CAMAG TLC SCANNER 4 WORLD LEADER IN PLANAR CHROMATOGRAPHY

Transcription

1 CAMAG TLC SCANNER 4 WORLD LEADER IN PLANAR CHROMATOGRAPHY

2 The TLC Scanner 4 s the most advanced workstaton for denstometrc evaluaton of TLC/HPTLC chromatograms. It can also be used for denstometrc measurements of other planar objects. All functons of the scanner are controlled by the software wncats. Only the postonng of the object to be scanned s performed manually. If desred the operator can swtch on the nternal llumnaton to control or adjust the scan start poston. Optmal settngs for electronc amplfcaton are automatcally selected for measurements n absorpton and fluorescence mode. Perfect evaluaton wth wncats The well structured and easy to use wncats software controls and montors all functons of the scanner and processes data up to the fnal result. The wncats standard program for the TLC Scanner 4 features: Short measurement tmes Measurement of up to 36 tracks wth up to 100 substances per track Integraton ether wth automatc or manual baselne correcton / peak assgnment Automatc or easy manual assgnment of substance names to peaks Automatc recordng of spectra of all detected or selected peaks Prntout or export of color graphcs of measurement data Report prntout of the entre analyss ncludng all measurement data and mages of the TLC/HPTLC plate

3 A number of wncats program optons allow the user to ft the evaluaton system to the respectve need. The followng optons are avalable: Quanttatve evaluaton supports the followng calbraton modes: sngle level calbraton, mult level calbraton wth lnear or nonlnear regresson usng nternal or external standards. Statstcs are avalable as relatve standard devaton (cv) or confdence nterval (c). Sub-component evaluaton (ncluded n the opton Quanttatve Evaluaton ) can be used to quantfy unknown peaks by relatng them to the man component as s prescrbed by European or US pharmacopoeas ( Related Compounds ). Dual-wavelength scan The chromatogram s scanned at two ndvdually selectable wavelengths. Durng ntegraton the sgnal from the second wavelength s subtracted from that of the frst wavelength to elmnate matrx effects. Dual-wavelength scannng s also useful for the quanttaton of ncompletely resolved peaks. Mult-wavelength scan The chromatogram can be scanned automatcally at up to 36 dfferent wavelengths. Ths permts multple measurements between 190 and 900 nm n order to acheve optmum selectvty. For quanttaton, data from the optmum wavelength scan of each fracton can be selected. Ths software functon s unque n Thn- Layer Chromatography! Scanner qualfcaton (Selftest) Ths opton offers automatc montorng of the mechancal, optcal and electronc functons of the scanner. Results are evaluated, documented and saved. If approprate, lamp postons and monochromator algnment can be automatcally adjusted. Track optmzaton Each track of a chromatogram s scanned several tmes wth small lateral offset. From ths data the optmum track followng the peak maxma s calculated and used for quanttaton. Thereby dstorted chromatograms can be correctly evaluated. Spectrum lbrary Enables the user to create hs own lbrary f les, whch provdes the only way the user can save and compare spectra recorded on dfferent plates. Note The TLC Scanner 4 wth wncats s complant wth the requrements of GMP/GLP and can be IQ/OQ qualfed. If you want to use the nstrument n a 21 CFR Part 11 envronment, the opton 21 CFR Part 11 complance ready s requred for each wncats workstaton. CAMAG TLC Scanner 4 2 3

4 CAMAG TLC SCANNER 4 Key features Measurement of reflecton, ether n absorbance or fluorescence mode Object formats up to mm Spectral range from 190 to 900 nm Automatc start of all lamps: deuterum, halogen-tungsten, and hgh pressure mercury lamp Data step resoluton µm Scannng speed mm/s Spectra recordng speed up to 100 nm/s Automatc selecton of electronc amplfcaton Rapd data transfer Planar chromatogram Analog curve Calbraton functon

5 User-frendly operaton All functons of the scanner are controlled by wncats software. The electronc amplfcaton s set automatcally. In fluorescence mode ths s performed by a quck scan, whch measures the complete plate, e.g. 18 tracks n 20 seconds, and determnes the optmzed settngs from the result. In absorbance mode only one track s quck scanned ; ether track 1 (automatc mode) or a track of choce (manual mode). The 16 bt A/D converter ensures optmum resoluton of the sgnal. Ths s partcularly useful for the measurement of both low absorbance and weak fluorescence sgnals. Scanner operaton wth wncats software s convenent and easy to learn. The object, here a cm HPTLC plate, s smply postoned on the scannng table. The coordnates are dsplayed durng manual postonng of the stage and can be transferred nto the program by mouse-clck. CAMAG TLC Scanner 4 4 5

