Uv

interest in developing a routine high-performance liquid chromatographic system that is capable of separating unknown and complex extracts with the best possible resolution of individual nonvolatile components. A system has been developed employing liquidsolid adsorption chromatography with gradient elution by the application of a series of solvents of rising polarity. A series of solvent reservoirs is connected via a programmable multiple port rotary valve to a solvent mixing chamber containing primary solvent. A high-pressure reciprocating pump draws solvent from this system at the same time, causing a solvent gradient to be generated by pulling a secondary solvent from one of the reservoirs into the mixing chamber. Solvent combinations are changed at appropriate intervals by switching the rotary valve position. Sample introduction is effected by loop or septum injection. The column is a high-efficiency 250 x 4.2mm prepacked column containing microparticulate silica. The detectors incorporated into the system are a combined, variable wavelength ultraviolet and fluorescence photometer and a transport ionization detector.

There remains some doubt concerning the optimum number of solvents to be employed in the multiple solvent, gradient elution programme. Scott and Kucera [264] recommend a 12-solvent series, while Snyder [265] proposes a six-solvent series to cover the entire polarity range adequately.

11.7.4 Ion-exchange chromatography

Among the limited applications of ion-exchange chromatography to the separation of organics in water are the separation of organics in sewage effluents [266-268] and the determination of chlorinated biphenyls [269]. Pill et al. [268], using high-resolution ion-exchange chromatography, identified and quantified 56 and 13 organics, respectively, in primary and secondary sewage effluents. Many more unidentified compounds were present.

11.7.5 Gel filtration chromatography

Gel filtration chromatography using Sephadex G100 as column packing, and ultraviolet detectors, has been used in studies carried out on the elution of humic acid [270] and in characterization studies on secondary sewage effluents [271] and in organic substances in river waters [272].

11.7.6 Preconcentration

A limited amount of work has been carried out on freeze concentration methods for organics in non-saline waters. The method has been applied to the preconcentration of amino acids, carbohydrates, citric acid and phenylalanine [273] giving virtually complete recovery of these substances at concentrations up to 0.2mg L-1. The technique has been used in conjunction with Sephadex gel chromatography to characterize organic components in the molecular weight range 200- 5000 [274] in secondary sewage effluents. The filtered samples were concentrated using a freeze-drying unit with methyl alcohol as the bath fluid. The organic carbon recovery in the freeze-dried samples was between 92 and 96%.

11.8 Mineral waters 11.8.1 Preconcentration

Korenman et al. [275] preconcentrated organics from mineral waters by passage through a bed of Polysorb-1 presaturated with 1.5-2.0 parts of namyl acetate per weight of sorbent. The extracted organics were eluted with diazotized sulphanilic acid and determined photometrically. Infrared and ultraviolet spectroscopy, gas chromatography and luminescence analysis were all used to detect and identify extracted organics.

11.9 Ultrapure waters

11.9.1 High-performance liquid chromatography

Reust and Meyer [284] have shown that high-performance liquid chromatography on reversed-phase columns with ultraviolet detection is a useful tool for the determination of the amount of organic impurities present in ultrapure water. A sufficiently sensitive analysis of different types of ultrapure water requires sample volumes of several hundred millilitres combined with a very low detection wavelength. In this work sample volumes of 480mL were analysed at a detection wavelength of 210nm. Using these experimental conditions it was possible to detect and eliminate several contamination sources in the preparation and handling procedures of ultrapure water.

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