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 Small Molecules MW < 3000 Large Molecules MW > 3000 80~120A 공극크기 ( ze) 300A 컬럼의공극크기는분석하려는분자의크기에따라결정됩니다. 일반적으로크기가작은분자들은확산이잘되는 80~120A 공극크기를사용하며펩타이드나단백질같은크기가큰분자들은 300A 의컬럼을사용합니다. Eclipse Plus C18 초기선택컬럼의고정상 300SB-C18 대체적으로분석을많이하는중간극성및비극성시료는머무름시간의최대화를위하여 C18 컬럼으로시작합니다. C18 컬럼에서분리능이좋지않거나, 단백질과같이큰분자량및 C18 로분석하기어려운매우비극성의물질을분석할경우, C8 과같은짧은탄소길이의컬럼을사용합니다. 빠른 LC 분석의경우, Poroshell120 EC-C18 이나 Eclipse Plus HT/HD 컬럼 (UHPLC 전용 ) 을사용합니다. Small Molecules MW < 3000 Large Molecules MW > 3000 일반분석 빠른분석 일반분석 빠른분석 Eclipse Plus C18 4.6 x 150 mm, 3.5μm 제품번호 959963-902 Poroshell 120EC-C18 4.6 x 100 mm, 2.7μm 제품번호 695975-902 ZBAX HD Eclipse Plus C18 2.1 x 50 mm, 1.8μm 제품번호 959757-902 ZBAX 300SB-C18 4.6 x 150 mm, 5μm 제품번호 883995-902 ZBAX 300SB-C18 4.6 x 50 mm, 3.5μm 제품번호 865973-902 Poroshell 300SB-C18 2.1 x 75 mm, 5μm 제품번호 660750-902 일반분석 빠른분석 극성물질분석 ZBAX Eclipse Plus / XDB - 분석법개발을위한첫번째선택 - 일반적분석에적합한컬럼 Pursuit - 1~10 까지사용가능 - 높은 ing 량 Poroshell 120 5page - 일반LC기기로 UHPLC의결과를얻을수있는컬럼 - 낮은압력에서빠른분석이가능 HT / HD - 빠른분석용컬럼 - UHPLC 전용컬럼 3page StableBond (SB) 3page Extend C18 3page - 낮은 범위와높은온도에견딜수있는컬럼 - 다양한고정상제공 - 높은 (11.5) 에서도안정함 ( 2-11.5) - 비극성물질간의분리도향상 3page - 극성물질분석에적합 5page Agilent 역상컬럼 PLP-S - 1-14 까지사용가능 - Polymer 지지체 기본구조 - 작은분자에서큰분자까지 ze (100A ~4000A ) 에따라분리 Bonus-P 4page SB-Aq 4page Polaris 4page - Triple Endcapping으로인한 Peak Tailing 감소 - C18 고정상이아님 - 100% 수용성이동상사용가능 - SB 컬럼보다극성물질분석에적합 - 낮은 에서긴컬럼수명 - 고정상의손상없이재현성있는결과획득 - 수용성용매에사용가능 - 극성물질분리에적합 - 분석물질의극성도에따른선택이다양함 - 높은수용성이동상에서안정 - 극성물질분리에적합 4page 이동상 에따른컬럼선택 lica 장점 : 분리도가좋음 단점 : 높은 에서안정하지않음 Eclipse Plus & XDB : 2~9 StableBond (SB) : 1~6 Extend-C18 : 2~11.5 Pursuit : 1.5~10 (Diphenyl 의경우 1.5~7.5) Bonus-P : 2~8 SB-Aq : 1~8 Polaris : 1.5~10 Polymer 장점 : 높은 에서안정 단점 : 분리도가좋지않음 PLP-S : 1~14 2 Agilent LC & LC/MS 컬럼선택가이드
everse Phase Columns ZBAX Eclipse Plus/XDB ZBAX Eclipse Plus lica ZBAX Eclipse XDB 역상컬럼 everse Phase Columns 분석법개발을위한첫번째선택 염기성물질에서도좋은피크모양유지 QA/QC 테스팅으로재현성있는결과확보 1.8,3.5,5μm 입자크기제공으로일반분석에서빠른분석까지적용가능 특허된실리카제조공법으로분석결과향상 lica Improved Improved double endcapping 고밀도결합 (extra Densely Bonded) 으로컬럼수명연장 산성, 염기성, 중성물질분석시좋은피크모양 선택의폭이넓은컬럼사양 (Capillary, 일반분석, Prep 등 ) ze Temp. Limits ange* ZBAX Eclipse Plus C18 95A 160 m 2 /g 60 C 2.0-9.0 Double 9% ZBAX Eclipse Plus C8 95A 160 m 2 /g 60 C 2.0-9.0 Double 7% ZBAX Eclipse PAH 95A 160 m 2 /g 60 C 2.0-8.0 No 14% ZBAX Eclipse Plus Phenyl-Hexyl 95A 160 m 2 /g 60 C 2.0-8.0 Double 9% ze Temp. Limits ange* ZBAX Eclipse XDB-C18 80A 180 m 2 /g 60 C 2.0-9.0 Double 10% ZBAX Eclipse XDB-C8 80A 180 m 2 /g 60 C 2.0-9.0 Double 7.6% ZBAX Eclipse XDB-Phenyl 80A 180 m 2 /g 60 C 2.0-9.0 Double 7.2% ZBAX Eclipse XDB-CN 80A 180 m 2 /g 60 C 2.0-8.0 Double 4.3% ZBAX 80A StableBond lica 1 정도의낮은 와높은온도 (80-90 C) 에도긴컬럼수명과일관된재현성 SB-AQ 컬럼은 100% 수용성용매사용가능 Peptide 와 Protein 분석용으로 SB300 제공 다양한고정상으로여러가지분석에응용가능 =Isopropyl, Isobutyl 1=C18, C8, CN, C3 etc. H H 1 1 1 ze Temp. Limits ange* ZBAX SB-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-C3 80A 180 m 2 /g 80 C 1.0-8.0 No 4% ZBAX SB-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 ZBAX 80A Extend-C18 lica 높은 범위 ( 11.5) 까지사용가능 독특한결합구조로높은 에서도안정 LC/MS 를이용한 Peptide 분석에응용 폴리머베이스컬럼보다더좋은분리도와피크모양제공 lica Support C18 C18 Column: ZBAX Extend-C18 773450-902 4.6 x 150 mm, 5 μm Mobile Phase: 75% Methanol / 25% 50 mm Pyrrolidine Buffer, 11.5 Flow ate: 0.5 ml/min Temperature: 40 C Detector: UV, 215 nm 1 2 3 4 5 1. Doxepin 2. Imipramine 3. Nortriptyline 4. Amitriptyline 5. Trimipramine ze Temp. Limits ange* ZBAX Extend-C18 80A 180 m 2 /g 60 C 2.0-11.5 Double 12.5% 0 2 4 6 8 10 12 14 16 Time (min) 그림 1. 