6 Lamp selector Entrance lens system Monochromator entry slt Monochromator gratng THE OPTICAL SYSTEM: FEATURES AND TECHNICAL DETAILS Mrror The Optcal System Any of the three lght sources, hgh pressure mercury lamp, deuterum lamp, or halogen-tungsten lamp can be postoned n the lght path by a motor drve. The sgnal of the measurng photomultpler s contnuously offset aganst the sgnal of the reference photomultpler. Ths compensates for lamp agng and short-tme fluctuatons. It also reduces the warm-up tme requred to reach lamp stablzaton. All components of the optcal system, lamps, monochromator, scannng stage, and photomultpler are mounted on one sturdy metal support. Ths ensures hgh precson of the detector sgnal. For scannng at wavelengths below 200 nm t s advsable to flush the monochromator wth ntrogen. The scanner s equpped to do ths. A monochromator bandwdth of 5 nm or 20 nm can be selected. 5 nm bandwdth s used for spectra recordng, mult-wavelength scannng, and always when spectral selectvty s requred. 20 nm bandwdth offers hgher lght ntensty (mproves the sgnalto-nose rato and thus the reproducblty of the measurement) and enables measurement of several fractons wth slghtly dfferent absorpton maxma n one scan. The lens system wth nm transmsson range features automatc postonng for mcro and macro slt szes. Ths ensures that the lght energy avalable wth small slts n the mcro poston s almost the same as that for the correspondng slt n the macro poston, whch s four tmes larger. The lght beam strkes the object at rght angle. The photomultpler s algned at an angle of 30. Lght sources Deuterum lamp, usable contnuum nm Halogen-tungsten lamp, usable contnuum nm Hgh-pressure mercury lamp, lne spectrum nm The lamp, whch s postoned n the lght path, s automatcally gnted. All lamps are current stablzed. Plot lamp and compartment llumnaton The slt s automatcally llumnated wth vsble lght when the compartment llumnaton s swtched on. The scannng compartment s llumnated wth a 4 watt fluorescent tube UV 254 nm whch the user can replace by a UV 366 nm or a whte lght tube. Optcal system Apochromatc suprasl-fluorte lens system, transmsson range nm, astgmatc entry lens for optmal slt llumnaton; automatc swtchng between mcro and macro poston for optmal lght ntensty. Monochromator Concave holographc gratng, 1200 lnes/mm, bandwdth selectable 5 or 20 nm, wavelength range nm; monochromator drven by stepper motor, reproducblty of wavelength settng better than 0.2 nm, accuracy better than 1 nm; connector for flushng wth ntrogen. Maxmum speed of spectra recordng 100 nm/s, postonng at 200 nm/s.

7 Dsk wth slt apertures Lens system, can be postoned for mcro and macro slt mage Mrror Reference photomultpler Beam spltter Measurng photomultpler Scannng object Secondary flter Motor-drven flter wheel wth three automatcally selected flters for the elmnaton of second order wavelengths; 400 nm cut-off flter for fluorescence measurements; three postons for user selected flters. Scannng slt Revolvng dsk wth 20 fxed apertures; length of slt mages selectable between 0.2 and 12 mm, wdth between 0.1 and 1.2 mm n 42 combnatons. Detector Two matched broad band photo multplers, mult alkal type, spectral senstvty nm. Stage drve Independent n both drectons by stepper motors, mcro step drven for smooth movement; reproducblty of postonng better than 50 µm n Y-drecton, better than 100 µm n X-drecton; maxmum scannng speed 100 mm/s, postonng at 150 mm/s. Mans voltage 115 V and 230 V selectable; 50/60 Hz; maxmum energy use 180 W (tungsten and mercury lamp gnted). A/D converter 16 bt, 2-channel A/D converter, 100 ms per double converson. Connectons/nterfaces Seral nterface RS232 for communcaton to the computer, EquLnk for connecton to wncats software. Dmensons Wdth = 590 mm, depth = 650 mm, heght = 367 mm; net weght 39 kg. CAMAG TLC Scanner 4 6 7

