402 Dilutor Dispenser User’s Guide
402 Dilutor Dispenser User’s Guide LT801310/C - ©2003 Gilson SAS All rights reserved November 2003
Contents Table of Contents 0 Safety 1 Introduction Notational Conventions .................................................................. 1-2 Overview .......................................................................................... 1-3 Unpacking ...................................................................................... 1-5 Warranty ......................................................................................... 1-6 Customer Service ........................................
Upgrading to Dual-Syringe Model ........................................... 3-10 Removing the Cover ............................................................. 3-11 Connecting the Flat Cable .................................................... 3-11 Inserting the Module ............................................................ 3-11 4 Operation - Standard Use Essential Steps .................................................................................. 4-2 Standard Parts..............................
Changing the Piston Syringes and Piston Noses .................... 6-7 Changing the 100 and 250 µL Piston Noses ...................... 6-7 Changing the 500 µL Piston Noses ...................................... 6-9 Changing the 1 mL Piston Noses ....................................... 6-10 Changing the 25 mL Piston Noses ..................................... 6-11 Changing the 5 mL and 10 mL Piston Noses ................... 6-11 7 Troubleshooting Mechanical Problems ..........................................
0 Safety Safety Read this section carefully before installing and operating the instrument. For safe and correct use of the instrument, it is essential that both operating and service personnel follow generally accepted safety procedures as well as the safety instructions given in this document, the 402 Dilutor-Dispenser User’s Guide. The instrument described in this document is a syringe operated pump for transferring liquids.
Safety Individual components: parts of the valve, the Tee junction, and the syringes (except for the 25 mL syringe) may be autoclaved after dismounting (121°C, 0.1 MPa, 20 min). 1 This instrument1 must not be sterilized, using an autoclave, or any other mechanical device. When you need to clean an instrument, use one of the three following methods: 1 - a clean dry cloth, 2 - a cloth dampened with water, 3 - a cloth dampened with soapy water.
Introduction 1 The following chapter describes the instrument, the service and warranty conditions.
1 Notational Conventions Introduction Notational Conventions The conventions in this User’s Guide are as follows. Used in the stand-alone mode, this instrument is referred to as ‘402 Dilutor-Dispenser’ or simply 402. Specific keys to press on the Gilson Keypad are represented in bold capitals, for example: ‘key in the value and press ENTER’; where ENTER is a key on the keypad.
Introduction The 402 Dilutor-Dispenser, is used to transfer liquids between vials according to user defined protocols (Methods). You program the 402 Dilutor-Dispenser via the keypad, which includes an LCD screen, for communication purposes. Overview Overview 1 You may store a total of 99 linkable Methods using the keypad. Each Method consists of one or more of the following tasks: Dilute, Dispense, Mix, Pipette, and Rinse.
1 Overview Introduction Good Laboratory Practice (GLP) is facilitated by the following: 씮 codes to protect the program files against unauthorized modification, 씮 counters for the number of syringe and valve operating-cycles, 씮 checking protocol for the hydraulic specifications (IQ/OQ procedures), 씮 correction of the syringe-piston stroke to compensate for environmental factors (temperature, viscosity, etc.).
Introduction Your 402 Dilutor-Dispenser is shipped in a box together with its various accessories. The original box and packing material should be kept in case the equipment has to be returned to your Gilson distributor. Unpacking Unpacking 1 You should check off the parts received in the box against the parts list (refer to Appendix A), at the time that you receive the shipment, even if the 402 is not to be used immediately.
1 Warranty Introduction 1-6 Warranty If the 402 Dilutor-Dispenser does not appear to function correctly, first verify the electrical connections are correct and that the instrument is switched ON. Contact your Gilson distributor for technical advice or possibly a service visit. Any service required will be given within the warranty conditions assured by your Gilson distributor.
Introduction Gilson and its worldwide network of authorized distributors provide you with four types of assistance: sales, technical, applications and service. Customer Service personnel are able to serve you more efficiently if you provide the following information: Customer Service Customer Service 1 • The serial number and model number of the equipment involved. • The installation procedure you used. • A concise list of the symptoms.
Introduction 1-8 1
Description 2 The following chapter describes the mechanics and hydraulics of the Dilutor-Dispenser. Two models of 402 Dilutor-Dispenser are available: Single-syringe and Dual-syringe. Each model comes with a keypad, a probe, and accessories. You order and fit the syringe(s), from those described on page 2-6, most suited to your application. Your 402 is delivered with the appropriate modules fitted.
2 Single-syringe Model Description Single-syringe Model The Single-syringe model has a Valve module at the left, and a blanking plate at the right. The Valve module includes one motor to operate the syringe and another motor to switch the valve. It is a liquid transfer station that can be used to deliver up to 25 mL in one stroke, and down to 1 µL (typical value), with high accuracy.
2 Description The Dual-syringe model is fitted with a Valve module at the left and a Tee-junction module at the right. The modules are linked by plastic tubing. Tubing Clip (Self-adhesive) Bottle Clip for Tubing Cable Support Dual-syringe Model Dual-syringe Model Valve Module ON/OFF indicator Clamp for Probe Front Panel The front panel holds the hydraulic components and is fitted with an ON/OFF indicator.
2 Keypad and Probe Description Keypad and Probe Keypad The keypad includes an LCD display (two 24character lines), membrane touch-keys, and a connecting cord. Numeric keys: the keypad has the numbers 0 to 9, a decimal point key, and the Enter key. Five Soft-keys: the function of each key depends on the screen that is displayed (e.g. Next, Prev, Chge...). STOP key: Press this key to stop the current action (Method or direct manipulation).
