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Knauer PLATINblue AS-1 User Manual

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PLATINblue
Autosampler AS-1
User Manual
V6980A
UHPLC

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  • Page 1 PLATINblue Autosampler AS-1 User Manual V6980A UHPLC...

  • Page 2: Table Of Contents

    Air Segment with μl-Pick-Up Injection ......KNAUER AS-1User manual V6980A, version 1.2...

  • Page 3: Fig. 47: Configuration Window For Chromgate

    Operation Qualification Report ........KNAUER AS-1User manual V6980A, version 1.2...
  • Page 4 Table of Figures ..........KNAUER...

  • Page 5: Intended Use

    Usage is possible in laboratories of universities and research insti- tutions and in industrial and manufacturing routine laborato- ries in the following areas:  Biochemistry analyses  Chemical analyses  Food analyses  Pharmaceutical analyses  Environmental analyses KNAUER AS-1User manual V6980A, version 1.2...

  • Page 6: Features

    Since UHPLC generally uses columns with an inner diameter of 2 mm, we recommend using an injection volume of 1-5 μl for these applications. Optional Device Compo- KNAUER offers the device in two variations: nents  Autosampler without temperature control ...

  • Page 7: Eluents

    Fomblin®  Methanol  Phosphate buffer solu- tions (0.5 M)  Toluol  Ammoniated dilute solution  Acetic dilute acid (10- 50%), at 25° C  Dilute sodium hydrox- ide (1M)  Water KNAUER AS-1User manual V6980A, version 1.2...

  • Page 8: Safety For Users

     National and international regulations pertaining to labora- tory work  Original spare parts, tools, and eluents made or recom- mended by KNAUER  Good Laboratory Practice (GLP)  For development of methods and validation of devices: Pro- tocol for the Adoption of Analytical Methods in the Clinical...

  • Page 9: Definition Of Personal And Material Damages

    Type of damage Category Explanations Personal and material DANGER! Lethal or very serious injuries can occur. damages WARNING! Serious injuries can occur. CAUTION! Moderate injuries can occur. Device defect can occur. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 10: Symbols And Signs

    EU directives and comes with a dec- laration of conformity from the manufacturer. Warning signs Heavy lifting hazard, danger to the back Electrostatic-discharge hazard Danger of stab wounds HIgh-voltage hazard Mandatory signs Wear protective gloves. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 11: Installation

    1 x large solvent bottle Optional: Temperature control Checking the Scope of Delivery 1. Check whether the device and accessories are complete. 2. If a part is missing, inform the technical support of KNAUER. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 12: Unpacking And Setup

    You have various options to contact the Technical Support: Phone +49 30 809727-111 +49 30 8015010 E-mail support@knauer.net You can make your requests in English and German. Location Requirements Requirements The location for the device must meet the following require- ments: ...

  • Page 13: Unpacking

    6. Check for damages caused during transportation. In case you notice any damage, contact the technical support. 7. Set-up the device in its location. 8. Remove the adhesive tape from the door of the autosampler that was used as transportation protection. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 14: Adjusting The Height With Additional Feet

    PLATINblue P-1 high/pressure pump Separate feet Fig. 2 PLATINblue AS-1 with elevated additional feet CAUTION! Damage to the back when lifting or carrying the device alone! Together with the coolunit, the autosampler weighs approx. 21 kg without packaging. Therefore, the autosampler should be transported and set up by two persons.

  • Page 15: Fig. 3: Platinblue As-1 Autosampler Device Foot

    Installation Fastening the device feet Steps Figure 1. Lie the autosampler on its side. Fig. 3 PLATINblue AS-1 Auto- 2. Using an Allen wrench sampler device foot , loosen the Allen screw on the sepa- rate foot a few turns.

  • Page 16: Startup

    Front View of the Device Opening the door • Position your hands on both sides of the door and open. As shown in the figure, push the door inwards. Fig. 7 Push the door into the interior KNAUER AS-1User manual V6980A, version 1.2...

  • Page 17: Front View With Sample Compartment

    Legend Valve Syringe Needle guide Collecting container Flushing solution bottle Removing the cooler cover Tube connector for waste liquid Tube connector for Fig. 10 Front panel of the device with sample compartment condensed water KNAUER AS-1User manual V6980A, version 1.2...

  • Page 18: Rear View Of The Device

    LAN into your company network. The description applies to the operating system Windows® and all conventional routers. Note To set up a LAN, we recommend to use a router. The following steps are necessary: KNAUER AS-1User manual V6980A, version 1.2...