8 Manual Input Horzontal Developng Chamber Immerson Devce Statonary Phase Nanomat Developng Chamber Spray Sample Preparaton Defnton Sample Applcaton Chromatogram Development Dervatzaton Samples Automatc TLC Sampler ATS 4 Automated Multple Development AMD 2 Software Controlled Standards Lnomat 5 Automatc Developng Chamber ADC 2 wncats SOFTWARE wncats for all steps of nstrumental Thn-Layer Chromatography CAMAG has been developng software for denstometrc evaluaton of TLC/HPTLC chromatograms snce wncats s the result of a unque ntegrated software concept coverng all steps of Instrumental Thn-Layer Chromatography. The user can combne the ndvdual modules to create a complete soluton that meets all requrements wth respect to nstrument control, data acquston, evaluaton and documentaton. The modular desgn of wncats allows selectng or dsablng the ndvdual steps of Thn-Layer Chromatography accordng to the task at hand. wncats features The wncats standard program for the TLC Scanner 4 supports: Short measurement tmes Measurement of up to 36 tracks wth up to 100 substances per track Integraton ether wth automatc or manual baselne correcton / peak assgnment Automatc or easy manual assgnment of substance names to peaks Automatc recordng of spectra of all detected or selected peaks Prntout or export of color graphcs of measurement data Report prntout of the entre analyss ncludng all measurement data and mages of the TLC/HPTLC plate The program offers a unque combnaton of features: wncats combnes hgh performance wth easy handlng wncats offers a two-stage on-screen help system, a bref nfo lne and a detaled explanatory help text wncats features a structured data management and hgh data safety wth easy access to network and project drectores wncats stores the complete set of parameters used together wth all data from scannng raw data to documentaton n one sngle fle, and prnts everythng f desred wncats comples wth the rules of GMP/GLP and 21 CFR part 11

9 Image Import Image Annotaton Detecton Evaluaton Documentaton Report TLC Scanner Scanner Selftest Dual-Wavelength Scan Quanttatve Evaluaton Spectrum Lbrary TLC Vsualzer Image Enhancement Image Comparson 21 CFR Part 11 (FDA Regulaton) Mult-Wavelength Scan Track Optmzaton Modular desgn The modular desgn of wncats allows selectng or dsablng the ndvdual steps of Thn-Layer Chromatography accordng to the task at hand. Workflow Usng the explorer type vew the operator at all tmes remans n control over all actve steps of the workflow. These are: Selecton of plate materal and ts pre-treatment Defnton of samples, standards, and the calbraton method f applcable Sample applcaton Chromatogram development Dervatzaton (pre and/or post chromatographc) Detecton Spectra recordng Quanttatve evaluaton Documentaton wncats program optons for TLC Scanner 4 Quanttatve evaluaton page 12 Spectrum lbrary page 14 Track optmzaton page 14 Dual-wavelength scan page 15 Mult-wavelength scan page 15 Scanner selftest page CFR Part 11 complance ready page 16 CAMAG TLC Scanner 4 8 9

10 TLC SCANNER 4 AND wncats Data nput For routne analyss wncats starts off wth a method fle contanng all relevant data for the current task such as nstruments, parameters, etc. Ths method can be valdated for GxP work. Measurement The TLC Scanner 4 scans the plate and transfers raw data back to wncats where baselne correcton and peak recognton s performed automatcally n the background. From ths method an analyss fle s generated and parameters unque to the actual analyss,.e. sample names, amounts, etc. are adapted. After all steps n the method leadng up to denstometrc evaluaton are completed, the plate s postoned on the stage and scannng s started. Slt dmensons, scannng speed, lght source, wavelength, etc. have already been defned wthn the wncats method. A dsplay of ntegrated peaks wth substance names. Ths dsplay mode enables manual baselne correcton and peak ntegraton by mouse-clck. Data nput of scan parameters Spectra lst

