Agilent LC컬럼 선택가이드

Agilent LC & LC/MS 컬럼선택가이드 올바른컬럼선택이더나은분석결과를제공합니다. 영화과학과 Agilent 가제안하는정확한컬럼선택가이드를통해 최고의분석결과를만들어보세요 역상 everse Phase Ion- Exchange Special Application 순상 Normal Phase GPC/SEC BioSolution

Agilent 역상컬럼선택가이드 SmallMolecules MW<000 LargeMolecules MW>000 80~120A 공극크기 () 00A 컬럼의공극크기는분석하려는분자의크기에따라결정됩니다. 일반적으로크기가작은분자들은확산이잘되는 80~120A 공극크기를사용하며펩타이드나단백질같은크기가큰분자들은 00A 의컬럼을사용합니다. EclipsePlusC18 초기선택컬럼의고정상 00SB-C18 대체적으로분석을많이하는중간극성및비극성시료는머무름시간의최대화를위하여 C18 컬럼으로시작합니다. C18 컬럼에서분리능이좋지않거나, 단백질과같이큰분자량및 C18 로분석하기어려운매우비극성의물질을분석할경우,C8 과같은짧은탄소길이의컬럼을사용합니다. 빠른 LC 분석의경우,Poroshell120EC-C18 이나 EclipsePlusHT/HD 컬럼 (UHPLC 전용 ) 을사용합니다. SmallMolecules MW<000 LargeMolecules MW>000 일반분석 빠른분석 일반분석 빠른분석 Eclipse Plus C18 4.x150mm,.5μm 제품번호 9599-902 Poroshell 120EC-C18 4. x 100 mm, 2.μm 제품번호 9595-902 ZBAX HD Eclipse Plus C18 2.1 x 50 mm, 1.8μm 제품번호 9595-902 ZBAX00SB-C18 4. x 150 mm, 5μm 제품번호 88995-902 ZBAX00SB-C18 4. x 50 mm,.5μm 제품번호 859-902 2.1 Poroshellx 5 mm, 00SB 5μm-C 18 제품번호 050-902 일반분석 ZBAXEclipsePlus/XDB - 분석법개발을위한첫번째선택 - 일반적분석에적합한컬럼 Pursuit - 1~10 까지사용가능 - 높은 ing 량 page StableBond(SB) page ExtendC18 page - 낮은 범위와높은온도에견딜수있는컬럼 - 다양한고정상제공 - 높은 (11.5) 에서도안정함 ( 2-11.5) - 비극성물질간의분리도향상 page - 극성물질분석에적합 빠른분석 극성물질분석 Poroshell120 5page - 일반LC 기기로UHPLC 의결과를얻을수있는컬럼 - 낮은압력에서빠른분석이가능 HT/HD - 빠른분석용컬럼 - UHPLC 전용컬럼 5page Agilent 역상컬럼 PLP-S - 1-14 까지사용가능 - Polymer 지지체 기본구조 - 작은분자에서큰분자까지 (100A ~4000A ) 에따라분리 Bonus-P 4page SB-Aq 4page Polaris 4page - Triple Endcapping 으로인한 Peak Tailing 감소 - C18 고정상이아님 - 100% 수용성이동상사용가능 - SB 컬럼보다극성물질분석에적합 - 낮은 에서긴컬럼수명 - 고정상의손상없이재현성있는결과획득 - 수용성용매에사용가능 - 극성물질분리에적합 - 분석물질의극성도에따른선택이다양함 - 높은수용성이동상에서안정 - 극성물질분리에적합 4page 이동상 에따른컬럼선택 장점 : 분리도가좋음 단점 : 높은 에서안정하지않음 - EclipsePlus&XDB :2~9~ StableBond(S :1~ B) Extend-C18 :2~11.5 Pursuit :1.5~10(Diphenyl 의경우 Bonus-P 1.5~.5) :2~8 SB-Aq :1~8 Polaris :1.5~10 Polymer 장점 : 높은 에서안정 단점 : 분리도가좋지않음 - PLP-S : 1~14 2 AgilentLC&LC/MS 컬럼선택가이드

everse Phase Columns ZBAX Eclipse Plus/XDB 역상컬럼 everse PhaseColumns ZBAX EclipsePlus ZBAX EclipseXDB 분석법개발을위한첫번째선택 염기성물질에서도좋은피크모양유지 QA/QC 테스팅으로재현성있는결과확보 1.8,.5,5μm 입자크기제공으로일반분석에서빠른분석까지적용가능 특허된실리카제조공법으로분석결과향상 C H CH Improved double end capping 고밀도결합 (extradenselybonded) 으로컬럼수명연장 산성, 염기성, 중성물질분석시좋은피크모양 선택의폭이넓은컬럼사양 (Capillary, 일반분석, Prep 등 ) CH C H Temp. Limits ange* ZBAXEclipse PlusC18 95A 10 m 2 /g 0 C 2.0-9.0 Double 9% ZBAXEclipse PlusC8 95A 10 m 2 /g 0 C 2.0-9.0 Double % ZBAXEclipsePAH 95A 10 m 2 /g 0 C 2.0-8.0 No 14% ZBAXEclipse PlusPhenyl-Hexyl 95A 10 m 2 /g 0 C 2.0-8.0 Double 9% Temp. Limits ange* ZBAXEclipseXDB-C18 80A 180 m 2 /g 0 C 2.0-9.0 Double 10% ZBAXEclipseXDB-C8 80A 180 m 2 /g 0 C 2.0-9.0 Double.% ZBAX EclipseXDB-Phenyl 80A 180 m 2 /g 0 C 2.0-9.0 Double.2% ZBAX EclipseXDB-CN 80A 180 m 2 /g 0 C 2.0-8.0 Double 4.% ZBAX 80A StableBond 1 정도의낮은 와높은온도 (80-90 C) 에도긴컬럼수명과일관된재현성 SB-AQ 컬럼은 100% 수용성용매사용가능 Peptide 와 Protein 분석용으로 SB00 제공 다양한고정상으로여러가지분석에응용가능 =Isopropyl, Isobutyl 1 =C18,C8,CN,C etc. ZBAX 80A Extend-C18 1 H H 1 1 Temp. Limits ange* ZBAXSB-C18 80A 180 m 2 /g 90 C 0.8-8.0 No 10% ZBAX SB-C8 80A 180 m 2 /g 80 C 1.0-8.0 No 5.5% ZBAX SB-C 80A 180 m 2 /g 80 C 1.0-8.0 No 4% ZBAXSB-Phenyl 80A 180 m 2 /g 80 C 1.0-8.0 No 5.5% ZBAX SB-CN 80A 180 m 2 /g 80 C 1.0-8.0 No 4% ZBAX SB-Aq 80A 180 m 2 /g 80 C 1.0-8.0 No proprietary 높은 범위 (11.5) 까지사용가능 독특한결합구조로높은 에서도안정 LC/MS 를이용한 Peptide 분석에응용 폴리머베이스컬럼보다더좋은분리도와피크모양제공 C18 C18 Support Column: ZBAXExtend-C18 450-902 4.x150mm,5μm Mobile Phase: 5% Methanol / 25% 50mM Pyrrolidine Buffer, 11.5 Flow ate: 0.5mL/min Temperature: 40 C Detector: UV,215nm 1 2 4 1. Doxepin 2. Imipramine. Nortriptyline 4. Amitriptyline 5. Trimipramine 5 Temp. Limits ange* Carbo ZBAXExtend-C18 80A 180 m 2 /g 0 C 2.0-11.5 Double 12.5% 0 2 4 8 10 12 14 1 Time(min) 그림 1. 