UV-detector

The concentrations of benzoic acid, aspartame, caffeine, and saccharin in a variety of beverages are determined in this experiment. A Gig column and a mobile phase of 80% v/v acetic acid (pH = 4.2) and 20% v/v methanol are used to effect the separation. A UV detector set to 254 nm is used to measure the eluent s absorbance. The ability to adjust retention times by changing the mobile phase s pH is also explored. 

This experiment describes the quantitative analysis of the asthma medication Quadrinal for the active ingredients theophylline, salicylic acid, phenobarbital, ephedrine HGl, and potassium iodide. Separations are carried out using a Gi8 column with a mobile phase of 19% v/v acetonitrile, 80% v/v water, and 1% acetic acid. A small amount of triethylamine (0.03% v/v) is included to ensure the elution of ephedrine HGl. A UV detector set to 254 nm is used to record the chromatogram. 

The use of an amperometric detector is emphasized in this experiment. Hydrodynamic voltammetry (see Chapter 11) is first performed to identify a potential for the oxidation of 4-aminophenol without an appreciable background current due to the oxidation of the mobile phase. The separation is then carried out using a Cjg column and a mobile phase of 50% v/v pH 5, 20 mM acetate buffer with 0.02 M MgCl2, and 50% v/v methanol. The analysis is easily extended to a mixture of 4-aminophenol, ascorbic acid, and catechol, and to the use of a UV detector. 

R-propranolol), Tenormim (oL-atenolol) and Lopressor (oL-metaprolol). The mobile phase was 90 10 (v/v) acetonitrile and water. A UV detector set to 254 nm is used to obtain the chromatogram. 

In this experiment students analyze an artificial RNA digest consisting of cytidine, uridine, thymidine, guanosine, and adenosine using a Cjg column and a mobile phase of 0.4% v/v triethylammonium acetate, 5% v/v methanol, and 94.6% v/v water. The chromatogram is recorded using a UV detector at a wavelength of 254 nm. 

Caffeine in tea and coffee is determined by CZE using nicotine as an internal standard. The buffer solution is 50 mM sodium borate adjusted to pH 8.5 with H3PO4. A UV detector set to 214 nm is used to record the electropherograms. 

High performance Hquid chromatography (hplc) may be used to determine nitroparaffins by utilizing a standard uv detector at 254 nm. This method is particularly appHcable to small amounts of nitroparaffins present, eg, in nitro alcohols (qv), which caimot be analyzed easily by gas chromatography. Suitable methods for monitoring and deterrnination of airborne nitromethane, nitroethane, and 2-nitropropane have been pubUshed by the National Institute of Occupational Safety and Health (NIOSH) (97). Ordinary sorbant tubes containing charcoal are unsatisfactory, because the nitroparaffins decompose on it unless the tubes are held in dry ice and analyzed as soon after collection as possible. 

In hplc, detection and quantitation have been limited by availabiHty of detectors. Using a uv detector set at 254 nm, the lower limit of detection is 3.5 X 10 g/mL for a compound such as phenanthrene. A fluorescence detector can increase the detectabiHty to 8 x 10 g/mL. The same order of detectabiHty can be achieved using amperometric, electron-capture, or photoioni2ation detectors. 

Simultaneous quantification of the herbicides atra2ine, sima2ine, terbut5la2ine, propa2ine, and prometryne and their principal metabohtes has been reported in natural waters at 3—1500 ng/L concentration (104). The compounds were enriched on graphiti2ed carbon black and analy2ed with hplc and a diode array uv detector. 

All the chlorophenols can be separated usiag C g-grafted siUca columns. In NH2 grafted columns, the elution depends on the An electrochemical detector ia oxidation mode, more sensitive than uv detectors, is generally used to detect very low quantities, especially ia analy2iag 2,4,5-trichlorophenol. 

We have developed the method for quantitative analysis of urinary albumin with CE. A capillary electrophoresis systems Nanophor 01 (Institute of Analytical Instmmentation, Russian Academy of Sciences, Saint-Petersburg) equipped with a UV-detector was used to determine analyte. Separation was achieved using 45 cmx30 p.m I.D. fused silica capillary column with UV-detection at 214 nm. 

A capillary electrophoresis systems CAPEE 103R ( Eumex Etd, Saint-Petersburg) equipped with a UV-detector was used to separate and quantify analytes. 

For selective estimation of phenols pollution of environment such chromatographic methods as gas chromatography with flame-ionization detector (ISO method 8165) and high performance liquid chromatography with UV-detector (EPA method 625) is recommended. For determination of phenol, cresols, chlorophenols in environmental samples application of HPLC with amperometric detector is perspective. Phenols and chlorophenols can be easy oxidized and determined with high sensitivity on carbon-glass electrode. 

Experimental part was provided by device Model Knauer-Compact with UV-detector (b=3 mm) at 250 nm and column Spherisorb-ODS-2 (250x4,6 mm). Sample volume was 1-2 p.1 injected by Reodyne 7725. Concentration range was 0.4-0.5 mg/ml for solutions of studied substances in DMSO. The organic modificator concentration range was 75-85 % w for methanol and 40-60 % w for acetonitrile in eluent (flow rate -1 ml/min). 