2 Description Valve The two-position valve, which is fitted to the lefthand side of the 402, has three ports. It switches the syringe port to either the reservoir or to the probe side (see opposite figure). The valve, which has a 0.5 sec rotation period, is driven by a stepper motor. It is designed to withstand a million rotations, when used with water. The liquid contact surfaces of the valve, which is autoclavable, are in PEEK and ceramic (121°C, 0.1 MPa, 20 min).
2 Hydraulic Components Description 0.1 mL 0.25 mL 0.5 mL 1 mL There are seven syringe sizes available for the 402, all except the 25 mL are autoclavable. The lifetime of all pistons (see Chapter 6), when they are used with water at a pressure of up to 0.3 MPa is over 30000 cycles (each cycle contains 2 strokes: one up and one down). The available syringe sizes are: Part Number Description 25025341 25025342 25025347 25025343 0.1 mL 0.25 mL 0.
Installation 3 This chapter explains how to install the fuses, syringes and hydraulics according to your system requirements. This chapter explains how to add the extra module to upgrade your 402 to a dual syringe pump.
3 Assembling the Probe Installation Assembling the Probe For Standard Use Access Point In standard use the probe is used with the 2 mL tapered transfer-tubing (see opposite figure). 1. Thread the tapered end of the transfer-tubing through the plastic ‘tunnels’ in each of the 5 premounted cable supports (see figure below). Probe Probe Nozzle 2. Lay the tubing on the probe’s groove such that about 4 cm of the tapered end protrudes from the end of the probe.
3 Installation Special adapters for the probe D200 tips, pack of 10 D1000 tips, pack of 10 Transfer-tubing (outlet) non-tapered, FEP (2 mL) 1. Lay the tubing on the probe’s groove such that the end with the small washer is just at the orifice of the probe. 2. Push the tubing into the groove in the probe at the 3 round access points using the rounded end of a pencil (for example). Do not press on the tubing with your fingernail. 3. Screw the tip-holder onto the end of the probe. 4.
3 Connecting the Tubing Installation Connecting the Tubing Tubing connectors should be hand-tightened, tools should not be used as their use will damage the connector’s thread. Single-syringe Model Valve Inlet Tubing A length of Fluoride Ethylene Propylene (FEP) tubing, supplied with the 402, is used to connect the inlet port to the solvent reservoir. Screw the connector into the inlet port (left-hand) of the valve and put the other end into the reservoir.
3 Installation Connecting the Tubing After assembling the probe, the other end of the tubing is installed by screwing the connector into the right-hand port of the valve. When you use Gilson Diamond Tips, the special non-tapered tubing, included in the kit, must be fitted. Dual-syringe Model Valve Inlet Tubing A length of Fluoride Ethylene Propylene (FEP) tubing, supplied with the 402, is used to connect the inlet port to the solvent reservoir.
Connecting the Tubing Installation 3 The transfer tubing connects the outlet port (righthand) of the Tee-junction to the probe. A 2 mL tapered transfer-tubing is supplied as standard ; 1 mL, 5 mL, and 10 mL are available as additional accessories. Self-adhesive clips are provided for holding the transfer tubing securely against the side of the 402 (see previous page). The 1 mL transfer tubing is recommended for transferring volumes smaller than 1 mL.
3 Installation Electrical Connections Electrical Connections The rear panel of the 402 Dilutor-Dispenser houses: - sector block containing the power socket, fuse drawer and ON/OFF switch, - keypad connector, - probe connector, - GSIOC connector. Sector Block The sector block contains the power socket, fuse holder and ON/OFF switch. Your 402 can be connected to a 100-120 VAC or a 220-240 VAC supply. In either case, two 1A type “T” slow blow fuses must be installed (reference 6730104006).
3 Electrical Connections Installation Connecting the Power Cord Make sure that the ON/OFF switch is in the OFF (down) position. Connect the power cord to the power socket at the rear of the instrument and then to the power supply source. When the ON/OFF switch is put to the ON position, the instrument initializes; the valve motor rotates the spindle and stops with the valve set to the outlet position. The piston motor raises the piston to the upper position.
3 Installation Outlet Inlet The 402 Dilutor-Dispenser is supplied without syringes. Order a syringe (or syringes), appropriate to your Method, for your 402 Dilutor-Dispenser. Mount the syringe on the valve or Tee-junction. Assembling a Syringe The syringe and its piston are supplied assembled. However, if you remove the syringe from the piston, ensure that the piston and the piston nose are free from dust and contamination before reinserting the piston into the syringe.
3 Upgrading to Dual-syringe Model Installation 3-10 Upgrading to Dual-syringe Model The single syringe 402 can be upgraded to improve the versatility of the system. To upgrade to the Dual-syringe 402, you must install a Tee-junction module (kit reference F4015010) at the right-hand-side of the 402. You must first install the syringe module (includes motor etc.) and then fit the Tee-junction.
3 Installation Upgrading to Dual-syringe Model Removing the Cover 1) Undo the four screws (with two washers) at the base of the 402 that secure the top cover. Place the 402 in the upright position, pull the sides of the cover outwards, and then lift off the cover. 2) Undo the two nuts inside the front panel (near the top) that secure the blanking plate, then remove the blanking plate.
3 Upgrading to Dual-syringe Model Installation Inserting the Module Do not try to force the module into position as this may affect the alignment of the component parts. You should proceed carefully as follows. 1) Tilt the top of the module inwards and carefully insert the upper part of the module into the front aperture. When the syringe drive motor is inside the aperture, tilt the module into the upright position.
