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1 Systems And especially: SPE Module Visit us at

2 Systems General e Description Highlights Modules Structure and Availability FREESTYLE TM BASIC FREESTYLE TM SPE 2

3 FREESTYLE General Description The FREESTYLE Systems are fully automated sample preparation systems for unattended processing of food, feed, environmental and pharmaceutical samples. They can be upgraded with different modules. They can be modified by choosing different trays, racks etc. 3

4 FREESTYLE Highlights 1/4 Robotic platform design No general decision for a certain platform or a certain configuration needed: d Customer just buys what he needs right at the moment Any change of usage means: customer has to purchase more modules or racks and integrate them in a method. No need of teaching the system! No need to change the robotic setup by a service engineer! 4

5 FREESTYLE Highlights 2/4 Customer has to make only one general decision which cannot be modified in future: Customer has to decide about the size of the robotic system available with usable widths of 490 mm, 740 mm or 1240 mm! The system with 740 mm usable width is our recommended standard size. Shown here with GPC module 5

6 FREESTYLE Highlights 3/4 Modular Design All modules can be used in any combination or individually When customer purchases any FREESTYLE configuration the system can be upgraded/extended in any direction afterwards No hardware changes / modifications needed when using different FREESTYLE modules (once they are installed) Each module shows unmatched advantages; most of the advantages will be protected by patents (already applied for) 6

7 FREESTYLE Highlights 4/4 The Allows intuitive iti handling and easy usage in daily routine Indicates most of the logical failures done by the customer choosing the wrong parameters. Always included in the purchase price of modules. Installation is extremely easy. 7

8 FREESTYLE Summary The FREESTYLE Systems offer a wide range of applications and offer huge advantages in nearly every single step compared to competitors like GILSON and others. This is a first time that a SPE method can be transferred from manual handling to automation without loosing any single step. 8

9 FREESTYLE Structure and Availability All combinations will be possible by October 2011 GPC (Clean-up) SPE (Handling of IAC & SPE columns) EVA (Automated evaporation) TACS (BASIC & GPC and EVA) available Sept Oct Oct FREESTYLE BASIC (dilution, pipetting etc.) Upgrade by modules upgrade: October

10 BASIC = Part of any FREESTYLE system 10

11 FREESTYLE BASIC Robotic System Special valve for all applications Needle loop Syringe pump Electric devices & electronic control Wash port (needle) Needle (with double function + (not shown) 11

12 FREESTYLE BASIC FREESTYLE BASIC consists of x-y-z robotic system for sample injection onto the column and fraction collection (combined autosampler / fraction collector module) Windows-based (Windows 7 ) instrument control software via PC 12

13 FREESTYLE BASIC Cont. +++ Special valve which allows easy upgrade - for sample clean up via GPC, - handling of SPE-columns etc. and - automated evaporation High precision syringe / 10 ml - sample volume between min. 0.1 and 10 ml -sample aspiration speed individually stored in the method Calibrated sample loop with 5.0 ml (available up to 10.0 ml) 13

14 FREESTYLE BASIC Main FREESTYLE BASIC functions Handling of Solvents with a needle Handling of Nitrogen Planned function / software update will come till end of 2011 Transfer of liquids Dilution (incl. calculation) Pipetting Mixing Adding of external solvents 14

15 FREESTYLE BASIC Unique rack design Racks are individually positioned by hooking them into a toothed th plate No tools needed No base plate needed Whenever customer needs an additional rack: just buy it, no need to teach the robotic system Racks can be directly mixed on the robotic area by the customer according to needs; no change of base plate needed! d! 15

16 FREESTYLE BASIC frame = rack tray with vials Frame can be used for different kinds of vials Tray can be taken out of frame and prepared independently 16

17 FREESTYLE BASIC Racks User-defined combination of any flask sizes from 1 ml to 1 L Up to 720 x 1 ml vials or 24 x 1 L round bottom flasks Manifold applicability of flask types, e.g. 1 ml (GC-vials), 4 / 16 / 60 / 100 / 250 / 500 ml, 1 L flasks, bottles and round bottom flasks as well as test tubes, e. g. 16 x 100 mm or special vials for the TurboVap system Processing of septa-sealed sealed vials 17

18 FREESTYLE BASIC These sample numbers are possible to run* usable space / type of vial 740 mm 1240 mm 60 ml bottles ml bottles ml bottles ml bottles L bottles ml TurboVap ml TurboVap ml round bottom flask ml round bottom flask ml round bottom flask L round bottom flask test tubes (100 x 16mm) test tubes (75 x 12mm) *Th The extract tis provided din 16 ml-vials! 18