염기성이동상사용시유기산분석의예 Pursuit / Pursuit Xs 1~10 까지사용가능 높은 ing 량으로높은 Capacity 다양한사양보유 lica ze ange Volume Ligand Coverage Pursuit C18 200A 200 m 2 /g 1.5-10 Yes 12.9% 11 ml/g 3.5 μmol/m 2 Pursuit C8 200A 200 m 2 /g 1.5-10 Yes 7.4% 11 ml/g 3.8 μmol/m 2 Pursuit Diphenyl 200A 200 m 2 /g 1.5-8.0 Yes 7.3% 11 ml/g 2.8 μmol/m 2 Pursuit PFP 200A 200 m 2 /g 1.5-10 Yes 6.3% 11 ml/g 3.4 μmol/m 2 Pursuit PAH 200A 200 m 2 /g 1.5-10 Yes 11 ml/g Pursuit Xs C18 100A 440 m 2 /g 1.5-10 Yes 22% 11 ml/g 2.9 μmol/m 2 Pursuit Xs C8 100A 440 m 2 /g 1.5-10 Yes 15% 11 ml/g 3.7 μmol/m 2 Pursuit Xs Diphenyl 100A 440 m 2 /g 1.5-8.0 Yes 14.6% 11 ml/g 2.6 μmol/m 2 Pursuit Xs 100A 440 m 2 /g 1.5-10 Yes 11 ml/g Pursuit Xs Ultra C18 100A 440 m 2 /g 1.5-10 Yes 23.2% 11 ml/g 3.2 μmol/m 2 Pursuit Xs Ultra C8 100A 440 m 2 /g 1.5-10 Yes 15% 11 ml/g 3.7 μmol/m 2 Pursuit Xs Ultra Diphenyl 100A 440 m 2 /g 1.5-8.0 Yes 14.6% 11 ml/g 2.6 μmol/m 2 3
ZBAX Bonus-P Triple endcapping 으로인한 Peak tailing 감소 역상분리에독특한고정상 극성물질분석에적합 lica 100% 수용매 1 1 1 1 1 PG PG PG Column: ZBAX Bonus-P 883668-901 4.6 x 150 mm, 5 μm Mobile Phase: 60% 25 mm Phosphate Buffer, 7.0:40% ACN Flow ate: 1.5 ml/min Temperature: 23 C Toluene Plate Height, cm 0.0018 0.0016 0.0014 Competitive alkyl/ amide Bonus-P 1 0.0012 SB-Aq 낮은 에서매우긴컬럼수명 고정상의손상없이재현성있는결과를얻을수있음 매우극성물질분리에적합 Polaris ze 극성물질분리에적합 분석물질의극성도에따라다양한선택가능 Temp. Limits ange* H H 1 1 1 ZBAX Bonus-P 80A 180 m 2 /g 60 C 2.0-9.0 Triple 9.5% ze Temp. Limits ange* ZBAX SB-Aq 80A 180 m 2 /g 80 C 1.0-8.0 No proprietary lica lica ze 100% 수용매 100% 수용매 End capped Volume = Isopropyl Liquid Coverage Polaris C18-A 180A 200 m 2 /g 13.8% Yes 1.1 cm 3 /g 3.9 μmol/m 2 Polaris C8-A 180A 200 m 2 /g 7.4% Yes 1.1 cm 3 /g 4.8 μmol/m 2 Polaris C18-Ether 180A 200 m 2 /g 12.1% Yes 1.1 cm 3 /g 3.3 μmol/m 2 Polaris C8-Ether 180A 200 m 2 /g 7.1% Yes 1.1 cm 3 /g 4.5 μmol/m 2 Polaris Amide C18 180A 200 m 2 /g 15% Yes 1.1 cm 3 /g 4.4 μmol/m 2 Polaris NH2 180A 200 m 2 /g 5.5% Amide 1.1 cm 3 /g 3.8 μmol/m 2 Polaris -A 180A 200 m 2 /g N/A N/A 1.1 cm 3 /g N/A Column: ZBAX SB-Aq 883975-914 4.6 x 150 mm, 5 μm Mobile Phase: 5% MeH/95% water (0.1% TFA) Flow ate: 2.0 ml/min Temperature: 35 C Detector: UV, 254 nm PLP-S 0.0010 0 5,000 10,000 15,000 20,000 25,000 30,000 35,000 Column Volume of Purge 그림 2. 타사비교자료 (Triple endcapping 으로인해 7 에서도안정함 ) mau 160 120 80 40 0 0 2 Time (min) 그림 3. SB-Aq 를이용하여수용성비타민분석의예 Polymer 지지체기본구조 (USP L21) 작은분자에서큰분자까지 size(100a -4000A ) 에따라분리 Column Specifications ange 1-14 Buffer Content Unlimited rganic Modifier 1-100% Temperature Limits 200 C Maximum Pressure 5-8 μm: 3000 psi (210 bar) / 3 μm: 4000 psi (300 bar) PLP-S Applications ze 100A 300A 1000A 4000A Polymer Application Small molecules/synthetic biomolecules ecombinant peptides/proteins Large proteins DNA/high speed 1 2 3 4 1. Thiamine 2. Nicotinic Acid 3. Pyridoxine 4. Niacinamide 여러가지고정상에따른결과분석응용에맞추어최적의고정상을찾으세요. 1. indole 2. pindolol k=1.00 SB-Phenyl ptimal selectivity for the beta blocker Pindolol. k=6.00 0 0.5 1 1.5 2 2.5 3 3.5 k=0.75 k=4.25 0 0.5 1 1.5 2 2.5 3 3.5 m min SB-CN min Columns : ZBAX HT StableBond 4.6 x 50 mm 1.8 μm Mobile Phase : A: 5 Na Acetate (USP method) B: MeCN, (70:30) Temperature : 25 C Flow : 1.5 ml/min. Detection : 219, 4 nm, 0.2 s response time, semi-micro flow cell Sample : USP pindolol resolution solution k=0.25 k=7.75 SB-C18 0 0.5 1 1.5 2 2.5 3 3.5 min 4 Agilent LC & LC/MS 컬럼선택가이드
