to load multiple samples automatically. The sample pump includes an integrated pressure sensor that protects the column and media from overpressure.
2 | The NGC Instrument Detailed Attributes Flow rate delivery: 0.01–100 ml/min Pressure range: 0–1,450 psi (0–10 MPa) Operating viscosity range: 0.35–10 cP Sample Pump LEDs The sample pump module is fitted with LEDs. Blue LEDs indicates the pump is in use. How the Sample Pump Works The reciprocating pistons in the sample pump synchronize to deliver continuous flow with low pulsation. The sample pump module also has an emergency Pause button and a Purge button.
Pumps Tip: You can pause the system through ChromLab using either the touch screen or the ChromLab computer. You can also set timed purging of the sample pump through ChromLab, which will set the inject valve to Waste for a set amount of time. For more information, see the NGC Chromatography Systems and ChromLab Software User Guide.
2 | The NGC Instrument Valves All NGC systems include a sample inject valve. Your system might also include one or more of the following valves: Buffer blending valve Buffer inlet valve Sample inlet valve Outlet valve Column switching valve All valves (except the buffer blending valve) are motorized rotary valves with a number of defined inlet and outlet ports. As the rotary valve turns, the flow path for the valve changes. The active ports are identified by LEDs.
Valves Sample Inject Valve The sample inject valve enables the system to load a specific, predetermined volume of sample onto a column. When the sample pump is installed, the sample inject valve enables the system to easily switch between manual loop filling, automated loop filling, and direct sample injection onto a column without replumbing the fluidic connections. You can use the sample pump to load sample either directly onto a column or into a sample loop.
2 | The NGC Instrument Ports on the Sample Inject Valve Port Function Inject Manually load sample using a syringe Column Outlet to top of column Pump Inlet from the system pump Loop E Inject: Inlet for buffer Load: Outlet to waste Waste 1 Outlet to waste (from the sample pump and sample loop) Sample pump Inlet from the sample pump Loop F Inject: Outlet to column Load: Inlet for the sample Waste 2 38 Outlet to waste (from the system pump) | NGC Chromatography Systems and C
Valves Sample Inject Valve LEDs The sample inject valve module is fitted with LEDs. Solid green LEDs indicate the flow is from the system pump. Solid blue LEDs indicate the flow is from the sample pump. Blinking green LEDs indicate the line to plumb using the Point-to-Plumb™ feature. How the Sample Inject Valve Works The sample inject valve enables sample to be loaded onto the column.
2 | The NGC Instrument Valve Position Explanation Manual Load Loop/System Pump to Column Directs system pump flow directly toward the column Aligns the sample loop with manual injection port so that sample can be manually filled into the loop using a syringe.
Valves Valve Position Explanation Sample Pump Direct Inject/System Pump Waste Directs system pump flow to waste (W2) Directs sample pump flow directly onto the column to load large volumes of sample Chromatogram x-axis based on sample pump flow rate Directs system pump flow directly onto the column Directs sample pump flow into the loop to load the loop with sample.
2 | The NGC Instrument Buffer Blending Valve The buffer blending valve is used for automatic online buffer preparation. The buffer blending valve blends buffers to a specific pH using standard acid and base buffer solutions. This enables automated pH scouting without manually preparing buffers at various pH values. The buffer blending valve titrates the appropriate amounts of the acid and base stock solutions to produce buffers at the required pH values.
Valves Detailed Attributes Operating pressure up to 15 psi Instrument Guide | 43
2 | The NGC Instrument Ports on the Buffer Blending Valve Port Function Q1–Q4 Buffer inlet ports Outlet A Outlet to system pump A or to buffer inlet valve A Outlet B Outlet to system pump B or to buffer inlet valve B Buffer Blending Valve LEDs The buffer blending valve module is fitted with LEDs. 44 A single green LED indicates the active inlet port is open. Multiple green LEDs indicate the buffer blending valve is cycling between the inlet ports.
Valves How the Buffer Blending Valve Works The buffer blending valve operates in two modes: buffer blending and gradient. Buffer Blending Mode Acid, base, water, and salt are blended together in the required proportions to form a buffer of specified pH over a linear salt gradient, or a pH gradient at a specified salt concentration.
2 | The NGC Instrument Inlet Valves The inlet valves automate the use of multiple buffers and samples during method development. In ChromLab, you can program the inlet valves to Switch between buffers during a method run Automate sample selection and loading by way of the sample pump Switch between cleaning solutions and buffers during system wash and preparation The inlet valve has eight defined inlet ports and one outlet port.
Valves Inlet Valve Detailed Attributes Maximum operating pressure: 500 psi Optional air sensors indicate the end of buffer or sample injection See Air Sensors on page 80 for specific information about the air sensors. Tip: In ChromLab, you can customize the names of ports on each inlet valve. For example, you might change the name to reflect the sample or buffer that is associated with that position. For more information, see the NGC Chromatography System and ChromLab Software User Guide.
2 | The NGC Instrument Ports on the Inlet Valves Buffer Inlet Valve Port Function 1–7 Buffer inlets 8/BB 8: Inlet for systems without the buffer blending valve BB: Reserved inlet for systems with the buffer blending valve Out Outlet port Sample Inlet Valve One sample inlet valve in fluidic scheme 48 Port Function 1–8/BB Sample inlets Out Outlet port to sample pump | NGC Chromatography Systems and ChromLab Software
Valves Two sample inlet valves (S1 and S2) in fluidic scheme Valve Port Function S1 and 1–7 Sample inlets 8/BB Inlet from sample inlet valve 2 S2 S1 S2 Sample inlet S1 Out S2 Outlet to sample pump Outlet to S1 Inlet Valve LEDs The inlet valve modules are fitted with LEDs. Solid green LEDs indicate the flow is from the system pump. Solid blue LEDs indicate the flow is from the sample pump. Blinking green LEDs indicate the line to plumb using the Point-to-Plumb feature.
2 | The NGC Instrument How the Inlet Valves Work The function of an inlet valve is determined by the type of pump to which it is connected to. Inlet valves connected to the system pumps are used for buffers and are referred to as buffer inlet valves. Inlet valves connected to the sample pump are used for samples and are referred to as sample inlet valves. Buffer Inlet Valve The NGC systems support the use of up to two buffer inlet valves.
Valves Outlet Valve The outlet valve enables the system to direct the flow to the BioFrac™ fraction collector, to waste, or to another user-defined location. The outlet valve has 12 defined outlet ports and one inlet port. The active ports are identified by LEDs. Green LEDs indicate that the flow through the port is from the system pumps. Blue LEDs indicate that the flow is from the sample pump. The outlet valve enables the system to collect large-volume fractions.
2 | The NGC Instrument Detailed Attributes Maximum operating pressure is 500 psi Ports on the Outlet Valve One outlet valve in fluidic scheme 52 Port Function 1 Outlet to the diverter valve on the fraction collector (if included in the fluidic scheme) Outlet to waste (if the fluidic scheme does not include a fraction collector) 2–12 Outlet ports In Inlet port from the conductivity monitor or pH monitor | NGC Chromatography Systems and ChromLab Software
Valves Two outlet valves (O1 and O2) in fluidic scheme Valve Port Function O1 1 Outlet to the diverter valve on the fraction collector (if included in the fluidic scheme) Outlet to waste (if the fluidic scheme does not include a fraction collector) O1 2–11 Fraction outlets 12 Outlet to O2 inlet port O1 In Inlet port from the conductivity monitor or pH monitor O2 1–12 Fraction outlets O2 In Inlet from O1 Outlet Valve LEDs The outlet valve modules are fitted with LEDs.
2 | The NGC Instrument How the Outlet Valve Works The NGC system supports the use of up to two daisy-chained outlet valves. This configuration is ideal for collecting large-volume fractions. The inlet port on the first outlet valve (O1) connects to the outlet port of either the conductivity monitor or the pH detector. Port 1 on O1 connects to the diverter valve on the fraction collector or directs the flow to waste.
Valves Column Switching Valve The NGC system supports up to three column switching valves. Each column switching valve is used to connect up to five columns simultaneously, supporting up to 15 columns. This enables automated column scouting, and the use of different columns without connecting and disconnecting the columns. The valve includes an internal bypass mode, which enables buffers to bypass the connected columns when necessary (for example, when priming or cleaning the system).
2 | The NGC Instrument Detailed Attributes 56 Maximum operating pressure: 3,650 psi Dual pressure sensors for precolumn and delta pressure detection Internal bypass capability Reverse flow capability | NGC Chromatography Systems and ChromLab Software
Valves Ports on the Column Switching Valve Port Function Top 1–5 Ports to connect to the top of columns Bottom 1–5 Ports to connect to the bottom of columns In Inlet port from the system pump through precolumn pressure sensor Out Outlet port through the postcolumn pressure sensor to the UV detector Instrument Guide | 57
2 | The NGC Instrument Column Switching Valve LEDs The column switching valve modules are fitted with LEDs. Solid green LEDs indicate the flow is from the system pump in the forward direction. Blinking green LEDs indicate the flow is from the system pump in the reverse direction. Solid blue LEDs indicate the flow is from the sample pump in the forward direction. Blinking blue LEDs indicate the flow is from the sample pump in the reverse direction.
Valves The section Flow Paths of the Column Switching Valve explains the specific flow paths. The column switching valve has integrated precolumn and postcolumn pressure monitors. The precolumn pressure monitor measures the pressure that the system exerts on the column (precolumn pressure). The postcolumn pressure monitor measures the pressure at the column outlet and provides the delta pressure across the column.
2 | 60 The NGC Instrument Select Bypass to bypass the flow past all ports. Select Reverse Flow Direction to reverse the flow so that it flows from the bottom to the top of the column.
Mixer Mixer The mixer module consists of a mixer assembly and an integrated system pressure sensor. This module homogenizes the buffers proportioned by the two system pumps and the buffer blending valve (if present). The pressure sensor, located in this module, monitors the NGC system pressure. The mixer module is located between the two system pumps (A and B). It is plumbed so that flow from the two system pumps 1. Enters at the mixer assembly base 2. Exits the mixer assembly top 3.
2 | The NGC Instrument During operation, the system pressure sensor continuously measures and records the pressure. If the system pressure exceeds user- or system-specified pressure limits, the system pumps shut down. Alternatively, when you select the option in ChromLab to control the flow to avoid overpressure, the NGC system reduces the flow rate until the system is within safe operation pressure limits. See F10 Pump on page 30 and F100 Pump on page 30 for specific pressure limitations.
Mixer Detailed Attributes The mixer is composed of a base motor, a stirring plate, and a hollow mixer barrel in which the buffers are mixed. An optional mixer barrel extender is available to adapt the mixer volume to the flow rate used. Bio-Rad offers several different mixer chamber sizes for different flow rates. Table 2 and Table 3 list the recommended barrel sizes for different flow rates for each type of system pump. To change the barrels, see Replacing the Mixer Components on page 186.
2 | The NGC Instrument Mixer LEDs The mixer module is fitted with LEDs. A left green LED indicates the flow is from system pump A. A right green LED indicates the flow is from system pump B. Both LEDs light when both pumps are running. Blinking green LEDs indicate the line to plumb using the Point-to-Plumb feature. Detectors All NGC systems include a detector module that contains a conductivity monitor and either a single-wavelength UV detector or a multi-wavelength UV/Vis detector.
