Agilent ZORBAX Eclipse XDB-C8 RRHT Threaded Column Datasheet
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1. Acidic generally are best separated at pH 3 with this column However Eclipse XDB C8 is especially suited for separating basie compounds when intermediate pH 4 8 must be used to maintain compound stability or to obtain desired band spacing selectiv ity For optimum results and long term reproducibility the use of 10 50 mM buffers is always recommended when separating ionizable compounds For many basic compounds basic modifiers such as triethylamine will not be required to achiove efficient separations with symmetrical peaks For very strongly basie compounds the use of 10 20 mM triethylamine or 5 10 mM dimethyl octylamine is recommended when using any silica based reversed phase column especially when highly robust longterm reproducible methods are required Column Care Samples that contain particulate matter may plug the column inlet frit and should be filtered before injection into the column IF solvent flow appears to be restricted unusually high column back pressure check first to see that solvent flow is unobstructed up to the column inlet If the column has the restriction there may be particulate matter on the inlet frit An attempt should be made to remove any inlet debris by back flushing 25 30 mL of mobile phase through the column If this fails to return the column to near its original backpressure the column should be replaced To remove strongly retained materials from the column flush t2. a variety of LC instru ments The 2 1 and 3 0 mm ID columns will support 20 000 psi 1300 bar operation and 4 6mm ID columns will support 16 000 psi 1000 bar operation Opening columns may compromise these pressure limits Chromatographie performance has not been tested above 600 bar Because of the small particle size dry ZORBAX packings are respirable Columns should only be opened in a well ventilated area Operational Guidelines The direction of flow is marked on the column While it is not harmful to the column reverse flow should be avoided except to attempt removal of inlet blockage see Column Care section These columns are packed and assembled for high pressure ap to 600 bar use Disassembling the Column will degrade column performan Eclipse XDB C8 is compatible with water and all common organic solvents Avoid use of this column below pH 2 or above pH 8 optimum lifetime and performance are obtained at pH 3 8 Maximum operating pressure is 600 bar 9000 psi Maximum operating temperature is 60 C NOTE Eclipse XDB columns are designed for high stability over a wide pH range However all silica based packings have some solubility in pH gt 6 aqueous mobile phases Therefore when using silica based columns under conditions of pH gt 6 maximum column lifetime is obtained by operation at low temperatures lt 40 C using low buffer concentra tions in the ran
3. General Description Eclipse XDB C8 RRHT threaded columns are specially designed for higher pressure operation up to 600 bar and are packed with a high performance microparticulate C8 packing for high speed reversed phase HPLC Eclipse XDB C8 uses the technologies of eXtra Dense Bonding XDB of organo sitane ligands and double endeapping to protect the ultra pure Type B silica support from dissolution in mobile phases of intermediate and higher pH Eclipse XDB C8 is especially useful for the separation of acidic basie and other highly polar compounds by reversed phase liquid chromatography Eclipse XDB C8 packing is made by first chemically bonding a dense monolayer of dimethyl a octyisilane stationary phase to a specially prepared ultra high purity 299 905 Si0 ZORBAX Rx SIL porous silica Support This spectal ZORBAX silica support Type B is designed to reduce or eliminate strong adsorp tion of basic and highly polar compounds The bonded phase packing is then doubly endcapped using proprietary reagents and procedures to obtain maximum deactivation of the silica surface This dlonsely covered deactivated column packing can be used for acidic and neutral samples but is especially suited for separating basic compounds that produce poor peak shapes on most columns The extra dense GURE Eclipse XDB C8 RANT QC Chromatogram Operating Conditions Column Else XDE C8 RRHT 4 6 mm ID x 50 mm 1 8 yn threaded olun