Operation - Standard Use 4 This chapter describes how to operate the 402 in standard use. Chapter 5 contains more detailed and advanced information, together with less frequently used options. You are advised to read the first section of Chapter 5, ‘Correct Use of the 402’, before proceeding. To use your 402 Dilutor-Dispenser, you must first install the hardware (syringe, probe, keypad, etc.) suitable to your Method, and then inform the 402’s software of the installation, this is called ‘Configuration’.
4 Essential Steps Operation - Standard Use Essential Steps Here is a summary of the things you must do, and the order in which you should do them. 1) Check that you have received all standard parts. 2) Connect the keypad. 3) Fit the fuses and connect the power supply cord. 4) Assemble and connect the probe (including the support). 5) Connect the tubings. 6) Switch on the power supply. 7) Enter the configuration details using the keypad. 8) Fit the syringe or syringes.
4 Operation - Standard Use Essential Steps If you have a Dual-syringe 402, install the tubings as described in Chapter 3, page 3-5. You must connect 4 pieces of tubing: inlet (reservoir), Valve to Tee-junction, transfer (probe), and waste. Switch-on 1) Switch on your 402 (the power switch is on the back panel): the indicator light on the front panel comes on, the keypad is illuminated, and briefly displays messages giving the software version numbers, Initialization Screen 1, for example.
4 Essential Steps Operation - Standard Use Configuration (Conf) You only have to enter the Configuration details one-time; the 402 keeps this information in its memory, even after you switch off. The next time you switch on the 402, the Configuration and all Methods previously saved as files will be available to modify or run. Configuration Screen The software must be given details about syringe sizes, flow rates, etc.
4 Operation - Standard Use Essential Steps If you want to change the configured volume, press Chge (until you reach the correct value); when the volumes are the same press ENTER, press the ESC key to return to the Configuration Screen, or press Next, after which you see the Air Gap Volume Screen. Key in the ‘Air gap volume’, press ENTER to confirm then press the ESC key to return to the Configuration Screen (see Chapter 5, for a definition of Air Gap).
4 Essential Steps Operation - Standard Use Next is used to go to the next screen (see Air Flow Screen), Prev is used to go to the previous screen (see Resv flow Screen). Resv Flow Screen Air Flow is the rate in mL/min. that air is drawn into the transfer tubing when aspirating air. Key in the required value, then press ENTER. Press the ESC key to return to the Configuration Screen or press Next to go to the Resv Flow Screen. Resv (Reservoir) Flow is the rate in mL/min.
4 Operation - Standard Use 1) First you must lower the piston rod(s), as follows: - press the Conf key to access the Configuration Screen: - press the Syr soft-key (ensure that the size is correct before continuing), - press the Exch soft-key. ‘Please wait ...’ appears on the screen whilst the piston rod(s) descend; then ‘Exchange the Syringe(s)’.
4 Essential Steps Operation - Standard Use In-run Prompts You can defer some parameters (volumes and flow rates) by entering a question mark (?) in place of the value. You enter the actual value after pressing the RUN key. This feature is particularly useful when optimizing parameters. In the following example, the aspirate flow rate (Asp Flow) is submitted as an unknown. Then, each time the Method is run, you can submit a different value (6, 12, 18).
4 Operation - Standard Use Press Chge until you see ‘Source: PROBE’, press ENTER to confirm, then press Next. Dispense Screen (1) Dispense Screen (2) Press Chge until you see ‘Fraction vol: SAME’, press ENTER to confirm, then press Next. Key in the number of fractions (up to 99 are possible) to dispense (3 in this example), press ENTER to confirm, then press Next. Key in the ‘Common vol:’ (20), press ENTER to confirm, then press Flow.
4 Essential Steps Operation - Standard Use Prime the 402 (Man) Manipulation Screen Initialization Screen 2 Prime is used to thoroughly clean out all fluid pathways with fresh solvent and to clear bubbles from the tubings. Press the Man soft-key, the Manipulation Screen appears. Press Prime, Initialization Screen 2 appears. [Measure and Titrate are described in Chapter 5.] You should prime the 402 before each run, to ensure that all fluid pathways contain fresh solvent and no air bubbles.
4 Operation - Standard Use Essential Steps ‘Homing ...’ appears on the screen until the syringe(s) have returned to their uppermost (home) positions, during which time any liquid contained in the transfer-tubing is expelled from the probe-end. After this the Action Screen reappears. Press ESC to return to the start-up screen for the current file. Whenever you see ‘Homing’ on the screen, the syringe piston(s) are returned to their uppermost position(s).
4 Operation - Standard Use Essential Steps Ensure that the probe-end of the transfer-tubing is in a suitable waste container, then press OK. ‘Homing ...’ appears on the screen until the syringe(s) have returned to their home positions, during which time any liquid contained in the transfer-tubing is expelled from the probe-end. After this the Action Screen reappears. Press ESC to return to the start-up screen for the current file.
4 Operation - Standard Use (iii) Put the tip of the transfer-tubing into the sample then press the command button to aspirate liquid. The In Run Screen (4a) command light is steady and you see alternately In Run Screens 4a and 4b. (iv) Put the tip of the transfer-tubing into a recipient vessel then press the command button to dispense the fraction, the command light is steady. Repeat for the other fractions. The command light is steady and you see alternately In Run Screens 5a and 5b.
Essential Steps Operation - Standard Use 4-14 4 Press ‘Abort’ to confirm that you wish to stop and you are returned to the Run Screen. Alternatively, you can bring the 402 to a halt by switching off the power, in which case you must ensure that the probe end of transfer-tubing is in a suitable waste container before switching on again.
Operation - Advanced Use 5 This chapter explains in details how liquids are aspirated and dispensed using the 402 Dilutor-Dispenser and gives in depth details of various software functions.