Detectors Single-Wavelength UV Detector and Conductivity Monitor Multi-Wavelength UV Detector and Conductivity Monitor Instrument Guide | 65
2 | The NGC Instrument Single-Wavelength UV Detector LEGEND 1 Conductivity monitor 2 LED display 3 UV flow cell 4 UV detector housing 5 Point-to-Plumb™ indicator Detailed Attributes Two integrated LED UV bulbs LED operating usage: approximately 5,000 hours Note: If the reference output is below 0.9 volts you might need to replace the LED bulbs depending on the requirements of the experiment.
Detectors Monitors UV absorbance one wavelength at a time 255 nm 280 nm (default) Tip: Use ChromLab to change the absorbance wavelength values. For more information, see the NGC Chromatography Systems and ChromLab Software User Guide.
2 | The NGC Instrument Multi-Wavelength UV/Vis Detector LEGEND 1 Conductivity monitor 3 UV/Vis detector housing 2 UV/Vis detector flow cell Detailed Attributes Contains one tungsten and one deuterium lamp Lamp operating usage: approximately 2,000 hours for the tungsten bulb as well as the deuterium bulb For information about changing the UV lamps, see Replacing the MultiWavelength UV/Vis Detector Lamps on page 209.
Detectors Monitors up to four wavelengths in the UV/Vis range of 190–800 nm Tip: Use ChromLab to change the absorbance wavelength values. For more information, see the NGC Chromatography Systems and ChromLab Software User Guide. UV absorbance range: 0–3,000 mAU UV linearity: ±5% Three interchangeable flow cells with path lengths of 2 mm, 5 mm (default), and 10 mm; see Table 4 for specific information For information about changing the flow cells, see Replacing the UV Flow Cell on page 196.
2 | The NGC Instrument How They Work UV Detectors ChromLab recognizes the connected optic module (that is, either the single-wavelength UV or multi-wavelength UV/Vis module). The lamps turn on when the NGC instrument powers up. At the start of the method run, the system determines if the monitor is ready. The warmup time for the LED lamp in the single-wavelength detector is negligible. The tungsten and deuterium lamps in the multi-wavelength detector require 60–90 min to warm up.
Detectors The Backpressure Regulator A backpressure regulator can be added to the flow path to generate backpressure and prevent air bubbles from forming in the UV flow cells when medium pressure columns are used. (The NGC systems ship with a 40 psi backpressure regulator.) Bio-Rad recommends that the backpressure regulator be plumbed between the conductivity monitor and the pH monitor to avoid exceeding the 70 psi limit of the pH probe.
2 | The NGC Instrument pH Detector The pH detector includes an integrated flow cell in which to insert a pH electrode. The pH of the solution flowing across the pH electrode is measured in real time. This is very useful for applications in which knowledge of the buffer pH is critical for a successful separation and/or the stability of the sample (for example, in antibody purifications). The pH can be temperature compensated.
Detectors Detailed Attributes Flow cell volume: 90 μl (200 μl including internal valve passageways) Electrode maximum operating pressure: 70 psi pH reading range: 0–14 Accuracy: 0.1 pH units from 2–12 Slope: 80–120% Offset: ±60 mV Note: Store the electrode in a pH electrode storage buffer or pH 7 standard buffer. For information about changing the pH probe, see Replacing the pH Probe on page 218.
2 | The NGC Instrument Port Description In Inlet from the conductivity monitor Out Outlet to the fraction collector Cal In Calibration inlet reserved for calibrating the pH valve Cal Out Outlet to calibration waste How the pH Detector Works The integrated pH electrode enables inline pH monitoring during a run. ChromLab calculates and displays temperature-compensated pH values.
Detectors Changing the Flow Paths of the pH Valve During a manual run, you can change the flow path of the pH valve by accessing its dialog box in ChromLab. To change the flow path of the pH valve 1. Touch or click the pH Valve module in the fluidic scheme to access its dialog box. 2.
2 | The NGC Instrument Tubing, Loops, Columns, and Fittings This section lists the accessories supported for use with the NGC systems. Tubing Table 5. NGC system tubing details Description Diameter Scope of Use Maximum Pressure Rating Mounted at Delivery PEEK tubing, orange OD: 1/16" High pressure tubing, recommended for flow rates less than 40 ml/min 7,000 psi Yes 7,000 psi Yes ID: 0.
Tubing, Loops, Columns, and Fittings Sample Injection Loops Table 6.
2 | The NGC Instrument Fittings The NGC chromatography systems ship with 10/32 fittings for column attachment and 1/4-28 fittings that accommodate a large variety of commercially available columns. Table 7.
LEDs and LED Screens LEDs and LED Screens The modules on the NGC system are fitted with light emitting diodes (LEDs) that are located next to ports on the valves and on the inlet and outlet tubing connections for pumps and all detectors (including the UV detectors and the air sensors).
2 | The NGC Instrument Sensors Air Sensors The NGC chromatography systems can be fitted with an external air sensor module that can hold up to four air sensors. This module can be daisy chained to an extension module that can hold an additional four air sensors, providing a total of eight air sensors. The air sensors detect air, indicating the end of sample or buffer, and trigger appropriate events through ChromLab software.
Sensors The air sensors are inserted into ports on the air sensor module. ChromLab detects the presence or absence of air sensors in the air sensor module. If you have fewer than four air sensors in the module, you must cover the unoccupied ports with a protective blank cover that Bio-Rad provides to prevent liquid from entering into the module’s electronic circuitry.
2 | The NGC Instrument Air Sensor LEDs The air sensor module is fitted with LEDs. Green LEDs indicate the module is connected. Blue LEDs indicate air in the tubing. Prime the lines to remove air. See Priming and Purging the Systems on page 118 for more information. Air Sensor Tubing Length and Bubble Size The response of the air sensor module is highly dependent on the bubble size and flow rate, and requires a minimum tubing length between the sensor outlet and the pump inlet.
Sensors Pressure Sensors The NGC systems can have up to four integrated pressure sensors: System pressure sensor — located in the mixer module Precolumn and postcolumn pressure sensors — located in the column switching valve These pressure sensors protect the column and media from overpressure. One pressure sensor measures the pressure before the column to protect the column hardware. The other sensor measures the pressure after the column and calculates the pressure difference over the media bed.
2 | The NGC Instrument NGC Touch Screen The NGC system’s touch screen is the main interface between the NGC system computer and the ChromLab computer. From the touch screen’s pull-down menu, you can perform all system control functions such as starting a manual run, starting and monitoring a method run, or calibrating the system. The touch screen displays a chromatogram viewer and the fluidic scheme.
Buffer Tray Buffer Tray The buffer tray on the top of the instrument can accommodate up to six 1 L bottles. Note: Ensure that the drainhole is connected to tubing so that any potential leaks are properly drained.
2 | The NGC Instrument Product Configurations The NGC chromatography system is available in four standard configurations. This section lists the modules available for each configuration. You can add or remove modules to an existing configuration to customize the system. The configuration at your site might differ slightly from any of the standard configurations. Note: All NGC systems are available with either the 10 ml/min or the 100 ml/min system pumps. Table 9.
Product Configurations Table 9. NGC chromatography system configurations, continued Module NGC Quest NGC Scout NGC Discover NGC Discover Pro Fourth expansion tier Sample inlet valve 1 Outlet valve 1 *Only the multi-wavelength UV/Vis detector is available in the NGC Discover series.
2 | 88 The NGC Instrument | NGC Chromatography Systems and ChromLab Software
3 Preparing the Instrument The NGC™ instruments ship preassembled with the modules necessary to perform chromatographic separations. The instruments require minimal postinstallation setup to prepare them to run simple gradient separations. This chapter explains how to prepare the NGC systems for a method run.
3 | Preparing the Instrument Module Review The NGC systems are shipped in one of three standard configurations. Preparing the NGC systems differs slightly depending on the configuration. Tip: For information about adding, removing, or moving modules, see Replacing or Repositioning Modules on the NGC Instruments on page 227. Standard NGC System Configurations The standard NGC system configurations are depicted in the images that follow.
Module Review NGC Discover™ Chromatography System NGC Discover Pro Chromatography System Instrument Guide | 91
3 | Preparing the Instrument NGC Systems Accessories Kit Each NGC system ships with an accessories kit. The contents of the kit are specific to the system. Verify the contents against those listed in the sections that follow. The accessories kits contain the tubing, loops, and fittings necessary to complete the setup of the shipped configuration.
NGC Systems Accessories Kit NGC Scout Accessories Kit The kit for the NGC Scout system includes all the accessories in the NGC systems kit as well as Tube #2 (40 psi backpressure regulator to pH valve inlet) pH waste tubing pH calibration port inlet tubing Colored tubing for the buffer blending valve NGC Discover Accessories Kit The kit for the NGC Discover system includes all the accessories in the NGC Scout kit as well as Pump head wash inlet tubing (2) for the sample pump Pump hea
3 | Preparing the Instrument Preparing the Buffer and Waste Bottles After you plumb the instrument, prepare buffer and waste bottles. Buffer Bottles Place buffer bottles on the bench next to the instrument or in the buffer tray on top of the instrument. The tray can accommodate up to six 1 L bottles. Waste Bottles Place waste bottles on top of the bench or on the floor below the instrument. Ensure that the waste containers are large enough to hold all buffer that will flow into them.
Connecting the Sample Injection Loops Connecting the Sample Injection Loops The NGC chromatography systems support a variety of injection loop types and sizes, including static loops and dynamic loops. Table 6 on page 77 lists the supported loops. Note: You might have already connected a loop if you completed all of the steps in the section Plumbing and Priming the NGC Chromatography Systems in the NGC Chromatography Systems and ChromLab Software Installation Guide.
3 | Preparing the Instrument Inserting Check Valves into the Sample Inject Valve All NGC systems ship with two check valves in ports W1 and W2 on the sample inject valve. In addition, the sample pump ships with two extra check valves. The check valves apply positive backpressure to the waste ports on the sample inject valve and ensure against any leaks. Each check valve has an arrow to indicate the direction of flow.
Preparing the Detectors 4. Connect the tubing to the check valves. Preparing the Detectors The NGC systems ship with the conductivity monitor and the UV detectors installed, calibrated, and ready for use. These modules require no preparation. The pH module (if installed) requires you to install and calibrate the pH probe. See the section Replacing the pH Probe on page 218 for information about installing a new pH probe. UV Lamps The UV detector ships with the UV lamps installed.
3 | Preparing the Instrument Changing the Mixer Barrel The mixer module is used to blend buffers from two to four separate solutions. It is important to use a mixer with a barrel size that is based on the flow rate of separation. Bio-Rad offers several different mixer barrel sizes for different flow rates. The F10 systems ship with the 263 μl mixer base and top assembly. When using a higher flow rate, the mixer volume of this base unit can be expanded by adding a 750 μl or 2 ml barrel.
Changing the Mixer Barrel Table 11. Recommended NGC system mixer barrel sizes — with buffer blending valve Barrel Size Flow Rate, ml/min F10 Pump F100 Pump 0.1–1.99 n/a 2.00–5.99 <5.00 2 ml 6.00–11.99 5.00–11.99 5 ml 12.00–20.00 12.00–39.99 12 ml n/a 40.00–80.00 263 μl (Base and top) 750 μl (Base and top) To insert or change the mixer barrel To insert or change the mixer barrel, see Replacing the Mixer Components on page 186.
