Agilent Guide to Capillaries Reagents Supplies for CE CEC Manual
Contents
1. Description Unit Part No Long life Deuterium lamp 8 pin with RFID tag for G7100 5190 0917 Deuterium lamp for G1600 2140 0585 Electrode assembly standard for G1600 only G1600 60007 Electrode assembly short for G1600 only 61600 60033 Electrode assembly standard for G7100 only G7100 60007 Electrode assembly short for G7100 only G7100 60033 Electrode O ring silicone 5 pk 5062 8544 Electrolyte bottle 500 mL 9300 1748 Electrolyte bottle 100 mL for G7100 only 5042 6478 Electrolyte bottle cap 9300 1747 Bottle sealing O ring 0905 1163 Glass filter solvent inlet 20 um 5041 2168 Filter frit adapter 3 mm 4 pk 5062 8517 Bottle cap plug G1600 23223 Air filter 5 um 3150 0619 Pre puncher G1600 67201 Screws for pre puncher insulation plate holding 10 pk G1600 62402 5183 4669 590 3003 Accessories Description Part No CE accessory kit G7100 68705 Includes electrode tool screwdriver fuses air filter glass frit vials and caps Includes 50 ym ID capillaries L 64 5 Standard L 64 5 Extended Light Path L 48 5 Standard and alignment interfaces red and green Rack for 12 mm 2 ml vials holds 50 vials per rack 5 pk 9301 0722 CE column cutter 5183 4669 Diamond blade replacement kit for CE column cutter 5183 4670 Capillary tubing cutter 4 pk 5181 8836 Window Etching Tool The window etching tool is designed for fast convenient and reproducible preparation of2. Cross linked and Bonded uSIL Capillaries USIL FC and pSIL DNA Capillaries with Windows A series of coated capillaries specifically designed for CE which are prepared by cross linking and bonding a novel proprietary flourocarbon FC polymer USIL FC capillaries are chemically inert hydrophobic and stable from pH 2 5 10 0 These capillaries are a must have for clEF protein peptide and carbohydrate separations as well as replaceable gel CE applications such as oligonucleotides DNA fragments and PCR product separations UV Adsorption 260 nm uSIL DNA capillaries are also coated with Time min an FC polymer but have a 75 um ID to accommodate the viscosity of entangled polymer solutions All SIL capillaries are batch tested to ensure the highest performance and reproducibility Analysis of Allelic ladder with pSIL DNA D USIL WAX Capillaries with Windows USIL WAX features a modified polyethylene oxide hydrophilic coating made through a special cross linking and bonding process The coating effectively masks active silanol sites offering exceptional efficiency resolution peak shape and reproducibility The highly stable coating and near zero EOF of uSIL WAX makes the capillary ideal for CE MS and protein and peptide separations from pH 2 5 Bulk uSIL DB Capillaries The uSIL DB coated capillaries are available as LSIL DB 1 and ySIL DB 17 In combination with a cellulose based buffer syst
3. Guide to Capillaries Reagents and Supplies for CE and CEC Maximize your CE and CEC System and experience greater laboratory uptime Our measure is your success products applications software services Soa Agilent Technologies Your Link to Success Agilent Technologies Capillary Electrophoresis Capillaries Reagents Solutions Kits and Supplies Agilent Technologies a world leader in capillary electrophoresis CE technology is committed to helping you improve the quality of analyses by offering application solutions simplifying the use of software and instrumentation and offering unsurpassed technical support Capillary Electrophoresis CE offers fast separations with exceptional efficiency and resolution for charged substances such as biomolecules low molecular weight basic or acidic drugs and ions separations that are often difficult to achieve with HPLC CE also excels where sample amounts are very limited and requires much less buffer than liquid or ion chromatography Used in standalone mode as the separations component of a CE MS or as a complementary orthogonal technology to LC The Agilent 7100 CE system delivers best in class analytical performance the industry s broadest selection of detectors and plug and play compatibility with all of Agilent s 6000 Series mass spectrometers Table of Contents CE Method Development
4. CE System Start up and Test Kits Chemical test kits and validation packages functionality The 00 PV kit is only part are available to help comply with regulatory of the validation services available from and quality standards The Installation Agilent Technologies When implemented Qualification 10 Chemical Kit and Hardware by qualified Agilent personnel our service Start Up Kits which are shipped with new packages can be used to help validate your instruments are useful for rapidly verifying Agilent CE system system functionality For rigorous testing the Operational Qualification 0Q Performance Verification PV Kit can be used to verify DAD noise drift linearity and wavelength accuracy and replenishment Temperature stability DAD function Voltage stability Wavelength accuracy Linearity Noise and drift Injector reproducibility and linearity Replenishment functionality Operational Qualification Performance Verification 00 PV EJ Sample p hydroxyacetophenone Buffer 20 mM Borate pH 9 2 Capillary 40 cm L 48 5 cm 50 um ID Injection 250 mbar s Voltage 30 kV Temperature 20 C Detector 192 4nm 10 and 00 PV test method CE System Start up and Test Kits Description Part No CE Installation Qualification IQ Kit 5063 6514 Includes buffer 20 mM borate pH 9 3 100 mL test sample 4 hydroxy acetophenone 2 mL capillary conditioning solution 0 1 N sodium h