염기성이동상사용시유기산분석의예 Pursuit / PursuitXs ange Volume Ligand Coverage Pursuit C18 200A 200 m2/g 1.5-10 Yes 12.9% 11 ml/g.5 μmol/m2 1~10 까지사용가능 높은 ing 량으로높은 Capacity 다양한사양보유 CH C H Pursuit C8 200A 200 m2/g 1.5-10 Yes.4% 11 ml/g.8 μmol/m2 Pursuit Diphenyl 200A 200 m2/g 1.5-8.0 Yes.% 11 ml/g 2.8 μmol/m2 Pursuit PFP 200A 200 m2/g 1.5-10 Yes.% 11 ml/g.4 μmol/m2 Pursuit PAH 200A 200 m2/g 1.5-10 Yes 11 ml/g Pursuit Xs C18 100A 440 m2/g 1.5-10 Yes 22% 11 ml/g 2.9 μmol/m2 Pursuit Xs C8 100A 440 m2/g 1.5-10 Yes 15% 11 ml/g. μmol/m2 Pursuit Xs Diphenyl 100A 440 m2/g 1.5-8.0 Yes 14.% 11 ml/g 2. μmol/m2 Pursuit Xs 100A 440 m2/g 1.5-10 Yes 11 ml/g Pursuit Xs Ultra C18 100A 440 m2/g 1.5-10 Yes 2.2% 11 ml/g.2 μmol/m2 Pursuit Xs Ultra C8 100A 440 m2/g 1.5-10 Yes 15% 11 ml/g. μmol/m2 Pursuit Xs Ultra Diphenyl 100A 440 m2/g 1.5-8.0 Yes 14.% 11 ml/g 2. μmol/m2

1 1 ZBAX Bonus-P Triple endcapping 으로인한 Peak tailing 감소 역상분리에독특한고정상 극성물질분석에적합 1 1 1 1 1 PG PG PG Column: ZBAXBonus-P 888-901 4.x150mm,5μm Mobile Phase: 0%25mM PhosphateBuffer,.0:40%ACN Flow ate: 1.5mL/min Te mperature: 2 C Toluene Plate Heigh t, cm 0.001 8 Competitive alkyl/ amid e Bonus-P 0.001 0.001 4 SB-Aq Temp. Limits 낮은 에서매우긴컬럼수명 고정상의손상없이재현성있는결과를얻을수있음 매우극성물질분리에적합 ange* H H 1 1 ZBAXBonus-P 80A 180 m 2 /g 0 C 2.0-9.0 Triple 9.5% Column: ZBAXSB-Aq 8895-914 4.x150mm,5μm Mobile Phase: 5%MeH/95%water (0.1% TFA) Flow ate: 2.0 ml/min Te mperature: 5 C Detector: UV,254nm 0.001 2 0.001 0 mau 10 120 80 0 5,000 10,000 15,000 20,000 25,000 0,000 5, 000 Column Volume ofpurge 그림 2. 타사비교자료 (Tripleendcapping 으로인해 에서도안정함 ) 1 2 1. Thiamine 2. Nicotinic Acid. Pyridoxine 4. Niacinamide =Isopropyl 40 Temp. Limits ange* ZBAX SB-Aq 80A 180 m 2 /g 80 C 1.0-8.0 No proprietary 4 0 0 2 Time (min) 그림.SB-Aq 를이용하여수용성비타민분석의예 Polaris PLP-S Polymer 극성물질분리에적합 분석물질의극성도에따라다양한선택가능 End capped Volume Liquid Coverage Polaris C18-A 180A 200 m 2 /g 1.8% Yes 1.1 cm /g.9 μmol/m 2 Polaris C8-A 180A 200 m 2 /g.4% Yes 1.1 cm /g 4.8 μmol/m 2 Polaris C18-Ether 180A 200 m 2 /g 12.1% Yes 1.1 cm /g. μmol/m 2 PolarisC8-Ether 180A 200 m 2 /g.1% Yes 1.1 cm /g 4.5 μmol/m 2 PolarisAmide C18 180A 200 m 2 /g 15% Yes 1.1 cm /g 4.4 μmol/m 2 Polaris NH2 180A 200 m 2 /g 5.5% Amide 1.1 cm /g.8 μmol/m 2 Polaris-A 180A 200 m 2 /g N/A N/A 1.1 cm /g N/A Polymer 지지체기본구조 (USPL21) 작은분자에서큰분자까지 size(100a -4000A ) 에따라분리 Column Specifications ange 1-14 Buffer Content Unlimited rganic Modifier 1-100% TemperatureLimits 200 C MaximumPressure 5-8 μm: 000 psi (210 bar) / μm: 4000 psi (00bar) PLP-SApplications 100A 00A 1000A 4000A Application Small molecules/syntheticbiomolecules ecombinantpeptides/proteins Largeproteins DNA/highspeed 여러가지고정상에따른결과분석응용에맞추어최적의고정상을찾으세요. SB-Pheny k=1.00 1. indole ptimal selectivity for the beta blocker l Pind olol. 2. pindolo l k=.00 0 0.5 1 1.5 2 2.5.5 min m k=0.5 k=4.25 0 0.5 1 1.5 2 2.5.5 SB-C N min Columns: ZBAXHT StableBond4.x50mm1.8μm Mobile Phase : A: 5 Na Acetate (USP method) B:MeCN, (0:0) Temperature: 25 C Flow : 1.5mL/min. Detection : 219,4 nm,0.2s responsetime,semi-micro flowcell Sample: USPpindololresolutionsolution k=0.25 k=.5 SB-C1 8 0 0.5 1 1.5 2 2.5.5 min 4 AgilentLC&LC/MS 컬럼선택가이드