19 FREESTYLE BASIC Safety features are important for handling such a system All liquid containing tubing is located outside the instrument, thus easily accessible and strictly separated from any electric wiring Plausibility check of software input Collisions recognition with the possibility to proceed without loss of sample Possibility of immediate emergency stop in case of danger or to prevent any dangerous situation 19

20 FREESTYLE BASIC Standards of development and production & safety guidelines Developed and produced at highest level according to Machine Directive 2006/42/EC of the European Parliament and of the Council of 17 May 2006 on machinery, and amending Directive 95/16/EC (recast), including CE certification. Developed and produced according to ISO 9001:2008. CE labelled safety guidelines 20

21 SPE = FREESTYLE BASIC + SPE module SPE (Handling of IAC & SPE columns) FREESTYLE BASIC (dilution, pipetting pp etc.) 21

22 Content Instrument overview 22

23 FREESTYLE SPE 23

24 FREESTYLE SPE SPE module consists of: Second z-axis Holder for SPE-cartridges (different dimensions for different SPE formats e. g. 1mL, 3mL, 6mL, LCTech glass columns etc) allows to transfer manual steps to automation SPE module can be used with any other module for FREESTYLE System (e. g. high end evaporation module EVA) 24

25 FREESTYLE SPE Highlights Positive pressure for all steps in which liquids are used One rack for all kinds of columns: reusable adapters allow an easy to change and mixed setup for all kinds of columns, e. g. SPE columns, LCTech glass columns and IAC-columns Columns can be moved to any place in the robotic area No hardware change by running different application (when the modules were installed) Standard three solvents, upgrade option up to six solvents All kind of manual SPE-steps can be transferred into an automated process Powerful software (Computer needed) 25

26 FREESTYLE SPE Overview FREESTYLE SPE = FREESTYLE BASIC + additional parts + second z-axis + software upgrade (SPE handling) + ports for washing and handling (waste position) + electric devices & electronic control Solvents going g to waste 26

27 Second z-axis: Manipulation Tool for SPE 1st z-axis (FREESTYLE BASIC) 2nd z-axis (handling of SPE column) solvent can go through this z-axis as well Both z-axes are connected through a valve and syringe pump Holder for SPE column SPE column 27

28 Second z-axis: Manipulation Tool for SPE Power tool (second z-axis) Adapter can be modified easily for any tools which h should be handled d SPE column or any other tool Hermetically sealed connection (air & liquid) 28

29 Second z-axis: Manipulation Tool for SPE Power tool (second z-axis) Adapter can be modified easily held by the tool of 2nd z-axis SPE column or any other toolinsert in the adapter 29

30 Content Instrument overview 30

31 Steps that can be chosen Steps chosen by the user Parameters for the single steps, each one can be easily changed by moving the bar via mouse Calculated time for this step Input and output vials 31

32 FREESTYLE SPE is structured according the necessary steps for IAC-/SPE handling only used for IAC columns different modes acc. sample volume / application different modes 32

33 FREESTYLE SPE Method consists of an arrangement of steps in a way like customer wants (nearly without any limitation). Each step offers the possibility of individual id programming The software allows to transfer all kinds of manual steps to an automated process 33

34 FREESTYLE SPE Flexibility everywhere ALL IN ONE: Samples can be mixed in one sample list, e. g.: - (closed) IAC & SPE & different formats of both - different solvents - different methods Easy changing of the priority in sample list 34

35 FREESTYLE SPE Method Development Move next step from the basic tools with drag & drop to second column The default button remains in the basic tools and can be choosen again The collected steps in the second column build the method. Each button there can be individually changed and adopted by using the bars. 35

36 FREESTYLE SPE Method Development The whole method can be stored Very nice tuning of the method, because there are a lot of parameters for the different steps, e. g - flow rate - volume - repetition of single steps, - waiting time - fractionating ( eluting step) - aspiration speed, etc. Nearly no limitation for usage of bottles, vials, ground joint flasks, rotary evporator flasks, TurboVAP, test tubes for sample and eluate. Customer can chose whatever he wants 36

37 FREESTYLE SPE Emptying (special step for IAC columns) 37

38 FREESTYLE SPE Conditioning 38

39 FREESTYLE SPE Different modes for the loading step Example Load : Transfer Sample-Aliquot over sample loop (recommended for samples > 2 ml) Loading mode can be selected by drop-down d menu 39

40 FREESTYLE SPE Different modes for the loading step Choose the mode from the pull-down menu in step Load 1 Standard: using the needle loop aspiration through the needle and push out through SPE cartridge; this mode should be used for volumes above 2 ml (can go up to 250 ml) Quantitative transfer possible (incl. washing step of sample reservoir) 2 Sample transfer via needle aspiration in the needle and dispensing the sample on top of the SPE-cartridge (recommended for small samples up to < 2 ml) 3 Sample transfer by aspiration through h the tip of the SPE cartridge (for small samples with max 1.0 ml volume) available via software upgrade from October 40