The peaks shown were obtained using a low dispersion UV detector (cell volume, 1.4 pi) in conjunction with a sample valve with a 1 pi internal loop. All tubes were of... 

A low volume (0.2 pi) Valeo sample valve was employed with one end of the open tube connected directly to the valve and the other connected directly to the sensor cell of the detector. The UV detector was the LC 85B manufactured by Perkin Elmer, and specially designed to provide low dispersion with a sensor volume of about 1.4 pi. The total variance due to extra-column dispersion was maintained at... 

A computer program was compiled to work out the ray-tracing of UV detector of high performance capillary electrophoresis at the investigation of 5 and 6 (98MI59). The capacity factor of 5 at different temperature and at different mobile phase compositions was experimentally determined in bonded-phase chromatography with ion suppression (98MI15). 

Figure 1.2 Chromatogram of coal-tar oil obtained by using the following conditions column, Waters Spherisorb PAH 5 mm in 250 p.m id X 30 cm fused silica column oven temperature, 100°C UV detector wavelength to 254 nm mobile phase, 100 to 300 bar CO2 and 0.10 to 1.00 p.L min methanol over 30 minutes.

Figure 1 The gel-chromatogram of the obtained product from the alkylation reaction of toluene with EC under various reaction conditions T, K = 273 (1), 293 (2), 313 (3), 333 (4). The ratio of toluene epichlorohydrin 0.5 1 (5) 1 1 (6), 2 1 (7), and 5 1 (8). (-) refractometer detector (—) UV-detector.

No account of UV detectors would be complete without mention of the diode array (multichannel) detector, in which polychromatic light is passed through the flow cell. The emerging radiation is diffracted by a grating and then falls on to an array of photodiodes, each photodiode receiving a different narrow-wavelength band. A microprocessor scans the array of diodes many times a... 

The dead point is obtained by including in the sample a trace of an unretained solute or, more often, one of the components of the mobile phase. For example, when using a methanol water mixture as the mobile phase, the dead point is obtained from the elution of a pure sample of methanol. The pure methanol can often be monitored, even by a UV detector, as the transient change in refractive index resulting from the methanol is sufficient to cause a disturbance that is detectable. 

As a result of its highly polar character, silica gel is particularly useful in the separation of polarizable materials such as the aromatic hydrocarbons and polynuclear aromatics. It is also useful in the separation of weakly polar solute mixtures such as ethers, esters and in some cases, ketones. The mobile phases that are commonly employed with silica gel are the n-paraffins and mixtures of the n-paraffins with methylene dichloride or chloroform. It should be borne in mind that chloroform is opaque to UV light at 254 nm and thus, if a fixed wavelength UV detector is being used, methylene dichloride might be a better choice. Furthermore, chloroform is considered toxic and requires special methods of waste disposal. Silica gel is strongly deactivated with water and thus, to ensure stable retentive characteristics, the solvent used for the mobile phase should either be completely dry or have a controlled amount of water present. The level of water in the solvent that will have significant effect on solute retention is extremely small. The solubility of water in n-heptane is... 

If the mixture to be separated contains fairly polar materials, the silica may need to be deactivated by a more polar solvent such as ethyl acetate, propanol or even methanol. As already discussed, polar solutes are avidly adsorbed by silica gel and thus the optimum concentration is likely to be low, e.g. l-4%v/v and consequently, a little difficult to control in a reproducible manner. Ethyl acetate is the most useful moderator as it is significantly less polar than propanol or methanol and thus, more controllable, but unfortunately adsorbs in the UV range and can only be used in the mobile phase at concentrations up to about 5%v/v. Above this concentration the mobile phase may be opaque to the detector and thus, the solutes will not be discernible against the background adsorption of the mobile phase. If a detector such as the refractive index detector is employed then there is no restriction on the concentration of the moderator. Propanol and methanol are transparent in the UV so their presence does not effect the performance of a UV detector. However, their polarity is much greater than that of ethyl acetate and thus, the adjustment of the optimum moderator concentration is more difficult and not easy to reproduce accurately. For more polar mixtures it is better to explore the possibility of a reverse phase (which will be discussed shortly) than attempt to utilize silica gel out of the range of solutes for which it is appropriate. 

The pressure sensitivity of a detector will be one of the factors that determines the long term noise and thus can be very important. It is usually measured as the change in detector output for unit change in sensor-cell pressure. Pressure sensitivity and flow sensitivity are to some extent interdependent, subject to the manner in which the detector functions. The UV detector, the fluorescence detector and the electrical... 

The UV detector is the most popular and useful LC detector that is available to the analyst at this time. This is particularly true if multiwavelength technology is included in the genus of UV detectors. Although the UV detector has definite limitations, particularly with respect to the detection of non-polar solutes that do not possess a UV chromaphore, it has the best combination of sensitivity, versatility and reliability of all the detectors so far developed for general LC analyses. 

There are two types of UV detector the fixed wavelength detector and the multi-wavelength detector. A diagram of a fixed wavelength UV detector is shown in figure 5. 

The popularity of the UV detector, the electrical conductivity detector and the fluorescence detector motivated Schmidt and Scott (5,6) to develop a trifunctional detector that detected solutes by all three methods simultaneously in a single low volume cell. 

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