HPLC A Praactical User''''''''S Guide Part 1 ppt
Tài liệu HPLC A practical user`s guide pptx
... solvent.8 ADVANTAGES AND DISADVANTAGES OF HPLC 1ADVANTAGES ANDDISADVANTAGES OF HPLC 3The first things we need to understand are how an HPLC system works, itsbest applications and advantages over ... remember that HPLC is both an analytical and a preparative tool. Often the preparative capabilities of the HPLC are over-looked. While normal analytical injections contain picogram to nanogramquantities, ... tool.Twenty-five years in HPLC, first as a user, then in fieldsales and application support for HPLC manufacturers, and finally working as a teacher and consultant has shown me that the average user wants an instru-ment...
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HPLC A Practical User’s Guide P1 doc
... remember that HPLC is both an analytical and a preparative tool. Often the preparative capabilities of the HPLC are over-looked. While normal analytical injections contain picogram to nanogramquantities, ... tool.Twenty-five years in HPLC, first as a user, then in fieldsales and application support for HPLC manufacturers, and finally working as a teacher and consultant has shown me that the average user wants an instru-ment ... linear solvent gradients and a peristaltic pump is used to feedsolvent to the column head. Packing materials generally made of silica gel,alumina, and agarose are available to allow separation...
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HPLC A Practical User’s Guide P2 potx
... equilibrated andstandardized, we are ready to carry out an HPLC separation on a real sample.We might add an internal standard (if necessary, to correct for injectionvariations), dilute our sample ... internal stan-dard to both our sample and our known standard mixture, we can calculatepeak heights or areas relative to that of the internal standard. Variations inthe injection size of the sample ... laboratories are mixed bed, activatedcharcoal, and ion exchange systems that produce water on demand. Thesesystems usually have a couple of ion-exchange cartridges and one activatedcharcoal filter...
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HPLC A Praactical User''''S Guide Part 1 ppt
... Data Acquisition / 13 2 10 .4 Solvent Conservation / 13 3III HPLC UTILIZATION 13 5 11 Preparative Chromatography 13 7 11 .1 Analytical Preparative / 13 8 11 .2 Semipreparative / 13 9 11 .3 “True” Preparative ... solvent.8 ADVANTAGES AND DISADVANTAGES OF HPLC CONTENTSPREFACE xiI HPLC PRIMER 1 1 Advantages and Disadvantages of HPLC 3 1. 1 How It Works / 4 1. 1 .1 A Separation Model of the Column / 5 1. 1.2 Basic ... “True” Preparative / 13 9 12 Sample Preparation and Methods Development 14 3 12 .1 Sample Preparation / 14 3 12 .1. 1 Deproteination / 14 4 12 .1. 2 Extraction and Concentration / 14 5 12 .1. 3 SFE (Cartridge...
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HPLC A Praactical User''''S Guide Part 2 doc
... of a packing material, sampleclean-ups, and broad separations of classes of materials. They are available inalmost any type of packing available for HPLC separations: partition, ionexchange, ... Service and support have always been the differentiating factorsthat really separate and define the best companies. 2. 1.4 Brand Names and Clones A single supplier working on an O.E.M. basis has always ... such as a caronal charged aerosol detector or a mass spectrometer andinterface module, will dramatically increase the system price. In 20 04, I talkedto a laboratory director who had just purchased...
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HPLC A Praactical User''''S Guide Part 3 doc
... peak height times the half peakwidth. The areas of each peak are summed to give a total area. Dividing thisinto the area of each peak gives a relative area percentage for each peak. Likepeak ... 1 to 8 for analytical separations and 4 to 12 forpreparative. a s range from 1 to 2; at a = 1, peaks completely overlap, muchabove a = 2 and the separation can be made in a separatory funnel. ... equilibrated andstandardized, we are ready to carry out an HPLC separation on a real sample.We might add an internal standard (if necessary, to correct for injectionvariations), dilute our sample...
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HPLC A Praactical User''''S Guide Part 4 pdf
... use a technique called sequentialanalysis (Fig. 5.6). For example, we can make a size separation, then take a sizefraction and do a partition separation. This is commonly used in separating a complex ... Anionic packing materials havean affinity for anions (negatively charged ions) and have positively chargedfunctional groups on their surfaces, usually organic amines. Cationic packingsattract cations ... inducible charges. At one pH they areuncharged, at a different pH they are charged. Examples are organic acids,which are uncharged at pH 2.0, but form cations at pH 6.5,and organic primaryamines,...