역상컬럼 everse Phase Columns Poroshell 120 lica 일반 HPLC 에서도 UHPLC 의결과를얻을수있습니다! 1.7μm의 Solid Core에 0.5μm의 Porous ut-layer의고정상 독특한고정상으로 Sub-2μm 컬럼에비해최대 90% 효율, 50% 정도의낮은압력 ( Sub-2μm : 2μm 보다작은입자크기 ) 3.5μm 컬럼보다약 2 배높은효율 최대 600bar까지사용가능하여일반 LC 와 UHPLC에모두이상적 12가지고정상으로다양한선택성 ze Temp. Limits ange EC-C18 120A 60 C 2.0-8.0 Double 10% EC-C8 120A 60 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 60 C 3.0-11.0 Double Proprietary HPH-C8 100A 60 C 3.0-11.0 Double Proprietary Phenyl-Hexyl 120A 60 C 2.0-8.0 Double 9% SB-Aq 120A 80 C 1.0-8.0 No Proprietary Bonus-P 120A 60 C 2.0-9.0 Triple 9.5% HILIC 120A 60 C 0.0-8.0 No N/A EC-CN 120A 60 C 2.0-8.0 Double 3.5% PFP 120A 60 C 2.0-8.0 Yes 5.1% Solid Core 1.8 µm totally porous size 120 Å, ideal for small molecules Agilent Poroshell 120 2.7 µm 1.7 µm 1.8 µm 2.7 µm 0.5 µm 0.646 0.826 1.148 1.518 2.016 2.223 695975-302 959964-302 Mobile phase: 60% Acetonitrile, 40% water Flow rate: 0.58 ml/min N = 25053, Press = 182 bar Injection volume: 4 μl N = 27295, Press = 386 bar Temperature: 26 Detection: DAD g = 254,4 nm ef = 360,100 nm Sample preparation: LC checkout sample P/N 5188-6529 spiked 2.5 5 7.5 10 2.5 5 7.5 10 12.5 min w/ 50 μl 2 mg/ml thiourea in water/acetonitrile (65:35) 2.816 4.132 6.273 9.753 0.649 0.865 1.275 1.756 2.418 2.705 3.494 5.281 8.21 8 13.041 그림 4. 독특한고정상입자로 1.8μm 컬럼에비해 90% 효율과 50% 정도의낮은압력의결과를보여주는 Poroshell 120 컬럼 HD & HT lica 빠르고정확한분석을위한선택 최고성능의 UHPLC 전용으로분석을더욱정확하고빠르게 HT : apid esolution High Throughput - 1.8μm / 600bar 까지 HD : apid esolution High Definition 1.8μm / 1200bar 까지 ze area ange ZBAX Eclipse Plus C18 95A 160 m 2 /g 2.0-9.0 Yes ZBAX Eclipse Plus C8 95A 160 m 2 /g 2.0-9.0 Yes ZBAX Eclipse XDB-C18 80A 180 m 2 /g 2.0-9.0 Yes ZBAX Extend-C18 80A 180 m 2 /g 2.0-11.5* Yes ZBAX StableBond SB-C18 80A 180 m 2 /g 1.0-8.0* No ZBAX StableBond SB-C8 80A 180 m 2 /g 1.0-8.0* No ZBAX StableBond SB-Phenyl 80A 180 m 2 /g 1.0-8.0* No ZBAX StableBond SB-CN 80A 180 m 2 /g 1.0-8.0* No ZBAX StableBond SB-Aq 80A 180 m 2 /g 1.0-8.0* No ZBAX Eclipse PAH 95A 160 m 2 /g 2.0-8.0 No ZBAX HILIC Plus 95A 160 m 2 /g 0.0-8.0 No * HD 컬럼의고정상은계속업데이트됩니다. 1. Kathon 1A 2. Kathon 1B 3. Carbendazim 4. 1,2-Benzisothiazol-3(2H)-one 5. 2-Phenoxyethanol 6. Benzoic Acid 7. Methyl Paraben 1 Pressure 1017 bar 2 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 그림 5. ZBAX HD Eclipse Plus C18 2.1х 50mm, 1.8μm 컬럼을이용하여방부제 7 종분석의예 3 4 A: H 2 (0.05 % TFA) B: ACN (0.04 % TFA) Time 0.0 95/5 A/B Time 1.0 55/45 A/B Time 1.1 0/100 A/B Flow ate: 1.7 ml/min 1 μl injection 50 ppm std. 30 C DAD: 275 nm (0 min) 225 nm (0.46 min) 255 nm (0.67 min) 5 6 7 min 5
