Interferences matrix

Matrix interferences in AFS are due mostly to light scatter from particles present in the atomizer, and molecular fluorescence of matrix compounds. Matrix interferences may be reduced by preparing sample-like standards. Matrix effects may also be corrected by wavelength modulation and by using the double-beam technique. Wavelength modulation is usually performed with a piece of quartz or a filter. 

The use of an ICP as an excitation source makes it possible to correct interference due to light scatter by means of several different techniques. The double-beam technique is possible because ion and resonance lines can be obtained by the ICP which do not appear in cooler atomizers. In this technique, non-resonance lines near the resonance line of the analyte are employed. 

To avoid spectral interferences when non-dispersive AFS instruments are used, line-like radiation sources are required. Spectral interferences using narrow line sources are uncommon, while when using continuum sources they are quite common. Corrections can be made by wavelength or amplitude modulation, which can be performed with filters. Different wavelengths are separated from each other by filters whose advantage is their low price and good spectral transmission. 

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Compensating for an interference in the sample s matrix is more difficult. If the identity and concentration of the interferent are known, then it can be added to the reagent blank. In most analyses, however, the identity or concentration of matrix interferents is not known, and their contribution to S stead, the signal from the interferent is included as an additional term... 

When the identity of the matrix interference is unknown, or when it is impossible to adjust the flame to eliminate the interference, then other means must be used to compensate for the background interference. Several methods have been developed to compensate for matrix interferences, and most atomic absorption spectrophotometers include one or more of these methods. 

When possible, a quantitative analysis is best conducted using external standards. Unfortunately, matrix interferences are a frequent problem, particularly when using electrothermal atomization. Eor this reason the method of standard additions is often used. One limitation to this method of standardization, however, is the requirement that there be a linear relationship between absorbance and concentration. 

Accuracy When spectral and chemical interferences are minimized, accuracies of 0.5-5% are routinely possible. With nonlinear calibration curves, higher accuracy is obtained by using a pair of standards whose absorbances closely bracket the sample s absorbance and assuming that the change in absorbance is linear over the limited concentration range. Determinate errors for electrothermal atomization are frequently greater than that obtained with flame atomization due to more serious matrix interferences. 

Furthermore, the extent to which we can effect a separation depends on the distribution ratio of each species in the sample. To separate an analyte from its matrix, its distribution ratio must be significantly greater than that for all other components in the matrix. When the analyte s distribution ratio is similar to that of another species, then a separation becomes impossible. For example, let s assume that an analyte. A, and a matrix interferent, I, have distribution ratios of 5 and 0.5, respectively. In an attempt to separate the analyte from its matrix, a simple liquid-liquid extraction is carried out using equal volumes of sample and a suitable extraction solvent. Following the treatment outlined in Chapter 7, it is easy to show that a single extraction removes approximately 83% of the analyte and 33% of the interferent. Although it is possible to remove 99% of A with three extractions, 70% of I is also removed. In fact, there is no practical combination of number of extractions or volume ratio of sample and extracting phases that produce an acceptable separation of the analyte and interferent by a simple liquid-liquid extraction. 

The problem with a simple extraction is that the separation only occurs in one direction. In a liquid-liquid extraction, for example, we extract a solute from its initial phase into the extracting phase. Consider, again, the separation of an analyte and a matrix interferent with distribution ratios of 5 and 0.5, respectively. A single liquid-liquid extraction transfers 83% of the analyte and 33% of the interferent to the extracting phase (Figure 12.1). If the concentrations of A and I in the sample were identical, then their concentration ratio in the extracting phase after one extraction is... 

Frequently, preconcentration of an analyte is necessary because the detector used for quantitation may not have the necessary detectabiUty, selectivity, or freedom from matrix interferences (32). Significant sample losses can occur during this step because of very small volume losses to glass walls of the recovery containers, pipets, and other glassware. 

Mercury generally is found in low and trace concentrations. So there is need to determine Hg in ranges corresponding to various types of water samples. Detection levels of Hg can be improved by the use of vapour generation technique. This technique allows to sepai ate the analyte from the sample matrix and so to overcome the matrix interference. The fluorescence technique, with its high sensitivity and linearity, in combination with vapour generation, provides for a possibility to detect Hg in parts per trillion per liter regions. 

The trends begun with the general introduction of FTIR technology will undoubtedly continue. It is safe to say that the quality of the data being produced far exceeds our ability to analyze it. In fact, for many current applications, the principle limitations are not with the equipment, but rather with the quality of the samples. Thus, the shift from qualitative to quantitative work will proceed, reaching high levels of sophistication to address the optical and matrix interference problems discussed above. 

Full quantitation is accomplished in the same manner as for most analytical instrumentation. This involves the preparation of standard solutions and matching of the matrix as much as possible. Since matrix interferences are usually minimized in ICPMS (relative to other techniques), the process is usually easier. 

ICP-OES is a destructive technique that provides only elemental composition. However, ICP-OES is relatively insensitive to sample matrix interference effects. Interference effects in ICP-OES are generally less severe than in GFAA, FAA, or ICPMS. Matrix effects are less severe when using the combination of laser ablation and ICP-OES than when a laser microprobe is used for both ablation and excitation. 

W. Pennincks, P. Vankeerberghen, D.L. Massart and J. Smeyers-Verbeke, A knowledge-based computer system for the detection of matrix interferences. Atomic Absorption Spectrometric Methods, J. Anal. Atom. Spectrom. inch Atomic Absorption Spectrom. Updates, 10 (3) (1995) 207-214. 