3 | Preparing the Instrument Connecting the Air Sensor Module The air sensors enable the ChromLab™ software to detect air bubbles and automatically trigger appropriate events. The air sensor module has a magnetic backing and can attach to the side of the NGC instrument. The module can hold up to four air sensors. Two types of air sensors are available that differ in their internal diameter.
Connecting the Air Sensor Module Air Sensor Tubing Length and Bubble Size The response of the air sensor module is highly dependent on the bubble size and flow rate, and requires a minimum tubing length between the sensor outlet and the pump inlet. Table 12 lists recommended tubing lengths for various flow rates and bubble sizes. Table 12.
3 | Preparing the Instrument Attaching and Activating Air Sensors This section explains how to attach the air sensor module and the air sensors. It also explains how to activate the air sensors after they are attached. Note: Bio-Rad recommends that all buffers be filtered and degassed prior to use. To attach the air sensor module and air sensors 102 | 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC system. 2.
Connecting the Air Sensor Module 5. 6. To connect an air sensor to the system pumps: a. Connect tubing from the buffer bottle or from the outlet port on the buffer inlet valve to the port on top of an air sensor. b. Connect tubing from the port on the bottom of the air sensor to the inlet port on either system pump. To connect air sensors to the buffer blending valve: a. Connect tubing from each buffer bottle to the port on the top of each air sensor that will be used. b.
3 | Preparing the Instrument Connecting External Devices to the NGC Instrument Tip: If you do not plan to connect the C-96 autosampler or other external devices to the NGC instrument you can skip this section. This section explains how to connect the signal import module (SIM) to the NGC instrument, and how to connect external devices to the SIM. For information about connecting the C-96 autosampler to the SIM, see Appendix D, Connecting the C-96 Autosampler to NGC Systems on page 289.
Connecting External Devices to the NGC Instrument Importing Analog Signals to the NGC Instrument You can import analog (voltage) signal from up to two external devices and convert it to digital NGC data via the SIM. The SIM imports and converts the signal to the appropriate trace units through its SIM 1 and SIM 2 ports.
3 | Preparing the Instrument 3. 106 | Insert the other end of the cable into one of the peripheral connector ports on the side of the NGC instrument.
Connecting External Devices to the NGC Instrument Connecting External Devices to the SIM WARNING! These procedures require you to work with exposed wires. Ensure that your systems are powered off and unplugged before performing this task. This section explains how to connect different external devices to the SIM. Before connecting the device, locate the connector ports that you will use on the back of the SIM.
3 | Preparing the Instrument 3. Loosen all three screws on the top of the connector. Tip: These screws adjust the terminals that hold the wires in place. The screws do not dislodge from the connector. 4. Locate the analog output cable that ships with your detector. 5. If connectors are attached to both ends of the cable, remove the connector that does not insert into your detector. Tip: Alternatively, contact the manufacturer for a suitable cable. 108 | 6.
Connecting External Devices to the NGC Instrument 9. On the analog output cable, determine the positive and negative wires. 10. Insert the positive wire into the left terminal on the connector and tighten the screw. Note: Take care not to overtighten or fray the wire.
3 | Preparing the Instrument 11. Insert the negative wire and one end of the jumper wire into the middle terminal on the connector and tighten the screw. 12. Insert the other end of the jumper wire into the right, or ground, terminal on the connector and tighten the screw.
Connecting External Devices to the NGC Instrument 13. Insert the connector into the SIM 1 or SIM 2 port on the back of the SIM. 14. Refer to the chapter System Controls in the NGC Chromatography Systems and ChromLab Software User Guide for information about how to enable signal import from the device.
3 | Preparing the Instrument To connect an external device to the DAC A and DIGITAL OUT ports 1. On the back of the SIM, firmly grasp the DAC A connector and pull it straight out of its port. Note: ChromLab supports solely the DAC A port. When connecting an analog instrument to the SIM, use the DAC port on the right (DAC A). 2. (Optional) If you want the NGC instrument to trigger the device to start measuring data, also remove the DIGITAL OUT connector and perform the steps that follow. 3.
Connecting External Devices to the NGC Instrument 9. Insert the positive and ground wires into the appropriate terminals on the DAC A connector and tighten the screws, taking care not to fray the wires. 10. Insert the trigger wire into terminal 2 on the DIGITAL OUT connector and tighten the screw, taking care not to fray the wire. Trigger wire Ground Positive wire wire 11. Insert the connectors into their ports on the back of the SIM. 12.
3 | Preparing the Instrument Starting the NGC Instrument Caution: The NGC system runs under high pressure. Follow the safety precautions established for the laboratory at all times. To start the NGC instrument Locate and press the soft power switch on the side of the NGC instrument. Tip: Starting the system and the internal software might take some time.
Starting the NGC Instrument Connecting the NGC System to ChromLab Software Note: For information about installing ChromLab, see the NGC Chromatography Systems and ChromLab Software Installation Guide. To connect the NGC system to ChromLab software Do one of the following on the computer running ChromLab (known as the ChromLab computer in this document): Double-click the ChromLab desktop icon. Select ChromLab on the Start menu.
3 | Preparing the Instrument Verifying Plumbing with the Point-to-Plumb Feature The Point-to-Plumb™ feature in ChromLab helps you when making changes to the plumbing or to verify that the instrument is plumbed correctly for the fluidic scheme that you want to use. You can access the Point-to-Plumb feature on the touch screen menu or through the Tools menu on the System Control tab in ChromLab.
Verifying Plumbing with the Point-to-Plumb Feature To verify the plumbing path 1. With both the Point-to-Plumb screen and the front of the instrument in view, touch or click a segment along the gray path. The segment you selected turns green. At the same time, the LEDs on the front of the instrument indicate the selected plumbing path. 2. Verify that the tubing is connected at the ports indicated by the LEDs, to replicate the Point-to-Plumb display. If it is not correct, replumb it. 3.
3 | Preparing the Instrument Priming and Purging the Systems This section explains how to prime your system before running the pumps, and how to purge your system of air bubbles or rapidly exchanging buffer solutions. Note: This section assumes that you primed the pump head wash system when you installed the system. See Plumbing All NGC Systems on page 265 for more information. Priming and purging tasks are specific to the NGC system configuration.
Priming and Purging the Systems Priming and Purging the NGC Quest System Caution: Before priming or purging the system, ensure that any attached columns are offline. The priming process occurs at a high flow rate and can significantly harm or destroy attached columns. Priming and purging the NGC Quest system requires access to ChromLab. You can perform the steps on the ChromLab computer or on the NGC touch screen. Important: The instrument’s buffer lines cannot be primed when the system is under pressure.
3 | Preparing the Instrument 6. Remove the syringe from the priming port and discard the liquid into waste. 7. Repeat steps 2–6 for system pump B. To purge the system pumps 120 | 1. On the ChromLab computer or the touch screen, start ChromLab in Manual mode. 2. Set the fluidic scheme to either NGC Quest or NGC Quest Plus. 3. In the fluidic scheme, touch or click the Sample Inject Valve module to open its dialog box. 4. Set the sample inject valve to System Pump Waste.
Priming and Purging the Systems 5. In the fluidic scheme, touch or click the System Pump module to open its dialog box. 6. In the System Pump dialog box: a. Set Flow Rate to 10 ml/min. b. (Optional) Set Duration to at least one min. c. Click Start.
3 | Preparing the Instrument Priming and Purging the NGC Scout System Priming the NGC Scout system is a manual procedure and involves priming the buffer blending valve starting from the Q4 port to Q1. You can purge one line at a time, or all lines sequentially. These procedures require access to ChromLab. Tip: You can perform the steps on the NGC touch screen. Priming the NGC Scout System Note: This section assumes that you have not introduced buffer into the lines.
Priming and Purging the Systems 5. In the System Pump dialog box, select Priming on the Mode dropdown list. Note: When in Priming mode, the sample inject valve switches the system pump flow to waste (W2). After the process is completed, the sample inject valve switches the system pump to manually load the column through the loop. By default, all ports are closed. The Closed radio button is selected. 6. Select Q4 as the port to open and prime. 7.
3 | Preparing the Instrument Purging the NGC Scout System To purge the buffer blending valve 1. 2. Open the System Pump dialog box and select Priming in the Mode dropdown list. To purge an individual line of air bubbles, proceed to step 2 that follows. To purge all lines of air bubbles sequentially, proceed to step 3 on page 125. To purge an individual line of air bubbles and fill it with buffer: a. Select a port to open. b. (Optional) Change the default flow rate.
Priming and Purging the Systems 3. To purge all lines of air bubbles and fill them with buffer: a. Click All. b. (Optional) Change the default flow rate. The default flow rate is set to 20 ml/min. c. Click Start. The system pumps and the buffer blending valve start operating at the specified flow rate, exchanging the solution in the inlet lines and expelling any trapped bubbles, cycling sequentially through the ports Q1 to Q4.
3 | Preparing the Instrument Priming and Purging the NGC Discover System Priming the NGC Discover system is a manual procedure and involves priming the buffer blending valve starting from the Q4 port to Q1 then the buffer inlet valve starting from buffer inlet A1. Purging the NGC Discover system is an automated procedure and involves removing air from one or all buffer lines as well as the buffer inlet valves and the system and sample pumps. Both procedures require access to ChromLab.
Priming and Purging the Systems 4. In the fluidic scheme, touch or click the System Pump module to open its dialog box. 5. In the System Pump dialog box, select Priming on the Mode dropdown list. Note: When in Priming mode, the sample inject valve switches the system pump flow to waste (W2). After the process is completed, the sample inject valve switches the system pump to manually load the column through the loop. 6. Select Q4 as the port to open and prime.
3 | Preparing the Instrument 7. Insert a 30 ml syringe into the priming port on system pump A. 8. Turn the priming port valve counter-clockwise one full turn to open the seal. 9. Slowly withdraw the plunger to withdraw liquid from the port until air no longer enters the syringe (approximately 10 ml) and all tubing is filled with liquid. 10. Turn the priming port valve clockwise to close the port. 11. Remove the syringe from the priming port and discard the liquid into waste. 12.
Priming and Purging the Systems To prime the buffer inlet valves 1. Immerse the tubing for the inlet valves in their respective buffer containers. 2. On the ChromLab computer or the touch screen, start ChromLab in Manual mode. 3. In the System Pump dialog box, select the first inlet port to prime, for example Buffer A 1. 4. Insert a 30 ml syringe into the priming port on system pump A. 5. Turn the priming port valve counter-clockwise one full turn to open the seal. 6.
3 | Preparing the Instrument 8. Remove the syringe from the priming port and discard the liquid into waste. 9. Repeat steps 4–8 for the remaining buffer inlet ports. 10. To prime system pump B, select buffer B1 and perform steps 3–9 on system pump B. To prime the sample pump 130 | 1. Immerse the inlet tubing for the sample pump in the appropriate container. 2. Insert a 30 ml syringe into the priming port. 3. Turn the priming port valve counter-clockwise one full turn to open the seal. 4.
Priming and Purging the Systems To prime the sample inlet valve (if available) 1. Immerse the tubing for the inlet valve in the appropriate container. 2. On the ChromLab computer or the touch screen, start ChromLab in Manual mode. 3. In the fluidic scheme, touch or click the Sample Pump module with the sample inlet valve to open its dialog box. 4. In the Sample Pump dialog box, select the first sample inlet port to prime.
