Quantification linearity

Limit of Detection Identification Quantification Linear Range Range... 

The features to be determined to validate a procedure are repeatability (within-run) and intermediate (between-run) precision, limit of detection, limit of quantification, linearity of the response of the assay, specificity of the assay and whether there are any interferences, its trueness, the vmcertainty associated with an individual result with a stated limit of confidence, and appropriate reference ranges. While examining these topics further information relevant to reagent stability, necessary frequency of recalibration, suitable IQC protocol, and overall assay weakness will be obtained. 

Validation of the HPLC method was performed. The external standard method was used to calibrate the chromatographic system for protein quantification. Linearity between the concentration of casein fraction, P-lactoglo-bulin, and a-lactalbumin in bovine and the UV absorbance at 280 nm was maintained over the concentration ranges of 0.001-0.2, 0.004-0.6, and 0.002-0.2 g/100 ml, respectively. Repeatability of the injection was performed by 10 consecutive injections of bovine, ovine, and human milks. The relative standard deviation (RSD) values for peak areas were all below 2.3%. Recovery studies were carried... 

Limit of Detection identification Quantification Linear Range Range (LOD) (LOi) (LOQ) (LOLR) (LOR)... 

In Raman spectroscopy the intensity of scattered radiation depends not only on the polarizability and concentration of the analyte molecules, but also on the optical properties of the sample and the adjustment of the instrument. Absolute Raman intensities are not, therefore, inherently a very accurate measure of concentration. These intensities are, of course, useful for quantification under well-defined experimental conditions and for well characterized samples otherwise relative intensities should be used instead. Raman bands of the major component, the solvent, or another component of known concentration can be used as internal standards. For isotropic phases, intensity ratios of Raman bands of the analyte and the reference compound depend linearly on the concentration ratio over a wide concentration range and are, therefore, very well-suited for quantification. Changes of temperature and the refractive index of the sample can, however, influence Raman intensities, and the band positions can be shifted by different solvation at higher concentrations or... 

To quantify the concentration of a colorant, one must consider that linearity between the colorant concentration and the fluorescence emission intensity exists only at very low concentrations. The reason for deviation from linearity may be reabsorption of the emission light by other fluorophores or formation of dimers. If no extraction and controlled dilution of the fluorescent colorant are performed, the colorant quantification will be only qualitative. 

The accuracy and precision of carotenoid quantification by HPLC depend on the standard purity and measurement of the peak areas thus quantification of overlapping peaks can cause high variation of peak areas. In addition, preparation and dilution of standard and sample solutions are among the main causes of error in quantitative analysis. For example, the absorbance levels at of lutein in concentrations up to 10 mM have a linear relationship between concentration and absorbance in hexane and MeOH on the other hand, the absorbance of P-carotene in hexane increased linearly with increasing concentration, whereas in MeOH, its absorbance increased linearly up to 5 mM but non-linearly at increasingly higher concentrations. In other words, when a stock solution of carotenoids is prepared, care should be taken to ensure that the compounds are fuUy soluble at the desired concentrations in a particular solvent. 

The adsorption TLC operating in the linear range of the adsorption isotherm (sometimes dubbed as the liuear adsorptiou TLC or simply as the linear TLC) is utilized for purely analytical purposes (which iuclude establishing of a qualitative composition of a given mixmre of analytes, often followed by their quantification in the examined sample with aid of the calibration plot approach). In order to introduce certain amount of rationale to the linear adsorption TLC (and enable... 

Linear, exponential, or quadratic calibration curves may be used to quantitate the amount of analyte in each sample. Quantification of each analyte is made independently. 

Famoxadone, IN-JS940, and IN-KZ007 residues are measured in soil (p-g kg ), sediment (p-gkg ), and water (pgL ). Quantification is based on analyte response in calibration standards and sample extract analyses determined as pg mL Calibration standard runs are analyzed before and after every 1 samples in each analytical set. Analyte quantification is based on (1) linear regression analysis of (y-axis) analyte concentration (lagmL Q and (x-axis) analyte peak area response or (2) the average response factor determined from the appropriate calibration standards. The SLOPE and INTERCEPT functions of Microsoft Excel are used to determine slope and intercept. The AVERAGE and STDEV functions of Microsoft Excel are used to determine average response factors and standard deviations. 