4. a Matile phase 60 Acetontle 40 Water Flow rate 20 mL min Tamara bint 23 C Detector UV 254 nm Sample volume 2 pL Peak ety lac 2 Phen Sumt 200 y o ug ml tnnt 1 t Cknitrobenzene ANagiahalene Agilent ZORBAX Eclipse XDB C8 RRHT Threaded Column Datasheet coating of bonded phase and exhaustive endeapping simultaneously deactivate the silica surface from deleterious interactions with samples and also protects the silica support from dissolution in intermediate and higher pH The uniform spherical Eclipse XDB C8 particles are based on ZORBAX Rx SIL silica support that has a nominal surface area of 180 m g and a controlled pore size of 80A Columns are loaded to a stable uniform bed density using a proprietary high pressure slurry loading technique to give maximum column efficiency Column Characteristics A typical Quality Control test chromatogram for a 1 8 pm Eclipse XDB C8 RRHT 4 6mm ID 50 mm threaded column is shown in Figure 1 The actual QC test and performance of your column is described on the Column Performance Report enclosed with Safety Considerations All points of connection in liquid chromatographic systems are potential sources of leaks Users of liquid chromatographic equipment should be aware ofthe toxicity or flammability oftheir mobile phases These RRHT assembled columns are mechanically stable and have been tested to very high pressures to assure safe lab operation on
5. ge of 0 01 to 0 02 M Column stability at pH gt 6 is also enhanced by avoiding phosphate and carbonate buffers ref H A Claessens MLA van Straten and J J Kirkland J Chromatogr A 728 1996 259 Columns should not be maintained at neutral or elevated pH or elevated temperature when not in use Mobile Phase Selection The bonded stationary phase is nonpolar in nature and is best used with mobile phases such as methanol water or acetonitrile water mixtures Increasing the amount of organie component usually reduces the retention time of the sample Due to the relatively high viscosity of recommended mobile phases inereased efficiency can be achieved with the use of column temperatures in the range of 40 60 C however best column lifetime is achieved with operation at lt 40 C Gradient elution techniques for this packing often use 5 methanol or acetonitrile as the initial solvent and 100 methanol or acetonitrile as the final solvent Additional informa tion on solvent selection may be found in Chapters ix and Seven Introduction to Modern Liquid Chromatography Second Edition L R Snyder and J J Kirkland John Wiley amp Sons 1979 and Chapters Six Seven and Eight Practical HPLC Method Development Second Edition LR Snyder J L Glajeh and J J Kirkland John Wiley amp Sons 1997 Applications Ectipse XDB C8 can be used with basic neutral or acidic compounds fonizable compounds basic
6. he column with stronger less polar solvents Solvents such as methanol acetonitrile or a 95 5 mixture of dichloromethane and methanol should remove most highly retained compounds In extreme cases dimethyl sulfoxide or dimethylfor mamide at low flow rates may also be used for this purpose When switching between solvents with vastly different polarities it may be necessary to first purge the column with a mutually miscible solvent such as isopropanol Since columns have 3 Sinch end nuts a short 3 8 inch wrench should be used to attach the columns to the instrument to avoid any additional tightening of the end fittings Over tightening the end fittings will cause damage and require column replacement Storage Recommendations Long term storage of siliea based bonded phase columns should be in a pure organic solvent preferably an aprotie liquid such as 100 acetoni trile If the column was previously used with a buffered mobile phase the buffer should first be removed by purging the column with 20 30 column volumes of a 50 50 mixture of methanol or acetoni trile and water followed by 20 20 column volumes of the pure solvent Before storing the column the end fittings should be tightly capped with end plugs to prevent the packing from drying out Columns may be safely stored for short periods in most mobile phases However to protect equipment it is desirable to remove salts from the instrument and column by purging t
7. he column with the same mobile phase without the buffer for example using 60 40 ACN H O to remove a 60 40 ACN 0 02 M phosphate buffered mobile phase Re equilibration is rapid with the original mobile phase when using this approach and any danger of corrosion from the salts is eliminated Agilent Ordering Information For more information on our products visit our Agilent Technologies home page on the World Wide Web agilent com chem supplies For Technical Support in the US and Canade call 1 800 227 9770 or call your local Agilent sales office Agilent Technologies Printed in USA 03 10 2008 Pat No 20100006 A
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Agilent ZORBAX Eclipse XDB C8 RRHT Threaded Column Datasheet