3 | Preparing the Instrument 132 | 5. Insert a 30 ml syringe into the sample pump’s priming port. 6. Slowly withdraw the plunger to withdraw liquid from the port until air no longer enters the syringe (approximately 10 ml). 7. Remove the syringe from the priming port and discard the liquid into waste. 8. Repeat steps 4–7 for the remaining sample inlet ports.
Priming and Purging the Systems Purging the NGC Discover System To purge the buffer blending valve 1. Open the System Pump dialog box and select Priming on the Mode dropdown list. Note: When in Priming mode, the sample inject valve switches the system pump flow to waste (W2). After the process is completed, the sample inject valve switches the system pump to manually load the column through the loop. To purge an individual line of air bubbles, proceed to step 2 on page 134.
3 | Preparing the Instrument 2. To purge an individual line of air bubbles and fill it with buffer: a. Select the port to open. b. (Optional) Change the default flow rate. The default flow rate is set to 20 ml/min. 3. c. Click Start. d. Repeat steps 2a–c for any other port to purge. To purge all lines of air bubbles and fill them with buffer: a. Select All. b. (Optional) Change the default flow rate. The default flow rate is set to 20 ml/min. c. Click Start.
Priming and Purging the Systems To purge the buffer inlet valves 1. Open the System Pump dialog box and select Priming on the Mode dropdown list. Note: When in Priming mode, the sample inject valve switches the system pump flow to waste (W2). After the process is completed, the sample inject valve switches the system pump to manually load the column through the loop. 2. Select the first buffer port to purge, for example Buffer A 1. 3. (Optional) Change the default flow rate.
3 | Preparing the Instrument To purge the sample pump 136 | 1. In the fluidic scheme, touch or click the Sample Pump module to open its dialog box. 2. In the Sample Pump dialog box, set Flow Rate to 10 ml/min.
Priming and Purging the Systems 3. Click Change Position to set the sample inject valve to Sample Pump Waste and close the dialog box. 4. (Optional) When an air sensor is present, in the Sample Pump dialog box, select Enable Air Sensor to use end-of-sample detection to stop the pump. 5. Click Start. To purge the sample inlet valve (if available) 1. In the fluidic scheme, touch or click the Sample Pump module with the sample inlet valve to open its dialog box.
3 | Preparing the Instrument 2. 138 | In the Sample Pump dialog box, do one of the following: If only one sample inlet valve is present, select S1 Port 8 as the port to purge. If two sample inlet valves are present, click Valve 2 and select S2 Port 8 as the port to purge. 3. Set Flow Rate to 10 ml/min. 4. Click Change Position to set the sample inject valve to Sample Pump Waste and close the dialog box. 5.
Preparing the System and Sample Pump Piston Washing Systems Preparing the System and Sample Pump Piston Washing Systems Note: Place the washing systems reservoir for the system and sample pumps at the same height or above the level of the pump heads to avoid siphoning of the solution back to the reservoir. If the washing system is not thoroughly primed and devoid of air, recirculation might not occur at flow rates below 0.3 ml/min for the F10 pump piston washing system 3.
3 | Preparing the Instrument Preparing the Columns Before a method can be run, the column must be connected to the instrument. Plumb the column between the sample inject valve and UV detector or the column switching valve (if installed). You can mount columns using the column clips included in the accessories kit. Small columns can be mounted either on the front or the side of the instrument using the cartridge holders, which are capable of holding 1–10 ml cartridges.
Preparing the Columns Connecting Column Clamps to the NGC Instrument The NGC instrument has indented metallic strips on both the right and left sides on which to mount column clamps, air sensors, and the fittings tightener. The column clamps can be placed at any position on the strips. The column clamps are adjustable and can accept a variety of column sizes up to 35 mm in diameter. Tip: For long columns use two column clamps to keep the columns from tipping. Caution: Do not overtighten the clamps.
3 | Preparing the Instrument Connecting Tubing to the Columns Note: Ensure that the system has been purged of air and that the tubing is filled with buffer before connecting the columns. Review and perform the tasks in Priming and Purging the Systems beginning on page 118. To connect tubing to the columns 1. 142 | Connect tubing from one of the following modules to the top of the column: Sample inject valve column port Column switching valve inlet port (1–5) 2.
Preparing the Columns Washing the Columns Cleaning a column requires access to ChromLab. You can perform this task from the ChromLab computer or the NGC touch screen. This procedure explains how to manually wash columns through the System Control tab in ChromLab. Alternatively, you can wash columns in the following phases in the Method Editor: System Preparation, Column Activation, or Column Wash. See the NGC Chromatography Systems and ChromLab Software User Guide for more information about these phases.
3 | Preparing the Instrument e. 144 | Click Start. 4. Run the buffer solution through the column for 3–5 column volume (CV) (or manufacturer’s specification). 5. Run degassed deionized water through the column for 3–5 CV (or manufacturer’s specification). 6. Run the manufacturer’s recommended reagent through the column for 3–5 CV (or manufacturer’s specification). 7. Run degassed deionized water through the column for 10 CV (or manufacturer’s specification). 8.
Calibrating the NGC Instrument Calibrating the NGC Instrument The NGC systems include integrated pressure and temperature sensors. The pressure sensors monitor the system pressure to protect the column and media from overpressure. The temperature sensor in the conductivity monitor enables the system to provide real-time temperature compensation for conductivity and pH measurements. The measured temperature appears as a trace on the chromatogram.
3 | Preparing the Instrument Calibrating the Pump Flow Rate Prime and purge the system pumps with water before starting the calibration procedure. Calibrate the flow rate with water. Note: Ensure that the column is offline before performing this task. To calibrate the pump flow rate 1. Do one of the following: On the ChromLab computer, select the System Control tab, click Tools > Calibrate, and select Pump Flow Rate on the dropdown list.
Calibrating the NGC Instrument 3. Follow the steps in the Calibration dialog box. 4. Verify that Status (at the bottom of the dialog box) is Calibrated. Calibrating the Conductivity Monitor Flow conductivity standard from any commercial supplier through the conductivity flow cell until a stable reading is obtained. To calibrate the conductivity monitor 1.
3 | Preparing the Instrument 148 | NGC Chromatography Systems and ChromLab Software
A Maintaining the Instrument Regular cleaning and maintenance ensures that your NGC™ system continues to perform optimally. This appendix provides a maintenance schedule and explains proper care for the NGC instrument. WARNING! Disconnect power to the NGC system before servicing any module. Do not attempt to service any component on the NGC instrument unless noted in this manual. Contact Bio-Rad for service requests.
A | Maintaining the Instrument Cleaning the Outer Surfaces of the NGC Instrument During normal operation, spills and splashes can cause residue to form on the module faceplates and fluidic lines. Use a damp cloth to clean the faceplates. Avoid using excessive moisture around the power switch and connector outlets located on the lower right and back sides of the unit. Clean the buffer tray. Rinse away any residue with water and empty through the drainage hole.
Storing the NGC Instrument Storing the NGC Instrument Tip: You can use the System CIP/Storage phase in ChromLab to store the fluidic lines and system components. See the NGC Chromatography Systems and ChromLab Software User Guide for more information. To store the NGC instrument overnight, note the following precautions: Thoroughly flush all valves with water to remove buffer salts. If the NGC instrument is in a cold environment, keep it powered on to prevent condensation.
A | Maintaining the Instrument Recommended Maintenance Schedule Table 14 lists recommended maintenance tasks for the NGC instrument. Table 14. NGC instrument maintenance schedule Interval Component Action pH monitor Calibrate the pH monitor. Start ChromLab software and select Tools > Calibrate and choose pH from the dropdown list. Daily Store the pH electrode in storage solution when not in use. See Table 15 for recommended storage solutions.
Recommended Maintenance Schedule Table 14. NGC instrument maintenance schedule, continued Interval Component Action Sample pump seals and check valve Replace the seals and check valves using the pump head maintenance kit. System pump seals and check valve Replace the seals and check valve using the pump head maintenance kit. Post-inject valve tubing Replace all tubing. Inline filter Replace if installed. Bottle filter Replace or clean if installed.
A | Maintaining the Instrument Table 14. NGC instrument maintenance schedule, continued Interval Component Action Tubing and connectors Replace all tubing and connectors. Pressure monitors Calibrate the monitors. Start ChromLab software and select Tools > Calibrate. Complete the calibration steps in the dialog box. pH electrode Check the values for slope and offset. Start ChromLab software and select Tools > Calibrate and choose pH from the dropdown list.
Recommended Cleaning and Storage Solutions Recommended Cleaning and Storage Solutions Table 15 lists the recommended cleaning and storage solutions for the proper care of the NGC instrument. Table 15. Cleaning and storage solutions for the NGC instrument Component Solution to use Outside surface Water or window cleaner on lint-free towelette Caution: Do not spray water or other cleaning solutions directly onto the surface of instrument. See Cleaning the Outer Surfaces of the NGC Instrument on page 150.
A | Maintaining the Instrument System and Sample Pumps The NGC system’s gradient pump system consists of two dual-piston pumps (system pumps A and B). The pumps form linear salt gradients over a set volume or period of time at a controlled flow rate. The NGC system’s sample pump is a dual-piston pump used to load large sample volumes onto a column or to fill large sample loops. The system pumps and the sample pump are also equipped with a piston wash system.
System and Sample Pumps Pump Head Assembly LEGEND 1 Outlet check valve housing 2 Pump head small washer 3 Pump head seal 4 Inlet check valve housing 5 Check valve capsule 6 Pump head housing Instrument Guide | 157
A | Maintaining the Instrument Piston Wash Assembly LEGEND 1 Fitting 2 Piston base 3 Piston 4 Wash housing large washer 5 Wash housing seal 6 Wash housing inlet check valve 7 Wash housing O-ring 8 Wash housing (flow direction up) 9 Wash housing outlet check valve 158 | NGC Chromatography Systems and ChromLab Software
System and Sample Pumps Disassembling the Pumps The system and sample pumps can be disassembled to clean or replace their components. This section explains how to Remove the pump heads Remove the wash housing Remove the piston Removing the Pump Heads Caution: Flush any hazardous material from the system with water or an appropriate cleaning solvent. Remove the buffer and wash system inlet tubing from their bottles to avoid siphoning fluids during disconnection. To remove the pump heads 1.
A | Maintaining the Instrument 3. Use the 3/16" hex key that is provided in the fittings kit to remove the hex bolts on the front of the pump head housing. Caution: Avoid pulling the pump head housing off at an angle or rocking or twisting the pump head housing, as this can cause the piston to break. 4. 160 | Carefully separate the pump head from the pump face plate. Pull the pump head straight toward you.
System and Sample Pumps 5. 6. Locate the wash housing O-ring and the pump head small washer. If you do not see them on the separated pump head: The O-ring might be attached to the wash housing. The small washer might have remained on the pump piston. Remove and store the O-ring and small washer in a safe place such as a petri dish or other small container. Tip: You can remove the O-ring from its groove with a pair of tweezers if necessary. Note: Do not lose or damage the O-ring or small washer.
A | Maintaining the Instrument 9. (Optional) Inspect and clean the piston. Note: To replace the piston, you must remove the wash housing. For more information, see Removing the Wash Housing on page 169. To clean the pump heads 1. Remove the pump head inlet and outlet check valves as explained in To remove the pump head check valves on page 162. 2. Immerse the entire pump head in an ultrasonic cleaner. 3. Sonicate the pump head with a standard cleaning solution for about 5 min, or as necessary. 4.