5. detection windows on fused silica capillaries The polyimide coating is removed without destroying the inner polymeric coating It contains 3 glass blocks with fine grooves controlling precisely the size of the windows Description Part No Window etching tool 3 pk 590 3003 Basic Capillary Electrophoresis Troubleshooting Symptom Unstable Current Variable or no current Possible Cause Air bubble formed in capillary Solution s Flush capillary ramp voltage to limit initial heating and or degas buffers Clogged capillary Flush capillary with absorbing solutiion such as NaOH A step on the base line shukd be observed when viewing the online signal at 200 nm If still plugged flush manually with syringe or high pressure gas Broken capillary Replace capillary No or incorrect solution in buffer vials Fill change buffer vials Large volume injection Normal situation Current should stabilize during analysis Basic Capillary Electrophoresis Troubleshooting Unstable Baseline Spikes in baseline Precipitates in buffer Filter buffer through 0 2 or 0 45 um filter Micro air bubbles in buffer Degas buffer by ultrasonication or vacuum Precipitation of sample Verify that sample components are sufficiently soluble in buffer Noisy baseline Optical slit in capillary interface is occluded Clean slit with methanol or water View under magnifier Aging deuter
6. 1311 UPAGE 3 3 T 3 C PAGE pd A o 39 ag g9 Oligonucleotide standard for yPAGE 3 and uPAGE 5 kits 75 191 3211 uPAGE 5 5 T 5 C PAGE pd A gt 5 39 40 69 Oligonucleotide standard for pPAGE 3 and uPAGE 5 kits 75 191 5211 Note The PAGE capillaries are not pre aligned for the G1600A CE and G7100 CE systems To cut them to the correct length use the CE column cutter P N 5183 4669 To create Detection Window use the Window Etching Tool P N 590 3003 PAGE Buffer Solutions and Oligo Standards Kit as defined by type of pPAGE capillary Part No PAGE Tris borate and urea buffer for uPAGE 10 4 x 237 mL 590 4005 PAGE Tris borate and urea buffer for yPAGE 3 and pPAGE 5 4 x 237 mL 590 4001 HPAGE pd A os 39 40 60 Oligonucleotide standard for yPAGE 3 and uPAGE 5 3 x 50 uL 590 4000 zp Standard Bare Fused silica Capillaries Fused silica capillaries are the heart of CE Pre aligned capillaries from Agilent Technologies are designed and optimized for ease of use and reliability All capillary 2504 ends are cut to a smooth mirror like finish In addition the polyimide outer coating is removed from the ends These processes ensure minimal sample adsorption and help maintain sharp peak shapes All capillaries have a pre made detection window and a built in alignment stopper which allows rapid and precise insertion in the alignment interface mAU 200nm ilil CE
7. 4 CE Method Optimization 5 CE Solution Kits 6 Inorganic Anion Solutions Kit 7 Cation Solutions Kit 8 Organic Acids Solutions Kit 9 Forensic Anions Solutions Kit 10 uPAGE Solutions Kits for High Resolution DNA Fragments Analysis 10 Standard Bare Fused Silica Capillaries 12 Extended Light Path Bubble Cell Bare Fused Silica Capillaries 13 Universal Bare Fused Silica Capillaries 14 Poly Vinyl Alcohol PVA Coated Capillaries 15 CEP Coated Capillaries 17 Cross linked and Bonded SIL Capillaries 17 18 Capillary Electrochromatography CEC Capillaries 19 Alignment Interfaces and Capillary Cassette 20 High Sensitivity Detection Cell 21 22 CE MS Accessories 23 CE Standards amp Reagents 24 25 CE System Start up and Test Kits 26 27 Instrument Parts and Supplies 28 29 Basic Capillary Electrophoresis Troubleshooting 30 31 CE Method Development CE Method Optimization YES NO MEKC NO CZE CE Solution Kits Agilent continues to introduce new CE solutions kits designed to simplify many of your applications e Inorganic anions e Cations e Organic acids e Forensic anions e uPage kits for high resolution DNA fragment analysis These kits include all you need to begin your CE analyses including buffers capillaries conditioning solutions test samples methods and detailed descriptions Each kit is designed to take advantage of the automation of the Agilent CE system to make your time in the laboratory more effici
8. 50 pm ID with Electroosmotic flow maintains the plug flow in the bubble Optical slits matched to the zone geometry maintain resolution CZE analysis of a tryptic digest of carbonic anhydrase using a standard capillary id 50 pm compared with an Agilent Extended Light Path Capillary Universal Bare Fused Silica Capillaries These are capillaries with a window and 75 cm effective length and 363 um OD fitting into any CE instrument To cut them to the correct length we recommend using P N 5183 4669 CE column cutter a Universal Bare Fused Silica Capillaries ID pm Total Length cm Effective Length cm Part No 20 100 75 190 0431 50 100 75 190 0131 75 100 75 190 0231 100 100 75 190 0331 Bulk Fused Silica Capillaries 363 pm OD Capillary ID pm Total Length m Part No 20 5 160 2660 5 50 5 160 2650 5 75 5 160 2644 5 Poly Vinyl Alcohol PVA Coated Capillaries PVA coated capillaries contain a permanently adsorbed layer of poly vinyl alcohol This coating minimizes hydrophobic and electrostatic solute wall interactions and eliminates electroosmotic flow EOF Using a proprietary deposition process the PVA coating is stable over a wide pH range even under basic conditions from 2 5 to 9 5 This stability allows use of a wide range of common CE buffers Because the silica surface is covered many proteins and amines can be analyzed without the peak tailing found with uncoated capilla