  • Page 19: Configuring The Lan Settings

    HPLC system separate from the company network. Note You will find patch cables for each device and the router in the accessories kit. To connect the router to a WAN, an additional KNAUER AS-1User manual V6980A, version 1.2...

  • Page 20: Configuring The Router

    1. Check that the IP-address range of the router and of the com- pany network do not overlap. 2. In case of an overlapping, change the IP-address range of the router. 3. Use the patch cable to connect the router WAN port to the company network. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 21: Controlling Several Systems Separately In A Lan

    4. If the router is integrated into a company network, pull out the patch cable from the WAN port.  Can the devices communicate with the computer, even though the router is disconnected from the com- pany network? KNAUER AS-1User manual V6980A, version 1.2...
  • Page 22  Has this been successful? 7. Replace the patch cable to the device with that no connection could be established.  Has this been successful? KNAUER AS-1User manual V6980A, version 1.2...

  • Page 23: Pasa™ Loop Injection Principle

    3 x loop volume for loops up to 100 μl The sample loss per injection is the sum of the x-time overfilling of the sample injection and the flush volume set for the employed needle. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 24: Full Loop Injection

    INJECT position. Sample needle and air needle are inserted into the vial. The air needle creates pres- sure, which prevents air and steam bubbles from developing. Fig. 14 Full loop injection: Initial conditions KNAUER AS-1User manual V6980A, version 1.2...

  • Page 25: Fig. 15 Full Loop Injection: The Needle And Sample Lines Are Flushed

    Full loop injection: The needle and sample lines are flushed 3. The valve switches to LOAD position in order to transport the sample material to the inlet of the sample loop. Full loop injection: Valve switches to LOAD position Fig. 16 KNAUER AS-1User manual V6980A, version 1.2...

  • Page 26: Needle Flushing

    The flush volume must be at 30 μl in case of a standard needle and an injection with air segment; otherwise, 35 μl need to be flushed. You may need to insert a higher flush volume for KNAUER AS-1User manual V6980A, version 1.2...

  • Page 27: Partial Loop Fill

    Increase the flush volume and reduce the syringe speed to achieve better results for highly viscous samples. Legend Needle Sample loop Sample Buffer tube Eluent Fig. 20 Air segment for partial loop fill Flush segment Air segment KNAUER AS-1User manual V6980A, version 1.2...

  • Page 28: Fig. 21: Scheme Of Initial Position For Partial Loop Fill

    The partial loop fill is processed automatically. Functional schematic Explanations 1. The valve switches to the INJECT position. Tube and capillaries are full of flush solution. Fig. 22 Partial loop fill: Valve position "Inject" KNAUER AS-1User manual V6980A, version 1.2...

  • Page 29: Fig. 23 Partial Loop Fill: Aspirate Sample

    Partial loop fill: Valve position "Load" 4. The valve switches to the INJECT position and the pump transports the sam- ple and the eluent to the column. Fig. 25 Partial loop fill: Inject into the column KNAUER AS-1User manual V6980A, version 1.2...

  • Page 30: Fig. 26 Partial Loop Fill: Aspirate Flushing Solution

    Fig. 26 Partial loop fill: Aspirate flushing solution 6. The syringe injects the flushing solution into tube and capillary. Both sample and eluent get flushed. Fig. 27 Partial loop fill: Flush sample and eluent KNAUER AS-1User manual V6980A, version 1.2...

  • Page 31: Microliter Pick-Up Injection

    Fig. 28 Microliter pick-up injection: Initial conditions compatible. 3. The valve switches to LOAD. A washing fluid segment is aspirated into the sample loop. Fig. 29 Microliter pick-up injection: Sample line is filled with eluent KNAUER AS-1User manual V6980A, version 1.2...

  • Page 32: Fig. 30 Microliter Pick-Up: Sample Is Aspirated

    Fig. 30 Microliter pick-up: Sample is aspirated 5. The sample is aspirated from the sample vial according to the amount of programmed injection volume. Fig. 31 Microliter pick-up: The injection volume is aspirated. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 33: Fig. 32: Microliter Pick-Up: The Sample Is Transported Through

    HPLC flow of the mobile phase. The sample is transported to the column. The analysis starts. Fig. 33 Microliter pick-up: The sample is transported to the column. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 34: Air Segment With Μl-Pick-Up Injection

    Legend Needle Transport segment Sample loop Sample Air segment Fig. 34 Microliter pick-up injection with air segment (A), with- out air segment (B) Buffer tube Eluent KNAUER AS-1User manual V6980A, version 1.2...