11 Evaluaton By clckng the rght mouse key on an analog curve the dsplay can be swvelled and tlted, and addtonal nformaton can be dsplayed. Analog curves can be dsplayed ether ndvdually or n a 3D dagram. Both can be prnted n color together wth the analyss report. Recordng of spectra wncats can automatcally record spectra as soon as all peak postons are known. Spectra can be dsplayed ndvdually or overlad n one dagram. Spectra can be measured from 190 to 900 nm. If the emsson range of the deuterum lamp s exceeded, the scanner automatcally swtches to the halogen-tungsten lamp. Both lamps reman gnted. All 3D dagrams can be swvelled and tlted for easy evaluaton. Excerpts from a wncats report (can be personalzed by user) CAMAG TLC Scanner

12 QUANTITATIVE EVALUATION Quanttatve evaluaton s based on the comparson of peak heghts or peak areas of the unknowns wth those of calbraton standards chromatographed on the same plate. Dependng on the task at hand sngle-level or mult-level calbraton can be selected. Calbraton modes Sngle-level calbraton s sutable for analyses where the concentraton of the unknowns shall be checked wthn narrow lmts. Snglelevel calbraton requres reduced calbraton efforts. Mult-level calbraton s used when the target values are expected n a comparatvely wde range. wncats offers a choce of four calbraton modes, the selecton s made based on whch mode gave the best result durng method valdaton. Lnear regresson s a useful functon when the calbraton range s narrow or the absolute amounts per fracton are small, whch s often the case by fluorescence scannng. Non-lnear regresson becomes necessary when a wder calbraton range s needed and/or the absolute amounts per fracton are hgh. Polynomal regresson s sutable for calbraton over a wde concentraton range. Mchaels-Menten regresson s sutable for calbraton over a wde concentraton range and for hgh amounts of substances. The Mchaels-Menten functon type 1 passes through the orgn. In cases where ths behavor s not sutable, Mchaels-Menten type 2 may be chosen. Flexble nput screens for calbraton data wncats offers a wde choce for data nput: from enterng amount per fracton to enterng amounts from balance readngs or from stock solutons. Accordng to the European and the US Pharmacopoea, related compounds can be quantfed by comparson wth small amounts of the man component n case ther dentfcaton s not requred. Ths man-/ subcomponent evaluaton s part of the standard quanttatve evaluaton program. Identty and purty checkng by spectra comparson The software opton quanttatve evaluaton ncludes dentty and purty checkng of substances by spectra comparson. For dentty checkng spectra recorded at the peak maxma are compared wth those of standard substances. The user can defne a lmt for the correlaton coeffcent, or let wncats calculate denttes by statstcal crtera. For purty checkng spectra are recorded at peak maxmum and at both slopes. These spectra are then compared by wncats ether to match user specfcatons or accordng to statstcal crtera. Orderng nformaton Quanttatve chromatogram evaluaton

13 Applcaton example Analyss of 25 water-soluble food dyes n food HPTLC s the most effectve, rapd and cost-eff cent soluton for the analyss of water-soluble food dyes n food. Up to 20 runs can be chromatographed under dentcal condtons (by use of the Automatc Developng Chamber ADC2) wthn 12 mnutes (see CBS 105 or 25 food dyes (splt n 3 mxtures, applcaton volumes of 1, 2, and 3 µl) and 6 food samples (applcaton volumes of 2 µl; energy drnk (ED), yoghurt (Jog), frut drnk (FD), bakery nk formulaton (BI)) Mult-wavelength scan of mxture 1 (superposton of absorpton curves at 6 wavelengths). Calbraton functon of the red food dye E122 (x) and determnaton n frut drnk, energy drnk and bakery nk formulaton (+). Calbraton functon of the blue food dye E131 (x) and determnaton n the yoghurt sample (+). CAMAG TLC Scanner

14 SPECTRUM LIBRARY TRACK OPTIMIZATION Recordng, dsplayng and comparng spectra of substances chromatographed on one plate s ncluded n the standard Scanner software. In order to compare spectra of substances chromatographed on dfferent plates or to compare spectra wth those of a spectra collecton, the software opton spectrum lbrary s requred. The spectrum lbrary can be used durng substance assgnment for valdaton of assgned substances as well as for dentfcaton of unknown fractons. Durng the search process for dentfcaton the spectrum lbrary shows a ht lst of the closest substances ncludng ther spectra. Wth ths software opton a dstorted chromatogram can be corrected. Each track s scanned several tmes wth small lateral offset. The number of scans and the dstance n between s selected accordng to the requrements of the partcular chromatogram. When all scans of one track are completed, the software selects the maxmum sgnal for all peaks. Only those data are then used for the calculaton of the result from the corrected chromatogram. Note: When used wth chromatograms wth proper track algnment, the results obtaned wth or wthout track optmzaton are about the same In case of dstorted chromatograms, results obtaned wth track optmzaton are comparable to those of a good chromatogram scanned wthout track optmzaton Orderng nformaton Spectrum lbrary Orderng nformaton Track optmzaton