9. extended light path capillaries and the high sensitivity detector cell Capillary cassette Description G7100 CE G1600 CE Part No Part No Capillary cassette G7100 60002 G1600 60002 Note Only use G71000 60002 cassette in G7100 and G1600 6002 cassette in G1600 Never mix cassettes Alignment Interfaces Optical filter for DAD Description G7100 CE G1600 CE Part No Part No Optical filter for DAD G7100 62700 G1600 62700 260 nm for DNA analysis with polyacrylamide filled capillaries and oligonucleotide analysis te and interfaces accept High Sensitivity Detection Cell The Agilent High Sensitivity Detection Cell Dramatically Enhances Sensitivity and Linearity for the Agilent CE Capillary Electrophoresis System The Agilent high sensitivity detection cell a technological leap which extends sensitivity by an order of magnitude provides a solution to sensitivity limitations often encountered in CE This improvement will substantially increase the utility of CE for impurity analysis of chiral drugs biologicals and compounds of environmental interest among others The high linear range allows quantification of both lt 0 1 impurities and the main component in one run This is useful for all impurity determinations and is especially useful for determining chiral excess The High Sensitivity Detection Cell for the Agilent CE system not only improves detection sensitivity more than 10
10. fold over standard capillaries but also extends linearity beyond 2000 mAU and provides unsurpassed spectral fidelity These improvements are a result of a proprietary micromachined design which increases the detection pathlength from 75 um to 1200 um while dramatically reducing stray light The High Sensitivity Detection Cell has a design comprised of a fused silica cell body and removable capillaries The light path through the cell is made from black fused silica which significantly minimizes stray light and defines the aperture for the diode array spectrometer In addition the reflective interior functions as a lightpipe insuring almost 100 transmission of light which entered the cell These properties A result in enhanced linearity and unsurpassed j spectral fidelity with the diode array detector ED Characteristics of the Agilent High Sensitivity Detection Cell e 10 fold increase in signal to noise e Detector linearity beyond 2000 mAU for accurate quantitative analysis e Decoupled design allows replaceable capillaries and reduced cost operation ___ High Sensitivity Cell with 75 pm id capillary e Special capillary geometry insures 75 pm id standard capill maintenance of peak symmetry Pepe Ea S Eep ay e Full diode array spectral capabilities e Design fits all Agilent CE instruments Agilent High Sensitivity Detection Cell vs 75 pm Standard Capillary for the CZE separation of
11. naphthalene sulfonic acids High Sensitivity Detection Cell G1600 CE Part No Description G1600 68723 High sensitivity cell kit Includes detection cell 75 um ID inlet capillary 72 cm and outlet capillary 8 5 cm pair capillary cassette fittings 3 fitting screws with seals 2 fitting caps cleaning solution CE Partner CD ROM CE cell fitting kit 61600 63200 Includes 3 fitting screws 2 fitting caps Replacement detection cell G1600 60027 Cell cleaning fluid 1L 5062 8529 Capillary Kits for High Sensitivity Detection Cell Description Effective Length cm G1600 CE Part No 75 um capillary kit with 8 5 cm outlet 56 G1600 68716 72 G1600 68715 88 G1600 68714 PVA coated 75 um capillary kit with 8 5 cm outlet 56 G1600 68319 os CE MS Accessories The CE MS adapter kit simplifies coupling the Agilent CE system with MS systems equipped with an electrospray ionization ESI source Integral to this kit is the CE MS cassette which completely thermostats the capillary until it exits the CE system The cassette offers multiple capillary paths which vary the capillary length A methods development configuration uses on line diode array detection and MS For rapid or routine MS analysis the detector can be bypassed to decrease the total capillary length and reduce analysis time The CE MS adapter kit can be used with the complete Agilent 6000 Series mass spectrometers or virtually any el
12. phosphates walls and detergents or sample adsorption Solution s Condition capillary and allow sufficient equilibration time Replace capillary Caused by conditioning capillary at high or low pH and employing a low or high pH runnning buffer Hysteresis of wall charge Changes in buffer pH changes due to electrolysis Avoid pH differences Allows sufficient equilibration time Replenish buffer composition P Buffer evaporation Tightly cap buffer vials and reduce carousel temperature Conditioning solution waste flushed into outlet reservoir Use separate vial to collect waste Conditioning solutiion carried over into buffer vial First dip capillary in separate buffer or water vial Buffer reservoirs not level Generation of laminar flow Level liquid in reservoirs If not replenishing buffer do not use inlet vial for flushing capillary Different silanol content of capillary batches Different wall charge and variations in EOF Measure EOF and normalize Temperature changes Changes in viscosity and EOF Use system with capillary thermostating Poor Peak Area Reproducibility Sudden application of high Heating thermal expansion of buffer and expulsion of voltage sample Ramp separation voltage or inject buffer plug after sample Sample evaporation Increasing sample concentration and peak area Cap vials and or reduce temperature of sample carousel Instrumenta