순상컬럼 Normal-Phase Columns ZBAX Normal-Phase Columns lica ze ange ZBAX x-sil 80A 180 m 2 /g 0-8.0 No ZBAX Eclipse XDB-CN 80A 180 m 2 /g 2.0-8.0 Yes 4.3% ZBAX CN 70A 300 m 2 /g 2.0-7.0 Yes 7% ZBAX NH 2 70A 300 m 2 /g 2.0-7.0 Yes 4% Column : ZBAX CN, 880952-705, 4.6 x 250 mm, 5 μm Mobile Phase : A : Heptane B : 2-Methoxyethanol/Isopropanol (50/50) Flow ate : 2 ml/min Temperature : 50 C Detector : 278 nm Sample : ctylphenoxy (polyethylene oxy) Ethanol Surfactant (n= 10) ZBAX x-sil 고순도실리카 ( > 99.995% ) 1.8μm, 5μm 입자크기제공 HILIC mode 로사용가능 극성물질분석에응용 USP L3 ZBAX Eclipse XDB-CN x-sil 의고순도실리카사용 염기성물질분석에적합 USP L10 0 5 10 15 Time (min) 20 ZBAX CN ZBAX SIL 의실리카에 Cyanopropyldimethylsilane 처리 lica 컬럼보다내구성이강함 USP L10 ZBAX NH 2 Weak anion-exchange 컬럼으로사용 비타민 A 와 D 분석에적합 USP L8 ZBAX HILIC Plus lica 순상고정상과역상이동상의만남 극성물질의머무름최대화를위해사용 LC/MS 응용에적합 EPA Method 1694 에적합 ze ange Non-bonded silica 95A 160 m 2 /g 0-8.0 6 Agilent LC & LC/MS 컬럼선택가이드
Ion Exchange Columns ZBAX Ion Exchange : SAX & SCX 이온교환컬럼 Ion Exchange Columns lica ZBAX SAX Column : Quaternary amine 작용기로수용성물질 / ( ) 이온성물질분석에적합 ZBAX SCX Column : Sulfonic acid 작용기로염기성, 수용성물질, 바이오물질 / (+) 이온성물질분석에적합 고효율과빠른분석제공 ze ange Functionality Max Pressure ZBAX SAX 70A 300 m 2 /g 2.0-7.0 Quaternary amine 350 bar ZBAX 300SCX 300A 50 m 2 /g 2.0-7.0 Sulfonic acid 350 bar Hi-Plex Polymer 응용 : 단당, 이당과올리고당, 유기산분석 USP L17, L19, L34, L58 에적합 물과희석된산용매사용가능 다양한 Application 제공 Conditions Sample : Column : rganic acids Agilent ( 제품번호 PL1170-6830) Mobile phase : 0.1 M H 2 S 4 Flow rate : 0.6 ml/min Temperature : 50 C Detector : I Conditions Sample : Sugars and sugar alcohols Column : Agilent Hi-Plex Ca (Duo) ( 제품번호 PL1F70-6850) Sample size : 10 mg/ml Mobile phase : 100% DI H 2 Flow rate : 0.4 ml/min Injection volume : 10 μl Temperature : 85 C Detector : I Temperature ange 1 1 2 3 2 4 5 6 0 min 28 3 4 5 6 Flow ate (ml/min) 그림 6. 컬럼을이용한유기산 9 종분석의예 0 min 30 그림 7. Hi-Plex Ca 컬럼을이용한단당류및다당류분석의예 7 7 8 8 9 9 Eluent Hi-Plex Ca 80-90 C 0.6 Water Hi-Plex Ca USP L19 80-90 C 0.3 Water 70-90 C 0.6 Water for carbohydrates 60-70 C 0.6 Water for organic acids 40-60 C 0.6 Dilute Acid Hi-Plex Ca (Duo) 80-90 C 0.6 Water Hi-Plex K 80-90 C 0.6 Water Hi-Plex Na (cto) 80-90 C 0.6 Water, Sodium Hydroxide Hi-Plex Na 80-90 C 0.3 Water 1. xalic acid 2. cis-aconitic acid 3. Tartaric acid 4. Malic acid 5. Lactic acid 6. Formic acid 7. Fumaric acid 8. Propionic acid 9. Butyric acid 1. affinose 2. Sucrose 3. Lactulose 4. Glucose 5. Galactose 6. Fructose 7. ibitol 8. Mannitol 9. Sorbitol Application USP Methods Specifying L17 Media USP Methods Specifying L19 Media USP Methods Specifying L34 Media USP Methods Specifying L58 Media Mono- and Disaccharides Anomer Separations rganic Acids Alcohols Adulteration of Food and Beverages Food Additives Dairy Products Sweetened Dairy Products Confectionery Fruit Juice Wine Wood Pulp Hydrolysates (cellulose/hemi-cellulose) Fermentation Monitoring ligosaccharides Samples with High Salt Content (molasses) ligosaccharides <Dp5 with Monosaccharides Corn Syrups ecommended Column Hi-Plex Ca and Hi-Plex Ca (Duo) Hi-Plex Na and Hi-Plex Na (cto) Hi-Plex Ca Hi-Plex Na (cto) Hi-Plex Ca Hi-Plex Ca Hi-Plex K Hi-Plex Ca and Hi-Plex Ca and Hi-Plex Ca and Hi-Plex Ca and Hi-Plex Ca Hi-Plex Na Hi-Plex Na (cto) Hi-Plex Ca (Duo) Hi-Plex Na 7