5 Correct Use of the 402 Operation - Advanced Use Correct Use of a 402 Single-syringe Model • Samples and reagents are aspirated from the probe side. • Liquid aspirated from the probe side may not enter the syringes. Thus, the tubing that connects the syringe to the probe, called transfer tubing, has to be carefully chosen to accommodate the volumes of samples and reagents defined in the Method. • Liquids should be separated by an air gap to minimize contamination.
5 Operation - Advanced Use Correct Use of the 402 • If a drop of liquid is present at the end of the tubing after dispensing, you must include this drop in the result vial. • The larger capacity syringe must always be mounted on the left-hand side. • Only the left-hand syringe aspirates the liquid from the reservoir (diluent and rinsing liquid).
5 Correct Use of the 402 Operation - Advanced Use Aspirating Large Volumes via the Probe (c) Left-hand Syringe Aspirating from Probe From Tee From Probe 1. The valve switches to connect the lefthand syringe to the Tee-junction and probe. 2. The piston of the right-hand syringe remains in the top position to prevent liquid from entering the syringe. 3. The left-hand piston descends to aspirate the specified volume of sample liquid.
5 Operation - Advanced Use Software Software You must first enter the configuration details for your 402 Dilutor-Dispenser. Then you may do one of three things: (i) Perform a direct manipulation (Prime, Measure, Titrate). (ii) Run a prefabricated Method (files 90 - 94). (iii) Create your own Method. A Method consists of a file saved in the 402’s memory, which can hold up to 99 files. Each file will contain one or more of the following tasks: Dilute (Dil), Dispense (Disp), Pipette (Pip), Rinse, and Mix.
5 Operation - Advanced Use Software 4 Press the Next soft-key to go to the following screen. 5 Press the Chge soft-key to choose an option or to select from a list of possible values. 6 Press the ESC key to return to the level of the previous screen or menu. 7 Press CLEAR to remove the value (before pressing ENTER) and return to the previous value. 8 Press Deflt to select the default value or option.
5 Operation - Advanced Use Software If you want to change the configured volume, press Chge (until you reach the correct value); when the volumes are the same press ENTER, press the ESC key to return to the Configuration Screen, or press Next, after which you see the Air Gap Volume Screen. Key in the ‘Air gap volume’, press ENTER to confirm then press the ESC key to return to the Configuration Screen.
5 Operation - Advanced Use Software When you specify left and right syringe capacities (left must be > right), you must enter compatible volumes. An error message is displayed if you try to enter an incompatible volume, in which case you may need to change the volume of the other syringe, before entering the volume of the one that provoked the error message. Syringe Correction Screen When you turn the syringe correction factor ON, a plus sign (+) appears after the file number, as a reminder.
5 Operation - Advanced Use Software Counters Under normal use you should expect to fit new (or refurbished) syringes after 30000 cycles with water. This feature is made easy by the 402, which automatically counts the number of operating cycles performed by the syringe(s) and valve. (A cycle is completed when the syringe leaves its home position.) You should only need to reset these counters after changing one or more of these items. To do this, press Count, the Syringe Counter Screen appears.
5 Software Operation - Advanced Use Syringe Correction Screen to access the required task and its parameters, (ii) if you have entered a syringe correction factor in the configuration process, the Syringe Correction Screen appears first, after which you select ON or OFF using the Chge soft-key, press ENTER, and then Next, after which the Task Selection Screen appears; you can now access the screens for the task and enter the task related parameters (see page 5-15).
5 Operation - Advanced Use Press ESC to return to the New File Screen, or press Dir to access the file directory. New File Directory Screen New File Screen Software When you select Yes, the file is deleted from the directory and you see the New-File Directory Screen. Copy a File This action applies to the file currently selected; so to copy another file you must first key in the number of the file to be copied and press ENTER. Press Copy, the Copy Screen appears.
5 Software Operation - Advanced Use Locked File Screen To unlock this file you must key in the code that was used to lock the file. You may not edit a locked file, so the Edit soft-key is replaced by the View soft-key, which allows you to browse through the contents of that file, without making any changes. However, you are permitted to copy a locked file. After copying the original file remains locked but the copied file is unlocked automatically by the software.
5 Operation - Advanced Use Software Press Abort, when you want to stop priming; Initialization Screen 2 reappears. Ensure that the probe-end of the transfer-tubing is in a suitable waste container, then press OK. ‘Homing ...’ appears on the screen until the syringe(s) have returned to their uppermost (home) positions, during which time any liquid contained in the transfer-tubing is expelled from the probe-end. After this the Action Screen reappears.
5 Software Operation - Advanced Use Measurement Screen After completely aspirating the liquid being measured, you see (for example) the Measurement Screen. Procedure 1) Aspirate all the liquid contained in the source vial, plus an air cushion, to ensure that you have aspirated all the liquid. You should press the command button to stop aspirating when you can see the air/liquid interface in the transfer tubing. 2) Press the Asp key to reverse the direction of flow; (Asp is replaced by Disp).
5 Operation - Advanced Use • Press Titrate, Titrate Screen (1) appears. Select either reservoir or probe as the source using the Chge soft-key, then press ENTER to confirm your choice. • Press the Next soft-key, key in the volume to aspirate, then press ENTER, Titrate Screen (2) appears. Normally, you would aspirate sufficient liquid for several titrations.
Software Operation - Advanced Use 5 Keys The use of the soft-keys is as follows: - Prev is used to go to the previous task (or screen within a task), - Next is used to go to the next task (or screen within a task), - Add is used to add a task to the Method (after the task displayed), - Param is used to access the parameters for the current task, - Ins is used to insert a task (before the task displayed), - Del is used to remove the task currently displayed.
