Flow standardization reagents

Fig. 3.9 Scheme of typical flow system I, II, III, IV solutions (e.g., sample, standard, reagents), PI and P2 pumps, Z valve, MP sample preparation module, M mixing coil, DET detection system... 

Determination of orthophosphate in phosphatic rock digests and hydrochloric acid in the commercial reagent. The sample was injected into the flowing standard solution, and different fluid elements, each with a specific sample-to-reagent volumetric ratio, were considered for application of SAM [355]. 

The reduction of the yellow-colored Mo(VI) complex to the blue-colored Mo(V) complex is a slow reaction. In the standard spectrophotometric method, it is difficult to reprodudbly control the amount of time that reagents are allowed to react before measuring the absorbance. To achieve good precision, therefore, the reaction is allowed sufficient time to proceed to completion before measuring the absorbance. In the FIA method, the flow rate and the dimensions of the reaction coil determine the elapsed time between sample introduction and the measurement of absorbance (about 30 s in this configuration). Since this time is precisely controlled, the reaction time is the same for all standards and samples. 

Flow rate The limitations associated with the volume of flow cell can be overcome by accurately controlling the flow rate of each stream entering into the manifold. This experimental parameter controls the residence time of the chemiluminescent solution within the cell and can be easily optimized by the operator. How rates are directly proportional to the rate of the CL reaction. As the rate of the reaction increases, the flow rate should be increased but, at the same time, consumption of reagents increases. The flow rate also affects the shape and the height of the peak as well as the measurement rate (number of sample or standard solutions injected per hour). 

FIA has also found wide application in pharmaceutical analysis.214,215 Direct UV detection of active ingredients is the most popular pharmaceutical analysis application of FIA. For single component analysis of samples with little matrix interference such as dissolution and content uniformity of conventional dosage forms, many pharmaceutical chemists simply replace a column with suitable tubing between the injector and the detector to run FIA on standard HPLC instrumentation. When direct UV detection offers inadequate selectivity, simple online reaction schemes with more specific reagents including chemical, photochemical, and enzymatic reactions of derivatization are applied for flow injection determination of pharmaceuticals.216... 

Abstract A preconcentration method using Amberlite XAD-16 column for the enrichment of aluminum was proposed. The optimization process was carried out using fractional factorial design. The factors involved were pH, resin amount, reagent/metal mole ratio, elution volume and samphng flow rate. The absorbance was used as analytical response. Using the optimised experimental conditions, the proposed procedure allowed determination of aluminum with a detection limit (3o/s) of 6.1 ig L and a quantification limit (lOa/s) of 20.2 pg L, and a precision which was calculated as relative standard deviation (RSD) of 2.4% for aluminum concentration of 30 pg L . The preconcentration factor of 100 was obtained. These results demonstrated that this procedure could be applied for separation and preconcentration of aluminum in the presence of several matrix. 

HPLC analysis of furosine (-peak II) in hydrolyzates of non-exposed- (bottom), buffer-exposed (middle), and glucose-exposed (top) dentin samples. Dentin was not reduced prior to hydrolysis. Only the relevant parts of the chromatograms are shown. Amino acids are visualized after post-column labelling with a fluorescent dye. I lysine, II furosine. III homoarginine (internal standard). Column Merck Polyspher AA-NA 120 x 4.6 mm flow 0.2 ml/min gradient pH 5.0 -10.2 postcolumn reagent 0.2 ml/min fluorescence Xgx 330 nm, 440 nm 100-yl injections in buffer pH 2. 

Ninhydrin assay. Our method to determine reactivity towards ninhydrin was a modification of a method described previously (Moore and Stein, 1954 Moore, 1968). Briefly, dried samples were dissolved in 0.10 ml 0.1 M acetic acid and mixed with an equal volume of ninhydrin reagent (Sigma). After 15 minutes in a boiling water bath, samples were diluted with 0.80 ml ethanol/water (1 1 by vol.) and measured for absorbance at 550 nm on a flow-through spectrophotometer (Vitatron). Standards containing 0-0.6 mM leucine in 0.1 M HAc were included. Values were therefore calculated as leucine-equivalents. 

An impurities analytical procedure should be described adequately so that any qualified analyst can readily reproduce the method. The description should include the scientific principle behind the procedure. A list of reagents and equipment, for example, instrument type, detector, column type, and dimensions, should be included. Equipment parameters, for example, flow rate, temperatures, run time, and wavelength settings, should be specified. How the analytical procedure is carried out, including the standard and sample preparations, the calculation formulae, and how to report results, should be described. A representative chromatogram with labeled peak(s) should be included in the procedure. 

Figure 3.10 — Flow manifolds for implementation of flow-through biosensors. (A) Flow injection merging-zones manifold for the bioluminescence detennination of ATP. ATP standards (30 fiL) and luciferin (30 fiL) are injected into the buffered carrier streams, each pumped at 0.7 mL/min and synchronously merged 12.5 cm downstream. Distance from merging point to immobilized enzyme coil, 2.2 cm. (Reproduced from [59] with permission of Elsevier Science Publishers). (B) Completely continuous flow manifold for the determination of NADH. (Reproduced from [71] with permission of the Royal Society of Chemistry). (C) Segmented-flow manifold for the determination of L-(+)-lactate. (Reproduced from [65] with permission of Marcel Dekker, Inc.). (D) Single-channel flow injection manifold with immobilized reagent for the detennination of glucose. (Reproduced from [77] with permission of Elsevier Science Publishers).

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