Poroshell 120 역상컬럼 everse PhaseColumns 일반 HPLC 에서도 UHPLC 의결과를얻을수있습니다! 1.μm 의 Solid Core 에 0.5μm 의 Porous ut-layer 의고정상 독특한고정상으로 Sub-2μm 컬럼에비해최대 90% 효율, 50% 정도의낮은압력 ( Sub-2 μm : 2μm 보다작은입자크기 ).5μm 컬럼보다약 2 배높은효율 최대 00bar 까지사용가능하여일반 LC 와 UHPLC 에모두이상적 12 가지고정상으로다양한선택성 BondedPhase Temp.Limits ange EC-C18 120A 0 C 2.0-8.0 Double 10% EC-C8 120A 0 C 2.0-8.0 Double 5% SB-C18 120A 90 C 1.0-8.0 No 8% SB-C8 120A 80 C 1.0-8.0 No 5.5% HPH-C18 100A 0 C.0-11.0 Double Proprietary HPH-C8 100A 0 C.0-11.0 Double Proprietary Phenyl-Hexyl 120A 0 C 2.0-8.0 Double 9% SB-Aq 120A 80 C 1.0-8.0 No Proprietary Bonus-P 120A 0 C 2.0-9.0 Triple 9.5% HILIC 120A 0 C 0.0-8.0 No N/A EC-CN 120A 0 C 2.0-8.0 Double.5% PFP 120A 0 C 2.0-8.0 Yes 5.1% Solid Core 1.8 μm totally porous Agilent Poroshell 120 2. μm 1. μm size 120A, ideal for small molecules 1.8 μm 2.μm 0.5 μm 0.4 0.82 1.148 1.518 2.01 2.22 제품번호 9595-02 제품번호 95994-02 Mobile phase: 0%Acetonitrile, 40%water Flowrate: 0.58 ml/min N = 2505, Press = 182 bar Injectionvolume: 4 μl N = 2295, Press = 8 bar Temperature: 2 Detection: DADg=254,4nm ef=0,100nm Samplepreparation:LCcheckoutsample P/N5188-529 spiked 2.5 5.5 10 2.5 5.5 10 12.5 min w/50μl2mg/mlthiourea in water/acetonitrile(5:5) 2.81 4.12.2 9.5 0.49 0.85 1.25 1.5 2.418 2.05.494 5.281 8. 21 8 1.041 그림 4. 독특한고정상입자로 1.8μm 컬럼에비해 90% 효율과 50% 정도의낮은압력의결과를보여주는 Poroshell 120 컬럼 HD & HT 빠르고정확한분석을위한선택 최고성능의 UHPLC 전용으로분석을더욱정확하고빠르게 - HT: apidesolutionhighthroughput- 1.8 μm / 00bar 까지 - HD: apidesolutionhighdefinition 1.8 μm / 1200bar 까지 * HD 컬럼의고정상은계속업데이트됩니다. BondedPhase area ange ZBAX Eclipse PlusC18 95A 10m 2 /g 2.0-9.0 Yes ZBAX Eclipse PlusC8 95A 10m 2 /g 2.0-9.0 Yes ZBAX EclipseXDB-C18 80A 180m 2 /g 2.0-9.0 Yes ZBAXExtend-C18 80A 180m 2 /g 2.0-11.5** Yes ZBAX StableBondSB-C18 80A 180m 2 /g 1.0-8.0* No ZBAX StableBond SB-C8 80A 180m 2 /g 1.0-8.0* No ZBAX StableBond SB-Phenyl 80A 180m 2 /g 1.0-8.0* No ZBAX StableBondSB-CN 80A 180m 2 /g 1.0-8.0* No ZBAX StableBond SB-Aq 80A 180m 2 /g 1.0-8.0* No ZBAX EclipsePAH 95A 10m 2 /g 2.0-8.0 No ZBAX HILICPlus 95A 10m 2 /g 0.0-8.0 No 1.Kathon1A 2. Kathon1B.Carbendazim 4. 1,2- Benzisothiazol-(2H)-one 5. 2-Phenoxyethanol. BenzoicAcid. Methyl Paraben 1 Pressure 101 bar 2 그림 5. ZBAX HD Eclipse Plus C18 2.1х 50mm, 1.8μm 컬럼을이용하여방부제 종분석의예 0 0.1 0.2 0. 0.4 0.5 0. 0. 0.8 min 4 A: H2 (0.05 % TFA) B:ACN(0.04%TFA) Time0.0 Time1.0 Time1.1 5 95/5A/B 55/45A/B 0/100A/B Flow ate: 1.mL/min 1μLinjection 50ppm std. 0 C DAD:25nm(0min) 225nm(0.4min) 255nm(0.min) 5

순상컬럼 Normal-PhaseColumns ZBAX Normal-Phase Columns ange ZBAXx-SIL 80A 180 m 2 /g 0-8.0 No ZBAX EclipseXDB-CN 80A 180 m 2 /g 2.0-8.0 Yes 4.% ZBAXCN 0A 00 m 2 /g 2.0-.0 Yes % ZBAXNH 2 0A 00 m 2 /g 2.0-.0 Yes 4% Column: ZBAXCN,880952-05,4.x 250mm, 5 μm MobilePhase : A: Heptane B :2-Methoxyethanol/Isopropanol (50/50) Flow ate: 2mL/min Temperature : 50 C Detector: 28nm Sample: ctylphenoxy (polyethyleneoxy) EthanolSurfactant(n=10) ZBAX x-sil 고순도실리카 ( >99.995% ) 1.8μm, 5μm 입자크기제공 HILICmode 로사용가능 극성물질분석에응용 USPL ZBAX Eclipse XDB-CN x-sil 의고순도실리카사용 염기성물질분석에적합 USPL10 0 5 10 15 Time(min) 20 ZBAX CN ZBAXSIL 의실리카에 Cyanopropyldimethylsilane 처리 컬럼보다내구성이강함 USPL10 ZBAX NH 2 Weakanion-exchange 컬럼으로사용 비타민 A 와 D 분석에적합 USPL8 ZBAX HILIC Plus 순상고정상과역상이동상의만남 극성물질의머무름최대화를위해사용 LC/MS 응용에적합 EPAMethod194 에적합 ange Non-bondedsilica 95A 10 m 2 /g 0-8.0 AgilentLC&LC/MS 컬럼선택가이드