System and Sample Pumps 2. Unscrew each IN check valve housing and pull it down out of the pump head. You should have removed a total of two IN check valve housings and four check valve capsules. If one of the capsules remains in the pump head, use tweezers to remove it. 3. Turn the pump head housings upside down with the OUT check valve on the bottom. 4. Unscrew each OUT check valve housing and pull it out of the pump head.
A | 164 | Maintaining the Instrument 3. Insert two check valve capsules into each IN check valve housing so the arrow on each capsule points in the direction of flow (away from the cap on the IN check valve). 4. Insert two check valve capsules into each OUT check valve housing so the arrow on each capsule points in the direction of flow (toward the cap on the OUT valve).
System and Sample Pumps To install the pump head check valve assemblies 1. To determine the inlet and outlet pump head housing ports, place the two pump head housings side by side with the flat sides touching. 2. Insert an IN check valve assembly into the bottom port of each pump head housing and hand tighten firmly. 3. Turn the pump head upside down so that the IN check valve is on top.
A | Maintaining the Instrument 4. 166 | Insert an OUT check valve assembly into the open port on each pump head and hand tighten firmly.
System and Sample Pumps To replace the pump head seal 1. Insert the seal removal end of the small seal tool in the seal cavity on the pump head. The seal removal end is long and thin. It has a barbed tip to catch the seal. 2. Pull the seal out of the head. 3. Place the new seal on the other end of the seal tool. The seal insertion end is shorter than the removal end. The tip is smooth, not barbed like the removal end. 4. Carefully align the seal with the seal cavity on the pump head. 5.
A | Maintaining the Instrument 6. 168 | Remove the seal tool and inspect the pump head to verify that the exposed small seal surface is lower than the surrounding pump head housing.
System and Sample Pumps Removing the Wash Housing To remove the wash housing 1. Complete the steps in the section Removing the Pump Heads on page 159. 2. Disconnect inlet and outlet tubing from both the left and right wash housings. 3. Observe the black arrow on the side of each wash housing. The arrows indicate the direction of wash flow. On the left housing, the arrow points upward. On the right housing, the arrow points downward.
A | Maintaining the Instrument Removing the Piston Caution: Use extreme caution when removing the piston. The sapphire piston can easily break if dropped or bent. To remove the piston 1. Complete the steps in the sections Removing the Pump Heads on page 159 and Removing the Wash Housing on page 169. 2. Remove the wash housing seal from the piston if it did not detach with the wash housing.
System and Sample Pumps 5. Gently grasp the piston by its metal base and carefully lift the piston straight up to dislodge it from its carrier. The left side of the following image shows a completely disassembled pump panel.
A | Maintaining the Instrument 6. Rinse the piston and examine it for damage. 7. Store the piston in the same container as the large washer and the wash housing seal. To replace the wash housing seal 1. Insert the seal removal end of the large seal tool in the seal cavity on the pump head. The seal removal end is long and thin. It has a barbed tip to catch the seal. 2. Pull the seal out of the head. 3. Place the new seal on the other end of the seal tool.
System and Sample Pumps To replace the wash housing check valves 1. Identify the left and right wash housings. The wash system has two wash housings for each pump. The left housing has check valves connected at the inlet and outlet ports. It is installed on the pump with the arrow (flow direction) pointing up. The right wash housing has empty fittings connected at the inlet and outlet ports. The right housing is installed with the arrow (flow direction) pointing down.
A | Maintaining the Instrument To identify the left and right housings after you remove them from the pump, remove a fitting on one of the wash housings. If the fitting is a check valve then the housing is the left housing. If the fitting is hollow then the housing is the right housing. Tip: You might have labeled the wash housings Left and Right in the section To remove the wash housing on page 169. 2. Locate one inlet and one outlet piston wash check valve for the left wash housing.
System and Sample Pumps Reassembling the Pumps This section explains how to Insert the piston Install the wash housing Install the pump heads Inserting the Piston Caution: Use extreme caution when inserting the piston. The sapphire piston can easily break if dropped or bent. To insert the piston 1. Holding the metal piston base, carefully slide the base into its carrier until it is securely seated. 2. Slide the large washer onto the piston. Push it back as far as it will go.
A | Maintaining the Instrument Installing the Wash Housing Before installing the wash housing, ensure that The large washer is installed on the piston and pushed all the way back The wash housing seal is installed in the wash housing To insert the wash housing 1. If you removed the left wash housing check valve see To replace the wash housing check valves on page 173. 2. Reinstall the wash housing O-rings if you removed them. Ensure the O-rings are fully inserted into the O-ring grooves. 3.
System and Sample Pumps 4. Position the left wash housing with its arrow pointing up and carefully slide it into place over the piston. 5. Position the right wash housing with its arrow pointing down and carefully slide it into place over the piston. 6. Connect a pump wash tubing to the top left and right wash housing ports. 7. Connect an inlet tube to the bottom left wash housing inlet port. 8. Connect a priming tube to the bottom right wash housing port. 9.
A | Maintaining the Instrument Installing the Pump Heads Note: When installing new pump heads, ensure that you install two pumps of the same flow rate (that is, either two F10 pumps or two F100 pumps). Do not mix the pump heads. To install the pump heads 1. If you removed the inlet and outlet check valves from the pump head, insert the valves and firmly tighten each one. For more information about replacing the check valves, see To remove the pump head check valves on page 162. 178 | 2.
System and Sample Pumps 6. Identify the pump head assemblies. To identify the left and right pump heads, position both pump heads so the front of the pump head faces you and the check valve OUT ports are visible. The left pump head has a straight edge on its right side. The right pump head has a straight edge on its left side. Caution: Do not force the pump head into place. If the pump head does not slide onto the housing easily, pull out the pump head and examine it for any obstruction. 7.
A | Maintaining the Instrument 8. Push on the center of the pump head to seat it firmly against the pump face. 9. Using the hex key, firmly tighten the hex bolts (alternating from one bolt to the other) while centering the pump head. 10. Reconnect the buffer inlet and outlet tubing and tighten with a fittings tool. 11. Prime the system. See To plumb and prime the system pump wash lines on page 267 or To plumb the sample pump on page 268 as appropriate.
System and Sample Pumps Replacing the Priming Manifold Check Valve The priming manifold check valve has one check valve capsule. This procedure explains how to replace both the priming manifold check valve and its capsule. To replace the priming manifold check valve 1. Unscrew the priming manifold outlet port check valve housing (labeled OUT).
A | 182 | Maintaining the Instrument 2. Using a pair of tweezers, remove the check valve capsule from inside the port. 3. Locate a new check valve capsule for the priming manifold check valve. 4. Carefully observe the capsule. The arrow on the side of the capsule indicates the flow direction. Ensure that you insert the capsule with the arrow pointing in the direction of flow (upward).
System and Sample Pumps 5. Using the tweezers, insert the new check valve capsule into the priming manifold outlet port. Note: Ensure that the capsule is upright before inserting the check valve housing.
A | Maintaining the Instrument 6. 184 | Carefully insert the OUT check valve housing in the priming manifold outlet port and hand tighten firmly.
System and Sample Pumps Conditioning New Seals New seals should be conditioned before use. To condition the seals, run the pump with wetted seals under controlled flow rate and pressure conditions. This allows the seals to seat properly and proof against leaks. Important: Use only organic solvents to condition new seals. Buffer solutions and salt solutions should never be used to condition new seals. Recommended solvents are HPLC-grade methanol and IPA or aqueous mixtures of these solvents.
A | Maintaining the Instrument Mixer Replacing the Mixer Components The NGC system’s mixing chamber volume can be adjusted so that it is optimized for the flow rate being used. See Table 2 on page 63 for supported flow rates.
Mixer Instrument Guide | 187
A | Maintaining the Instrument Disassembling the Mixer Use this procedure to replace any of the components of the mixer. Important: Before disassembling the mixer, flush any hazardous material from the NGC system. Ensure that the pumps are not running and bleed residual pressure from the system. To disassemble the mixer 188 | 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC instrument. 2.
Mixer 3. Twist the mixer assembly clockwise approximately 1/8 turn and lift it upward to remove it from the socket. 4. Tilt the mixer assembly over a container to drain any residual fluid from the mixer. 5. Using the hex key that is provided in the fittings kit, remove the four hex bolts on the bottom of the mixer base.
A | Maintaining the Instrument 6. 190 | Separate the mixer top from the mixer base and barrel extension (if used).
Mixer To change the O-rings 1. Locate the O-ring in the groove on the mixer base and the mixer barrel extension (if used). 2. With your finger, pry the O-ring out of the groove. 3. Insert the replacement O-ring in the groove. Note: If you are using only the mixer body, then only one O-ring is required. If a mixer barrel extender is used, two O-rings are required.
A | Maintaining the Instrument To replace or exchange the mixer barrel Tip: If you are replacing the mixer barrel, store the current mixer barrel and the hex bolts in your accessories kit or another safe place. 1. Locate the replacement mixer barrel. 2. Ensure that the O-ring on the mixer base is in its groove and that the stir bar is properly seated. 3. With the O-ring on the mixer base facing up, place the required mixer barrel extension on the mixer base with its O-ring facing up.
Mixer To clean the mixer body 1. Wipe the inside of the mixer body with a lint-free towel. 2. Soak and sonicate the following components in a bath of mild detergent for about 15 min: Mixer barrel extension O-rings Magnetic stir bar Tip: The stir bar is magnetic. To remove it, turn the mixer base upside down and gently tap it on the benchtop or tabletop.
A | Maintaining the Instrument Reassembling the Mixer Tip: To reinsert the magnetic stir bar, place it on a tabletop and carefully place the mixer base over it. Ensure that the magnetic stir bar lies flat in the mixer base. To reassemble the mixer 1. Place the mixer top onto the top of the mixer base assembly. Note: Ensure that the four bolt holes on the bottom of the mixer top align with the four bolt holes on the top of the mixer base assembly. 2.
Mixer 3. Using the hex key, firmly tighten the hex bolts. Alternate by tightening opposite bolts (tighten Bolt 1, then Bolt 3; Bolt 2, then Bolt 4). 4. Insert the mixer assembly into the socket and fasten it by twisting it counterclockwise until it clicks. 5. Reconnect the tubing to the top, bottom, and sides of the mixer and ensure that they are secure. 6. Restart the NGC instrument.
A | Maintaining the Instrument The Detectors Replacing the UV Flow Cell The NGC systems ship with a 5 mm UV flow cell installed. Additional flow cells for analytical (10 mm) and preparative (2 mm) applications are available. This section explains how to change or replace the UV flow cell. The procedure is the same for both the single-wavelength UV and multi-wavelength UV/Vis detectors. To replace the UV flow cell 1.
The Detectors Multi-Wavelength UV/Vis Flow Cell 4. Firmly grasp the bolts on the flow cell and lift the flow cell from its socket.
A | Maintaining the Instrument Multi-Wavelength UV/Vis Flow Cell 5. Verify that the square gasket is removed with the flow cell. Tip: If the gasket is not on the flow cell, look inside the socket. The rubber gasket might have remained in the groove inside the socket. Remove the gasket and place it onto the flow cell that you removed. 6. 198 | Store the flow cell with the attached gasket and bolts in a safe, clean place for future use.
The Detectors 7. 8. Locate the flow cell. The flow cell should include Two bolts One square rubber gasket Place the flow cell into the socket on the UV monitor. Note: Ensure that the arrow on the top of the flow cell is pointing to the right, and that the two bolts on the flow cell align with the bolt holes in the socket. The guides on the bottom of the flow cell help to prevent inserting the flow cell incorrectly.
