Dynamic chromatographic detectors

Figure 15-7. Detection limits and dynamic range of some chromatographic detectors.

At first glance HSGC and fats and oils may look incompatible. Despite the fact that the majority of substances related to lipids are involatile, some niches exist where HSGC fits in perfectly. In general, a major weakness of static HSGC - a lack of sensitivity when compared with the dynamic variant - can be overcome by cryofocusing. Such systems are now commercially available. This allows one to present to the gas chromatographic detectors the same vapour mixture as one smells on a sample, except at concentrations which are detectable not only by the human nose but also by the detectors commonly used in GC. 

Sulfur compounds play a major role in determining the flavor and odor characteristics of many food substances. Often sulfur compounds are present in trace levels in foods making their isolation and quantification very difficult for chromatographers. This study compares three gas chromatographic detectors the flame photometric detector, sulfur chemiluminescence detector and the atomic emission detector, for the analysis of volatile sulfur compounds in foods. The atomic emission detector showed the most linearity in its response to sulfur the upper limit of the linear dynamic range for the atomic emission detector was 6 to 8 times greater than the other two detectors. The atomic emission detector had the greatest sensitivity to the sulfur compounds with minimum detectable levels as low as 1 pg. 

An evaluation of gas chromatographic detectors typically involves a study of dynamic range, mimimum detectable level and selectivity. Dynamic range is one of the major response characteristics of detectors. It is the range of sample... 

The flame ionization detector (FID) is the most popular detector because of its high sensitivity and wide linear dynamic range [1]. The FID is an ionization detector that exhibits a nearly universal response to all organic compounds. The sensitivity, stability, excellent linear range, ease of operation and maintenance, along with wide applicability and low cost has made this detector the most popular gas chromatographic detector in use today [2]. 

Another dynamic measurement is the LCEC technique which can be thought of, simpHsticaHy, as EIA using a chromatographic column positioned between the sample injection port and the detector. Bioanalytical systems (BAS) of West Lafayette, Indiana, specializes in instmmentation for LCEC. Their catalogs come with extensive bibhographies covering a variety of appHcations. 

So far the plate theory has been used to examine first-order effects in chromatography. However, it can also be used in a number of other interesting ways to investigate second-order effects in both the chromatographic system itself and in ancillary apparatus such as the detector. The plate theory will now be used to examine the temperature effects that result from solute distribution between two phases. This theoretical treatment not only provides information on the thermal effects that occur in a column per se, but also gives further examples of the use of the plate theory to examine dynamic distribution systems and the different ways that it can be employed. 

The 1/16" x 0.02" i.d. transfer line also functioned as a sample dilution device in other applications, a stainless steel column packed with glass beads has been found to be useful for dilution. This simple dynamic dilution technique has been used extensively in flow injection analysis.3 A refractive index detector is typically used to measure the sample transfer time. As shown in Figure 4, approximately 5 minutes is required to transfer the sample plug to the Rheodyne valve. As the apex of the sample band passes though the Rheodyne valve, the valve is activated and 1 pi injected onto the liquid chromatographic column. The sample transfer time was checked periodically over 1 year of operation and found to be stable. 

The dynamic range of protein expression represents a main obstacle since abundant proteins are seldom of interest and others such as transcription factors are only present in a few copies. There is no detector that is able to visualize all proteins at the same time so that prefractionation and the investigation of subproteomes is required. In fact, pre-MS sample preparation techniques exploiting electrophoretic, chromatographic, or chemical properties of the analyte are often the bottleneck of proteomics. 

With a modern variable-wavelength or PDA detector, it is safe to use a maximum absorbance of up to 1.0-1.5 absorbance units (AU). A typical UV detector today should have a baseline noise in the order of 10 micro absorbance units (10 AU). Therefore, modern UV detectors should have a dynamic range of 4-5 orders of magnitude. The sample concentration and injection volume can be adjusted to make it possible to quantify the major component and impurities at the 0.05% level in the same chromatographic run without changing detection wavelength, sample concentration, or injection volume. 

It is generally required that all methods allow for the monitoring of API and impurities/degradation products in the same chromatographic run, that run times per sample should not be too long, and that for precise and robust quantitative analyses, the separation of the peaks of interest should have target resolutions of >2.0. To allow for easy transfer, the detector response for the nominal concentration of the API (100%, w/w) should be about 75% of the qualified linear dynamic range of the detector. Methods should be temperature controlled (e.g., at 35°C,... 

Chromatographic approaches have been also used to separate nanoparticles from samples coupled to different detectors, such as ICP-MS, MS, DLS. The best known technique for size separation is size exclusion chromatography (SEC). A size exclusion column is packed with porous beads, as the stationary phase, which retain particles, depending on their size and shape. This method has been applied to the size characterization of quantum dots, single-walled carbon nanotubes, and polystyrene nanoparticles [168, 169]. Another approach is hydro-dynamic chromatography (HDC), which separates particles based on their hydro-dynamic radius. HDC has been connected to the most common UV-Vis detector for the size characterization of nanoparticles, colloidal suspensions, and biomolecules [170-172]. 

Nowadays, most chromatographic software is capable of calculating the detector noise and drift. Typically, the detector should be allowed to warm up and stabilize prior to the test. Temperature fluctuations should be avoided during the test. The noise and drift tests can be performed under static and dynamic conditions. For a static testing condition, the flow cell is filled with methanol, and no... 

For a detector to be of use in quantitative analysis, the signal output should be linear with concentration for a concentration-sensitive detector and with mass for a mass-sensitive detector. Some detectors have an additional time constant purposely introduced to remove the high-frequency noise. This should always taken into consideration, since a slow detector response can significantly broaden and attenuate chromatographic peaks relative to those actually sensed. Moreover, a versatile detector should have a wide linear dynamic range so that major and trace components can be determined in a single analysis, over a wide concenua-tion range. 

---