Ion ExchangeColumns ZBAX Ion Exchange :SAX & SCX 이온교환컬럼 Ion ExchangeColumns ZBAX SAXColumn: Quaternaryamine 작용기로수용성물질 /(-) 이온성물질분석에적합 ZBAX SCXColumn: Sulfonicacid 작용기로염기성, 수용성물질, 바이오물질 /(+) 이온성물질분석에적합 고효율과빠른분석제공 BondedPhase ange Functionality Max Pressure ZBAXSAX 0A 00m 2 /g 2.0-.0 Quaternaryamine 50 bar ZBAX00SCX 00A 50 m 2 /g 2.0-.0 Sulfonic acid 50 bar Hi-Plex Polymer 응용 : 단당, 이당과올리고당, 유기산분석 USPL1,L19,L4,L58 에적합 물과희석된산용매사용가능 다양한 Application 제공 ded hase Temperature ange Flowate(mL/mn) Flow ate (ml/min) Eluent Eluent Application on Hi-Plex Ca 80-90 C 0. Water USPMethodsSpecifyingL1Media Hi-PlexCaUSPL19 80-90 C 0. Water 0-90 C 0. Water Hi-PlexHforcarbohydrates 0-0 C 0. Water Hi-PlexHfororganicacids 40-0 C 0. DiluteAcid Hi-Plex Ca(Duo) 80-90 C 0. Water Hi-Plex K 80-90 C 0. Water Hi-PlexNa(cto) 80-90 C 0. Water, SodiumHydroxide Hi-PlexNa 80-90 C 0. Water Conditions Sample : Column: rganic acids Agilent ( 제품번호 PL110-80 Mobilephase: 0.1MH2S4 Flowrate: 0. ml/min Temperature : 50 C Detector: I Conditions Sample : Column: Sample size : Mobile phase: Flowrate: Injection volume : 10 μl Temperature: 85 C Detector: I Sugarsandsugaralcohols Agilent Hi-Plex Ca (Duo) ( 제품번호 PL1F0-850) 10mg/mL 100%DIH 2 0.4 ml/min 1 2 그림.Hi-PlexH 컬럼을이용한유기산 9 종분석의예 1 4 5 0 min 0 2 45 0 min 0 그림.Hi-Plex Ca 컬럼을이용한단당류및다당류분석의예 8 8 9 9 1. xalic acid 2. cis-aconitic acid. Tartaric acid 4. Malic acid 5. Lactic acid. Formic acid. Fumaric acid 8. Propionic acid 9. Butyric acid 1. affinose 2. Sucrose. Lactulose 4. Glucose 5. Galactos e. Fructose. ibitol 8. Mannitol 9. Sorbitol USPMethodsSpecifyingL19Media USPMethodsSpecifyingL4Media USPMethodsSpecifyingL58Media Mono-andDisaccharides AnomerSeparations rganicacids Alcohols Adulteration of Food andbeverages FoodAdditives DairyProducts Sweetened DairyProducts Confectionery FruitJuice Wine Wood Pulp Hydrolysates (cellulose/hemi-cellulose) FermentationMonitoring ligosaccharides ecommended Column Hi-Plex Ca and Hi-Plex Ca(Duo) Hi-Plex Na and Hi-PlexNa(cto) Hi-Plex Ca Hi-PlexNa(cto) Hi-Plex Ca Hi-Plex Ca Hi-Plex K Hi-PlexCaand Hi-Plex Caand Hi-Plex Caand Hi-Plex Caand Hi-Plex Ca Hi-PlexNa SampleswithHighSaltContent(molasses) Hi-PlexNa(cto) ligosaccharides <Dp5 with Monosaccharides CornSyrups Hi-Plex Ca(Duo) Hi-PlexNa

5 9 9 P 5 4 P S 스페셜컬럼 SpecialColumns Special Application Columns ZBAX Carbohydrate Analysis 재현성있고효율적인당분석을위한컬럼 Aminopropylpacking material 로단당류와이당류분리에적합 4.X150mm 컬럼기준 50μL 정도많은량의시료주입가능 ID(efractiveindex detectors) 사용시추천 Poer ange* ZBAX CarbohydrateAnalysis 0A 00 m 2 /g 2.0-8.0 No.5% Xylose h a m n o s e Xylitol F r u c t o s e 1 2 4 5 X y l o s e h a m n o s e F r u c t o s e G l u / G a l G l u c o s e o s e M a l t o s e L a c t u l o s e S u c r L a c t o s e S u c r o s e Maltose Lactose a f f i n o s e a f f i n o s e 1 2 4 5 ZBAX Amino Acid Analysis(AAA) 응용 : 아미노산 24 종분리 PA 와 FMC 유도체화방법사용 표준물질과유도체화시약 Agilent 공급가능 Agilent Autosampler 를사용하여자동화된유도체화로쉽고간편함 Column Agilent ZBAX Eclipse AAA 4. 