13. LULAR LECES Pere T feece Signa aadddedds mait aa LLLA e T Oo ol N Szi CZE of a tryptic digest of recombinant human growth hormone using a standard fused silica capillary with 75 pm internal diameter Standard Bare Fused Silica Capillaries 2 pk ID pm Total Length cm Effective Length Color Code Part No a 50 33 24 5 Green G1600 63211 48 5 40 Green G1600 60211 64 5 56 Green G1600 61211 80 5 72 Green G1600 62211 75 33 24 5 Blue G1600 63311 48 5 40 Blue G1600 60311 64 5 56 Blue G1600 61311 80 5 72 Blue G1600 62311 112 5 104 Blue G1600 64311 100 33 24 5 Gray 61600 63411 48 5 40 Gray 61600 60411 64 5 56 Gray 61600 61411 80 5 72 Gray G1600 62411 1125 104 Gray 1600 64411 Helpful hint Different inner diameters of capillaries need to use different alignment interfaces to guarantee optimal detection The color coding of the capillary and the alignment interface allow you to easily match the correct interface with the capillary Extended Light Path Bubble Cell Bare Fused Silica Capillaries 25 pm with 125 pm bubble Electroosmotic flow maintains the plug flow in the bubble Optical slits matched to the zone geometry maintain resolution 10 46 47 Time min Time min Use Agilent Technologies extended light path capillaries bubble cell capillaries Analysis of cold medicine ingredients in a standard capillary
14. T and C values have smaller pores and are therefore more effective at resolving smaller smaller molecules PAGE 10 10 T O C capillaries provide high resolution capabilities for separation of antisense therapeutic agents primers and probes as well as nucleotides UPAGE 5 5 T 5 C allows single base resolution of oligonucleotides pd A ranging from 20 150 bases For your convenience PAGE capillaries and PAGE buffers can be purchased together or separately To achieve the highest reproducibility and provide optimal longevity use UPAGE buffer with PAGE capillaries PAGE Starter Kits Includes 3 PAGE capillaries 75 cm total length 50 cm effective length oligonucleotide standard and PAGE buffer Kit as defined by type of pPAGE capillary uPAGE 10 10 T 0 C PAGE pd A o _39 oligonucleotide standard for yPAGE 10 kit PAGE buffer 2 x 237 mL ID pm 100 Part No 92 1311 uPAGE 5 5 T 5 C HPAGE pd A os 39 40 60 Oligonucleotide standard for pPAGE 3 and uPAGE 5 kits PAGE buffer 2 x 237 mL 75 92 5211 UPAGE 3 3 T 3 C PAGE pd A 2530 40 60 Oligonucleotide standard for pPAGE 3 and uPAGE 5 kits PAGE buffer 2 x 237 mL 75 192 3211 PAGE Basic Kits Includes 3 PAGE capillaries 75 cm total length 50 cm effective length Kit as defined by type of pPAGE capillary uPAGE 10 10 T 0 C PAGE pd A o _39 oligonucleotide standard for yPAGE 10 kit ID um 100 Part No 191
15. ectospray MS platform The CE MS cassette completely thermostats the capillary until it exits the CE system Methods development configuration uses online diode array detection DAD and MS For rapid or routine MS analysis the DAD can be by passed to decrease the total capillary length and reduce analysis time The CE ESI MS Nebulizer Kit includes the electrospray needle and splitter assembly which allows the direct connection of the CE instrument with Agilent and other electrospray MS systems The CE ESI MS Nebulizer Kit needs the CE MS Adapter Kit for fully supporting the CE MS coupling Leu Enkephalin MW 555 6 Met Enkephalin MW 573 7 VYV MW 379 4 Angiotensin II MW 1046 2 ad Abundance nm 450000 25 350000 250000 150000 50000 4 MS TIC scan CE MS of 4 component peptide mixture 210 fmol Interfacing the capillary requires an electospray needle which is not included in this kit but in the CEESI MS Nebulizer Kit For coupling with non Agilent MS please contact the MS vendor The CE with tandem UV Vis and MS detection allows the analysis of complex mixtures Analyte mixtures are separated and the components detected via UV Vis absorption allowing preliminary identification based on peak elution time and or UV Vis spectra when compared to a standard The on line coupling to electrospray ionization mass spectrometry ESI MS then reveals unambiguous information on the solute s molecular weight and po
16. em SIL DB coated capillaries have been widely used in clEF applications PCR product and DNA fragment separation and many other CE applications which require reduced EOF P E st D a B 2 a 3 a 3 lt gt l Time min Analysis of Myoglobin tryptic digest using pSIL WAX pSIL FC and pSIL DNA Capillaries with Windows Capillary ID pm Total Effective Film Unit Part No Length Length Thickness C cm pm USIL FC 50 80 50 0 075 3 pk 194 8111 uSIL DNA 75 65 50 0 075 2 pk 199 2602 USIL WAX 50 100 75 0 1 2 pk 196 7203 USIL WAX 100 100 75 0 1 2 pk 197 7202 Bulk pSIL DB Capillaries Capillary ID mm Length m Film Thickness Part No nm DB 1 0 05 10 0 05 126 1012 DB 1 0 20 10 0 05 126 1013 DB 1 0 10 10 0 10 127 1012 DB 17 0 10 10 0 05 126 1713 DB 17 0 10 10 0 10 127 1712 DB 17 0 20 10 0 10 127 1713 Capillary Electrochromatography CEC Capillaries Capillary electrochromatography is a hybrid of CE and LC and can be performed in the Agilent CE system Using CE capillaries packed with LC stationary phases CEC offers the loadability and selectivity of LC and the high efficiency of CE Using the high pressure capabilities of the Agilent CE system both ends of the CEC capillary can be pressurized This process prevents outgassing upon application of high voltage and significantly extends capillary lifetime Use CEC to improve resolution o