  • Page 35: Air Needles

    Legend Deep Greiner Bio- ® microtiter plate with closure. Shallow Greiner Bio- ® microtiter plate ® Fig. 36 Standard air needle with Greiner Bio-One microtiter plates KNAUER AS-1User manual V6980A, version 1.2...

  • Page 36: Choosing The Correct Air Needle

    Protrusion length 50 mm, yellow 34-40 mm 56 mm, red 28-34 mm 62 mm, white (standard nee- 22-28 mm dle) 68 mm, blue 16-22 mm 74 mm, green 10-16 mm 80 mm, black 4-10 mm KNAUER AS-1User manual V6980A, version 1.2...

  • Page 37: Fig. 38: Selected Air Needles With Sample Vials

    Legend Deep Greiner Bio- ® microtiter plate with closure., 56 mm air needle Shallow Greiner Bio- ® microtiter plate, 80 mm air needle ® Fig. 39 Selected air needles with Greiner Bio-One microtiter plates KNAUER AS-1User manual V6980A, version 1.2...

  • Page 38: Calculation Example For Air Needle

    Do not use sample vials that are unclosed.  Only use air-tight closure seals to prevent air bubbles from forming and volatile components from evaporating.  Do not use sample vials with hard closures that the sample needle cannot pierce. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 39: Mixing And Diluting

    If this is preceded by an ADD ... to Destination command, mixing is performed in the destination vial. If this is preceded by an ADD ... to Sample command, mixing is performed in the sample vial. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 40: Sample Positions In Mix Method

    Processing in Columns When column processing is used, the following positions are possible for the sample, destination, reagent A and reagent B: Fig. 40 Sample positions in columns KNAUER AS-1User manual V6980A, version 1.2...

  • Page 41: Processing In Rows

    Mixing and Diluting Processing in Rows When row processing is used, the following positions are possi- ble for the sample, destination, reagent A and reagent B: Fig. 41 Sample positions in rows KNAUER AS-1User manual V6980A, version 1.2...

  • Page 42: Capillary And Hose Connections

    Connector for tube buffer to syringe Plastic capillary to injection needle 1 2 3 4 5 6 Sample loop Fig. 43 Connectors of special injection valve Connection from stain- less steel capillary to pump KNAUER AS-1User manual V6980A, version 1.2...

  • Page 43: Connecting The Syringe

    Legend Tube guide for elu- Tube guide for flush- ing solution Collecting container with hole for tube guide (flushing fluid) Fig. 45 Tube guide for flushing solution tube KNAUER AS-1User manual V6980A, version 1.2...

  • Page 44: Connecting The Drainage Tubing

    Make sure that the drainage tubing is not pinched so that the liquid can flow away. Legend Tube for waste liquid Tube for condensed water Adapter for drain- age tube Fig. 46 Connecting the drainage tubing KNAUER AS-1User manual V6980A, version 1.2...

  • Page 45: Connecting The Autosampler To Other Devices

    Chromatography Software You control the autosampler directly with software, e. g. ® ChromGate from KNAUER. • Using the LAN connection at the rear panel of the device, connect the autosampler directly to the network. Checking and Configuring the Autosampler Parameters...

  • Page 46: Autosampler Device Software

    Needle flushing can be controlled using the chromatography software or PLATINblue Autosampler AS-1 Service Manager . Note KNAUER recommends using a mixture of water and isopropanol (80%/20%) or the mobile phase as the flushing solution. ® Needle Flushing with ChromGate 1.

  • Page 47: I/O Connection

    Defining the closed-contact Output  Inject Marker: The closed-contact output is activated when the injection valve switches from LOAD to INJECT.  Alarm: The closed-contact output is activated when there is an autosampler fault. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 48: Configuration Of I/O Connection (9 Pins)

    HPLC or UHPLC pump, flow rate of 1 ml/min  UV detector (data rate 50 Hz, flow cell: 10 mm path length)  Chromatography software  Eluent: 90 % water, 10 % methanol (HPLC quality) KNAUER AS-1User manual V6980A, version 1.2...

  • Page 49: Test No. 1: Reproducibility Of Sample Volume

    Needle wash: active 2 times  Air segment: yes  Headspace pressure: yes  Injections/vial: 9 (7 for microtiter plates)  Inj. volume: 2 μl  Vial position: 1A1. Start the single run with KNAUER AS-1User manual V6980A, version 1.2...