5 Operation - Advanced Use Minimum Maximum Air-gap Volume (in Configuration) Dilution Ratio (Dual-syringe) Dilution Ratio (Single-syringe) Number of Serial Dilutions Number of Reagents Number of Fractions Number of Cycles (mixing) Sample Volume (Vs) Reagent Volume (Vr) Diluent Volume (reservoir) Diluent Volume (via probe) Common Volume (reservoir) Common Volume (via probe) Total Volume (/) Total Volume (via probe) Extra Volume Final Volume (adjust via probe) Final Volume (adjust from reservoir) Flow Rate
5 Software Operation - Advanced Use Dilute If you specify a volume of sample greater than that of the righthand syringe, the left-hand will be used to aspirate the sample. Three types of dilution task are available ‘Normal’, ‘Unknown’, and ‘Serial’. Three modes are available for ‘Normal’ and ‘Serial’: Total Volume, Diluent Volume, and Dilution Ratio. The first action is to select the type of dilution task that you are going to use, by pressing the appropriate soft-keys.
5 Operation - Advanced Use Software Press Next, then key in the sample volume, press ENTER. Depending on the mode, you see one of three screens after pressing Next. Key in the value, press ENTER. Optionally, for any mode, you can display the variables calculated by the software by pressing Data. Optionally, you can modify the flow rates; press Flow, For any mode, key in a value for Asp flow, press ENTER, press Next, key in a value for Disp flow, press ENTER.
5 Operation - Advanced Use Software For each dilution in the series: Vt = Vs x Rd, Vd = Vt - Vs For example, after selecting Dilution Ratio Mode, you enter Sample Volume = 10 and Dilution Ratio = 3; the software calculates the Total Volume as 30 and the required Diluent Volume as 20. 1. When diluent comes from the reservoir (RESV); aspirate the required volume of diluent (for all dilutions, up to the capacity of the left-hand syringe).
Operation - Advanced Use Software Optionally, for any mode, you can display the variables calculated by the software by pressing Data. Optionally, you can modify the flow rates; press Flow, For any mode, key in a value for Asp flow, press ENTER, press Next; key in a value for Disp flow, press ENTER. Optionally, for any mode, you can mix the diluted liquid; press Mix. The volume used for mixing is 90% of the diluent volume. Key in number of cycles, press ENTER.
Software Operation - Advanced Use 5 Optionally, you can supply up to 12 reagents, press Reag, key in number of reagents, press ENTER. Press Next, then for each reagent, key in volume press ENTER. After the data are entered the Reag key changes to REAG. Press ESC until you see this screen (three times if you have added reagents) Press ESC again to reveal the Save Screen; select Yes, the Method is then saved, after which you see the Run Screen.
5 Operation - Advanced Use Software If for any reason you want to repeat the measurement (e.g. you need to select the LEFT syringe instead of the RIGHT), put tip of the transfertubing back into the original container, then press Redo to discharge the liquid. When you are ready to dilute the previously measured sample, press End. The command light is steady and you see alternately the next pair of screens.
5 Software Operation - Advanced Use volume of liquid, is dispensed, at the end of the process, into the waste. When you configure disposable tips in air mode, ‘Extra volume’ is replaced by ‘Air push volume’ in this task. 1. 2. 3. 4. Aspirate an air gap. Aspirate liquid via the probe. Dispense fractions via the probe. Dispense extra volume to waste. Typical Task Entry Key in the File # (e.g. 45), ENTER. Press Edit. Disp, then press Chge to switch source.
5 Operation - Advanced Use Software Press ESC (twice). Press ESC. Pipette You use this task to aspirate a series of fractions via the probe, then dispense the total volume aspirated into a recipient vessel. Each liquid aspirated is separated by an air gap (if air gap is configured as > 0). You may also aspirate an ‘Extra volume’ to ensure the accuracy of the first fraction. This ‘Extra volume’, which is aspirated with the first fraction, is dispensed at the end of the process, into the waste. 1.
5 Software Operation - Advanced Use Optionally at the first of the volume screens, press Flow. Key in a value for Asp flow, press ENTER. Press Next. Key in a value for Disp flow, press ENTER. Optionally at the first of the volume screens, press Extra, Key in ‘Extra volume’, press ENTER. Press ESC (twice). Press ESC. Rinse The purpose of this task is to minimize carry-over by flushing the transfer-tubing and syringes with solvent from the reservoir, between tasks.
Operation - Advanced Use Press Chge to switch modes. Press ENTER, then Next. Key in volume, press ENTER. Optionally, press Flow. • For AIR mode, key in a value for Disp flow, press ENTER. This mode uses the configured value for Asp. Flow (default = 4 mL/min). • For ASP/DISP mode only, key in a value for Asp flow, press ENTER, press Next, then key in a value for Disp flow, press ENTER. Press Next, then key in number of cycles. Press ENTER. Press ESC. Press ESC.
5 Software Operation - Advanced Use Key in the file number of the second Method to link(8), press ENTER, press Next. Key in the file number of the third Method to link(9), press ENTER, press Next. Key in the file number of the fourth Method to link(8), press ENTER, press Next. After a file is linked in this manner it can not be deleted. Press ESC to access the Save Screen, press Yes.