17. ent All kits are prepared using the same quality procedures as our buffers and are thoroughly tested and supported While the kits have been optimized for use with the Agilent CE system they may be used with virtually any commercial or home built CE system Formate Acetate Propionate n Butyrate n Pentanoate n Hexanoate n Heptanoate n Octanoate l 2 3 4 5 6 T 8 Separation of short chain carboxylic acids using the Organic Acids Solutions Kit Inorganic Anion Solutions Kit The Inorganic Anion Solutions Kit contains all components needed for the analysis of common inorganic anions such as chloride bromide iodide fluoride sulfate and phosphate Applications include the analysis of inorganic ions in e Ultra pure water e Waste water e High purity chemicals e Drug formulations e Pulp and paper solutions e Semiconductor solutions Using an indirect UV detection system optimized for small anions analyses are sensitive and rapid and provide an alternative to traditional ion chromatography The kit contains the buffer capillaries test mixture and instructions Thiosulfate Bromide lodide Chloride Sulfate Nitrite Nitrate Molybdate VI Azide Thiocyanate Chlorate Fluoride 1 2 3 4 5 6 7 8 Separation of common anions Inorganic Anion Solutions Kit Inorganic Anion Solutions Kit 5063 6511 Ultra pure CE water 500 mL 5062 8578 0 1 N sodium hydr
18. f solutes which are difficult to resolve by HPLC for hydrophobic solutes which cannot be solubilized in MEKC buffers or for reduced sample and solvent consumption compared to HPLC Helpful hint Isocratic conditions Capillary Electrochromatography of diuretic test mixture courtesy of Dr Melvin Euerby Astra Charnwood UK Standard Packed CEC Capillaries 2 pk Chlorothiazide Hydrochlorothiazide Chlorthalidone Hydroflumethiazide Bendroflumothiazide Bumetanide Buffer 50 mmol Na HPO pH 2 5 60 20 20 v v MeCN Buffer H 0 40 20 40 v v MeCN Buffer H 0 Description ID pm Total Length Effective Color Code Part No cm Length cm C18 3 um 100 33 5 25 Gray 5063 6512 100 48 5 40 Gray 0063 6513 C8 3 ym 100 33 5 25 Gray 5063 6535 100 48 5 40 Gray 2063 6540 Phenyl 3 um 100 335 25 Gray 5063 6536 100 48 5 40 Gray 9063 6541 The color coding of the capillary alignment stopper and the interface allows you to easily combine the correct alignment interface with the capillary Thiourea Methylparaben Ethylparaben Ptopylparaben Butylparaben Pentylparaben Naphthalene Biphenyl m po St e e ee oom Fluorene Phenanthrene 11 Anthracene 10 Capillary Electrochromatography of parabenes and aromatics CEC capillaries require an Agilent CE system with external gas supply capabilities 12min 12 Fluoranthene TD Alignment Interfaces and Capilla
19. id 25 pm and an Agilent Extended to improve sensitivity 3 to 5 fold over Light Path Capillary standard capillaries With extended light path capillaries the inner diameter is increased only at the detection window Extended Light Path Bubble Cell Bare Fused Silica Capillaries 2 pk onenng ne SENSUM oraora ID pm Total Effective Bubble Optical Color Code Part No diameter capillary and the low current Length Length Factor Path generation of a narrow one cm cm Length nm Resolution is not sacrificed when used with 25 48 5 40 5 125 Black 61600 60132 Agilent Technologies matching optical 64 5 56 5 125 Black 61600 61132 alignment interfaces 80 5 72 5 125 Black 61600 62132 50 43 5 35 3 150 Red 01600 60233 Through a computer controlled proprietary 48 5 40 3 150 Red 61600 60232 process the diameter is increased three to 64 5 56 3 150 Red 61600 61232 five times with a manufacturing precision 80 5 72 3 150 Red 61600 62232 better than 3 Take advantage of this 1125 104 3 150 Red 61600 64232 process to extend the detection pathlength 75 48 5 40 27 200 Yellow 61600 60332 of 25 um ID capillaries to 125 um 50 pm 64 5 56 27 200 Yellow 61600 61332 ta 0 Um and 2S pato 200 pi 80 5 72 27 200 Yelow 61600 62332 112 5 104 27 200 Yellow _ 61600 64332 Helpful hint Use narrow 25 and 50 um id bubble cell capillaries for highly conductive buffers without sacrificing sensitivity SD U J 50pm ID
20. ifies that the solution CZE Buffers for Charged Analytes is free of fluorescence active impurities 1 and 2 Rayleigh stray light of zero and Description Volume mL Part No first order 3 Raman stray light 50 mM sodium phosphate buffer pH 2 5 250 5062 8571 50 mM sodium phosphate buffer pH 7 0 250 5062 8572 50 mM sodium tetraborate buffer pH 9 3 250 5062 8573 20 mM sodium tetraborate buffer pH 9 3 100 8500 6782 CZE Buffers for Proteins Description Volume mL Part No 50 mM phosphate 0 05 hydroxyethyl cellulose buffer pH 2 5 250 8500 6786 150 mM phosphate 200 mM ammonium sulfate buffer pH 7 0 250 8500 6787 MEKC Buffers for Neutral and Charged Analytes CZE analysis of a peptide mixture using premade Description Volume mL Part No 50 mM sodium phosphate buffer pH 2 5 50 mM sodium tetraborate 100 mM sodium dodecyl sulfate buffer 250 5062 8574 pH 9 3 Dilute with 50 mM sodium tetraborate pH 9 3 P N 5062 8573 to reduce SDS concentration without affecting the tetraborate composition or pH Plating Bath Analysis Buffer Description Volume mL Part No Plating bath analysis buffer 250 5064 8236 PAGE Buffer Solutions and Oligo Standards PAGE Tris borate and urea buffer for PAGE 3 and pPAGE 5 4 x 237 mL 590 4001 PAGE Tris borate and urea buffer for pPAGE 10 4 x 237 mL 590 4005 HPAGE pd A os 39 40 60 Oligonucleotide standard for pPAGE 3 and pPAGE 5 3 x 50 pL 590 4000 Fa
21. in PVA Coated Capillaries for Agilent CE System Users ID pm Total Bubble Color Code Part No Length Factor Effective Length Optical Path Length CT cm non Agilent systems 50 64 5 56 0 50 Green 61600 61219 64 5 56 3 150 Red 61600 61239 125 21 5 0 50 Blue 61600 67219 Now the PVA is also available for use with 75 64 5 56 0 1200 61600 68319 the High Sensitivity Detection Cell for even 195 15 0 J5 Blue 61600 67319 further improved HPLC like sensitivity In 700 18 5 10 0 100 Gra 61600 60419 addition PVA coated capillaries are offered i 7 ner ae 64 5 56 0 100 Gray 61600 61419 fro CE MS applications The capillaries are Wor compatible with Borate buliers provided with normally positioned detection Note The PVA capillaries for CE MS have a blue alignment stopper matching the blue color code of the window to allow tandem UV Vis and MS alignment interface for MS UV Det The alignment stopper of the 50 um ID PVA capillary for CE MS has a detection for improved sample black dot for easy identification identification PVA Coated Capillaries for Non Agilent CE System Users ID pm Total Length Effective Bubble Factor Optical Path Part No cm Length cm Length pm 50 71 60 0 50 G160U 61219 71 60 3 150 G160U 61239 100 56 45 0 100 G160U 60419 71 60 0 100 G160U 61419 Not compatible with Borate buffers Note When extended pathlength capillaries are used in non Agilent s