스페셜컬럼 Special Columns Special Application Columns ZBAX Carbohydrate Analysis 재현성있고효율적인당분석을위한컬럼 Aminopropyl packing material 로단당류와이당류분리에적합 4.6 X150mm 컬럼기준 50μL 정도많은량의시료주입가능 ID(efractive index detectors) 사용시추천 hamnose Xylose Xylitol Fructose Glu/Gal Sucrose Maltose Lactulose Lactose affinose 1 2 3 4 5 6 7 ze ange* Xylose hamnose Fructose Glucose Sucrose Maltose Lactose affinose ZBAX Carbohydrate Analysis 70A 300 m 2 /g 2.0-8.0 No 3.5% ZBAX Amino Acid Analysis(AAA) 응용 : 아미노산 24 종분리 PA 와 FMC 유도체화방법사용 표준물질과유도체화시약 Agilent 공급가능 Agilent Autosampler 를사용하여자동화된유도체화로쉽고간편함 Column Agilent ZBAX Eclipse AAA 4.6 150 mm, 5 μm ( 제품번호 993400-902) Mobile Phase Column: ZBAX Carbohydrate Analysis, 843300-908, 4.6 x 150 mm, 5 μm Mobile Phase: 75% ACN/25% H 2 Flow ate: 2 ml/min Temperature: 30 C Detector: ID Det. Temp: 30 C Sample: (54 μg each), Inj. = 6.3 μl A: 40 mm Sodium phosphate(di-basic) 0.1% Phosphoric acid B: Acetonitrile / Methanol / DW = 45/45/10 Detector DAD 338nm (1차아미노산 ), 262nm (2차아미노산 ) FLD Ex : 340nm, Em : 450nm (1차아미노산 ) Ex : 266nm, Em : 305nm (2차아미노산 ) Flow rate 1.5 ml/min Temp. 40 Time (min) B (%) 0.0 0 1.9 0 18.1 57 18.6 80 22.3 80 23.2 0 26.0 0 제품명 제품번호 PA reagent, 10 mg/ml each in 0.4 M borate butter o-phthalaldehyde (PA) 5061-3335 and 3-mercaptopropionic acid, 6 x 1 ml ampoules FMC reagent, 2.5 mg/ml in acetonitrile, 9-fluorenylmethylchloroformate, 5061-3337 1 ml. 10 ampoules Borate Buffer 100mL 5061-3339 AdvanceBio Peptide Mapping 일관된 Lot 별재현성 2.7μm Poroshell 구조의고정상으로좋은분리도 HPLC, UHPLC 모두사용가능 600bar 까지사용가능 LC/MS 분석시최적의감도제공 짧은시간안에정확하게 Peptide Mapping 완성 Column: AdvanceBio Peptide Mapping, 2.1 x 250 mm, 2.7 μm, 651750-902 Flow ate: 0.4 ml/min Injection: 5 μl (2.0 mg/ml) Temp: 55 C Detection: 220 nm Gradient: A, water (0.1% FA); B, ACN (0.1% FA), 0-28 min, 3-45% B; 28-33 min, 45-60% B; 33-34 min, 60-95% B AdvanceBio Glycan Mapping HILIC 고정상으로재현성있고정확하게 Glycan Mapping 완성 좋은분리도와빠른분석가능 (1.8μm :<1200bar, 2.7μm Poroshell : <600bar) LC/MS 와 LC/FLD 에추천 1 차아미노산 mau 40 30 20 10 0 2 차아미노산 mau 40 30 20 10 0 1 2 3 4 5 6 7 Xylose hamnose Xylitol Mannitol Lactulose 1 2 3 4 5 6 7 Time (min) 그림 8. Carbohydrate 컬럼을이용한당과당알콜분석의예 DAD1 A, g=338,10 ef=390,20 (AMIN3\AA6-0003.D) 1.792 - ASP 3.790 - GLU 6.038 - ASN 6.304 - SE 6.936 - GLN 7.166 - HIS 7.503 - GLY 7.699 - TH 8.239 - AG 8.889 - ALA 그림 9. 24 종아미노산동시분석의예 10.027 - TY 11.494 - CY2 11.887 - VAL 12.092 - MET 12.437 - NVA 12.895 - TP 13.300 - PHE 13.493 - ILE 14.125 - LEU 14.586 - LYS affinose 0 2 4 6 8 10 12 14 16 18 DAD1 B, g=262,16 ef=324,8 (AMIN3\AA6-0003.D) 0.806 1.077 1.793 3.314 3.786 5.276 5.455 5.684 5.851 6.038 6.303 6.940 7.167 7.498 7.699 8.236 8.704 8.887 10.027 11.257 11.494 11.662 12.006 11.887 12.092 12.441 0 2 4 6 8 10 12 14 16 18 300 250 200 150 100 50 0 0 5 10 15 20 25 30 12.745 12.895 13.119 13.300 13.493 그림 10. Biosimilar EP 에대한 Peptide Mapping 의예 14.125 14.586 14.959 - HYP 16.650 17.060 - SA 17.654 - P min min 8 Agilent LC & LC/MS 컬럼선택가이드
GPC / SEC Columns & Standards 분자량에따라사용되는 GPC/SEC 컬럼 유기용매사용 PLgel MIXED 컬럼가이드 SEC 컬럼 ze Exclusion Columns 분자량 2,000~40,000,000 500~10,000,000 추천컬럼 PLgel 20 μm MIXED-A PLgel 10 μm MIXED-B GPC/SEC 컬럼 PL aquagel-h PLgel lexis PL HFIPgel 특징수용성용매사용높은분자량의고분자분석 HFIP 용매사용 200~2,000,000 200~400,000 Up to 30,0000 PLgel 5 μm MIXED-C PLgel 5 μm MIXED-D PLgel 3 μm MIXED-E PLgel Preparative PL apide PolarGel Plus Prep의용도로사용되는 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 6 Å 600,000 to 10,000,000 PLgel 5/10 µm 10 5 Å 60,000 to 1,700,000 PLgel 5/10 µm 10 4 Å 10,000 to 450,000 PLgel 5/10 µm 10 3 Å 500 to 60,000 PLgel 5/10 µm 50Å 