5 Operation - Advanced Use TASK/Parameter Method 92 Dispense Sample Source Fractions Nb of Fractions Common Volume Reservoir (RESV) Same 3 1000 Reservoir (RESV) Same 3 1000 Dispense Sample Source Fractions Nb of Fractions Fraction 1 Fraction 2 Fraction 3 Adjust Source Final Volume Probe Different 3 50 100 200 Reservoir (RESV) 1000 Probe Different 3 500 700 500 Reservoir (RESV) 1000 Pipette Fractions Nb of Fractions Fraction 1 Fraction 2 Fraction 3 Fraction 4 Different 4 50 80 100 200 Different 4
5 Software Operation - Advanced Use When you have finished, or wish to change programs, press STOP, to instantly halt the program. Alternatively, you can bring the 402 to a halt by switching off the power, in which case you must ensure that the probe end of transfer-tubing is in a suitable waste container before switching on again. Pressing STOP When you press STOP, the motion of the syringe(s) ceases and the Abort Screen appears. Press Proceed, when you are ready to continue with your Method.
5 Operation - Advanced Use Flow Rates Flow rates depend on the nature of the liquid (viscosity), the capacity of the syringe, and the internal diameter of tubing. For liquids transferred via the probe, it is possible to adjust the aspirate and dispense flow rates according to the nature of the liquid (e.g. viscous liquids must be aspirated slowly). The default value for ‘Air flow rate’ is 4 ml/min. Whatever the capacity of the syringe(s).
5 Additional Information Operation - Advanced Use Maximum Dispense Flow Rate (No Air Gaps) Tubing Type 1 mL 2 mL 5 mL 10 mL Micro Tips (liquid) Micro Tips (air) Large Tips (air) 100 250 Syringe Volume (µL) 500 1000 5000 4 4 4 4 4 4 4 10 10 10 10 10 10 10 20 20 20 20 20 20 20 Recommended Flow Rate with an Air Gap (10 µL) Syringe size 100 µL 250 µL 500 µL 1 mL 5 mL 10 mL 25 mL Flow rate (mL/min) 4 10 20 30 Asp: 30 - Disp: 50 Asp: 30 - Disp: 50 Asp: 30 - Disp: 50 40 40 40 40 40 40 40 120 120 120
Operation - Advanced Use A correction factor may be applied to all volumes of liquid to compensate for the effects of temperature (for example, when the 402 is used in a culture room) or for the effects of viscosity. This factor should be calculated from gravimetric assays and entered in the Configuration menu. With the Dual-syringe model, you may apply different corrections to each syringe.
5 Additional Information Operation - Advanced Use Gilson D200 tips are used for volumes from 1 µL to 78 µL (with a 100 µL syringe, minimum recommended air-push-volume is 20 µL). Gilson D1000 tips are used for volumes from 50 µL to 900 µL (minimum recommended air-push-volume is 100 µL). The liquid is ejected with an Air push volume of at least 20 µL when using a Gilson D200 tip and 100 µL when using a Gilson D1000 tip.
Maintenance 6 This chapter explains how to perform the routine maintenance procedures. The only maintenance operations necessary for the 402 DD are: • Cleaning the exterior of the instrument. • Cleaning of the valve, Tee, syringe assemblies and the tubings. • Replacement of the piston syringe after approximately 30 000 cycles.
6 Cleaning Maintenance Cleaning Cleaning the Exterior The instrument cover and front panel may be cleaned. Ensure that the instrument is switched OFF and the power cord is removed when you clean the instrument. When you need to clean the instrument, use one of the three following methods: 1) A clean dry cloth. 2) A cloth dampened with water. 3) A cloth dampened with soapy water. If a cloth dampened with soapy water is used to clean the instrument, only domestic soap may be used.
6 Maintenance Cleaning 1) Pull the axle away from the valve main body. 2) Remove the ceramic stator from the valve axle. 3) Tap the valve inner body against a clean, solid surface to remove the spring and PTFE spring cover. 4) Do not remove the inner ceramic surface from the valve main body. 5) The parts can be cleaned with a non-abrasive cloth dampened with alcohol, or by autoclaving them. 6) Dry the components using a lint-free cloth.
6 Cleaning Maintenance Cleaning the Syringe Assembly The 25 mL syringe is not autoclavable, all of the other size syringes are autoclavable. You must dismount the syringe and piston assembly from the instrument and then separate the syringe and piston assembly before cleaning or autoclaving them. 1) Dismount the Syringe assembly from the instrument. 2) Undo the piston securing screw to free the piston assembly from the piston operating rod.
6 Maintenance Autoclaving Autoclaving Certain parts of the 402 can be autoclaved (20 min. at 0.1 MPa, 121° C). They must be removed from the instrument for autoclaving. The autoclavable parts are: • The valve (disassembled) or the Tee-junction. • The syringe (except the 25 mL syringe). • The piston assembly. • PTFE and FEP tubing. Autoclaving the Tubing The tubing can be cleaned or autoclaved. This procedure prevents the end of the tubing from deforming.
6 Chaning the Syringe Piston Assembly Maintenance Item Part Number 100 µL piston kit F4015072 250 µL piston kit F4015073 500 µL piston kit F4015074 1 mL piston kit F4015075 5 mL piston kit F4015076 10 mL piston kit F4015077 25 mL piston kit F4015078 Changing the Syringe Piston Assembly For all syringe sizes, it is necessary to replace the piston assembly after approximately 30 000 cycles (using water).
6 Maintenance The piston nose should be replaced after the pump has performed approximately 30 000 cycles. All piston noses are replaceable, the process is relatively easy, but for most pistons (except the 25 mL pistons), special tools are required to mount the piston nose. Part Number 100 µL Piston: F4015062 Description F4015072 5 x piston noses, 2 x support collars and mounting tool (for 100 and 250 µL pistons).
