This section provides some general guidelines for the care, use and storage of hilic columns. Quickly and easily reversed hplc column develop methods based on compound type, usp methods, application, or desired column phase. Hilic is an increasingly popular alternative to reversedphase lc for challenging polar analytes because it provides better retention of watersoluble. Atlantis hilic silica columns are used in combination with high organic mobile phases 80% acetonitrile to provide retention of analytes that are simply too polar to be retained by reversephase chromatography. A robust hplc uhplc hilic column with neutral character. Hilic stationary phases thermo fisher scientific us. Column troubleshooting is more usually performed by reference to the chromatogram and chromatographic peak.
New agilent infinitylab poroshell 120 hilic chemistries combine innovative particle technology and bonded phases to improve retention and peak shape for polar analytes. Hydrophilic interaction chromatography hilic is therefore. Enhanced usability, traceability, and security with a preprogrammed id tag. If the column performance is poor after regenerating and cleaning, call your local waters office for additional support. Thermo fisher scientific understands the demands of chemical analysis in the pharmaceutical industry. Amino phase based on ultrapure silica gel purity 99 %, metal content waters. The acclaim mixed mode hilic1 columns stationary phase consists of a hydrophobic alkyl chain with a terminal diol group. Maintenance of spray stability is a key challenge in lcms analysis of. If this flushing procedure does not solve the problem, purge the column with 5. Choose from our wide selection of hplc and lc columns as well as gc columns for both your perkinelmer and non perkinelmer chromatography instrumentation. All of these situations are likely to result in your instrument going offline for maintenance.
Our separation and detection technologies, combined with experience and applications competence, provide. So you can quickly and efficiently separate challenging, highly charged polar compoundsand achieve superior analytical sensitivity in ms mode using msfriendly eluents. View the large range of highly specialized, high performance hplc columns designed for the analysis of small molecules. Shimpack velox hilic shimadzu shimadzu corporation. How to avoid common problems with hilic methods restek. This mode of hplc is now commonly referred to as hilic, hydrophilic interaction liquid chromatography. Hilic column general information halo description halo hilic is a highspeed, highperformance liquid chromatography column based on a new fusedcoretm particle design. Not a good type of mode hilic is a mode of chromatography, not a column. This unique bonded phase provides a highly polar stationary phase on which to perform hilic. Atlantis hilic silica columns are designed to retain very polar, organic molecules that are too polar to retain by rp. Atlantis hilic silica columns are designed to retain very polar, organic. Eliminating ionpairing reagents improves detection limits, method reproducibility and robustness, while reducing instrument maintenance due to harsh mobilephase environments. Hilic stands for hydrophilic interaction liquid chromatography.
Methods of concrete column repair for damages and cracks columns may be repaired by using one or more of the following methods. I am using a zichilic column to detect polar molecules, but i have some background peaks. Zwitterion hilic mnhilic nucleodur hilic is a special zwitterionic modified stationary phase based on ultra spherical nucleodur particles. Since 2003, waters has developed innovative stationary phases for hydrophilic interaction chromatography hilic to overcome the challenge of retaining and. Hydrophilic interaction liquid chromatography columns strongly retain compounds not retained under conventional reversed phase conditions, without the need. Agilent infinitylab poroshell 120 columns for hplc and uhplc more than 15 years ago, agilent introduced poroshell 300, the first commercial columns to use superficially porous particle technology. Chromatography and mass spectrometry training, subjects covered include, hplc, gc, spe, mass spec, hilic. Uhplc guard columns to further extend the life of your analytical column. Consider an ionexchange method for the sample cleanup using a mixedmode ionexchanger, followed by elution in high organic with injection onto the hilic column.
This is particularly true for gags, because the polarity of a given oligosaccharide increases with increasing length and sulfate content. Hydrophilic interaction chromatography hilic is an increasingly popular separation mode that can be used to improve the retention of challenging polar analytes. Therefore, it is important to monitor column back pressure regularly and solve the problem timely. It seems to me that waters has the best service and help for users in europe. The fusedcore particle provides a thin porous shell of highpurity silica surrounding a solid silica core. Thermo scientific product manual for acclaim hilic10 columns page of 23 06533902. Learn how high purity silica of this hplc column is used to retain and separate small, polar analytes specifically in hydrophilic interaction chromatography. Care, maintenance, and troubleshooting oftroubleshooting.
Note if above treatment fails to revive the column, the column should be replaced. The stationary phases used in hilic are quite diverse and users often have the misconception that hilic columns are interchangeable. Hilic an alternative separation technique for glycopeptides. Improved hilic lcms of heparinoids using a chip with postcolumn makeup flow. Improved hilic lcms of heparinoids using a chip with post. Fourteen commercially available particlepacked columns and a monolithic column for hydrophilic interaction liquid chromatography hilic were characterized in terms of the degree of.
So take off you lab coat, relax and let our web tool do the work. This particle design exhibits very high column efficiency due to the. An effective method development approach uses both an xbridge hilic column and an xbridge reversedphase column to screen. General purpose bare silica column for normal phase and hilic applications. Reduced need for sample preparation due to 2 m frits on 2.
Storing a column with buffer in the mobile phase is typically not recommended for a number of reasons, most have little to do with sorbent stability. Agilent zorbax hilic plus hplc columns data sheet column characteristics a typical quality control qc chromatogram for a 4. Highlighted features include high sample throughput, low solvent consumption, column ruggedness, high sensitivity, and lcms compatibility. Hilic uses hydrophilic stationary phases with reversedphase type eluents. The qc test with the performance of your column is shown on the column performance report enclosed with your column. Continuous use of hplc columns under high pressure can cause deterioration and overload of the equipment. Does anyone know why the hilic column stability is affected by the. The hamilton prpc18 is a new hplc column designed to provide highefficiency, reversedphase separations over an extended column life in nearly any mobile phase or ph. Ensure the best results from your chromatography analysis with our highperformance columns.
