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Agilent Technologies AdvanceBio Amino Acid Analysis, Руководство пользователя

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This booklet provides general information for the 

Agilent BioHPLC and AdvanceBio reversed-phase 

columns. For additional detailed information about 

your specific phase or family, see:

  

www.agilent.com/chem/biocolumnchoices 

Getting Started

A QC Column Performance Report, including a test chromatogram, 
is enclosed with every Agilent column. The QC test system has been 
modified from a standard system to minimize system dead volume, 
so it may vary from the system used in your lab. This allows a better 
evaluation of the column efficiency and assures a more consistent 
product. An optimized LC system will generate similar results to the 
chromatogram on your QC Performance Report.

If you have specific questions, contact the Technical Support team at 

www.agilent.com/chem/columnsupport

Содержание AdvanceBio Amino Acid Analysis

Страница 1: ...nceBio d Agilent BenutzeRINFORMATION zu Agilent BioHPLC und AdvanceBio S ulen Guida ALL USO per colonne Agilent BioHPLC e AdvanceBio Gu a de usuario para columnas Agilent BioHPLC y AdvanceBio Agilent...

Страница 2: ...atogram is enclosed with every Agilent column The QC test system has been modified from a standard system to minimize system dead volume so it may vary from the system used in your lab This allows a b...

Страница 3: ...ur Column Column Conditioning Every column is tested before shipment So for first use the shipping solvent must be replaced with eluent taking care that all components are miscible and soluble If mobi...

Страница 4: ...matographic performance and column lifetime and could be unsafe Other Operating Tips While generally not harmful to the column reverse flow should be avoided except to attempt removal of clogged frit...

Страница 5: ...use a precipitate to form or may not be fully miscible Agilent BioHPLC and AdvanceBio columns are compatible with all common eluents used for analysis of biomolecules ENGLISH Column Particle size Pres...

Страница 6: ...40 C max using low buffer concentrations in the range of 0 01 to 0 02 M Column Shipping solvent Compatibility AdvanceBio Peptide Mapping Mixture of acetonitrile and water Water and all common organic...

Страница 7: ...tarting Gradients Separations of biological molecules typically use gradient conditions increasing the amount of the organic component to achieve elution from the column Separations of peptides polype...

Страница 8: ...such samples It is recommended that samples are filtered before injecting them onto any column and all eluents are freshly prepared and filtered prior to use See www agilent com chem guards for more i...

Страница 9: ...ightly capped with end plugs to prevent packing from drying out 5 Discard column or consider stronger conditions e g 75 acetonitrile 25 isopropanol 6 Increase to 100 isopropanol 100 methylene chloride...

Страница 10: ...umns ZORBAX RRHD Use sample filtration or other sample prep as appropriate for your sample Learn more at www agilent com chem sampleprep Use certified lamps in your LC instruments for best performance...

Страница 11: ...st joint chaque colonne Agilent Le syst me de tests de CQ a t con u partir d un instrument standard modifi afin de minimiser les volumes morts celui ci peut donc varier du syst me utilis dans votre la...

Страница 12: ...s colonnes Chaque colonne est test e avant exp dition Par cons quent pour la premi re utilisation le solvant d exp dition doit tre remplac par un luant en veillant ce que l ensemble des composants soi...

Страница 13: ...t la dur e de vie de la colonne et pourrait s av rer dangereux Autres conseils de fonctionnement Bien que g n ralement non dangereux pour la colonne l inversion de flux doit tre vit e except pour tent...

Страница 14: ...rovoquer la formation d un pr cipit ou de ne pas tre enti rement miscible ne traverse la colonne Les colonnes BioHPLC et AdvanceBio d Agilent sont compatibles avec l ensemble des luants couramment uti...

Страница 15: ...dessous de pH 8 40 C au dessus de pH 8 AdvanceBio Oligonucleotide 3 0 11 0 65 C AdvanceBio Amino Acid Analysis PLRP S 1 0 14 0 200 C Colonne Solvant d exp dition Compatibilit AdvanceBio Peptide Mappi...