150 mm, 5 μm ( 제품번호 99400-902) Mobile Phase A: 40 mm Sodium phosphate(di-basic) 0.1% Phosphoric acid B: Acetonitrile / Methanol / DW =45/45/10 Detector DAD 8nm (1차아미노산 ), 22nm (2차아미노산 ) FLD Ex : 40nm, Em : 450nm (1차아미노산 ) Ex : 2nm, Em :05nm (2차아미노산 ) Flowrate 1.5 ml/min Temp. 40 Time (min) B (%) 0.0 0 1.9 0 18.1 5 18. 80 22. 80 2.2 0 2.0 0 제품명 제품번호 PA reagent, 10 mg/ml each in 0.4 Mborate butter o-phthalaldehyde(pa) 501-5 and -mercaptopropionicacid, x1mlampoules FMC reagent, 2.5 mg/ml in acetonitrile, 9-fluorenylmethylchloroformate, 501-1mL. 10ampoules BorateBuffer100mL 501-9 AdvanceBio Peptide Mapping 일관된 Lot 별재현성 2.μm Poroshell 구조의고정상으로좋은분리도 HPLC,UHPLC 모두사용가능 00bar 까지사용가능 LC/MS 분석시최적의감도제공 짧은시간안에정확하게 Peptide Mapping 완성 Column: ZBAX Carbohydrate Analysis,8400-908, 4.x150mm,5 μm Mobile Phase: 5% ACN/25% H2 Flow ate: 2 ml/min Temperature 0 C : Detector: ID Det.Temp: 0 C Sample: (54μg each),inj.=.μl Column: AdvanceBio PeptideMapping, 2.1 x250mm,2. μm,5150-902 Flow ate: 0.4mL/min Injection: 5μL(2.0mg/mL) Temp: 55 C Detection: 220nm Gradient: A, water(0.1%fa);b, ACN(0.1% FA), 0-28min,-45% B; 28-min,45-0% B; -4min,0-95% B AdvanceBio Glycan Mapping HILIC 고정상으로재현성있고정확하게 Glycan Mapping 완성 좋은분리도와빠른분석가능 (1.8μm :<1200bar,2.μmPoroshell : <00bar) LC/MS 와 LC/FLD 에추천 Glycan 표준물질및 Library Agilent 공급가능 1 차아미노산 mau 40 0 20 10 0 40 0 20 10 0 Xylose h a m n o s e Xylitol M a n n i t o l L a c t u l o s e 2 1 4 5 T i m e ( m i n ) 그림 8. Carbohydrate 컬럼을이용한당과당알콜분석의예 DAD1 A, g=8,10 ef=90,20 (AMIN \AA-000.D) 1.9 2 - AS.9 0 - GL U.0 8 - AS N.0 4 - SE.9 - GLN.1 - HI.50 - GL Y.9 9 - TH 0 2 4 8 2 차아미노산 mau DAD1 B, g=22,1 ef=24,8 (AMIN \AA-000.D) 0.80 1.0 1.9.1.8 5.2 5.455 5.8 5.85 4 1.08.0.940.1.498.9 8.2 9 - A G 8.88 9 - AL A 8.2 8.04 8.88 0 2 4 8 10 12 00 250 200 150 100 50 10.02 - TY 11.49 4 - CY2 11.88 - VA L 12.092- M ET 12.4- NVA 12.895 - TP 1.00 - PHE 1.4 9 - IL E 14.125 - LE U 14.58 - LYS a f f i n o s e 10 12 14 1 18 min 10.02 11.25 11.44 11.2 12.0011.88 12.092 12.441 12.4 12.89 1.11 5 1.0 0 9 1.49 14.12 그림 9. 24 종아미노산동시분석의예 0 5 10 15 20 25 0 0 그림 10. Biosimilar EP 에대한 Peptide Mapping 의예 14.959 - HY 14.58 1.50 1.00 - SA 1.54 - P 14 1 18 min 8 Agilent LC & L C/MS 컬럼선택가이드

GPC / SEC Columns & Standards 분자량에따라사용되는 GPC/SEC 컬럼 유기용매사용 PLgel MIXED 컬럼가이드 SEC 컬럼 Exclusion Columns 분자량 2,000~40,000,000 500~10,000,000 200~2,000,000 200~400,000 Up to 0,0000 추천컬럼PLgel 20 μm MIXED-A PLgel 10 μm MIXED-B PLgel 5 μm MIXED-C PLgel 5 μm MIXED-D PLgel μm MIXED-E GPC/SEC 컬럼 PL aquagel-h PLgel lexis PL HFIPgel PLgel Preparative PL apide PolarGel Plus 특징수용성용매사용높은분자량의고분자분석 HFIP 용매사용 Prep 의용도로사용되는 PLgel 빠른분석을위한선택수용성 / 유기용매모두적합최신기술로내구성향상 Column choices Polarity rganic solvents Polar/mixed solvents Aqueous solvents (PS) MW range (PEG/PE) g/mol g/mol 0 0 PLgel 10 μm 10 A 00,000 to 10,000,000 PLgel 5/10 μm 10 5 A 0,000 to 1,00,000 PLgel 5/10 μm 10 4 A 10,000 to 450,000 MW range PLgel 5/10 μm 10 A 500 to 0,000 PLgel 5/10 μm 50A 0 to 1,500 Columns PLgel /5/10 μm 100A 0 to 5,000 Agilent Plus Columns High pore volume for very high resolution PLgel 5/10 μm 500A 500 to 25,000 The latest technology in GPC for increased speed and resolution PLgel μm MIXED-E 0 to 25,000 Agilent PL apide Columns Special format for fast