Waters new hilicbased glycoprotein column enhances glycan. The hilicn stationary phase is based upon the ultrapure ace silica platform with a proprietary polyhydroxy bonded functionality. Supelco ondemand hplc and uhplc webinars and videos on theory, fundamentals. Hplc software and lc drivers for powerful and flexible instrument control. Care, maintenance, and troubleshooting oftroubleshooting of hplc columns columns and consumables edward kim apppp glications engineer january 17, 2008. The halo pentahilic column provides superior retention to silica hilic and amide hilic phases for acids and zwitterionic analytes figure 2 halo pentahilic uses proprietary bonding chemistry that includes five hydroxyl groups on the bonded ligand. The attached manual describes the acquity uplc beh columns and includes user recommendations.
Storage of hilic columns with buffer is recommended to decrease equilibration time. Prior to storage, buffered solutions used during hilic analysis should be washed out of the column and the hplc system. Simplify the switch to hilic with raptor hilicsi columns. The perkinelmer elite 608 columns specifically designed for the analysis of semivolatile pesticides. Optimizing hilicbased analyses of rapifluorms labeled. Application of hilic column for the determination of catechins. Understanding the possible hilic retention mechanisms involved for a particular columnsolventanalyte system is crucial in the process of selecting the right hilic stationary phase, leading to a successful application. Coming to the actual stability of the column, hilic columns may degrade in high.
Important goals in hplc are increasing speed and maintaining resolution. Chromatographic characterization of stationary phases for. The ace hilicn column is a bonded neutral character phase. Encasement or enlargement of the column cross section jacketing. Cathodic protection to stop reinforcing steel corrosion. Specific advice can be found in the appropriate product manual. Hydrophilic interaction chromatography or hydrophilic interaction liquid chromatography, hilic is a variant of normal phase liquid chromatography that partly overlaps with other chromatographic applications such as ion chromatography and reversed phase liquid chromatography. Now, infinitylab poroshell 120 columns have become critical tools for solving the latest challenges in the analytical world. Hydrophilic interaction chromatography hilic was first introduced by alpert in 1990 and since then has attracted increasing attention. I want to test my hilic column with different mobile phase %. Based on ultrapure silica gel zwitterionic hilic phases are ideal for the analysis of polar and ionizable compounds.
Oppositely, a plain silica column will act as a cation exchanger when ph is raised above 4, as the oh groups forms. The rugged beh particles wide usable ph range overcomes this ph limitation and results in long column lifetimes. Results and discussion the separation of a tryptic digest of infliximab was obtained with a 45 minute gradient and a total analysis time of 75 minutes, including column wash with high buffer content and reequilibration at initial conditions. Realkalization of the reinforcing steel to stop corrosion.
Cmms software helps you quickly address maintenance requests, improve facility conditions through routine maintenance, and manage multiple buildings with mobile work orders. By using an amide or amino bonded phase column, polar compounds can be retained by a normal phase or hydrophilic interaction chromatography retention mechanism. This article addresses common problems that analysts encounter when running hilic methods and details how to avoid them and successfully use the technique. Identifying the precise location of glycans within a protein structure is essential to delivering safe and effective.
Agilent infinitylab solutions for hplc and uhplc include instruments, columns, and supplies to help your lab achieve. Hydrophilic interaction chromatography, hilic, chromatographic characterization, structural selectivity, ion exchange interactions abstract. Hilic columns should be stored under hilic conditions. Whether youve run hilic methods before or are attempting to for the first time, there are significant differences between reversedphase and hilic methods that must be considered.
A suitable wash solution is 60% organic solvent in water unless otherwise specificed in product manuals. Figure 1 shows a uv chromatogram with an overlay of five. Unlike the highly aqueous mobile phases required for polar retention in rp separations, atlantis hilic silica columns employ highly volatile 80% organic mobile phases which are ideal for mass spectrometry ms response and sensitivity. Hilic seems to be a missing tool in the analysis of polar and ionized compounds, which are too strongly retained on polar stationary phases used in normal phase mode andor are poorly separated when reserved phase mode is. Hplc column maintenance autosampler contamination retention time variability in hplc troubleshooting hilic a quick guide to the top 10 things that must be avoided in your hplc analysis. If you do not find a reliable solution there, i recommend to use spe as the sample cleanup tool. Optimizing hilicbased analyses of rapifluorms labeled sialylated nglycans. Cation exchange hilic polysulfoethyl a similar to anion exchange hilic, the proper adjustment of ph determines the mode of operation. Whether youve run hilic methods before or are attempting to for the first time. Triazole column is an unique hilic phase that has cyclic amino groups. The raptor hilicsi column simpli fies the switch to hilic because it delivers rugged raptor performance.
Multi mode column for rp, np, hilic and ic, aminopropyl modification without endcapping, outstanding mechanical and chemical stability. Hilic and aqueous normal phase chromatography the terms hydrophilic interaction liquid chromatography hilic and aqueous normal phase anp. Maintenance teams help provide a safe and stable learning environment for students, teachers, and other staff. Our reversed phase hplc column match web tool allows you to develop reversed methods without having to go on the lab. Find raptor reliability modeling software related suppliers, manufacturers, products and specifications on globalspec a trusted source of raptor reliability modeling software information. Thermo scientific acclaim hilic10 product manual pdf. Methods of concrete column repair for cracks and damages. Troubleshooting for increased pressure introduction repeated analysis may increase back pressure. Agilent infinitylab poroshell 120 columns family agilent. Acquity uplc beh hilic columns offer advantages such as retention and separation of very polar basic compounds, alternate selectivity as compared.