  • Page 50: Method Parameters Of Uv Detector

    Inject 1, 2, 3, 4 and 5 μl of test solution b (250 ppm uracil, dissolved in water) respectively.  Position of sample vial: 1A3  Injection volume: 1 μl, 2 μl, 3 μl, 4 μl, 5 μl  Repetition: 3 KNAUER AS-1User manual V6980A, version 1.2...

  • Page 51: Operation Qualification (Oq)

    4. Click [Continue] to perform the function test. The chroma- tography software sets the flow rate to 1 ml/min and the wavelength of the detector to 274 nm. You can print out the test results. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 52: Operation Qualification Report

    6. Select [Exit] to end the function test of the device. Operation Qualification Report Legend Detailed information of the autosampler Detailed function test Date and fields for sig- nature and other information Fig. 51 Operation Qualification Report KNAUER AS-1User manual V6980A, version 1.2...

  • Page 53: Maintenance And Care

    You can make your requests in English and German. Maintenance Contract The following maintenance work on the device may only be per- formed by KNAUER or a company authorized by KNAUER and is covered by a separate maintenance contract: ...

  • Page 54: Leaks In The Capillary Screw Fittings

     During removal, consecutively loosen all screws by half a turn respectively, until they can be removed. Note Do not remove the screw in the cover plate hole diagonal to the valve! KNAUER AS-1User manual V6980A, version 1.2...

  • Page 55: Removing The Valve And Rotor Seal

    Disassembling the valve 4. Carefully remove the stator. 5. Remove the rotor seal from the rotor 6. Clean or exchange the rotor seal. Fig. 53 Removing the rotor seal Fig. 54 Rotor and rotor seal KNAUER AS-1User manual V6980A, version 1.2...

  • Page 56: Installing The Valve

    >2 mm to prevent the needle from contacting the bottom of the sample vial.  Tighten the screw fitting until it is finger-tight to prevent the plastic capillary from becoming blocked. Fig. 56 Sample needle set KNAUER AS-1User manual V6980A, version 1.2...

  • Page 57: Settings In Chromgate

    8. Choose Tray Home for Tray Position, choose Home for Syringe and click Set to apply the settings. 9. Choose [Close] to close the Service window. Legend Service window selected Needle Exchange selected ® Fig. 57 Needle exchange, presets in ChromGate KNAUER AS-1User manual V6980A, version 1.2...

  • Page 58: Exchanging The Sample Needle

    4. Remove the sample plates. 5. For Tray Position and Syringe Position, choose Needle Exchange and click Set to apply the settings. The needle moves to the replacement position. 6. Replace the air needle. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 59: Exchanging The Air Needle

    8. Insert the sample needle and fasten with the union nut. 9. Fasten the plastic capillary using the screw fitting on the injection valve. Legend Union nut Retaining nut Height adjustment screw Air needle Sample needle Fig. 59 Exchanging the air needle KNAUER AS-1User manual V6980A, version 1.2...

  • Page 60: Replacing The Sample Loop

    All smooth surfaces of the device can be cleaned with a mild, commercially available cleaning solution, or with isopropanol.  Clean collecting container and sample plates with a soft cloth.  To remove deposits, flush the drainage tube regularly using solvent. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 61: Environmental Protection

    (January, 2001), old devices manufactured by KNAUER are marked as waste electrical and electronic equipment: 160214 WEEE registration KNAUER as a company is registered by the WEEE number DE 34642789 in the German "Elektroaltgeräteregister" (EAR). The number belongs to category 8 under which fall all medical devices and laboratory equipments.

  • Page 62: Troubleshooting

    If the required degree of reproducibility is not achieved, check the following possible sources of error and instigate steps to eliminate them: Cause of fault Solution Air in liquid path Initialize the PLATINblue Autosam- pler AS-1. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 63 (steel, PEEK) or other flushing solutions. Empty sample contami- Use new empty sample. nated Cause unknown. Attempt to solve problem by using different solvents and eluents. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 64 7. If liquid escapes there, check the rotor seal. 8. If no liquid escapes there, check the HPLC with a manual valve. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 65: System Messages From Chromgate