separations Up to 10,000,000 PL apide H Up to 2,000,000 Poly PL apide M Up to 500,000 esi PL apide L Up to 25,000 Meso Up to,00 ligo PL apide F PLgel 5 μm MIXED-D 200 to 400,000 PLgel 5 μm MIXED-C 200 to 2,000,000 PLgel 10 μm MIXED-B 500 to 10,000,000 PLgel 20 μm MIXED-A 2,000 to 40,000,000 PolarGel-L 0 to 0,000 PolarGel-M 0 to 500,000 1,000 1,000 10,000 10,000 100,000 100,000 1,000,000 1,000,000 10,000,000 10,000,000 100,000,000 100,000,000 PL aquagel-h 20 5 μm 100 to 20,000 PL aquagel-h 0 8 μm 100 to 0,000 PL aquagel-h 40 8/15 μm 10,000 to 200,000 PL aquagel-h 50 8/15 μm 50,000 to 00,000 PL aquagel-h 0 8/15 μm 200,000 to 10,000,000 PL aquagel-h MIXED-H 8 μm,000 to 10,000,000 PL aquagel-h MIXED-M 8 μm 1,000 to 500,000 GPC/SEC 분석을위한고분자표준물 정확하고재현성있는 GPC/SEC 컬럼 Calibration 표준물질로 IS 9001:2000 인증을받은제품 좁은범위부터넓은범위까지다양한분자량의표준물질제공 키트뿐만아니라개별분자량별표준물질제공 Polymer Type Individual MW Calibration Kits EasiCal EasiVial Type of GPC/SEC Polystyrene rganic Polymethylmethacrylate rganic Polyethylene glycol (PEG) rganic/aqueous Polyethylene oxide (PE) rganic/aqueous Pullulan polysaccharide rganic/aqueous Polyacrylic acid Na salt Aqueous 9

바이오컬럼 BioColumns Biosolution Columns Primary Structure eversed-phase separations 소수성작용 Aggregates Fr agmentpattern Exclusion s eparations(sec) 분자크기분리 Biomolecule ChargeIsoforms Ion-Exchange s eparations(iex) 이온교환 Affinity ProteinA 친화력분석 eversed-phase Column for Protein & Peptides Poroshell 00 Biomolecules 분석을위한빠르고효율적인컬럼 1,000kDa 까지의 Protein 분석에효과적 독특한고정상구조로빠른분석가능 Temp.Limits* ange Poroshell 00SB-C18, C8,C 00A 90 C 1.0-8.0 No Poroshell00Extend-C18 00A ZBAX 00A ExtendC18 40 Cabove8 0 Cbelow8 넓은 범위에서매우효과적인고정상 ( 2-11.5) 높은 에서 Hydrophobic Peptides 의좋은분석결과도출 LC/MS 사용시 Ammonium-hydroxide 용매사용가능 2.0-11.0 Yes ZBAX 00A StableBond size00a 컬럼으로 Protein 과 Peptide 등거대분자에고효율역상분리 다양한고정상제공으로폭넓은응용지원 높은온도에서도재현성있고신뢰성있는데이터도출 Temp Limits* ange* ZBAX00SB-C18 00A 45 m 2 /g 90 C 1.0-8.0 No 2.8% ZBAX00SB-C8 00A 45 m 2 /g 80 C 1.0-8.0 No 1.5% ZBAX00SB-C 00A 45 m 2 /g 80 C 1.0-8.0 No 1.1% ZBAX00SB-CN 00A 45 m 2 /g 80 C 1.0-8.0 No 1.2% ZBAX00-Diphenyl 00A 45 m 2 /g 80 C 1.0-8.0 Yes 1.9% Temp Limits* ange* ZBAX00A Extend-C18 00A 45 m 2 /g 0 C 2.0-11.5 Double 4% Exclusion Agilent Bio IonExchange 재현성있고다양한선택이가능한 Agilent SEC 컬럼 AgilentBio SEC-& Bio-5: 다양한 size 선택가능 ProSEC 00S: 고농도의염조건에서 Globular Protein 분석에적합 ZBAXGF-250& GF-450: Protein 의 Preparative SEC 분석에적합 PL aquagel-h : 넓은분자량범위의 Biopolymer 분석에적합 다양한분야에서사용할수있도록제공 PEEK 재질과 StaninlessSteel 재질로제공 ID Particle SCX(Strongcation-exchange)-S0 H WCX (Weak cation-exchange) -C00H SAX(Stronganion-exchange)-N() 2.1 and 4. mm 1.,,5 and 10 μm Stability perating Temp Limit 2-12 80 C Flow ate 0.1-1.0 ml/min WAX (Weak cation-exchange) -N(C 2H 5) 2 Agilent Bio-monolith Protein A 세포배양에서 mab 농도의빠른검출을위해최적화된컬럼 유속에관계없이분리할수있는 Bio-monolith 기술로분석시간감소 10 Agilent LC & LC/MS 컬럼선택가이드