Страница 16: ...es biologiques utilisent g n ralement des conditions de gradient augmenter la quantit du composant organique permet d obtenir une lution de la colonne Les s parations de peptides de polypeptides et de...

Страница 17: ...est conseill de filtrer les chantillons avant de les injecter sur une colonne quelle qu elle soit et d utiliser des luants fra chement pr par s et filtr s Consultez le site www agilent com chem guards...

Страница 18: ...cher l ass chement du remplissage 5 Mettez la colonne au rebut ou envisagez des conditions plus fortes par ex 75 ac tonitrile 25 isopropanol 6 Augmentez jusqu 100 d isopropanol 100 de chlorure de m t...

Страница 19: ...PL rapide ZORBAX RRHD Utilisez la filtration ou d autres techniques de pr paration d chantillons selon le cas pour votre chantillon Pour en savoir plus consultez le site www agilent com chem samplepre...

Страница 20: ...Agilent BioHPLC AdvanceBio www agilent com chem biocolumnchoices www agilent com chem columnsupport...

Страница 21: ...1 8 m ZORBAX RRHD 600 bar 5042 8957 UHPLC 1200 bar 5067 4733 5042 8957 Agilent 1200 bar 5067 4733 10 20...

Страница 22: ...1 pH 2...

Страница 23: ...nceBio Oligonucleotide AdvanceBio Amino Acid Analysis AdvanceBio RP mAb 3 5 m ZORBAX 300SB 3 5 m 5 m 7 m 400 bar Poroshell 300 5 m ZORBAX RRHD 1 8 m 1200 bar PLRP S 3 m 275 bar 5 m 8 m 10 m 207 bar 10...

Страница 24: ...roshell 300 Extend C18 2 0 11 0 pH 8 60 C pH 8 40 C AdvanceBio Oligonucleotide 3 0 11 0 65 C AdvanceBio Amino Acid Analysis PLRP S 1 0 14 0 200 C 2 pH pH 6 pH 6 40 C 0 01 0 02 M AdvanceBio Peptide Map...

Страница 25: ...PLRP S 100 TFA FA pH chapter 11 pages 497 508 Introduction to Modern Liquid Chromotography Second Edition L R Snyder and J J Kirkland John Wiley Sons 1979...

Страница 26: ...1 8 m 0 5 m AdvanceBio Peptide Mapping 2 m 2 m www agilent com chem guards 1 8 m 1 2 10 20 3 100 4...

Страница 27: ...20 30 50 50 20 30 5 75 25 6 100 100 100 PLRP S 80 1 M NaOH 1 M HCI 20 1 8 m...

Страница 28: ...0 12 mm www agilent com chem lccapillaries ZORBAX RRHD www agilent com chem sampleprep 60 40 ACN H2O 60 40 ACN 0 02 M...

Страница 29: ...f r die QC Tests eingesetzte Chromatographiesystem ist ein im Hinblick auf minimales Totvolumen modifiziertes Standardsystem kann also von dem in Ihrem Labor verwendeten System abweichen Dies erm glic...

Страница 30: ...m Versand gepr ft Daher muss das Transportl sungsmittel vor dem ersten Einsatz durch Ihren Eluenten ersetzt werden Achten Sie darauf dass alle Komponenten mischbar und l slich sind Wenn in der mobilen...

Страница 31: ...chtigt die Chromatographieleistung und Lebensdauer der S ule und kann gef hrlich sein Weitere Tipps zur Bedienung In der Regel schadet das R cksp len der S ule zwar nicht es sollte aber dennoch vermi...

Страница 32: ...nicht bei erh htem pH Wert oder erh hter Temperatur aufbewahrt werden siehe Tabelle 2 Neue S ulen k nnen eine Mischung aus organischen L sungsmitteln und Wasser enthalten in der Puffersalze gel st sin...