22. ium lamp Use DAD test to measure lamp output and time on Replace if necessary Data acquisition rate too high Determine peak width and decrease acquisition rate if appropriate Improper reference wavelength Acquire UV spectrum during analysis Use lowest wavelength possible without impinging where sample absorbs Also use wide bandwidth Buffer absorbs at detection wavelength Use minimally UV absorbing buffers such as phosphate and borate especially bewlow 210 nm Drifting baselne Improper capillary alignment Re seat capillary cartridge in detector block Unequilibrated temperature Allow 10 20 minutes for equilibration after opening top cover Lamp recently ignited Allow 15 30 minutes for equilibration after igniting lamp Poor Peak Efficiency Broad peaks Skewed peaks Sample overloading Decrease sample injection or concentration Excessive Joule heating Mismatched sample buffer ion mobilities Reduce voltage buffer conductivity or capillary id Match mobilities or increase difference between buffer and sample conductivity Sample overloading Decrease samplle injection or concentration Tailing peaks Adsorption to capillary wall Use pH extremes high buffer concentrations polymer additives or coated capillary EJ Symptom Possible Cause Poor Migration Time Reproducibility Adsorption to capillary Changes in EOF caused by buffer especially
23. l limitations System rise time significant proportion of injection time Increasae injection time Poor Peak Area Reproducibility Sample carry over Extraneous injection Zero injection caused by simply dipping the capillary in the sample Extraneous injection Distorted peak shape tailing Non eluting sample Sample adsorption to capillary walls Use capillary with flat smooth injection end Remove polyimide from end of capillary Cannot be totally eliminated Increase injection amount to minimize effect Change buffer pH Increase buffer concentration Use additive such as cellulose or coated capillary Low signal to noise ratio Integration errors Optimize integration parameters Increase sample concentration Use peak height Temperature changes of Changes in viscosity and injection amount capillary environment Use system with capillary thermostatting ED www agilent com chem ce Information descriptions and specifications in this publication are subject to change without notice Agilent Technologies Inc 2009 Printed in the USA November 1 2009 5990 3822EN 5990 3822EN Agilent Technologies
24. nit Part No Cation Solutions Kit 5064 8206 Cation buffer 250 mL 5064 8203 Ultra pure CE water 500 mL 5062 8578 Bare fused silica capillary extended light path bubble factor 3 2 pk G1600 61232 50 um ID 56 cm long Cation test mixture 25 mL 5064 8205 Note The following part should be ordered separately for use with the Agilent CE System Alignment interface for 50 um ID extended light path capillary P N G1600 60230 Organic Acids Solutions Kit The Organic Acids Solution Kit is ideal for the analysis of short alkylchain carboxylic acids Employing an indirect UV detection agent optimized for organix acids the methodology is simple sensitive and provides accurate quantitative analysis Suited for the analysis of organic acids in a wide range of matrices it is especially useful for determination of organic acids in beverages and food Chloride Sulfate Oxalate Formate Malate Citrate Succinate pa ESN o a SO Pyruvate Acetate Lactate Phosphate Pyroglutamate Anions in beer Organic acids in beer and red wine Organic Acids Solutions Kit Chloride Sulfate Oxalate Formate Malate Citrate Succinate Pyruvate Acetate Lactate Phosphate Anions in red wine Includes 1000 ppm each of malate succinate and lactate Description Unit Part No Organic Acids Solution Kit 5063 6510 Organic acids buffer o 250ml 8500 6785 Ultra pure CE water 500 mL 5062 8578 1 0 N sodium hyd
25. onsuming buffer preparation process All Agilent buffers and reagents are designed to meet the stringent demands of CE Manufactured under GLP GMP conditions in 1S09001 facilities each is shipped with assay information and verification of purity Chemicals are all electrophoresis grade with nearly all ionic and organic impurities removed Solutions are prepared under Class 10 cleanroom conditions and prefiltered through 0 2 mm filters to ensure removal of particulates Superior quality control ensures reproducible results bottle to bottle and batch to batch EJ In addition to a set of kit buffers which are specially designed for dedicated applications Agilent offers a series of basic CZE buffers covering a broad pH range The product portfolio also includes special buffers for protein analysis and for Micellar Electrokinetic Chromatography MEKC Cleaning and conditioning solutions complete the offering Agilent Technologies Capillary Electrophoresis program includes premade buffers designed to reduce your time at the laboratory bench Ultra Pure CE Water Int EmL Ref rare Description Volume mL Part No 7 Ultra pure CE water 500 5062 8578 Capillary Conditioning Solutions Description Volume mL Part No 0 1 N sodium hydroxide 250 5062 8575 1 0 N sodium hydroxide 250 5062 8576 0 1 N phosphoric acid 250 5062 8577 The total fluorimetry spectrum of the 50 mM borate buffer pH 9 2 1 ver