A | Maintaining the Instrument 9. Tighten the bolts. If necessary, use the screwdriver. 10. Reconnect the 1/4-28 fittings from the tubing to the top and bottom of the flow cell and ensure that they are secure. 11. Reconnect the tubing lines to the conductivity monitor. 12. Restart the NGC instrument.
The Detectors Replacing the Single-Wavelength UV Detector LED This section explains how to replace the LED on the single-wavelength UV detector. Caution: It is strongly suggested that you wear gloves when you replace the LED. Do not touch the LED glass with bare hands, as oils from your skin will degrade the lamp over time. To replace the single-wavelength UV LED 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC instrument. 2.
A | 202 | Maintaining the Instrument NGC Chromatography Systems and ChromLab Software
The Detectors Tip: The door is tethered to the casing. It will hang to the side while you replace the lamps.
A | Maintaining the Instrument You see the LED module with three thumbscrews. The LED itself is on the other side of the board. 5. 204 | Completely loosen the top right thumbscrew.
The Detectors 6. Loosen the left thumbscrews, alternating one turn on the top left screw then one turn on the bottom left screw until they are completely loosened. Tip: You might need to loosen the left two screws with a flathead screwdriver. 7. Grasp the thumbscrews and pull the LED board straight forward out of the casing. WARNING! Use caution when removing the LED board. The LED might be hot to the touch after the board is removed.
A | Maintaining the Instrument The following image shows the LED housing without the LED board. 206 | 8. Dispose of the used LED in an appropriate waste receptacle. 9. Locate the new LED and remove it from its packing material.
The Detectors Front of LED Board Back of LED Board 10. Align the thumbscrews of the replacement LED with the holes on the housing and carefully insert the LED into the casing. 11. Tighten the three thumbscrews to secure the lamp into the casing. 12. Slide the door to the LED casing into place until the latch secures. 13. Reconnect the tubing lines to the top and bottom of the UV cell. 14. Reconnect the tubing lines to the conductivity monitor. 15. Restart the NGC instrument.
A | Maintaining the Instrument Resetting the Lamp Time Details The system tracks the lamp usage and displays the details on the Detector tab in the System Information dialog box in ChromLab. After you change the lamps, you need to reset the details in the system so that the details are current. To reset lamp time details 1. Start ChromLab. 2. Do one of the following: 3. On the ChromLab computer, from the System Control tab open File > System Information and select the Detector tab.
The Detectors Replacing the Multi-Wavelength UV/Vis Detector Lamps This section explains how to replace the deuterium lamp or tungsten lamp inside the multi-wavelength UV/Vis detector. The replacement lamp unit includes the lamp, cable connector, and plug. The lamp is replaced as a unit. Caution: It is strongly suggested that you wear gloves when you replace the lamps. Do not touch the lamp glass with bare hands, as oils from your skin will degrade the lamp over time.
A | Maintaining the Instrument The deuterium and tungsten lamps are both accessible. Tip: You might need to unscrew the cable connector for the tungsten lamp to access the collar to the cable connector for the deuterium lamp.
The Detectors 6. Remove the lamp: WARNING! Use caution when removing the lamps. The lamps might be hot to the touch after the module is removed. a. Using your fingers, loosen the collar on the cable plug. b. Pull the plug upward to disconnect the cable.
A | Maintaining the Instrument c. With a 2.5 mm hex key, remove the screws on the top and bottom of the lamp casing. d. Gently pull the lamp casing and lamp out of the socket. The following image shows a completely disassembled deuterium lamp unit. e. Dispose of the used lamp in an appropriate waste receptacle. 7. Locate the replacement lamp and remove it from its packaging. 8. Carefully insert the replacement lamp fixture into the socket. 9.
The Detectors 11. Insert the cable plug and tighten the collar. Tip: The deuterium lamp plug has seven connector pins, the tungsten lamp has five. 12. Replace the access door and tighten the screw with a Phillips screwdriver. 13. Slide the multi-wavelength UV/Vis module into its bay and fasten it in place by tightening the two screws. 14. Reconnect the tubing lines to the top and bottom of the UV cell. 15. Reconnect the tubing lines to the conductivity monitor. 16. Restart the NGC instrument.
A | Maintaining the Instrument Replacing the Conductivity Monitor The images in this section show the conductivity monitor on the single-wavelength UV detector module. Though the position of the conductivity monitor differs on the multi-wavelength UV/Vis module, the steps to replace the conductivity monitor are the same. To replace the conductivity monitor 214 | 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC instrument. 2.
The Detectors 4. Firmly grasp the detector and pull it toward you to detach it. 5. Dispose of the detector in an appropriate waste receptacle. 6. Locate the replacement detector and remove it from its packaging. 7. Align the two thumbscrews with the screw holes and the pin holes to the pins on the instrument. Tip: The thumbscrews are on the top of the detector. There are two guide bolts on the bottom of the detector. Caution: Do not bend or break the connector pins.
A | Maintaining the Instrument 8. Gently push the detector onto the connector pins on the instrument. 9. Tighten the thumbscrews to secure the detector in place. 10. Reconnect the tubing lines to the conductivity monitor. 11. Restart the NGC instrument. Note: The conductivity monitor will go through an automated calibration routine when you restart the system.
pH Probe pH Probe Storing the pH Electrode When not in use, store the pH electrode in storage solution. See Table 15 on page 155 for recommended storage solutions. Tip: Set the pH valve to Bypass mode until it is needed. This ensures the pH electrode remains in storage. To store the pH electrode 1. Start ChromLab software. 2. Choose Tools > Calibrate to open the Calibration dialog box. 3. Select pH on the Calibrate dropdown list. 4. Press Start to set the pH monitor to Calibration mode. 5.
A | Maintaining the Instrument To remove lipophilic organic contaminants Immerse the electrode in an organic solvent or a liquid detergent, for example Bio-Rad cleaning concentrate, catalog #161-0722 2% Contrad, catalog #176-4118 Replacing the pH Probe The replacement pH probe is packaged in storage solution to protect it during shipping. To replace the pH probe 218 | 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC instrument. 2.
pH Probe 3. Loosen the black collar on the pH probe. 4. To remove the pH probe, lift the probe out of the pH flow cell. Dispose of the probe in an appropriate waste receptacle. 5. Add a small amount of water (2 ml) to the pH flow cell. 6. Locate the replacement pH probe and remove it from the storage solution. 7. Ensure that the pH probe cord is threaded through the collar. 8. Inspect the probe to determine if the air bubble in the stem has moved into the bulb area.
A | Maintaining the Instrument Other Components Attaching an Expansion Tier to the NGC Instrument WARNING! Disconnect power to the NGC instrument before attaching an expansion tier. Do not attempt to service any component on the NGC instrument unless noted in this manual. Contact Bio-Rad for service requests. To attach the connector cable, you will need access to the back of the NGC instrument. If necessary, pull the NGC instrument away from the wall and rotate the instrument to gain access.
Other Components Tip: The buffer tray can remain inside the top. 4. Locate the package that contains the expansion tier and 52" (1.32 m) connector cable. Important: The cables that ship with third and fourth tiers are not interchangeable. You must use the cable that ships with the expansion tier. Inserting a third tier cable into the fourth tier connector ports will damage the cable and the ports. The catalog number for the 25" (63.5 cm) second-to-third tier connector cable is #100-24878.
A | Maintaining the Instrument Fourth expansion tier Previously attached third tier 222 | 7. Locate the connector cable that ships with the expansion tier. 8. View the connector ends. Each end is beveled. The direction of the bevel determines the tier into which to insert the connector: The end with the bevel slanting up inserts into the upper tier’s connector port. The end with the bevel slanting down inserts into the lower tier’s connector port.
Other Components For example: Instrument Guide | 223
A | Maintaining the Instrument 9. Firmly press the clips inward and insert the appropriate connector into the upper tier’s connector port. Tip: You hear a click when the connector is properly seated.
Other Components 10. Insert the other connector into the lower tier’s connector port. 11. Place the top, with the buffer tray, on top of the NGC instrument and place the buffer bottles into the buffer tray. 12. Insert tubing lines into the sample and buffer bottles.
A | Maintaining the Instrument 13.
Other Components Replacing or Repositioning Modules on the NGC Instruments WARNING! Disconnect power to the NGC instrument before removing or repositioning any module. Do not attempt to service any component on the NGC instrument unless noted in this manual. Contact Bio-Rad for service requests. WARNING! To reduce the chance of liquid seeping into the instrument, all open bays must be filled with the NGC Blank module (catalog #788-4005).
A | Maintaining the Instrument The following image shows an empty bay. 5. Store the module and the screws in a safe place for future use. Tip: If you received a replacement module from Bio-Rad, use the packaging provided with your shipment to return the damaged module to Bio-Rad. 6. (Optional) Convert the bay to single wide or double wide. See Converting Bays to Fit Modules on page 231. 7. 228 | Locate the module that you plan to install and remove it from its packaging.
Other Components 8. The frame of the bay has tracks on the top and bottom of the right side. The module has guide rails on the top and bottom right side. Align the rails on the module with the tracks in the bay to properly guide it into position. Top of Module 9. Bottom of Module Place the module into the open bay and gently push it in as far as it will go. Note: Each module has an alignment pin on the back to ensure that it aligns correctly with the main communication board. 10.
A | Maintaining the Instrument 13.
Other Components Converting Bays to Fit Modules Some modules fit into single-wide bays while others require double-wide bays (such as the system and sample pump modules and the UV and UV/Vis detector modules). Bays can be converted from one size to the other by adding or removing the center divider. The following image shows two adjacent, empty, single-wide bays.
A | Maintaining the Instrument To convert a single-wide bay to a double-wide bay 232 | 1. Complete steps 1–5 in the section To replace or reposition modules on the NGC instruments. 2. Gently lift the divider upward off of the lip to unhook it and then pull it out of the instrument. 3. Store the divider in a safe place for later use. 4. Complete steps 7–12 in the section To replace or reposition modules on the NGC instruments.
Other Components To convert a double-wide bay to a single-wide bay 1. Locate a divider. 2. Complete steps 1–5 in the section To replace or reposition modules on the NGC instruments on page 227. 3. Insert the top of the divider into its slot. 4. Gently bend the divider and slide the bottom of the divider up onto the lip. 5. Complete steps 7–12 in the section To replace or reposition modules on the NGC instruments.
A | Maintaining the Instrument Cutting Replacement Tubing Tip: Use the tubing cutter provided in the fittings kit. The cutter produces a flat, clean cut, which is important in making a fitting. To cut replacement tubing 1. Use the outer portion of the blade on the tubing cutter to cut the tubing to the approximate length. 2. Insert the tubing into the hole in the tubing cutter and cut the end of the tubing smooth and square. 3.
Other Components 4. Slide the fitting nut, lock ring, and the ferrule, in that order, onto the tubing. Note: The stainless steel lock ring is not in the fluid path, so biocompatibility is maintained. 5. Extend the tubing slightly beyond the end of the ferrule. 6. Use the fittings tightener (supplied in the fittings kit) to seat and tighten the ferrule onto the tubing. 7. a. Insert the assembly into one of the ports on the fittings tightener. b.