6 Changing the Piston Syringes and Piston Noses Maintenance 1. Remove the syringe from the pump as described in Chapter 4. Piston rod Piston button Allen screw Piston nose Nose holding tool 2. Slide the piston out of the syringe barrel. 3. Remove the piston nose from the piston rod by carefully pinching the piston nose with a pair of cutters and gently pulling the piston nose off the piston rod. Take care not to damage the end of the piston rod when removing the nose.
6 Maintenance 1. Remove the syringe from the pump as described page 6-4. 2. Slide the piston out of the syringe barrel. Part Number Description F4015064 5 x 500 µL piston noses and mounting tool kit 3. Remove the piston nose from the piston rod by carefully pinching the piston nose with a pair of cutters and gently pulling the piston nose off the piston rod. Take care not to damage the end of the piston rod when removing the piston nose. 4.
6 Changing the Piston Syringes and Piston Noses Maintenance Cover seal Valve screw Syringe body Before Mounting a 500 µL Syringe: To optimize the fitting of the 500 µL syringe on the Dilutor valve, the 500 µL syringe is supplied with a cover seal. The orientation of this seal must be respected to ensure that the connection with the syringe is airtight. (See the opposite figure). Changing the 1 mL Piston Noses You need the special tool for mounting the piston nose on the 1 mL piston.
6 Maintenance You don’t need a special tool for mounting the piston nose on the 25 mL piston. The nose screwfits onto the piston shaft. Part Number Description F4015068 1. Remove the syringe from the pump as described page 6-4. 1 x 25 mL piston nose. 2. Slide the piston out of the syringe barrel. It may be necessary to screw the piston securing screw back into the bottom of the piston and pull on the screw to remove the piston from the syringe barrel. 3.
6 Changing the Piston Syringes and Piston Noses Maintenance 6-12 5. Insert the new piston nose in the recess of the piston nose holder. Make sure that the aperture is facing outward and the O-ring is inside the seal. 6. Place the piston shaft in the piston shaft holder. 7. Screw the two parts of the tool together until tight. 8. Unscrew the tool, the piston nose should be correctly seated on the shaft. 9. Dip the piston nose in distilled water and slide the piston into the syringe. 10.
Troubleshooting 7 This chapter explains how to diagnose possible problems and how to overcome them.
7 Mechanical Problems Troubleshooting 7-2 Mechanical Problems Problem Possible Cause Solution Valve does not fit on to the 402. Valve axle not properly aligned with the motor drive. Ensure that the flat on the axle is aligned with the drive slot on the motor. Valve moves during aspirate or Valve mounting screw not tight. dispense. Check that the mounting screws, syringe and syringe mounting screw are all tight. Syringe does not move properly. Tubing blocked.
7 Troubleshooting Problem Possible Cause Solution Liquid leakage Connections incorrectly made. Tighten all tubing connections. Liquid leaving by the waste tube. Tubing blocked. Check all tubing for blockages. Ensure that the inlet filter is clear. Dirt trapped between the ceramic surfaces. Clean the valve. Refer to page 6-2.
7 Electrical Problems Troubleshooting 7-4 Electrical Problems Problem Possible Cause Solution Instrument does not respond. Power cord not connected. Check power cord is properly connected. 402 is not switched on. Check ON/OFF switch. Fuse blown. Check the fuses. See page 3-7.
Parts & Accessories Lists Appendix A Parts lists for the 402 Dilutor Dispenser, consisting of Standard Equipment, Additional Accessories, and Spare parts.
402 Single Syringe Parts & Accessories Lists Appendix 402 Single Syringe Part Number F410514 Qty Description 1 402 Dilutor Dispenser (Single-syringe) F4015016 1 Probe (standard) F40151403 1 Probe support 4011534125 1 Fixing screw for probe support 130221 1 Keypad F4015032 1 Tubing, tapered transfer (probe), FEP (2 mL, 1300 x 1.
Parts & Accessories Lists Part Number Qty Description F410515 1 402 Dilutor Dispenser (Dual-syringe) F4015016 1 Probe (standard) F40151403 1 Probe support 4011534125 1 Fixing screw for probe support 130221 1 Keypad F4015032 1 Tubing, tapered transfer (probe), FEP (2 mL, 1300 x 1.
Additional Accessories Parts & Accessories Lists Appendix Additional Accessories Part Number Description F4015015 25025341 Adapter kit for disposable tips 100 µL syringe 25025342 250 µL syringe 25025347 500 µL syringe 25025343 1 mL syringe 25025344 5 mL syringe 25025345 10 mL syringe 25025346 25 mL syringe F4015031 Tubing, tapered transfer (probe), FEP (1 mL), (1 x 2 x 1300 mm) F4015033 Tubing, tapered transfer (probe), FEP (5 mL), (1.5 x 2.
Parts & Accessories Lists Part Number Description F4015072 100 µL piston kit F4015073 250 µL piston kit F4015074 500 µL piston kit F4015075 1 mL piston kit F4015076 5 mL piston kit F4015077 10 mL piston kit F4015078 25 mL piston kit A Spare Parts Spare Parts Appendix Appendix A-5
Parts & Accessories Lists Appendix A-6 Appendix A
Chemical Compatibility Appendix B This chapter contains information about the chemical compatibility of the syringes and the tubing.
Chemical Compatibility Chemical Compatibility Appendix B Chemical Compatibility Materials in Contact with Liquids In the 402 Dilutor Dispenser flow-path, several materials are in contact with liquids: FEP PTFE/Ekonol PTFE Inlet tubing, Transfer tubing Piston nose of the syringes (except 25 mL) Piston nose of the 25 mL syringe Borosilicate glass Syringe Ceramic Valve PEEK Valve, Tee junction Compatibility The syringe assemblies and the tubings are compatible with a wide range of liquids: Appendix B-2
Technical Data Appendix C This chapter contains the Technical Data concerning the 402 Dilutor Dispenser.