    Troubleshooting System Messages from ChromGate ® ® The various system messages of the ChromGate chromatogra- phy software from KNAUER are explained below. The system messages are sorted alphabetically. System message Explanation  Autosampler is in run Quit the control software and mode.
  • Page 66 Error running user defines Switch the device off and on. program. Inform the Technical Support of the manufacturer in case the sys- tem message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 67 Error setting the prep. Switch the device off and on. mode. Inform the Technical Support of the manufacturer in case the sys- tem message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 68 Home sensor not de- Check whether there are visible activated. obstructions impairing the sample plate tray.  Switch the device off and on. Inform the Technical Support of the manufacturer in case the system message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 69 Injection valve or ISS unit Switch the device off and on. error. Inform the Technical Support of the manufacturer in case the sys- tem message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 70 %2f μl. The flush volume ware and correct entry. should be between 0 and %2f μl. Invalid input. Only val- Check parameters in control soft- ues with increments of ware and correct entry. %.2f allowed. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 71 Several SSV ware and correct entry. events have the same time. Invalid tray temperature Check parameters in control soft- %d ˚C. The temperature ware and correct entry. should be between 4 and 22 ˚C. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 72 Switch the device off and on.  Missing vial. Check position of needle unit.  Switch the device off and on.  Missing wash vial error. Check position of needle unit.  Switch the device off and on. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 73 Syringe dispenser unit Switch the device off and on. error. Inform the Technical Support of the manufacturer in case the sys- tem message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 74 Please select the correct tray in configuration dialog. ISS-A option not Check control software configura- installed on autosam- tion and correct entry. pler. Please switch off ISS-A option in configu- ration dialog. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 75 Check whether there are visible not reached. obstructions impairing the nee- dle unit.  Switch the device off and on. Inform the Technical Support of the manufacturer in case the system message repeats itself. KNAUER AS-1User manual V6980A, version 1.2...
  • Page 76 Check parameters in control soft- ware and correct entry. Wear-out limit reached. Switch the device off and on. If the system message appears again, notify KNAUER Technical Support. The valve must be replaced. Wrong loop volume. The Check parameters in control soft- largest loop volume for ware and correct entry.

  • Page 77: Technical Data

    Temperature range 4 – 40 ˚C; 39 – 104 ˚F Air humidity Below 90 % humidity (non-condens- ing) PLATINblue AS-1 UHPLC injections Up to 1000 bar Sample capacity Microtiter plates for max. 768 wells or sample plates for max. 96 vials ...

  • Page 78: Product Range

    Product Range Product Range Device and Accessories Name Order number PLATINblue AS-1, 1000 bar UHPLC autosampler A63502 with temperature control (cooling and heating), special valve (6/3 way valve), accessories PLATINblue AS-1, 1000 bar UHPLC autosampler, A63501 special valve (6/3 way valve), accessories...
  • Page 79 Air needle, yellow, 50 mm M20401 Air needle, red, 56 mm M20402 Air needle, blue, 68 mm M20403 Air needle, green, 74 mm M20404 Air needle, black, 80 mm M20405 Bottle holder P6051 Network cable A5255 KNAUER AS-1User manual V6980A, version 1.2...

  • Page 80: Legal Information

     Packaging and transport damage In the event of device malfunctions, directly contact the manu- facturer. Wissenschaftliche Gerätebau Dr. Ing. Herbert KNAUER GmbH Hegauer Weg 38 14163 Berlin, Germany Phone: +49 30 809727-0 Telefax:...

  • Page 81: Declaration Of Conformity

    Declaration of Conformity Declaration of Conformity Manufacturer name Wissenschaftliche Gerätebau and address Dr. Ing. Herbert KNAUER GmbH Hegauer Weg 38 14163 Berlin, Germany PLATINblue Order numbers: A63500; A63501; A63502 Autosampler AS-1 The device complies with the following requirements and prod- uct specifications: ...

  • Page 82: Abbreviations And Terminology

    Small bottles used in the sample plates of the autosampler. There are two different kinds of vials: sample vials and destination vials. Wells Deepening in mikrotiter plates, which are being used in the autosampler. KNAUER AS-1User manual V6980A, version 1.2...

  • Page 83: Table Of Figures

    Separate device foot ........16 Fig. 5: PLATINblue AS-1 Mounting device foot ..... 16 Fig. 6: Front View of the Device .
  • Page 84 Exchanging the air needle ....... . 60 KNAUER AS-1User manual V6980A, version 1.2...
  • Page 85 Wissenschaftliche Gerätebau © Dr. Ing. Herbert Knauer GmbH All rights reserved. The information in this document is subject to change without prior notice. Translation of the original German edition of this manual, version 1.2. 2013-03-15 Printed in Germany on environmentally friendly paper from sustainable forests.

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