Gas chromatography is commonly used to analyse mixtures for quantification. A wide variety of special detectors with adequate linear response ranges are available for quantification of various classes of compounds (cf. Table 4.14). Quantification by direct injection may be used to determine additives, residual monomers and solvents in product formulations, coated films, and solid materials [109]. On the other hand, reliable quantification by means of solid-injection PTV-GC, HS-GC and PyGC techniques is not always trivial. 

Also, if conversion of drug to active metabolite shows significant departure from linear pharmacokinetics, it is possible that small differences in the rate of absorption of the parent drug (even within the 80-125% range for log transformed data) could result in clinically significant differences in the concentration/ time profiles for the active metabolite. When reliable data indicate that this situation may exist, a requirement of quantification of active metabolites in a bioequivalency study would seem to be fully justified. 

A calibration procedure has to be validated with regard to general and specific requirements under which the calibration model has been developed. For this purpose, it is important to test whether the conditions represented in Fig. 6.6 are fulfilled. On the other hand, it is to assure by experimental studies that certain performance features (accuracy, precision, sensitivity, selectivity, specificity, linearity, working range, limits of detection and of quantification, robustness, and ruggedness, see Chap. 7) fulfil the expected requirements. 

MWNTs favored the detection of insecticide from 1.5 to 80 nM with a detection limit of InM at an inhibition of 10% (Fig. 2.7). Bucur et al. [58] employed two kinds of AChE, wild type Drosophila melanogaster and a mutant E69W, for the pesticide detection using flow injection analysis. Mutant AChE showed lower detection limit (1 X 10-7 M) than the wild type (1 X 10 6 M) for omethoate. An amperometric FIA biosensor was reported by immobilizing OPH on aminopropyl control pore glass beads [27], The amperometric response of the biosensor was linear up to 120 and 140 pM for paraoxon and methyl-parathion, respectively, with a detection limit of 20 nM (for both the pesticides). Neufeld et al. [59] reported a sensitive, rapid, small, and inexpensive amperometric microflow injection electrochemical biosensor for the identification and quantification of dimethyl 2,2 -dichlorovinyl phosphate (DDVP) on the spot. The electrochemical cell was made up of a screen-printed electrode covered with an enzymatic membrane and combined with a flow cell and computer-controlled potentiostat. Potassium hexacyanoferrate (III) was used as mediator to generate very sharp, rapid, and reproducible electric signals. Other reports on pesticide biosensors could be found in review [17],... 

A linear calibration curve for carvedilol in plasma was constructed over a range of 1 to 80 ng/mL. The correlation coefficient exceeded 0.999. Intra-day and inter-day coefficients of variation were 1.93 and 1.88%, respectively. The average carvedilol recovery was 98.1%. The limit of quantification was 1 ng/mL. This high-throughput method enabled the analysis of more than 600 plasma samples without significant loss of column efficiency. 

A linear calibration curve for epirubicin ranged from 0.50 to 100.0 ng/mL with a correlation coefficient of 0.999. Intra-day and inter-day coefficients of variation were less than 5.2 and 11.7%, respectively. Limit of detection and limit of quantification were 0.1 and 0.5 ng/mL, respectively. The extraction recoveries ranged from 89.4 to 101.2%. The validated method was successfully applied to the routine analysis of plasma samples from patients treated with epirubicin. 

GC is coupled with many detectors for the analysis of pesticides in wastewater. At the present time the most popular is GC-MS, which will be discussed in more detail later in this section. The flame ionization detector (FID) is another nonselective detector that identifies compounds containing carbon but does not give specific information on chemical structure (but is often used for quantification because of the linear response and sensitivity). Other detectors are specific and only detect certain species or groups of pesticides. They include electron capture,nitrogen-phosphorus, thermionic specific, and flame photometric detectors. The electron capture detector (ECD) is very sensitive to chlorinated organic pesticides, such as the organochlorine compounds (OCs, DDT, dieldrin, etc.). It has a long history of use in many environmental methods,... 

Figure 16.4. Quantification of A9-tetrahydrocannabinol (9THC) after TFA and 11-nor-9-carboxy-A9-tetrahydrocannabinol (THCCOOH) after PFP in blood using GC-MS/NCI. Drug-free blood was spiked with 9THC (a) and THCCOOH (b) to the concentrations of 0, 5, 10, and 50 or 20ng/mL and with ISs-9THC-D3 and THCCOOH-D3 to 20ng/mL. Monitoring ions were (m/z) 410.3 for 9THC and 572.3 for THCCOOH. The values of validation parameters, expressed in ng/mL, were LOD, 0.25 LOQ, 0.5 limit of linearity, 0.5 to 100 for both analytes [2].

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