41 FREESTYLE SPE Different modes for the loading step 4 Special method: sample from one vial transfered via needle loop and through a SPE column into a second vial. Using the SPE column as a for the whole sample; components which disturb the analysis are filtered out. It was specially developed for the clean up step for determination of the mineral oil hydrocarbon index (EN ISO :2000 or simply called H53). But can be used for other SPE application as well (e. g. for DON-analysis). Higher volumes (in this special case 60 ml) are used. Quantitative transfer possible (incl. washing step of sample reservoir) 41

42 FREESTYLE SPE Drying (with Nitrogen or ambient air) 42

43 FREESTYLE SPE Washing 43

44 FREESTYLE SPE Eluting 44

45 FREESTYLE SPE Four possibilities for the eluting step 1 Elution of a single solvent into one vial Elution 1 2 Elution of one solvent into several vials of the same size (up to 10). Each vial is filled to its maximum before the next vial is filled. Elution 1 45

46 FREESTYLE SPE Four possibilities for the eluting step 3 Elution of different solvents into different vials Here: sequence of three independent elution steps Elution 1 Elution 2 Elution 3 4 Elution of several solvents into one vial Elution 1/2/3 46

47 Content Instrument overview 47

48 Start N 2 Syringe pump Sample loop V Waste port Solvents Waste z1 y arm z2 SPE cartridge z Sample y x Eluate vial 48

49 Conditioning N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 49

50 Sample Aspiration N 2 Syringe pump Sample loop V Solvents y arm z2 Waste z1 z SPE cartridge x y Sample 50

51 Sample Loading N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 51

52 Column Washing N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 52

53 Column Drying N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 53

54 Elution N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Eluate vial 54

55 Content Instrument overview 55

56 Cleaning and Filtration SPE as CLEANING and FILTRATION step e.g. for Mineral Oil Hydrocarbon Index acc. EN ISO :2000 (simply py called H53) or other SPE-application (eg. DON clean up). Mineral Oil hydrocarbon index acc EN ISO :2000: A special glass column is used. This glass column is filled with Florisil and sodium sulfate and is used just as chemical to clean the whole sample from any moisture and components which can disturb the analysis. Ready-to-use columns w/ certificate are available from LCTech under the brand name ELUFIX 56

57 Cleaning and Filtration SPE as CLEANING and FILTRATION step e.g. for Mineral Oil Hydrocarbon Index acc. EN ISO :2000 (simply py called H53) or other SPE-application (eg. DON clean up). The special glass column filled with Florisil and sodium sulfate is just a to clean the whole sample from any moisture and components which can disturb the analysis. Special method: Sample from one vial transfered via needle loop and through a SPE column into a second vial This mode is used as a cleaning step for the whole sample; components which disturb the analysis are filtered out. Quantitative transfer possible (incl washing step of sample reservoir) 57

58 Cleaning and Filtration Start N 2 Syringe pump Sample loop V Waste port Solvents Waste z1 y arm z2 H53 cartridge z Sample y x Eluate vial 58

59 Cleaning and Filtration Conditioning Syringe pump Sample loop N 2 V Solvents z1 y arm z2 Waste z H53 cartridge y x Waste port 59

60 Cleaning and Filtration Sample Aspiration/Loading Syringe pump Sample loop N 2 V Solvents y arm z2 Waste z1 z H53 cartridge x y Sample 60

61 Cleaning and Filtration Column Drying Syringe pump Sample loop N 2 V Solvents z1 y arm z2 Waste z H53 cartridge x y Sample 61

62 Content Instrument overview 62

63 Special SPE Sometimes there is only a very small amount of sample available. This may occur eg. in forensic or pharmaceutical applications. Now standard cartridges can be used for small amounts of samples without any contamination of the instrument (sample goes only into the cartridge) and without loosing any sample. A precise volume can be aspirated through h the tip of a cartridge. Sample transfer by aspiration through the tip of the SPE Cartridge (software upgrade available in October) This mode of loading is recommended for small samples with max 1.0 ml volume. 63

64 Special SPE Start N 2 Syringe pump Sample loop V Waste port Solvents Waste z1 y arm z2 SPE cartridge z Sample y x Eluate vial 64

65 Special SPE Conditioning N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 65

66 Special SPE Sample Aspiration N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Sample 66

67 Special SPE Sample Aspiration N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Air 67

68 Special SPE Sample Loading N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 68

69 Special SPE Column Washing N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 69

70 Special SPE Column Drying N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Waste port 70

71 Special SPE Elution N 2 Syringe pump Sample loop V Solvents z1 y arm z2 Waste z SPE cartridge y x Eluate vial 71

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