Liquid chromatography definition

The need for a more definitive identification of HPLC eluates than that provided by retention times alone has been discussed previously, as have the incompatibilities between the operating characteristics of liquid chromatography and mass spectrometry. The combination of the two techniques was originally achieved by the physical isolation of fractions as they eluted from an HPLC column, followed by the removal of the mobile phase, usually by evaporation, and transfer of the analyte(s) into the mass spectrometer by using an appropriate probe. 

By definition, the e]q>erlmentally determined average mobile phase velocity Is equal to the ratio of the column length to the retention time of an unretalned solute. The value obtained will depend on the ability of the unretalned solute to probe the pore volume. In liquid chromatography, a value for the Interstitial velocity can be obtained by using an unretalned solute that Is excluded from the pore volume for the measurement (section 4.4.4). The Interstitial velocity Is probably more fundamentally significant than the chromatographic velocity in liquid chromatography (39). 

The general steps in developing an acceptable analytical method in liquid chromatography are summarized in Figure 4.26. Method development starts with a clear definition of the needs of the analysis. How many detectable components are present in the ample Are all peaks equally relevant In the first case all peaks must be resolved and the difficulty of providing the desired result will increase with the number of components in the sample. 

Complete the following definition of liquid chromatography by filling in the blanks. For each space, choose a word from the groups given below ... 

Liquid chromatography (LC) and, in particular, high performance liquid chromatography (HPLC), is at present the most popular and widely used separation procedure based on a quasi-equilibrium -type of molecular distribution between two phases. Officially, LC is defined as a physical method... in which the components to be separated are distributed between two phases, one of which is stationary (stationary phase) while the other (the mobile phase) moves in a definite direction [ 1 ]. In other words, all chromatographic methods have one thing in common and that is the dynamic separation of a substance mixture in a flow system. Since the interphase molecular distribution of the respective substances is the main condition of the separation layer functionality in this method, chromatography can be considered as an excellent model of other methods based on similar distributions and carried out at dynamic conditions. 

Matthijs, N., Perrin, C., Maftouh, M., Massart, D.L., Vander Heyden, Y. Definition and system implementation of strategies for method development of chiral separations in normal- or reversed-phase liquid chromatography using polysaccharide-based stationary phases. 7. Chromatogr. A 2004, 1041, 119-133. 

For each dmg substance, the maximum acceptable levels of the various impurities are described in the drug substance monograph or the specification included in the submissions to the regulatory authorities. In this chapter, the ICH Q6A [4] and Q6B [5] definition of specification is used. A specification consists of three parts the test (e.g. moisture content, impurities), references to the analytical procedure (e.g. high-performance liquid chromatography [HPLC], gas chromatography [GC]), and the acceptance criterion (e.g. not more than 0.50%). 

High-performance liquid chromatography (HPLC) has become a standard separation technique used in both academic and commercial analytical laboratories. However, there are several drawbacks to standard HPLC, including high solvent consumption, large sample quantity, and decreased detection sensitivity. Micro-HPLC (pHPLC) is a term that encompasses a broad range of sample volumes and column sizes (as shown in Table 3.1), but Saito and coworkers provided narrower definitions in their review based on the size of the columns. ... 

Using amylose tris-3,5-dimethylphenylcarbamate as the chiral selector in enantioselective high-performance liquid chromatography, micropreparative resolution of the DHA racemate was achieved and the chromatographic behaviour in enantio-GC could be defined by coinjecting these references of definite chirality (Fig. 17.4) [13]. 

The availability of stable isotope-labeled PA makes an accurate quantitative determination of this imino acid possible. A short high-performance liquid chromatography (HPLC) run prior to the mass spectrometer inlet will result in a discrete peak of PA. For the definitive diagnosis of AASA dehydrogenase deficiency, a simultaneous determination of AASA would be preferred. The absence of a commercially available labeled standard leaves this analysis in the experimental stage. 

The term A is related to the flow profile of the mobile phase as it traverses the stationary phase. The size of the stationary phase particles, their dimensional distribution, and the uniformity of the packing are responsible for a preferential path and add mainly to the improper exchange of solute between the two phases. This phenomenon is the result of Eddy diffusion or turbulent diffusion, considered to be non-important in liquid chromatography or absent by definition in capillary columns, and WCOT (wall coated open tubular) in gas phase chromatography (Golay s equation without term A, cf. 2.5). 

There are some modifications of the definitions and terms used in gas-liquid chromatography compared to those used in gas-solid chromatography. First, the phase ratio is described as... 

Introduction to fast performance liquid chromatography. Review Buffer preparation, Definitions of pH, Henderson-Hasselbalch equation, and Buffer calculations. 

LC/MS (Liquid Chromatography/Mass Spectrometry)—Chromatography system in which an HPLC is married to a mass spectrometric detector through an evaporated, ionizing interface. A variety of mass spectrometers are used to produce various LC/MS and LC/MS/MS configurations. MS detectors are universal, mass detectors that provide molecular weight information and can give a definitive identification of separated compounds. 

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