15 DUAL-WAVELENGTH SCAN AND MULTI-WAVELENGTH SCAN The software opton dual-wavelength scan serves for background correcton. The chromatogram s scanned wth two wavelengths, the measurng and the reference wavelength, whch each can be selected between 190 and 900 nm. As measurng wavelength one wll usually select the wavelength of the absorpton maxmum of the substance to be calbrated. The reference wavelength should be suffcently apart from ths maxmum absorbance n order to acheve good senstvty, however, not too far away as otherwse rregulartes n the layer wll not be relably compensated. A precedng recordng of the spectrum s helpful for selectng the rght combnaton of wavelengths. The opton mult-wavelength scan s a useful tool for the quanttaton of analyte mxtures wth components dfferng n absorpton maxma. The plate tracks can be scanned consecutvely at up to 36 dfferent wavelengths between 190 and 900 nm and the peak data s stored n one analyss fle. Durng the consecutve quanttatve evaluaton each component can now be automatcally evaluated at ts maxmum absorbance. Ths way the user s releved from manually evaluatng the same plate at dfferent wavelengths. For dentty checkng the 3D vew of the mult-wavelength scan s another very helpful tool. The 3D vews can be scaled, swvelled and tlted, and then coped to clpboard or saved as a btmap fle for use n other software, e.g. Word. Orderng nformaton Dual-wavelength scan Orderng nformaton Mult-wavelength scan CAMAG TLC Scanner

16 SCANNER SELFTEST 21 CFR PART 11 COMPLIANCE READY Wth ths opton an automatc selftest of the TLC Scanner 4 and a report can be generated. Ths opton s requred for complance wth the FDA regulaton regardng the recordng of electronc data and sgnatures. In the scanner selftest procedure the followng checks are performed: Wavelength accuracy of the monochromator Stage postonng Condton and algnment of all lamps Condton and algnment of the optcal system Condton of the electronc system The complete qualfcaton procedure can be carred out automatcally or manually n sectons. Certan defcences n lamp algnment and monochromator adjustment can be automatcally corrected. Features : Safety of data acquston and operaton by user dentfcaton wth password Secure storage of all results ncludng raw data ensures complete data ntegrty and trackng Documentaton of all actvtes n Hstory Log/Audt Tral for secure trackng of electronc sgnatures accordng to 21 CFR Part 11 requrements Example The stage wth the test pattern (straght lne) s moved multple tmes at 0.1 mm ncrements n Y drecton across the slt. The resultng analog curve s evaluated by the program. Ths test gves nformaton about: Unformty of slt llumnaton wth the respectve lamp, and Correct algnment of the lamp and the optcal system The result of each test s dsplayed on screen and can be prnted as part of the qualfcaton report. The report contans the target values and the values actually found together wth a pass/fal judgment. The opton scanner selftest s requred when the nstrument shall be operated n a GxP envronment. Orderng nformaton Scanner selftest Orderng nformaton CFR Part 11 complance ready

17 INSTRUMENT QUALIFICATION INSTRUMENT SERVICE For customers workng n a GMP/GLP envronment, CAMAG offers Installaton Qualfcaton (IQ) and Operaton Qualfcaton (OQ) as servce. The Installaton Qualfcaton (IQ) s performed at the ste and at the tme of nstallaton. It documents that all specfcatons and parameters comply wth the manufacturer s specf catons, envronmental parameters and safety requrements. The Operaton Qualfcaton (OQ) s ntally performed subsequent to IQ and s repeated at certan ntervals recommended by the manufacturer or defned by the customer. It documents that all modules of the qualf ed system functon properly wthn the specf ed operatng ranges. The Performance Qualfcaton (PQ) s an ongong process whch documents that the nstrument or system s sutable for the gven task. Thus only the user can perform PQ, employng hs substances, followng hs task descrpton and hs test procedures (SOPs). For all CAMAG nstruments for whch we offer IQ/OQ qualfcaton we also offer servce contracts. A servce contract ncludes: Preventve mantenance once per year or as requred Adjustments and tests Optonal performance of the qualfcaton (OQ) at the same tme CAMAG TLC Scanner