26. oxide 250 mL 5062 8575 1 0 N sodium hydroxide 250 mL 5062 8576 Bare fused silica capillary 50 um ID 72 cm long 2 pk 61600 62211 Inorganic anion test mixture 10 mL 5062 8524 Includes 1000 ppm each of fluoride chloride bormide nitrite sulfate and 2000 ppm phosphate Note The following part should be ordered separately for use with the Agilent CE System Alignment interface for standard 50 um ID capillary P N G1600 60210 Cation Solutions Kit The Cation Solutions Kit provides everything you need for the analysis of inorganic and Ammonium low molecular mass organic cations It is Potassium specially designed for the separation of Sodium alkali metal ions alkaline earth metal ions Calcium and alkyl amines in a wide range of Magnesium different matrices 12 Coffee 10 Each kit contains a cation buffer bare fused silica capillaries a cation standard CE grade water and a detailed description of the analysis method and most common applications including detection limits and reproducibility data The Cation 12 345 Solutions Kit and the separation methods Cation Standard 12 345 Cation Standard were developed to fit perfectly with the Agilent CE instrument and to support its Imin 4 Smin high automation capabilities The methods are very easy to perform and provide accurate and quantitative analyses Cations in coffee and energy drinks Energy Drink B Cation Solutions Kit Component U
27. ries In addition since EOF is eliminated cumbersome washing procedures are unnecessary and migration time reproducibility can be improved Each batch of PVA coated capillaries is rigidly tested by Agilent Technologies and includes a representative electropherogram to assure quality The color coding of the capillary alignment stopper and the alignment interfaces allow you to easily combine the correct interface with the capillary Capillaries for non Agilent CE system users have removable alignment stoppers without color code PVA capillaries can be used for a variety of applications including protein analysis at physiological pH isoelectric focusing and small anion analysis without the need for flow reversal agents in the buffer Untreated fused silica capillary PVA coated capillary Use of PVA capillaries to reduce protein adsorption Ce NS ok ew NS Cytochrome C Lysozyme Lactoglobulin A Lactoglobulin A Lactoglobulin B Benzidine Aniline o Toluidine p Cl Aniline 2 Aminonaphtaline 4 4 Methylene bis 2 chloroaniline 3 3 Dichlorobenzidine 2 Amino 5 azotoluene 2 Methyl 5 nitroaniline 9 CZE analysis of basic amines using PVA capillaries decomposition products of az The PVA coating is available in standard capillaries or in Agilent Extended Light Path Capillaries bubble cell capillaries for high sensitivity applications Both capillary types are available in greater lengths for use
28. roxide 250 mL 5062 8576 Bare fused silica capillary 75 um ID 72 cm long 2 pk G1600 62311 Organic acids test mixture 20 mL 8500 6900 Note The following part should be ordered separately for use with the Agilent CE System Alignment interface for 75 um ID capillary P N G1600 60310 Forensic Anions Solutions Kit This highly focused kit was developed specifically for the analysis of poisonous compounds such as cyanide azide selenate arsenate and arsenite In cases of poisoning analytical tools are needed to determine teh identity of the toxins quickly and accurately A rapid determination of anionic toxins in adulterated foods and beverages is possible using CE with indirect UV detection The forensic and other anions can be detected within 15 minutes with minimal sample preparation PAGE poly acrylamide gel filled capillaries are the most direct vehicles to transfer all of your applications from slab gel to CE enjoying the automation high speed high resolution and quantitative advantages of CE The capillaries are ideal for high resolution and quantitative advantages of CE THe capillaries are ideal for high resolution separations of oligonucleotides single stranded and double stranded DNA fragments polymerase chain reaction PCR products sequencing reaction products and oligosaccharides Cyanide Chloride Nitrite Nitrate Sulfate Azide Selenate de 2 3 4 5 6 8 Carbonate so Fl
29. ry Cassette Agilent Technologies alignment interfaces are an integral part of the Agilent diode array detection system These interfaces contain optical slits which are precisely matched to the capillary inner diameter for optimized sensitivity and linear detection range In combination with the capillary cassette alignment interfaces simplify capillary exchange protect the fragile detection window and ensure exact alignment of the window in the detector Quick change Capillary cassette G7100 60002 cassette allows capillary exchange in less than a minute Note The color code of the alignment interface must match with the color code of the capillary s built in alignment stopper Alignment Interfaces Description ID um Color Corresponding G7100 CE G1600 CE Code TELA Part No Part No Alignment interface for 59 Green Green G7100 60210 G1600 60210 standard capillary 75 Bhs Blue 67100 60310 G1600 60310 100 Gray Gray 150 Brown Brown Alignment interface for 25 Black Black G7100 60150 G1600 60150 one she Light 59 Red Red 67100 60230 61600 60230 ay capianes J35 Yelow Yellow G7100 60330 G1600 60330 CE MS alignment Blue Blue G7100 60400 G1600 60400 interface for 360 um Gray OD capillaries nonmetallic Note 75 100 and 150 um ID standard capillaries use the same interface color blue PVA coated 50 and 75 um ID capillary for CE MS use the same nonmetallic interface with color code blue for use with standard and