0 to 1,500 Columns PLgel 3/5/10 µm 100Å 0 to 5,000 PLgel 5/10 µm 500Å 500 to 25,000 PLgel 3 µm MIXED-E 0 to 25,000 The latest t technology ogy in GPC for increased ed speed ed and resolution MW range Agilent Plus Columns Agilent PL apide Columns High pore volume for very Special format for fast high resolution 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 3,300 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 60,000 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 30 8 µm 100 to 60,000 PL aquagel-h 40 8/15 µm 10,000 to 200,000 PL aquagel-h 50 8/15 µm 50,000 to 600,000 PL aquagel-h 60 8/15 µm 200,000 to 10,000,000 PL aquagel-h MIXED-H 8 µm 6,000 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 3 3 3 3 rganic Polymethylmethacrylate 3 3 3 rganic Polyethylene glycol (PEG) 3 3 3 rganic/aqueous Polyethylene oxide (PE) 3 3 3 rganic/aqueous Pullulan polysaccharide 3 3 rganic/aqueous Polyacrylic acid Na salt 3 3 Aqueous 9
바이오컬럼 Bio Columns Biosolution Columns Primary Structure eversed-phase separations 소수성작용 Biomolecule Aggregates Fragment Pattern Charge Isoforms ze Exclusion separations (SEC) Ion-Exchange separations (IEX) 분자크기분리 이온교환 Affinity Protein A 친화력분석 eversed-phase Column for Protein & Peptides Poroshell 300 Biomolecules 분석을위한빠르고효율적인컬럼 1,000kDa 까지의 Protein 분석에효과적 독특한고정상구조로빠른분석가능 ze Temp. Limits* ange Poroshell 300SB-C18, C8, C3 300A 90 C 1.0-8.0 No Poroshell 300Extend-C18 300A ZBAX 300A Extend C18 40 C above 8 60 C below 8 넓은 범위에서매우효과적인고정상 ( 2-11.5) 높은 에서 Hydrophobic Peptides 의좋은분석결과도출 LC/MS 사용시 Ammonium-hydroxide 용매사용가능 2.0-11.0 Yes ZBAX 300A StableBond size 300A 컬럼으로 Protein 과 Peptide 등거대분자에고효율역상분리 다양한고정상제공으로폭넓은응용지원 높은온도에서도재현성있고신뢰성있는데이터도출 ze Temp Limits* ange* ZBAX 300SB-C18 300A 45 m 2 /g 90 C 1.0-8.0 No 2.8% ZBAX 300SB-C8 300A 45 m 2 /g 80 C 1.0-8.0 No 1.5% ZBAX 300SB-C3 300A 45 m 2 /g 80 C 1.0-8.0 No 1.1% ZBAX 300SB-CN 300A 45 m 2 /g 80 C 1.0-8.0 No 1.2% ZBAX 300-Diphenyl 300A 45 m 2 /g 80 C 1.0-8.0 Yes 1.9% ze Temp Limits* ange* ZBAX 300A Extend-C18 300A 45 m 2 /g 60 C 2.0-11.5 Double 4% ze Exclusion 재현성있고다양한선택이가능한 Agilent SEC 컬럼 Agilent Bio SEC-3 & Bio-5 : 다양한 size 선택가능 ProSEC 300S : 고농도의염조건에서 Globular Protein 분석에적합 ZBAX GF-250 & GF-450 : Protein 의 Preparative SEC 분석에적합 PL aquagel-h : 넓은분자량범위의 Biopolymer 분석에적합 Agilent Bio Ion Exchange 다양한분야에서사용할수있도록제공 PEEK 재질과 Staninless Steel 재질로제공 ID Particle ze SCX (Strong cation-exchange) - S0 3 H WCX (Weak cation-exchange) - C00H SAX (Strong anion-exchange) - N( ) 3 2.1 and 4.6 mm 1.7, 3, 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 2 H 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 ze Exclusion Chromatography (SEC) Column Application Agilent Columns Application Agilent Columns Proteins and polypeptides ZBAX 300, 1.8 μm Peptides, proteins Agilent Bio SEC-3 HD 300SB-C18 HD 300SB-C8 Large biomolecules and samples with multiple molecular weight components Agilent Bio SEC-5 HD 300SB-C3 Globular proteins, antibodies ProSEC 300S HD 300-Diphenyl Proteins, globular proteins ZBAX GF-250/450 HD 300-HILIC ZBAX 300 StableBond 300SB-C18 Low MW polymers and oligomers, oligosaccharides, PEGs, lignosulfonates 2 or 3 PL aquagel-h PL aquagel-h 8 μm PL aquagel-h 20 5 μm 300SB-C8 PL aquagel-h MIXED-M 8 μm 300SB-C3 300SB-CN ZBAX 300 Extend-C18 Polydisperse