Страница 33: ...amid und Dimethylsulfoxid Poroshell 300 und 300 Extend Mischung aus Methanol und Wasser AdvanceBio Oligonucleotide Mischung aus Acetonitril und Wasser Wasser und alle gebr uchlichen organischen L sung...

Страница 34: ...ung von biologischen Molek len erfolgt in der Regel mithilfe von Gradientenelution Der Anteil der organischen Komponente wird dabei allm hlich erh ht um die Elution von der S ule zu erreichen Trennung...

Страница 35: ...or dem Injizieren zu filtern alle Eluenten frisch anzusetzen und vor dem Gebrauch zu filtrieren Weiterf hrende Informationen ber Vors ulen finden Sie unter www agilent com chem guards S ule reinigen u...

Страница 36: ...verschlossen werden um die S ulenpackung vor dem Austrocknen zu sch tzen 5 Verwerfen Sie die S ule oder w hlen Sie eine wirksamere L sungsmittelmischung z B 75 Acetonitril 25 Isopropanol 6 Erh hen Sie...

Страница 37: ...ch Minimierung der Kapillarl ngen zwischen den Komponenten Dadurch verringern Sie das Totvolumen und die Bandenverbreiterung Verwenden Sie f r Fast LC High Efficiency LC S ulen rote Kapillaren 0 12 mm...

Страница 38: ...di prova Il sistema di prova QC stato modificato partendo da una configurazione standard al fine di ridurre al minimo il volume morto pertanto pu differire dal sistema utilizzato nel tuo laboratorio...

Страница 39: ...Prima del trasporto ogni colonna viene sottoposta a test Quindi per il primo utilizzo il solvente di trasporto deve essere sostituito con eluente avendo cura che tutti i componenti siano miscibili e...

Страница 40: ...nna e inoltre potrebbe essere pericoloso Altri consigli operativi Anche se il flusso inverso generalmente non dannoso per la colonna dovrebbe tuttavia essere evitato se non nel tentativo di rimuovere...

Страница 41: ...cipitato oppure potrebbero non essere completamente miscibili tra loro Le colonne BioHPLC e AdvanceBio Agilent sono compatibili con tutti i comuni eluenti utilizzati per l analisi delle biomolecole Ta...

Страница 42: ...ido Poroshell 300 e 300 Extend Miscela di acetonitrile e acqua AdvanceBio Oligonucleotide Miscela di acetonitrile e acqua Acqua e tutti i comuni solventi organici Modificatori organici inclusi l HFIP...

Страница 43: ...parazione di molecole biologiche sono generalmente usate condizioni di gradiente aumentare la quantit di componente organico per ottenere eluizione dalla colonna Le separazioni di peptidi polipeptidi...

Страница 44: ...i si raccomanda l impiego di precolonne Si raccomanda inoltre di filtrare i campioni prima di iniettarli nella colonna e verificare che tutti gli eluenti siano preparati di fresco e filtrati prima del...

Страница 45: ...re se ritornata a condizioni normali in caso contrario 5 Eliminare la colonna oppure considerare condizioni pi estreme per es 75 acetonitrile 25 isopropanolo 6 Aumentare le concentrazioni 100 isopropa...

Страница 46: ...ZORBAX RRHD Impiega la filtrazione del campione o altre preparazioni secondo necessit per il campione considerato Per saperne di pi visita la pagina del sito www agilent com chem sampleprep Utilizza l...

Страница 47: ...e prueba para realizar el Control de Calidad QC se ha modificado a partir de un sistema est ndar a fin de minimizar el volumen muerto del sistema y por lo tanto podr a variar con respecto al sistema u...

Страница 48: ...las columnas se prueban antes de su env o Por lo tanto para el primer uso se debe reemplazar el disolvente de transporte con el eluyente para lo que es necesario tener en cuenta que todos los compone...