Hydraulics Technical Data Appendix C Hydraulics Dilution Ratio When using the dual syringe model equipped with a 25 mL syringe on the left and a 100 µL syringe on the right, the dilution ratio is 1/25000, but in order to stay within the RSD range a maximum ratio of 1/500 is recommended. Minimum Volume The minimum programmable transfer volume is 1 µL.
Technical Data Appendix Syringe capacity (µL) 100 250 500 1000 5000 10000 25000 Maximum Flow (mL/min) 4 10 20 40 120 120 120 Minimum Specified Volume (µL) 10 25 50 100 500 1000 2500 Nominal Volume (µL) 100 250 500 1000 5000 10000 25000 Hydraulics Syringes C Performance Specifications The following values are obtained using a defined protocol. If this protocol is followed, the obtained values must be inferior or equal to the listed figures.
Hydraulics Technical Data Appendix C The same specifications are obtained in the dispensing mode with liquid from the reservoir. See OQ Document for procedure and calculations. For 1 % syringe capacity, typical values are given. Syringe Volume capacity (µL) pipetted (µL) Accuracy % µL % RSD µL 100 1 5.0 0.05 3.0 0.03 250 2.5 3.0 0.075 2.0 0.05 500 5 3.0 0.15 2.0 0.1 1000 10 3.0 0.3 1.0 0.1 5000 50 3.0 1.5 0.8 0.4 10000 100 3.0 3 0.5 0.5 25000 250 3.0 7.5 0.
Technical Data Appendix C Mechanics Mechanics Valve Type: Three port, two position. Angle between ports: 120°. Valve drive: Stepper motor. Rotation time: 0.5 Second. Liquid contact materials: PEEK and ceramic. Lifetime: 1 000 000 cycles. Dead volume: 123 µL. Maximum Pressure: 0.8 MPa. Tee Junction Type: Three port, no moving parts. Diameter of channels: 1.5 mm. Length of channels: 14 mm. Angle between channels: 120°. Liquid contact material: PEEK. Dead volume: 51 µL.
Environmental Technical Data Appendix C Environmental Conformity to Norms Refer to separate leaflet LT801354 ‘Declaration of Conformity’. Storage Indoor use only. Installation: Category II. Altitude: operating storage Temperature range: operating storage up to 2000 m. up to 4600 m. 4°C to 40°C. -15°C to +40°C. Pollution degree 2. Humidity: Maximum relative humidity 80% for temperatures up to 31°C decreasing linearly to 50% relative humidity at 40°C. Power Requirements Frequency: 50 to 60 HZ.
Error Messages Appendix D This chapter contains the Error Messages concerning the 402 Dilutor Dispenser.
Error Messages Error Messages Appendix D Error Messages Error Message Cause/Possible Action Adjust volume too small In DISPENSE, you have, you have entered a value for “Final vol.” that is too small. “Final vol.” > volume of biggest fraction dispensed. Asp flow missing Re-enter Method, supplying the missing parameter, when prompted to do so (see DILUTE, DISPENSE, PIPETTE, and MIX tasks).
Error Messages Appendix Cause/Possible Action Parameters incompatible! Refer to page 5-16. Ratio impossible Dilution ratio calculated by software is not valid; check parameters using formulae given in page 5-18 (DILUTE task) and correct one or both of the other values. Ratio missing Re-enter Method, supplying the missing parameter, when prompted to do so (see DILUTE task). Reservoir not allowed Liquids must be aspirated using the probe.
Error Messages Appendix D-4 Appendix D
0 Index Abort 4-10, 4-11 Accessories list 4-5 Acetonitrile B-2 Air bubble free filling 4-10 Air Gap 4-4, 4-12, 5-2, 5-6, 5-17 Alcohol B-2 Alternative tubing 3-4 Autoclaving 6-5 parts 6-5 tubing 6-5 Autorefill 4-6, 5-9 Beeper 4-6, 5-9 Cable 3-8, 3-11 Chemical Compatibility B-2 Clamp 2-2, 3-2 Cleaning the instrument - exterior 6-2 the syringe assembly 6-4 the Tee junction 6-3 the tubing 6-5 the valve 6-2 CLEAR 2-4, 5-6 Clip 2-2, 2-3, 3-4, 3-5 Configuration 4-4 Conformity to Norms C-6 Connecting cable 3-8 k
Index Installation Keypad Large Volumes Liquid contact materials Link Lock 3-1 2-2, 3-8 5-4 B-2 5-27 5-11 Maintenance Autoclaving 6-5 Changing the piston assembly 6-7 Cleaning the instrument 6-2 Cleaning the syringe assembly 6-4 Cleaning the tubing 6-5 Cleaning the valve 6-2 Disassembling the valve 6-2 Dismounting the valve 6-2 Measure 5-13 Mechanical problems 7-2 syringes C-5 tee-junction C-5 valve C-5 Method Example 4-8 Method entering 4-7, 5-9 linking 5-27 prefabricated 5-29 storing 4-9, 5-10 Microswi
Index Titrate Troubleshooting electrical problems hydraulic problems mechanical problems Tubing connecting transfer valve inlet valve waste Typical Values 5-14 Union connector Unlock Unpacking Upgrading 6-5 5-11 1-5 3-10 Valve disassembly dismounting inlet tubing connections outlet connections Viscous liquids Voltage range selection Volumetric analyses Warranty 7-4 7-3 7-2 3-4 3-4 3-4 3-5 C-4 6-2 6-2 3-4 3-5 5-33 C-6 3-7 5-31, C-3 1-6 Index-3
Index Index-4
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