biopolymers, polysaccharides, cellulose derivatives 2 or 3 PL aquagel-h PL aquagel-h MIXED-H 8 μm PL aquagel-h 60/50/40 8 μm Peptides and proteins up to 1,000 kda, monoclonal antibodies and intact proteins Poroshell 300 300SB-C18 Very high MW polymers, hyaluronic acids, starches, gums PL aquagel-h 60/50/40 15 μm in series 300SB-C8 300SB-C3 300Extend-C18 Ion-exchange Column Small hydrophilic peptides in protein digests Peptides to DNA Poroshell 120 PLP-S 100 300 1000 4000 Small molecules/peptides/oligonucleotides PLP-S 100 ecombinant peptides/proteins PLP-S 300 Large proteins PLP-S 1000 DNA/high speed separation PLP-S 4000 Affinity Column Application Agilent Columns Monoclonal antibodies Agilent Bio MAb Peptides and proteins Agilent Bio IEX Proteins, peptides and deprotected synthetic oligonucleotides PL-SAX 1000 4000 Globular proteins and peptides PL-SAX 1000 Very large biomolecules/high speed PL-SAX 4000 Small peptides to large proteins PL-SCX 1000 4000 Globular proteins PL-SCX 1000 Very large biomolecules/high speed PL-SCX 4000 Application IgG(human and mouse), except for IgG class3 Agilent Columns Bio-Monolith Protein A Antibodies (IgG, IgM), plasmid DNA, viruses, phages and other macro biomolecules Bio-Monolith Bio-Monolith QA Bio-Monolith DEAE Bio-Monolith S 3 Agilent LC 컬럼의다양한사양및자세한 Application 은영화과학에문의해주세요. 02-2140-5460 Viruses, DNA, large proteins Bio-Monolith QA Plasmid DNS, bacteriophages Bio-Monolith DEAE Proteins, antibodies Bio-Monolith S 3 11
Agilent LC 컬럼 USP code USP USP Packing Materials Column L1 L3 L7 L8 L9 L10 L11 L13 L14 ctadecyl silane chemically bonded to porous silica or ceramic microparticles, 1.5 to 10 μm in diameter, or a monolithic rod Porous silica particles, 1.5 to 10 μm in diameter, or a monolithic silica rod ctylsilane chemically bonded to totally porous silica particles, 1.5 to 10 μm in diameter, or a monolithic silica rod An essentially monomolecular layer of aminopropylsilane chemically bonded to totally porous silica gel support, 3 to 10 μm in diameter Irregular or spherical, totally porous silica gel having a chemically bonded, strongly acidic cation-exchange coating, 3 to 10 μm in diameter Nitrile groups chemically bonded to porous silica particles, 3 to 10 μm in diameter Phenyl groups chemically bonded to porous silica particles, 1.5 to 10 μm in diameter Trimethylsilane chemically bonded to porous silica particles, 3 to 10 μm in diameter lica gel having a chemically bonded, strongly basic quaternary ammonium anion-exchange coating, 5 to 10 μm in diameter ZBAX Eclipse Plus C18 ZBAX Eclipse XDB-C18 ZBAX SB-C18 ZBAX x-c18 ZBAX Extend-C18 ZBAX DS ZBAX DS classic Pursuit Xs C18 Pursuit C18 Polaris C18-A Polaris C18-Ether Poroshell 120 EC- C18 Poroshell 120 SB- C18 Poroshell 120 HPH- C18 Poroshell 300 SB- C18 Poroshell 300 Extend- C18 Sep Tech ST60 C18 Sep Tech ST150 C18 Agilent Prep C18 ZBAX HILIC Plus ZBAX x-l ZBAX SIL Pursuit Xs Polaris -A Agilent Prep ZBAX Eclipse Plus C8 ZBAX Eclipse XDB-C8 ZBAX SB-C8 ZBAX X-C8 ZBAX C8 Pursuit Xs C8 Pursuit C8 Polaris C8-A Polaris C8-Ether Poroshell 120 EC-C8 Poroshell 120 SB-C8 Poroshell 120 HPH-C8 Poroshell 300 SB-C8 ZBAX NH 2 Polaris NH 2 ZBAX SCX ZBAX CN ZBAX SB-CN ZBAX Eclipse XDB-CN ZBAX Eclipse XDB Phenyl ZBAX Eclipse Plus Phenyl-Hexyl ZBAX Phenyl Poroshell 120 