Страница 49: ...n afectados lo que podr a ser inseguro Otros consejos operativos Aunque por lo general no resulta da ino para la columna se debe evitar el flujo inverso excepto si se intenta eliminar las part culas d...

Страница 50: ...elevada ni con un pH elevado consulte la Tabla 2 Las nuevas columnas pueden incluir una mezcla de disolventes org nicos y agua lo que puede contener sales de la soluci n tamp n Consulte la Tabla 3 par...

Страница 51: ...roshell 300 y 300 Extend Mezcla de metanol y agua AdvanceBio Oligonucleotide Mezcla de acetonitrilo y agua Agua y todos los disolventes org nicos comunes Modificadores como el HFIP y el TEAA AdvanceBi...

Страница 52: ...aciones de mol culas biol gicas usan por lo general condiciones de gradiente es decir se aumenta la cantidad de componente org nico para alcanzar la eluci n en la columna La forma m s habitual de real...

Страница 53: ...smo se recomienda que se filtren las muestras antes de inyectarlas sobre cualquier columna y que todos los eluyentes est n reci n preparados y se filtren antes de usarlos Consulte www agilent com chem...

Страница 54: ...eben tapar firmemente con tapones terminales para evitar que el empaquetado se reseque 4 Compruebe si la presi n vuelve a ser normal si no es as 5 Descarte la columna o tenga en cuenta condiciones m s...

Страница 55: ...i n de la longitud de los tubos entre los componentes de esta forma se disminuye el volumen de columna adicional y el ensanchamiento de banda Con las columnas LC r pidas o de alta eficacia utilice tub...

Страница 56: ...Agilent BioHPLC AdvanceBio www agilent com chem jp QC Agilent QC LC QC www agilent com chem jp...

Страница 57: ...1 8 m ZORBAX RRHD 600 bar p n 5042 8957 UHPLC 1200 bar p n 5067 4733 p n 5042 8957 Agilent 1200 bar p n 5067 4733 10 20...

Страница 58: ...1 pH 2...

Страница 59: ...2 7 m 600 bar AdvanceBio Peptide Plus AdvanceBio Oligonucleotide AdvanceBio Amino Acid Analysis AdvanceBio RP mAb 3 5 m ZORBAX 300SB 3 5 m 5 m 7 m 400 bar Poroshell 300 5 m ZORBAX RRHD 1 8 m 1200 bar...

Страница 60: ...FIP TEAA AdvanceBio Amino Acid Analysis PLRP S 7 1 N N 100 3 pH AdvanceBio Peptide Mapping 2 0 8 0 60 C AdvanceBio Peptide Plus ZORBAX RRHD 1 0 8 0 80 C ZORBAX 300SB C8 C3 CN ZORBAX 300SB C18 1 0 8 0...

Страница 61: ...100 PLRP S TFA FA pH L R Snyder and J J Kirkland Introduction to Modern Liquid Chromotography Second Edition Chapter 11 pages 497 508 John Wiley Sons 1979...

Страница 62: ...1 8 m 0 5 m AdvanceBio 2 m 2 m www agilent com chem jp 1 8 m 1 2 10 20 3 100 4...

Страница 63: ...20 30 50 50 20 30 5 75 25 6 100 100 100 PLRP S 1 M 1 M 80 20 1 8 m...

Страница 64: ...0 12 mm www agilent com chem jp LC ZORBAX RRHD www agilent com chem jp LC 60 40 ACN H2O 60 40 ACN 0 02 M...

Страница 65: ...Agilent BioHPLC AdvanceBio www agilent com chem biocolumnchoices Agilent www agilent com chem columnsupport...

Страница 66: ...1 8 ZORBAX RRHT ZORBAX RRHD Agilent 600 5042 8957 1200 5067 4733 UHPLC 5042 8957 Agilent 1200 5067 4733 10 20...