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HPLC A Praactical User''''S Guide Part 5 pot
... form as a pair of peaks or a badly tailing peak for eachcompound.Ion-pairing reactions can also be carried out using quaternary amines ascounter-charges for organic acid and organic phosphate ... I’m talking about a change that takes place gradually or all at once and remains high. Be awarethat gradient mixtures of some solvents like methanol and water go through a pressure maximum that ... as a tool that other chromatographershave used to produce separations at pH high enough to separate many aminesin their free amine form. Silica appears as a solid on evaporating fractions and,occasionally,...
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HPLC A Praactical User''''S Guide Part 6 potx
... covalently bonded phasesavailable on zirconium that can simulate silica C18separations with a- typepeak shifting, but bonded phases are available that provide truly unique sep-arations. Many ... monosaccharides and separate alcoholsugars from each other.8.1.2 Anionic: Weak and StrongIn strong anionic ion exchangers, the quaternary amine is attached to the silicasurface by an alkyl chain, and ... surface toward the mobile phase and are therefore moreavailable. Certain natural and synthetic dyes have been found to serve as sub-strate mimics for a class of enzymes call hydrogenases and have...
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HPLC A Praactical User''''S Guide Part 7 docx
... than the inte-grator to store a single chromatogram, however, it can use this raw data toreport tables of peak maxima, areas, and heights versus time and recalculate,redisplay, and compare ... reports each as a percent of the total.The computer handles data differently. It measures the voltage at a presettime interval and stores each displacement as a digital word, usually at a rateof ... thepeak area (summed from all the points from leaving baseline to finding itagain) and the maxima.At the end of the chromatogram,it reports these areasversus time, sums all areas, and reports each...
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HPLC A Praactical User''''S Guide Part 8 ppt
... necessary, we can increase to the next size largerloop without affecting resolution.1 38 PREPARATIVE CHROMATOGRAPHYTable 11.1 A Guide to Preparative Scale-upAnalytical Semipreparative PreparativeSize ... least important. Here we aregathering grams of material for biological testing and structural analysis, andas reaction intermediates. HPLC: A Practical User’s Guide, Second Edition, by Marvin ... ion-pairing reagent, pick the most nonpolar ion pairing reagent,extract, neutralize the target compounds charge, and back-extract with a polarsolvent.In a similar way, polar material that passes...
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HPLC A Praactical User''''S Guide Part 9 docx
... concentrations of the standardsare injected, areas are measured, and a calibration curve is platted. Unknownsamples are then injected, chromatograms run, and areas are calculated andcompared ... response factors are calculated relative to theinternal standards, and amounts of each unknown present are calculated fromthe standards calibration factors. Internal standards are usually used ... hydrocarbontype (chlordane) at 80% An/water UV, 220nm, the carbamate type (sevin) at40% An/water UV, 254nm, and the organic phospahate (malathion) at 50%An/water with UV, 192 nm or with a CAD....
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HPLC A Praactical User''''S Guide Part 10 pot
... macro programs.14.6.3 Databases: .DB2 FormatTo export data files into a database program, a database file format called.DB2 was developed in an early PC database, dBase II. Databases are madeup ... retained as a standard format. .WK files are Lotus-1,2,3 formats and .XLS are MicrosoftExcel formats that have become spreadsheet standards, allowing transfer ofdata, calculations, addresses, and ... research laboratories how transient and fragile their archived data filesreally are. It is critically important to have access to file translation from theseproprietary formats into a standard...
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HPLC A Praactical User''''S Guide Part 11 pot
... American Laboratory, LC/GC Mag-azine, the Journal of Liquid Chromatography, and Chemical Abstracts— HPLC Selects. Chemical Abstracts are also on-line as part of Dialog’scomputer database. I have ... Normal-Phase Chromatography.)Alpha (a) —(Separation or chemistry factor). A measure of separationbetween two peak maxima. Ratio of their k′ values.Attenuation—Measure of detector sensitivity. A larger ... nano-level LC/MS, but its potential has been slowto materialize.Chromatography A separation technique producing a qualitative record ofthe relative amounts of components, a chromatogram. HPLC...
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HPLC A Praactical User''''S Guide Part 12 docx
... sigma (s) width, 49Acidic phase silica, 93Alpha (a) 55 A/ D converter, 124 Affinity column, 59, 101Amine column oxidation, 75–76Amino acids, 162AMU (atomic mass unit), 183Analytical mode, 8Analog ... centrifugation.Saturation Column—Sacrificial column placed before the injector to protectthe main column from pH degradation.Seal—Wear surface that both lubricates and separates moving parts in the HPLC. ... Chromatography in Clinical Analysis, P.M. Kabra and L.J.Marton (eds), The Humana Press, Clifton, NJ, 1982, 466 pp.6. Basic Liquid Chromatography, E.L. Johnson and R. Stevenson, VarianAssociates,...
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