Agilent Bio Column 의다양한응용 eversed-phase Column ExclusionChromatography (SEC) Column Application Agilent Columns Application Agilent Columns Proteins and polypeptides ZBAX00, 1.8μm Peptides,proteins Agilent BioSEC- HD00SB-C18 HD00SB-C8 Large biomolecules andsamples with multiple molecular weight components Agilent BioSEC-5 HD00SB-C Globularproteins,antibodies ProSEC00S HD00-Diphenyl Proteins,globularproteins ZBAXGF-250/450 HD00-HILIC ZBAX 00 StableBond 00SB-C18 LowMWpolymersandoligomers, oligosaccharides, PEGs, lignosulfo nates 2orPLaquagel-H PLaquagel-H8μm PLaquagel-H205μm 00SB-C8 PLaquagel-HMIXED-M8μm 00SB-C 00SB-CN ZBAX 00 Extend-C18 Polydisperse biopolymers, polysaccharides, cellulose derivatives 2orPLaquagel-H PLaquagel-HMIXED-H8μm PLaquagel-H0/50/408μm Peptidesandproteinsupto 1,000kDA,monoclonalantibodies andintact proteins Poroshell00 00SB-C18 Very high MW polymers, hyalu ronicacids,starches,gums PL aquagel-h 0/50/40 15μm in series 00SB-C8 00SB-C 00Extend-C18 Ion-exchange Column Smallhydrophilicpeptides inproteindigests PeptidestoDNA Smallmolecules/peptides/oligonucleotides ecombinantpeptides/proteins Poroshell120 PLP-S 100 00 1000 4000 PLP-S100 PLP-S00 Largeproteins PLP-S 1000 DNA/high speedseparation PLP-S 4000 Affinity Column Application Monoclonalantibodies Peptidesandproteins Proteins, peptidesand deprotected syntheticoligonucleotides Globularproteinsandpeptides Very large biomolecules/highspeed Smallpeptidesto large proteins Globular proteins Very large biomolecules/highspeed Agilent Columns Agilent BioMAb AgilentBioIEX PL-SAX 1000 4000 PL-SAX1000 PL-SAX4000 PL-SCX 1000 4000 PL-SCX1000 PL-SCX4000 Application IgG(human andmouse), exceptforiggclass Agilent Columns Bio-Monolith ProteinA Antibodies(IgG, IgM), plasmiddna, viruses, phages and other macro bio molecules Bio-Monolith Bio-MonolithQA Bio-MonolithDEAE Bio-MonolithS Agilent LC 컬럼의다양한사양및자세한 Application 은영화과학에문의해주세요. 02-2140-540 Viruses,DNA,large proteins Plasmid DNS,bacteriophages Proteins,antibodies Bio-MonolithQA Bio-MonolithDEAE Bio-MonolithS 11

Agilent LC 컬럼 USPcode USP USP Packing Materials Column L1 ctadecyl silane chemically bonded to porous silica or ceramic microparticles, 1.5 to 10 μm in diameter, or a monolithicrod ZBAXEclipsePlusC18 ZBAXEclipseXDB-C18 ZBAX SB-C18 ZBAX x-c18 ZBAXExtend-C18 ZBAX DS ZBAXDSclassic PursuitXsC18 Pursuit C18 Polaris C18-A PolarisC18-Ether Poroshell120EC-C18 Poroshell120SB-C18 Poroshell120HPH-C18 Poroshell00SB-C18 USP USP Packing Materials Column L15 L1 L19 L20 L21 L22 L25 Hexylsilane chemically bonded to totally porous silica particles, to 10 μ min diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the hydrogen form, to 11 μm in diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in thecalcium form,9μmindiameter Dihydroxypropanegroups chemicallybondedto porous silicaparticles, to10 μmindiameter A rigid spherical styrenedivinylbenzene copolymer, 5 to 10 μm in diameter A cation-exchange resin made of porous polystyrene gel with sulfonic acid groups, about10μminsize Packinghavingthe capacityto separatecompoundswith a MWrange from 1,000 to 5,000 da (as determined by the polyethylene oxide), applied to neutral, anionic and cationic water-soluble polymers. A polymethacrylate resinbase, crosslinked withpolyhydroxylated ether (surface contained some residualcarboxyl functional gruops) wasfound suitable Metal C Hi-Plex Ca Hi-PlexCa(Duo) LiChrospherDiol PLgel PLP-S PLaquagel-H L Porous silica particles, 1.5 to 10 μm in diameter, or a monolithic silicarod Poroshell00Extend-C18 SepTechST0C18 SepTechST150C18 AgilentPrepC18 ZBAXHILICPlus ZBAX x-l ZBAX SIL L L4 Packinghavingthecapacityto separatedextronsby molecularsize overa range of 4,000 to 