Страница 67: ...Agilent 1 pH 2...

Страница 68: ...ptide Plus AdvanceBio Oligonucleotide AdvanceBio Amino Acid Analysis AdvanceBio RP mAb 3 5 ZORBAX 300SB 3 5 5 7 400 Poroshell 300 5 ZORBAX RRHD 1 8 1200 PLRP S 3 m 275 5 m 8 m 10 m 207 10 15 m 15 20 m...

Страница 69: ...de 2 AdvanceBio Amino Acid Analysis PLRP S 7 1 N N 100 3 pH AdvanceBio Peptide Mapping 2 0 8 0 60 C AdvanceBio Peptide Plus ZORBAX RRHD 1 0 8 0 80 C ZORBAX 300SB C8 C3 CN ZORBAX 300SB C18 1 0 8 0 90 C...

Страница 70: ...PLRP S TFA FA pH 11 497 508 Introduction to Modern Liquid Chromoto graphy Second Edition L R Snyder J J Kirkland John Wiley Sons 1979...

Страница 71: ...1 8 0 5 AdvanceBio Peptide Mapping 2 2 www agilent com chem guards 1 8 1 2 10 20 3 100 4...

Страница 72: ...50 50 20 30 20 30 5 75 25 6 100 100 100 PLRP S 80 1 M NaOH 1 M HCI 20 1 8...

Страница 73: ...H2O 60 40 60 40 0 02 0 12 www agilent com chem lccapillaries ZORBAX RRHD www agilent com chem sampleprep...

Страница 74: ...grama de teste Um sistema padr o foi modificado para gerar o sistema de teste de QC que minimiza o volume morto que pode diferir do sistema utilizado em seu laborat rio Este sistema permite avaliar me...

Страница 75: ...coluna Todas as colunas s o testadas antes do envio Portanto para a primeira utiliza o o solvente de envio deve ser substitu do por um eluente assegurando que todos os componentes se misturem e se di...

Страница 76: ...a vida til da coluna e pode n o ser seguro Outras dicas operacionais Embora geralmente n o seja prejudicial coluna deve se evitar o fluxo reverso exceto para tentar remover o frit entupido consulte a...

Страница 77: ...vel que possa formar um precipitado ou que n o esteja completamente misturada As colunas Agilent BioHPLC e AdvanceBio s o compat veis com todos os eluentes comuns para a an lise de biomol culas Tabela...

Страница 78: ...dimetilsulf xido Poroshell 300 e 300 Extend Mistura de metanol e gua AdvanceBio Oligonucleotide Mistura de acetonitrila e gua gua e todos os solventes org nicos comuns Modificadores incluindo HFIP e T...

Страница 79: ...l gicas geralmente emprega condi es de gradiente aumentando a quantidade do componente org nico para obter a elui o da coluna As separa es de pept deos polipept dios e prote nas s o realizadas mais fr...

Страница 80: ...amb m recomenda se filtrar as amostras antes de injet las em qualquer coluna e preparar e filtrar todos os eluentes momentos antes da utiliza o Acesse www agilent com chem guards para obter mais infor...

Страница 81: ...fechados com plugues para evitar que o empacotamento seque 5 Descarte a coluna ou considere condi es mais como por exemplo 75 de acetonitrila 25 de isopropanol 6 Aumente para 100 de isopropanol 100 d...

Страница 82: ...mize o desempenho do instrumento minimizando os comprimentos da tubula o entre os componentes para reduzir o volume extracoluna e o alargamento da banda Use a tubula o vermelha com 0 12 mm de di metro...

Страница 83: ...ormance for the separation and characterization of peptides and proteins The science behind AdvanceBio columns helps advance accuracy provide greater productivity and eliminate interferences that can...

Страница 84: ...gilent com chem columnsupport To place an order visit www agilent com chem wheretobuy Agilent offers a complete line of sample preparation products to support LC and LC MS applications To find parts a...

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