18 TECHNICAL DATA Lght sources Deuterum lamp, usable contnuum nm Halogen-tungsten lamp, usable contnuum nm Hgh-pressure mercury lamp, lne spectrum nm The lamp, whch s postoned n the lght path, s automatcally gnted. All lamps are current stablzed. Plot lamp and compartment llumnaton The slt s automatcally llumnated wth vsble lght when the compartment llumnaton s swtched on. The scannng compartment s llumnated wth a 4 watt fluorescent tube UV 254 nm whch the user can replace by a UV 366 nm or a whte lght tube. Optcal system Apochromatc suprasl-fluorte lens system, transmsson range nm, astgmatc entry lens for optmal slt llumnaton; automatc swtchng between mcro and macro poston for optmal lght ntensty. Monochromator Concave holographc gratng, 1200 lnes/mm, bandwdth selectable 5 or 20 nm, wavelength range nm; monochromator drven by stepper motor, reproducblty of wavelength settng better than 0.2 nm, accuracy better than 1 nm; connector for flushng wth ntrogen. Maxmum speed of spectra recordng 100 nm/s, postonng at 200 nm/s. Secondary flter Motor-drven flter wheel wth three automatcally selected flters for the elmnaton of second order wavelengths; 400 nm cut-off flter for fluorescence measurements; three postons for user selected flters. Scannng slt Revolvng dsk wth 20 fxed apertures; length of slt mages selectable between 0.2 and 12 mm, wdth between 0.1 and 1.2 mm n 42 combnatons. Detector Two matched broad band photo multplers, mult alkal type, spectral senstvty nm. Stage drve Independent n both drectons by stepper motors, mcro step drven for smooth movement; reproducblty of postonng better than 50 µm n Y-drecton, better than 100 µm n X-drecton; maxmum scannng speed 100 mm/s, postonng at 150 mm/s. Mans voltage 115 V and 230 V selectable; 50/60 Hz; maxmum energy use 180 W (tungsten and mercury lamp gnted). A/D converter 16 bt, 2-channel A/D converter, 100 ms per double converson. Connectons/nterfaces Seral nterface RS232 for communcaton to the computer, EquLnk for connecton to wncats software. Dmensons Wdth = 590 mm, depth = 650 mm, heght = 367 mm; net weght 39 kg.

19 ORDERING INFORMATION CAMAG TLC SCANNER 4 for scannng by absorbance and fluorescence, equpped for objects up to mm, wavelength range nm, complete wth deuterum lamp, tungstenhalogen lamp, and mercury vapor lamp, ncludng Equlnk ( ) to wncats, but wthout wncats lcense. wncats Software and optons: wncats lcense ncludng one year of nternet update servce Quanttatve chromatogram evaluaton Spectrum lbrary Track optmzaton Dual-wavelength scan Mult-wavelength scan Scanner selftest CFR Part 11 complance ready You can order the CAMAG TLC Scanner 4 as a complete system: CAMAG TLC Scanner 4, complete system CAMAG TLC Scanner 4, for scannng by absorbance and fluorescence, equpped for objects up to mm, wavelength range nm, complete wth deuterum lamp, tungsten-halogen lamp, and mercury vapor lamp Equlnk to establsh a 2-way communcaton between the scanner and wncats wncats lcense ncludng one year of nternet update servce wncats opton Quanttatve chromatogram evaluaton wncats opton Scanner selftest CAMAG TLC Scanner

20 CAMAG (Swtzerland) Sonnenmattstrasse 11 CH-4132 Muttenz 1 Phone Fax E-Mal nfo@camag.com CAMAG (Germany) Bsmarckstrasse D Berln Phone Fax E-Mal nfo@camag-berln.de CAMAG Scentfc (USA) 515 Cornelus Harnett Drve Wlmngton, NC Phone (800) Fax (910) E-Mal tlc@camagusa.com WORLD LEADER IN PLANAR CHROMATOGRAPHY SCAN4 10-E