Phenyl-Hexyl Pursuit Xs DiPhenyl Pursuit DiPhenyl ZBAX TMS ZBAX SAX IonoSpher A USP USP Packing Materials Column L15 L17 L19 Hexylsilane chemically bonded to totally porous silica particles, 3 to 10 μ m in diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the hydrogen form, 7 to 11 μm in diameter Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the calcium form, 9 μm in diameter L20 Dihydroxypropane groups chemically bonded to porous silica particles, 3 to 10 μm in diameter L21 L22 L25 L33 L34 L35 L43 L45 L50 L52 A rigid spherical styrenedivinylbenzene copolymer, 5 to 10 μm in diameter A cation-exchange resin made of porous polystyrene gel with sulfonic acid groups, about 10 μm in size Packing having the capacity to separate compounds with a MW range from 1,000 to 5,000 da (as determined by the polyethylene oxide), applied to neutral, anionic and cationic water-soluble polymers. A polymethacrylate resin base, crosslinked with polyhydroxylated ether (surface contained some residual carboxyl functional gruops) was found suitable Packing having the capacity to separate dextrons by molecular size over a 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 A zirconium-stabilized spherical silica packing with a hydrophilic (dioltype) molecular monolayer bonded phase having a pore size of 150A Pentafluorophenyl groups chemically bonded to silica particles by a propyl spacer, 5 to 10 μm in diameter Beta cyclodextrin bonded to porous silica particles, 5 to 10 μm in diameter Multifunction resin with reversed-phase retention and strong anionexhange functionalities. The resin consists of ethylvinylbenzene, 55% cross-linked with divinylbenzene copolymer, 3 to 15 μm in diameter, and a surface area of not less than 350 m 2 per g. Substrate is coated with quarternary ammonium functionalized latex particles consisting of styrene cross-linked with divinylbenzene Weak cation-exchange resin made of porous silica with sulfopropyl groups, 5 to 10 μm in diameter L53 Weak cation-exchange resin consisting of ethylvinylbenzene, 55% crosslinked with divinylbenzene copolymer, 3 to 15 μm diameter. Substrate is surface grafted with carboxylic acid and/or phosphoric acid functionalized monomers. Capacity not less than 400 μeq/column L56 L57 L58 L60 Propyl silane chemically bonded to totally porous silica particles, 3 to 10 μm in diameter A chiral-recognition protein, ovomucoid, chemically bonded to silica particles, about 5 μm in diameter, with a pore size of 120 A Strong cation-exchange resin consisting of sulfonated cross-linked styrene-divinylbenzene copolymer in the sodium form, about 6 to 30 μm in diameter Spherical, porous silica gel, 10 μm in diameter, the surface has been covalently modified with alkyl amide groups and endcapped 분석방법조정가이드 USP & FDA Parameter Max Spec 컬럼길이 ± 70% 컬럼 ID FDA ± 50% USP ± 25% 컬럼선속도유지 유속 ± 50% 주입량 최대한적게감도 / 재현성유지 입자크기 to 50% Metal C6 Hi-Plex Ca Hi-Plex Ca (Duo) LiChrospher Diol PLgel PLP-S PL aquagel-h ZBAX GF-250 Bio SEC-3 Bio SEC-5 ProSEC ZBAX GF 250 ZBAX GF 450 Pursuit PFP Poroshell120 PFP ChiraDex Chiral ZBAX 300SCX IonoSpher C Bio SAX ZBAX SB-C3 Ultron ES-VM Hi-Plex Na Hi-Plex Na (cto) Polaris Amide-C18 주요관공서의표준방법을 수정할경우기존분석법과 수정방법을동시에수행하여 정확성, 재현성등검증조건에 영향을주지않아야합니다. 제품상담 : 크로마토그래피팀 (T:02-2140-5460) AG BC 201409