30. ssibly also structure CE MS Adapter Kit Description CE MS Adapter Kit For interfacing the Agilent CE system with a mass spectrometer Includes parts below which can be ordered separately Part No G1603A CE MS interface cassette for G1600 and G7100 G1600 60013 CE MS alignment interface for 360 um OD capillaries nonmetallic for G1600 G1600 60400 CE MS alignment interface for 360 um OD capillaries nonmetallic for G7100 Bare fused silica 50 um ID 125 cm long 2 pk G7100 60400 G1600 67311 Interfacing the capillary requires an electrospray needle which is not included in this kit CE MS Sprayer Kit Description Unit CE MS Sprayer Kit Includes CE MS test sample 5 g quinine sulfate dihyrate and the parts listed below ES needle assembly Part No G1607A G1607 60041 CE ESI sprayer G1607 60001 Splitter assembly G1607 60000 PEEK ferrule 360 um for CE MS Sprayer 5022 2141 Nut fingertight fitting and ferrule 2 pk 0100 1543 Flex loc element 2 pk 1520 0401 Gasket 1 pk 61607 20030 lon kit ammonium acetate 5x5 mL 8500 4410 CE MS Capillaries Description Color Code Unit Part No Bare fused silica 50 um ID 125 cm long Green 2 pk G1600 67311 PVA coated capillary 50 ym ID 125 cm long Green 1 pk G1600 67219 PVA coated capillary 75 um ID 125 cm long Blue 1 pk G1600 67319 CE Standards amp Reagents Premade buffers help eliminate the time c
31. uoride 10 Arsenate 11 Phosphate 12 Glutamate 13 Acetate 14 Arsenite 15 Lactate Analysis of an Anion standard with the Forensic Aniona Analysis Kit Forensic Anions Solutions Kit Description Unit Part No Forensic Anions Solutions Kit 5 x 50 mL 5064 8208 Basic anion buffer 50 mL 5064 8209 Ultra pure CE water 500 mL 5062 8578 Bare fused silica capillary 50 um ID 104 cm long 2 pk 61600 64211 Inorganic anion test mixture 10 mL 5062 8524 Includes 1000 ppm each of fluoride chloride bormide nitrite sulfate and 2000 ppm phosphate Note The following part should be ordered separately for use with the Agilent CE System Alignment interface for standard 50 um ID capillary P N G1600 60210 UPAGE Solutions Kits for High Resolution DNA Fragments Analysis Single base pair separation of pd A ona3 T 3 C pPAGE capillary P N 1 E 3 m z c S B 3 a lt gt i oo 25 30 Time min UV Adsorption 260 nm mo Secondary peaks of identical base length as the primary peaks but retarded in electromigration due to loss of the terminal 5 phosphate group T 25 Time min Oligonucleotide samples with or without terminal 5 phosphate group uPAGE capillaries are available in three different pore sizes The size of the molecular sieving pores is controlled by the monomer concentration T and the degree of polymer cross linking C Gels with higher
32. ydroxide 100 mL CE Operational Qualification Performance Verification O0 PV Chemical Kit 5063 6515 Includes buffer 20 mM borate pH 9 3 100 ml test samples 0 1 0 5 1 0 and 5 0 mM 4 hydroxy acetophenone 2 ml ea capillary conditioning solution 0 1 N sodium hydroxide 100 ml test capillary L 48 5 cm 40 cm ID 50 ym diskette with methods sequence spectral library Note Method is supported for G1600 only CE 00 PV Chemicals Only Kit 5063 6520 Includes buffer 20 mM borate pH 9 3 100 mL test samples 0 1 0 5 1 0 and 5 0 mM 4 hydroxy acetophenone 2 mL ER Instrument Parts and Supplies Vials and Caps for CE Crimp snap top vial 1mL Polypropylene 100 pk 5182 0567 Clear wide opening crimp snap top glass vial 2 mL 100 pk 5182 9697 Clear wide opening crimp snap top glass vial 2 mL 500 pk 5183 4623 Amber wide opening crimp snap top vial write on spot 2 mL 100 pk 5183 4619 Crimp snap top vial polyethylene 250 uL 1000 pk 9301 0978 Snap caps PEO polyethylene olefin for chemical resistance 100 pk 5181 1507 Snap caps PEO polyethylene olefin for chemical resistance 500 pk 5181 1513 Snap caps PUR polyurethane for resealing 100 pk 5181 1512 Snap caps PUR polyurethane for resealing 500 pk 5042 6491 PUR caps are recommended to help prevent sample or buffer evaporation even after multiple injections Instrument Supplies
33. ystems loss of resolution may be found if the axial slit width is not reduced In Agilent systems the alignment interface contains properly matched slits to maintain resolution i N o a fo a a x T 16 17 Te T 18 min Analysis of meat proteins by c IEF using PVA capillaries a CEP Coated Capillaries CEP capillaries contain a permanently bonded polymer coating This CEP coating shields the silanol functionality of the capillary surface and helps prevent sample adsorption Additionally EOF is nearly eliminated making the capillary ideal for applications such as DNA separations with sieving polymer buffers Elimination of EOF also simplifies analysis of anions and organic acids by direct UV detection Without EOF reduction highly mobile ions such as nitrate can migrate in the opposite direction as the slower longer chain acids Restriction fragment separation 36 2645 bp The CEP coated capillary is stable from pH 2 to 8 It can be used with borate buffers CEP Coated Capillaries 2 pk fferi different surface functionality t ol a i Pren seats Ps ae ID um Total Length Effective Bubble Factor Optical Path Part No elp alleviate Sample adsorption Fac cm Length cm Length um batch of CEP coated capillaries is rigidly tested by Agilent Technologies and each a 80 5 12 0 1 61600 9218 capillary includes a representative electropherogram to assure quality
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