A | Maintaining the Instrument Installing the Backpressure Regulator The NGC systems ship with a 40 psi backpressure regulator in the accessories kit. If the column requires a high backpressure, you will have to install the backpressure regulator. Caution: Do not use the backpressure regulator with low pressure columns, as this will result in damage. To install the backpressure regulator 1. Install the backpressure regulator between the conductivity monitor and the pH detector (if one is in use).
Other Components To remove the backpressure regulator 1. Disconnect the tubing from the top and bottom of the installed backpressure regulator. 2. Store the backpressure regulator in a safe place. 3. Do one of the following: Attach a Tefzel union to the open tubing in place of the backpressure regulator. Replace the two open tubes with a single tubing line.
A | Maintaining the Instrument Repositioning the Touch Screen This procedure explains how to move the touch screen from one side of the NGC instrument to the other. To reposition the touch screen 1. On the touch screen, select Shut Down on the dropdown menu to exit ChromLab and shut down the NGC instrument. 2. Disconnect the monitor cable from the touch screen connector port on the back of the instrument. Unscrew the thumbscrews and firmly pull the cable out of the connector port. 3.
Other Components 4. Remove the side mount cover from the side of the instrument on which you plan to attach the touch screen. Note: The top section in this image is a third tier. (The NGC Discover™ system is a 3-tier system.) The touch screen is attached at the top of the second tier. If you have an NGC Quest™ or an NGC Scout™ system, you will not have a third tier. Locate the side mount cover at the top of the second tier. 5.
A | 240 | Maintaining the Instrument 6. Remove the display cable from the side and rear cable troughs on the instrument. 7. Using the hex key labeled Pumphead, remove the hex bolt from underneath the touch screen mount.
Other Components 8. Lift the touch screen display off the mounting post. 9. Place the touch screen monitor face down on a clean surface to prevent scratching. 10. Remove the touch screen mount and attach it to the other side of the NGC instrument.
A | Maintaining the Instrument 11. Using the 7/64 hex key provided in the accessories kit, remove the cover from the back of the touch screen monitor. 12. Using a Phillips screwdriver, remove the six screws that attach the touch screen to the pivot arm assembly and place them aside. 13. Lift the pivot arm assembly off of the touch screen. The mounting assembly should be in your right hand.
Other Components 14. Turn the pivot arm assembly around 180° so that the mounting assembly is now in your left hand and insert the assembly into its position on the back of the touch screen. 15. Using the six screws that you removed in step 12, reattach the pivot arm assembly to the touch screen with a Phillips screwdriver. 16. Reattach the cover to the back of the touch screen with the 7/64 hex key. 17.
A | Maintaining the Instrument 20. Route the display cable through the side and rear cable troughs on the instrument. 21. Locate the cable trough covers and slide mount cover that you removed in step 5 on page 239. 22. Slide the cable trough covers into place and secure them to the back of the instrument with 6-32 x 1¼" Phillips screws. 23. Insert the side mount cover on the side of the instrument from which you removed the touch screen. 24.
B Troubleshooting the Instrument This appendix lists potential problems and suggested solutions for the NGC™ chromatography systems. You can obtain more information about your system by registering for Consult Bio-Rad at www.bio-rad.com, where an online technical support service offers an extensive database of frequently asked questions (FAQs). Note: To troubleshoot other Bio-Rad instruments and devices, such as the BioFrac™ fraction collector and the C-96 autosampler, refer to their separate documentation.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution The pumps do not deliver the correct flow rate (continued). The fittings or tubing sizes might be wrong. Check system pumps A and B and the buffer blending or sample inlet valves: Note: It is important to use the recommended tubing and fitting sizes. Ensure that the inlet lines to system pumps A and B are tight. For information about tubing specifications, see Table 5, NGC system tubing details, on page 76.
System and Sample Pumps Symptom Possible Cause Possible Solution The pumps do not deliver the correct flow rate (continued). Air bubbles might be trapped in the pump heads, causing erratic liquid delivery. Inspect the inlet lines, ensure that the fittings are tight to prevent air from leaking into the system. Reprime the pumps to remove air that might be trapped in the system. Note: Always degas buffers before use by stirring them vigorously under vacuum for approximately 20 min.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution The pumps do not deliver the correct flow rate (continued). The pump seals or check valves might be fouled. Use an ultrasonic cleaner and a mild detergent solution and then rinse with water to recondition the pump seals and check valves. Perform CIP to clean the pump seals and check valves.
System and Sample Pumps Symptom Possible Cause Possible Solution Liquid continuously leaks from the pump head washout drain trough (continued). The check valves might need to be replaced. Replace badly clogged or damaged check valves. Note: Recalibrate the pumps after replacing check valves. In ChromLab, select Tools > Calibrate > Pump Flow Rate. Complete the calibration steps in the dialog box. The screws that secure the pump heads are not tight. Replace the wash housing small seal.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution The backpressure drops to zero or is much lower than expected (continued). The check valves might need to be cleaned or replaced. Perform CIP to clean the check valves. See the NGC Chromatography Systems and ChromLab Software User Guide for more information. Replace check valves. See Disassembling the Pumps on page 159. No pressure (psi) reading, or the psi value always reads zero. The pumps might not be running.
System and Sample Pumps Symptom Possible Cause Possible Solution Backpressure values are higher or lower than expected. The flow rate might have changed. In ChromLab, verify that the pump is set to deliver the correct flow rate. Note: Recalibrate after stopping the pumps. In ChromLab, select Tools > Calibrate > Pump Flow Rate. Complete the calibration steps in the dialog box. The column might need to be cleaned, or a frit might need to be replaced.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution Backpressure values are higher or lower than expected (continued). The inline filter is clogged. Inspect the inline filter and replace if necessary. Note: Recalibrate after stopping the pumps. In ChromLab, select Tools > Calibrate > Pump Flow Rate. Complete the calibration steps in the dialog box. Unintended increase in flow rate. In ChromLab, verify that the flow rate setting is correct.
UV or UV/Vis Detector UV or UV/Vis Detector Symptom Possible Cause Possible Solution The UV baseline is unstable or noisy. Air bubbles are trapped in the flow cell. The UV baseline shows a reproducible zig-zag or sawtooth trace. Note: Always degas buffers and solutions before use. Degas buffers by stirring vigorously under vacuum for approximately 20 min. Important: Use a heavy-wall, side-arm Erlenmeyer flask, as standard flasks may implode under vacuum.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution The UV baseline is unstable or noisy (continued). UV or UV/Vis lamps are dirty or are close to the end of their usable life. Check the lamp usage values in the System Information dialog box: 1. From the System Control tab in ChromLab open File > System Information > Detector tab. The UV baseline shows a reproducible zig-zag or sawtooth trace (continued). 2. Click Get Lamp Status.
UV or UV/Vis Detector Symptom Possible Cause Possible Solution Baseline noise continues when pumps are turned off. External environmental influences can cause noise spikes. If the spikes occur at regular intervals (for example, every 20– 30 sec), check for the presence of heating baths, drying ovens, or other heating devices on the same electrical circuit or in close proximity to the NGC system. Turn off these devices to see if the problem goes away. The UV trace will not zero.
B | Troubleshooting the Instrument Symptom Possible Cause Possible Solution The baseline is drifting. The eluant is nonhomogeneous perhaps due to poor mixing or flow rate variation. Check the flow rates of both system pumps. Recalibrate both system pumps. In ChromLab, select Tools > Calibrate > Pump Flow Rate. Complete the calibration steps in the dialog box. The column equilibration is slow. Certain ion exchangers are slow to re-equilibrate, especially when just sanitized.
UV or UV/Vis Detector Symptom Possible Cause Possible Solution Sample trace values are lower than expected. The lamps might be nearing the end of their usable life. Check the lamp usage values in the System Information dialog box. From the System Control tab in ChromLab open File > System Information > Detectors tab. If the lamp usage values are close to their limits (approximately 2,000 hours for the deuterium and tungsten lamps; approximately 5,000 hours for LEDs), change the lamps.
B | Troubleshooting the Instrument Conductivity Monitor Symptom Possible Cause Possible Solution Conductivity values are higher or lower than expected. The conductivity monitor might need calibration. Recalibrate the conductivity monitor. In ChromLab, select Tools > Calibrate > Conductivity Monitor. Complete the calibration steps in the dialog box. Note: The relationship between the conductivity reading and salt concentration is not linear. The curve will flatten out at higher salt concentrations.
pH Probe pH Probe Symptom Possible Cause Possible Solution The pH readings are higher or lower than expected. The pH probe might need to be calibrated. Recalibrate the pH probe. In ChromLab, select Tools > Calibrate > pH. Complete the calibration steps in the dialog box. The pH readings are noisy or unstable. Air bubbles might be present in the pH probe flow cell. Flush the system to remove air bubbles. Check the values for slope and offset.
B | Troubleshooting the Instrument Buffer Blending Valve Symptom Possible Cause Possible Solution Liquid leaks out from the drain plug at the bottom of the module. Liquid is building up inside the module. Shut down the system and call Bio-Rad Technical Support. Column Switching Valve Symptom Possible Cause Possible Solution Pressure sensors read > 0–1 psi when the NGC system is idle. With regular use, pressure sensor values may drift over time.
External Devices External Devices Symptom Possible Cause Possible Solution Unable to zero an external device connected to SIM 1 or SIM 2 ports on the signal import module (SIM). Possible voltage offset between the negative and ground input terminals on the SIM. Install a jumper between the negative and ground terminals on the SIM 1 or SIM 2 ports. See To connect an external device to the SIM 1 and SIM 2 ports on page 107. Incorrect readings from an external device connected to SIM 1 or SIM 2 ports.
B | Troubleshooting the Instrument 262 | NGC Chromatography Systems and ChromLab Software
C Plumbing the System With the exception of the inlet tubing, waste lines, and column, the NGC™ systems ship preassembled and preplumbed with the modules necessary to perform gradient separations. The inlet and waste lines should be plumbed during the installation process. However, as your needs change, you can reconfigure your NGC system by inserting and removing modules. After you reconfigure your system, you will have to replumb the modules.
C | Plumbing the System Plumbing NGC Systems Several plumbing tasks are required for all system configurations. Other tasks are specific to the configuration. This section explains how to plumb each instrument configuration. General Guidelines for Tubing Sizes The NGC systems use three sizes of tubing. Use the information in Table 18 to select the appropriate tubing. Table 18. NGC system tubing sizes Use Tubing Dimensions Tubing Material Fittings Vol/cm Prepump 1/8" OD x 0.
Plumbing NGC Systems Plumbing All NGC Systems Table 19 lists the tasks for plumbing all NGC systems. The tasks are explained in detail in the sections that follow. Table 19. Plumbing tasks for all NGC systems Task 1. 2. Inlet tubing to the pump head wash port on system pump A Pump head wash priming line to the pump head wash port on system pump B 3. 1/4-28 union to priming line on system pump B 4.
C | Plumbing the System System Pumps Wash Flow Sample Pump Wash Flow 266 | NGC Chromatography Systems and ChromLab Software
Plumbing NGC Systems To plumb and prime the system pump wash lines 1. Mount a tube holder to the right or left side of the instrument in either the upper or lower rack slots. 2. Fill a 50 ml tube with 20% isopropanol (IPA) and insert it in the tube holder. Tip: You can use 20% ethanol as an alternative. 3. Connect inlet tubing to the pump wash inlet port (left rear port) on system pump A. 4. Immerse the other end of the pump A wash inlet tubing in the tube of 20% IPA. 5.