500,000 da. It is spherical, silica-based, and processed to provide stability Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the lead form, about 9 μm in diameter ZBAX GF-250 Bio SEC- Bio SEC-5 ProSEC PursuitXs Polaris -A L5 A zirconium-stabilized spherical silica packing with a hydrophilic (dioltype) molecular monolayer bonded phase havingaporesizeof 150A ZBAXGF 250 ZBAXGF 450 L ctylsilane chemically bonded to totally porous silica particles, 1.5 to 10 μmin diameter,or a monolithicsilica rod Agilent Prep ZBAXEclipsePlusC8 ZBAXEclipseXDB-C8 ZBAX SB-C8 ZBAX X-C8 ZBAX C8 PursuitXsC8 Pursuit C8 Polaris C8-A PolarisC8-Ether Poroshell120EC-C8 Poroshell120SB-C8 L4 Pentafluorophenyl groups chemically bonded to silica particles by a propyl spacer,5to10μmindiameter L45 Betacyclodextrin bonded to porous silicaparticles, 5 to10μmindiameter L50 Multifunction resin with reversed-phase retention and strong anionexhange functionalities. The resin consists of ethylvinylbenzene, 55% crosslinked with divinylbenzene copolymer, to 15 μm in diameter, and a surface area of not less than 50 m 2 per g. Substrate is coated with quarternary ammonium functionalized latex particles consistingof styrenecross-linked with divinylbenzene L52 Weak cation-exchange resin made of porous silica with sulfopropyl groups, 5to10μmindiameter L5 Weak cation-exchange resin consisting of ethylvinylbenzene, 55% crosslinked with divinylbenzene copolymer, to 15 μm diameter. Substrate is surface grafted with carboxylic acid and/or phosphoric acid functionalized monomers.capacity notlessthan400μeq/column Pursuit PFP Poroshell120 PFP ChiraDex Chiral ZBAX 00SCX IonoSpher C Bio SAX Poroshell120HPH-C8 Poroshell00SB-C8 L5 Propyl silanechemically bonded to totally porous silica particles,to 10μm in diameter ZBAX SB-C L8 L9 L10 An essentially monomolecular layer of aminopropylsilane chemically bonded to totally poroussilica gel support, to 10 μmindiameter Irregular or spherical, totally porous silica gel having a chemically bonded, strongly acidic cation-exchange coating, to10μmindiameter Nitrile groups chemically bonded to porous silica particles, to 10μmindiameter ZBAX NH2 Polaris NH2 ZBAX SCX ZBAX CN ZBAX SB-CN L5 L58 L0 A chiral-recognition protein, ovomucoid, chemically bonded to silica particles,about5μmindiameter, withaporesizeof 120A Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the sodium form, about to 0 μm in diameter Spherical, porous silica gel, 10 μm in diameter, the surface has been covalently modified withalkylamide groups andendcapped Ultron ES-VM Hi-Plex Na Hi-PlexNa(cto) PolarisAmide-C18 ZBAXEclipseXDB-CN L11 Phenyl groups chemically bonded to porous silica particles, 1.5to10μmindiameter ZBAXEclipseXDBPhenyl ZBAXEclipsePlusPhenyl-Hexyl 분석방법조정가이드 USP & FDA L1 L14 Trimethylsilane chemically bonded to porous silica particles, to 10μmindiameter gelhavingachemically bonded,strongly basicquaternary ammoniumanion-exchangecoating,5to10μmindiameter ZBAX Phenyl Poroshell120Phenyl-Hexyl PursuitXsDiPhenyl Pursuit DiPhenyl ZBAX TMS ZBAX SAX IonoSpher A Parameter 컬럼길이 Max Spec ± 0% FDA±50% 컬럼 ID USP±25% 컬럼선속도유지 유속 ± 50% 주입량 최대한적게감도 / 재현성유지 입자크기 to 50% 주요관공서의표준방법을수정할경우기존분석법과수정방법을동시에수행하여정확성, 재현성등검증조건에영향을주지않아야합니다. 제품상담 : 크로마토그래피팀 (T:02-2140-540) AG BC 201409