For each group, one representative sample matrix has to be used for method validation. If the intended use is restricted to one of the crop groups, the method must be validated only for this group. On the other hand, the method has to be validated for all groups if the use is intended for a variety of crops that belong to two or more different groups. In addition, specific crops which are difficult to analyze due to matrix interference require individual method validation (e.g., hops, brassica varieties, bulb vegetables, herbs, tea). 

In contrast to the requirements for enforcement methods and to ensure sufficient quality of the generated data, validation data should be submitted for all types of crop samples to be analyzed. However, matrix comparability and a reduced validation data set may be considered where two or more very similar matrices are to be analyzed (e.g., cereal grain). A reduced sample set may also be acceptable (two levels, at least three determinations and an assessment of matrix interference) provided that the investigated samples belong to the same crop group as described in SANCO/825/00 (see also Section 4.2.1). 

These two terms (IDL and IQL) define only the limitations of the instmment. When analyzing real-life samples such as plant or animal tissue or even soil and ground water samples, matrix interference must be taken into consideration in order to define detection limits. This is because these real-life matrices are made up of hundreds (or even thousands) of compounds. These compounds may interfere in several ways in the detection and quantification of the analyte of interest. 

The value of Cl determined here is measured in terms of concentration and solution. This value does not take matrix interferences into account since RMSE was determined from calibration standards. Therefore, this value should be reported as the IDL. This value provides a good starting point for the next step, which is calculating the MDL. 

Although we speak generally of validated methods , only the performance of a method applied to a particular range of materials (matrices) is reported. The possibility of matrix interferences or the efficiency of cleanup steps may vary with matrix type. For that reason, methods should be validated in all matrix types, which differ significantly. In the context of the validation of enforcement methods by applicants, significant difference is not a well defined term. To avoid any dispute about completeness of validation, five material types had been selected for crops, which usually... 

At least one control water sample must be analyzed concurrently with the water samples to determine the presence of matrix interferences and/or background levels of the metabolites. Optima-grade bottled water is used as the matrix for the controls and the fortified samples for all wafer fypes, because obtaining ground and surface water specimens that are completely free of the metabolites is difficult. Our analyses of ground and surface waters have demonstrated the presence of low-level interferences in these matrices. Interferences from other pesticides are unknown, because none have been examined. However, none are expected due to the high level of specificity of the LC/MS/MS analysis. 

Two steps have been added to the original Pesticide Analytical Manual method to increase the stability of the trimethylsilyl derivatives and to clean up the final extract prior to GC analysis, namely the use of a sodium sulfate mini-column to dry the extract after derivatization and the use of a Florisil Sep-Pak cartridge to remove matrix interferences. The advantages of the current method are the simultaneous evaluation of the four analytes, reproducibility and low matrix interference. 

Sample preparation techniques vary depending on the analyte and the matrix. An advantage of immunoassays is that less sample preparation is often needed prior to analysis. Because the ELISA is conducted in an aqueous system, aqueous samples such as groundwater may be analyzed directly in the immunoassay or following dilution in a buffer solution. For soil, plant material or complex water samples (e.g., sewage effluent), the analyte must be extracted from the matrix. The extraction method must meet performance criteria such as recovery, reproducibility and ruggedness, and ultimately the analyte must be in a solution that is aqueous or in a water-miscible solvent. For chemical analytes such as pesticides, a simple extraction with methanol may be suitable. At the other extreme, multiple extractions, column cleanup and finally solvent exchange may be necessary to extract the analyte into a solution that is free of matrix interference. 

The analytical response generated by an immunoassay is caused by the interaction of the analyte with the antibody. Although immunoassays have greater specificity than many other analytical procedures, they are also subject to significant interference problems. Interference is defined as any alteration in the assay signal different from the signal produced by the assay under standard conditions. Specific (cross-reactivity) and nonspecific (matrix) interferences may be major sources of immunoassay error and should be controlled to the greatest extent possible. Because of their different impacts on analyses, different approaches to minimize matrix effects and antibody cross-reactivity will be discussed separately. 

A monoclonal antibody-based ELISA has been utilized to determine ceftiofur levels in milk. The authors noted that matrix interference occurred, but a 1 100 dilution lowered the interference, and a 1 1000 dilution eliminated the matrix interference. Because of the high dilution, samples could not be measured below l.Opgkg The assay measured ceftiofur, its major metabolite desfuroylceftiofur, and ceftiofur protein conjugates and has been utilized to measure residues in milk from cows treated with therapeutic doses of the drug. The results from the incurred residue correlated well with a previous study using radiolabeled ceftiofur, confirming the detection of a metabolite that was not detected by HPLC. 

Trace analysis of soil samples often requires post-extraction cleanup to remove coextracted matrix interferences. There are several difficulties that may arise during chromatographic analysis due to interferences present in sample extracts. To avoid these and other issues, one or more of the following cleanup techniques are often used. 

Bisphosphonic acids and their salts are analogs of pyrophosphate where the P-O-P linkage of the latter has been replaced by P-C-P. Bisphosphonates are known to inhibit bone resorption, and have attracted much attention as potential therapeutic agents. Bisphosphonates do not absorb or fluoresce, and sample matrix interferences can make detection difficult, especially in biological samples. Successful applications of IEC to bisphosphonate analysis have been described.173174... 

High selectivity (i.e. the ability to separate analytes from matrix interferences) is one of the most powerful aspects of SPE. This highly selective nature of SPE is based on the extraction sorbent chemistry, on the great variety of possible sorbent/solvent combinations to effect highly selective extractions (more limited in LLE where immiscible liquids are needed) and on the choice of SPE operating modes. Consequently, SPE solves many of the most demanding sample preparation problems. 

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