C | Plumbing the System 8. Attach a 30 ml syringe to the 1/4-28 female to male luer adaptor. 9. Withdraw the plunger on the syringe until liquid is visible in the syringe. 10. Detach the luer adaptor and syringe from the pump head wash priming tubing on pump B. 11. Locate the second inlet tubing with the 1/4-28 union that you set aside in step 7. 12. Connect the union to the pump head wash priming tubing on system pump B. 13. Immerse the other end of the inlet tubing in the tube of 20% IPA.
Plumbing NGC Systems Plumbing the Sample Inject Valve To plumb the sample inject valve 1. Connect waste tubing to waste ports Waste 1 and Waste 2 on the sample inject valve. 2. Connect an injection port to the center of the sample inject valve. 3. Connect the sample loop to ports Loop E and Loop F on the inject valve.
C | Plumbing the System Attaching the Backpressure Regulator If your column requires a high backpressure, install the backpressure regulator. To attach the backpressure regulator 1. Connect Tube #2 to the top of the conductivity monitor. 2. Connect the other end of Tube #2 to a 40 psi backpressure regulator. Note: The backpressure regulator has a raised arrow on its side. Ensure that the arrow on the regulator points in the direction of the flow.
Plumbing NGC Systems Plumbing the NGC Quest System Table 20 lists the tasks for plumbing the NGC Quest™ system. The tasks are explained in detail in the sections that follow. Note: Ensure that you complete the plumbing tasks in the section Plumbing All NGC Systems on page 265 before continuing. Table 20. Plumbing tasks for the NGC Quest system Tasks 1. Inlet tubing to the system pump A inlet manifold 2. Inlet tubing to the system pump B inlet manifold 3.
C | Plumbing the System NGC Quest Point-to-Plumb Connections 272 | NGC Chromatography Systems and ChromLab Software
Plumbing NGC Systems To plumb the system pumps 1. Connect inlet tubing to the inlet manifold of system pump A. 2. Connect inlet tubing to the inlet manifold of system pump B. 3. Immerse the open ends of both inlet tubing in a bottle of degassed, deionized water. To plumb the fraction collector 1. Connect Tube #4 to the open end of the backpressure regulator that you connected to the conductivity monitor in step 2 on page 270. 2.
C | Plumbing the System Plumbing the NGC Scout System Table 21 lists the tasks for plumbing the NGC Scout™ system. The tasks are explained in detail in the sections that follow. Note: Ensure that you complete the plumbing tasks in the section Plumbing All NGC Systems on page 265 before continuing. Table 21. Plumbing tasks for the NGC Scout system Tasks 1. Inlet tubing to ports Q1–Q4 on the buffer blending valve 2. Waste tubing to the calibration outlet port (Cal Out) on the pH valve 3.
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C | Plumbing the System To plumb the buffer blending valve Connect the inlet tubing to ports Q1–Q4 on the buffer blending valve. Note: The buffer inlet tubing is color-coded to identify the different buffer solutions. The color scheme for the tubing and ports is defined as follows: Tubing color Port Solution Green Q4 High salt buffer Yellow Q3 Water Blue Q2 Buffer base Red Q1 Buffer acid To plumb the pH valve 276 | 1.
Plumbing NGC Systems 3. Immerse the open end of the waste tubing in the waste container. 4. Connect Tube #2 from the 40 psi backpressure regulator that you connected in step 2 on page 270 to the inlet port (In) on the pH valve. 5. Connect Tube #4 to the outlet port (Out) on the pH valve. 6. Connect the other end of Tube #4 to the Common port on the BioFrac fraction collector diverter valve.
C | Plumbing the System Plumbing the NGC Discover System Table 22 lists the tasks for plumbing the NGC Discover™ system. The tasks are explained in detail in the sections that follow. Note: Ensure that you complete the plumbing tasks in the section Plumbing All NGC Systems on page 265 before continuing. Table 22. Plumbing tasks for the NGC Discover system Tasks 1. 2.
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C | Plumbing the System To plumb the buffer blending valve Connect the inlet tubing to ports Q1–Q4 on the buffer blending valve. Note: The buffer inlet tubing is color-coded to identify the different buffer solutions.
Plumbing NGC Systems To plumb the buffer inlet valves 1. Connect tubing from Outlet A on the buffer blending valve to port 8/BB on buffer inlet valve A. 2. Connect tubing from Outlet B on the buffer blending valve to port 8/BB on inlet valve B. 3. Connect tubing from the Out port on inlet valve A to the inlet manifold of system pump A. 4. Connect tubing from the Out port on inlet valve B to the inlet manifold of system pump B. 5.
C | Plumbing the System To plumb the pH valve 282 | 1. Connect the pump head wash priming tubing to the Cal In port on the pH valve. 2. Connect waste tubing to the Cal Out port on the pH valve. 3. Immerse the other end in the waste container. 4. Connect Tube #2 from the 40 psi backpressure regulator that you connected in step 2 on page 270 to the inlet port (In) on the pH valve. 5. Connect Tube #4 to the outlet port (Out) on the pH valve. 6.
Plumbing NGC Systems Plumbing the NGC Discover Pro System Table 23 lists the tasks for plumbing the NGC Discover Pro system. The tasks are explained in detail in the sections that follow. Note: Ensure that you complete the plumbing tasks in the sections Plumbing All NGC Systems on page 265 and Plumbing the NGC Discover System on page 278 before continuing. Table 23. Plumbing tasks for the NGC Discovery Pro systems Tasks 1.
C | Plumbing the System NGC Discover Pro Point-to-Plumb Connections 284 | NGC Chromatography Systems and ChromLab Software
Plumbing NGC Systems Plumbing Sample Inlet Valves Each sample inlet valve module includes all necessary tubing. The inlet tubing is approximately 5' (1.5 m) in length. Determine the length of inlet tubing you need and cut the tubing to the appropriate length. Note: NGC inlet valves can be used for buffer or sample selection. To plumb the sample inlet valves 1. If your system has only one sample inlet valve: a. Connect tubing from the Out port on the sample inlet valve to the sample pump inlet manifold.
C | Plumbing the System 2. 3. If your system has two sample inlet valves: a. Connect tubing from the Out port on sample inlet valve 1 (S1) to the sample pump inlet manifold. b. Connect tubing from the Out port on sample inlet valve 2 (S2) to port 8 on S1. c. Connect inlet tubing to ports 1–7 on S1 and to ports 1–8 on S2 as needed. Immerse the inlet tubing in sample containers. Plumbing Outlet Valves Each outlet valve module includes all necessary tubing. The tubing is approximately 5' (1.
Plumbing NGC Systems b. c. 2. 3. (Optional) Do one of the following: Connect Tube #4 from port 1 to the Common inlet port on the BioFrac fraction collector diverter valve. Connect Waste tubing to port 1 and immerse in a waste container. Connect outlet tubing to the remaining ports as needed. If your system has two outlet valves: a. Connect tubing from the Out port on the conductivity monitor or pH monitor to the In port on outlet valve 1 (O1). b.
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D Connecting the C-96 Autosampler to NGC Systems Using the C-96 Autosampler with NGC Systems The C-96 autosampler enhances NGC™ chromatography systems, providing automated, accurate, and reproducible sample injections. Its easy-to-install accessories allow for a wide range of injection volumes from 5 μl–10 ml. The signal import module (SIM) connects the C-96 autosampler to NGC systems. The SIM digitizes the analog signal from the C-96 autosampler and transmits the signal to the NGC system.
D | Connecting the C-96 Autosampler to NGC Systems 5. Locate the two cables that connect the autosampler to the SIM: 15-pin D connector cable (part #10027227) 9-pin D connector cable (part #10027224) These cables ship with the autosampler. 6. Attach the DIGITAL IN connector to the 15-pin D connector cable. Tip: The 15-pin D connector cable has three wires (red, black, and brown). a. From left to right, loosen the first, second, and last screws on the connector. b.
Using the C-96 Autosampler with NGC Systems 8. 9. On the back of the autosampler: a. Insert the 15-pin cable into the left (P1 RELAYS OUTPUTS) connector port and tighten the thumbscrews. b. Insert the 9-pin cable into the right (P2 INPUTS) connector port and tighten the thumbscrews. On the back of the SIM: a. Insert the P1 RELAYS OUTPUTS cable into the DIGITAL IN port. b. Insert the P2 INPUTS cable into the DIGITAL OUT port.
D | Connecting the C-96 Autosampler to NGC Systems Plumbing and Priming the C-96 Autosampler This section explains how to plumb the C-96 autosampler to the sample inject valve on the NGC instrument and to prime the autosampler after you plumb it. Before you plumb and prime, you must prepare the tubing. Use 1/16" PEEK tubing to plumb the autosampler to the sample inject valve.
Using the C-96 Autosampler with NGC Systems To prime and purge the C-96 autosampler Note: Before priming the autosampler, ensure that any column is offline. 1. Immerse the inlet tubing in a container of buffer or HPLC grade (filtered, degassed) or other high quality water. 2. Use a syringe to prime the pumps. See the section Priming and Purging the Systems on page 118 for specific information. 3. On the ChromLab™ computer or the touch screen, start ChromLab in Manual mode. 4.
D | Connecting the C-96 Autosampler to NGC Systems 294 | 7. In the fluidic scheme, touch or click the System Pump module to open its dialog box. 8. In the System Pump dialog box: a. Set Flow Rate to 5 ml/min. b. (Optional) Set Duration to at least 2 min. c. Click Start.
Using the C-96 Autosampler with NGC Systems Setting Up the C-96 Autosampler for Use in ChromLab Methods This section briefly explains how to set up the autosampler for use in a ChromLab method. For information about creating methods in ChromLab, see the NGC Chromatography Systems and ChromLab Software User Guide. To set up a method that uses the autosampler 1. In ChromLab create a method that includes the autosampler in the fluidic scheme.
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E Regulatory Information The NGC™ instrument has been tested and found to be in compliance with all applicable requirements of the following safety and electromagnetic compliance standards. The NGC instrument is labeled with the following compliance marks. Safety Compliance CE Mark: EN61010-1 Electrical Equipment for Measurement, Control, and Laboratory Use IEC 61010-1 Safety Requirements for Measurement, Control, and Laboratory Use, Part 1: General Requirements cTUVus Mark: UL STD No.
E | Regulatory Information Sample Loop PSI Ratings for U.S. and Canada The following pressure ratings are required specifically for U.S. and Canada applications. Bio-Rad recommends using these ratings to reduce the volume-pressure product of the column load liquid path in order to maintain compliance with UL61010-1:2012 and CSA C22.2 No. 61010-1-12. Table 24. Sample loop PSI ratings for U.S.
FCC Warning and Notes FCC Warning and Notes The NGC instrument complies with part 15 of the FCC Rules. Operation is subject to the following two conditions: (1) This device may not cause harmful interference, and (2) this device must accept any interference received, including interference that may cause undesired operation. It is possible that emissions from the NGC instrument may interfere with some sensitive appliances when placed nearby or on the same circuit as those appliances.
E | 300 | Regulatory Information NGC Chromatography Systems and ChromLab Software
F NGC Chromatography Systems Catalog Numbers Table 25.
F | NGC Chromatography Systems Catalog Numbers 302 | NGC Chromatography Systems and ChromLab Software
Bio-Rad Laboratories, Inc. Web site www.bio-rad.