Calibration plot

Normal calibration plot of hypothetical data from Table 5.1. 

To circumvent this need for calibration as well as to better understand the separation process itself, considerable effort has been directed toward developing the theoretical basis for the separation of molecules in terms of their size. Although partially successful, there are enough complications in the theoretical approach that calibration is still the safest procedure. If a calibration plot such as Fig. 9.14 is available and a detector output indicates a polymer emerging from the column at a particular value of Vj, then the molecular weight of that polymer is readily determined from the calibration, as indicated in Fig. 9.14. 

The worked out soi ption-photometric method of NIS determination calls preliminary sorption concentration of NIS microamounts from aqueous solutions on silica L5/40. The concentrate obtained is put in a solution with precise concentration of bromthymol-blue (BTB) anionic dye and BaCl, excess. As a result the ionic associate 1 1 is formed and is kept comparatively strongly on a surface. The BTB excess remains in an aqueous phase and it is easy to determinate it photometrically. The linear dependence of optical density of BTB solutions after soi ption on NIS concentration in an interval ITO - 2,5T0 M is observed. The indirect way of the given method is caused by the fact the calibration plot does not come from a zero point of coordinates, and NIS zero concentration corresponds to initial BTB concentration in a solution. 

The following polyvitamin prepai ations were analyzed Kal tsid (OAO Comfort Plus , Russia), Asvitol (OAO INC Marbiofarm , Russia), Pikovit (KRKA, d.d. The New Place, Slovenia), Yeast with vitamin C (000 EKKO Plus , Russia). Chromatographic experiment has been carried out using Silufol UV-254 (Kavalier, Czech Republic) and acetone - ethyl acetate - acetic acid - ethanol (3 5 1 1) - CTAB (2T0 M) as a mobile phase mixture. The linearity calibration plot, built in coordinate S = f (IgqAC), is valid in the interval 5-25 p.g. Correctness of the determination has been checked by photometry. The obtained results for the ascorbic acid determination are presented below. 

Eigure 3.3 shows the calibration plots for Zorbax PSM columns. (The calibration plots for silanized and unsilanized columns are comparable.) These calibration plots allow the chromatographer to select the appropriate columns for samples. Eor example, the Zorbax PSM 60 column provides resolution of... 

FIGURE 3.4 Calibration plots for Zorbax PSM Bimodal and Trimodal columns. 

Gels made in this way have virtually no usable porosity and are called Jordi solid bead packings. They can be used in the production of low surface area reverse phase packings for fast protein analysis and in the manufacture of hydrodynamic volume columns as well as solid supports for solid-phase syntheses reactions. An example of a hydrodynamic volume column separation is shown in Fig. 13.2 and its calibration plot is shown in Fig. 13.3. The major advantage of this type of column is its ability to resolve very high molecular weight polymer samples successfully. 

Figures 13.8 and 13.9 show the separation of polystyrene standards using a typical mixed-bed column and its calibration plot, respectively. The major advantages of using a large i.d. 10-mm column are low hack pressure and relatively short run times. As seen in Fig. 13.8,10 standards from toluene thru 8.4 X 10 MW can be resolved in a mere 21 min. Because of the large 10-mm i.d. columns, 1.5-ml/min flow rates give a linear velocity equivalent to that of only 0.9 ml/min using a 7.6-mm i.d. column. Also, the gel volume contained in one 10 mm i.d. X 500 mm column is 39.3 ml, whereas a 7.6 mm i.d. X 300 mm column contains only 13.6 ml of gel volume. This bulk volume factor, combined with the large pore volumes of gels, obtains essentially the same resolution as that obtained on three standard 7.6 X 300-mm columns in series, but in about one-half the usual time required using the smaller columns.

If the determination is a routine procedure, then a calibration plot will be available and it will be a simple matter to ascertain the concentration of the test solution in this case it is unnecessary to prepare a standard solution of the compound being determined. If a calibration plot is not available, then a series of solutions containing say 5.0, 7.5, 10.0 and 15.0mL of the standard solution are diluted to, say, 100 mL in graduated flasks. The absorbance of each of these solutions is measured and the results plotted against concentration. 

Procedure. To 100 mL of the neutral sample solution (containing not more than 0.4 mg nitrite) add 2.0 mL of solution A and, after 5 minutes, 2.0 mL of solution B. The pH at this point should be about 1.5. Measure the absorbance after 10 minutes in the wavelength region of 550 nm (yellow-green filter), in a spectrophotometer against a blank solution prepared in the same manner. Calculate the concentration of the nitrite from a calibration plot prepared from a series of standard nitrite solutions. 

Now record the second derivative spectrum of the Actifed solution, determine the long-wave peak heights for both components, and by comparison with the calibration plots of the individual components, deduce their proportions in the tablets. 

Fig. 19.11 Calibration plot for internal standard procedure for benzoic acid.

Use the adsorption theory (of Section 2-1-3) to explain why adsorptive stripping voltammetry results in nonlinear calibration plots. 

The measured potential is thus a linear function of pH an extremely wide (10-14 decades) linear range is obtained, with calibration plots yielding a slope of 59 mV per pH unit. The overall mechanism of the response is complex. The selective response is attributed to the ion-exchange properties of the glass surface, and in particular the replacement of sodium ions associated with the silicate groups in the glass by protons ... 

How would you extend the linear range of calibration plots based on the use of enzyme electrodes ... 

Figure L Calibration plot for nitrosation potential showing amount of t -nitroso-thiomorpholine formed vs, the square of the nitrogen dioxide concentration. Standard conditions of 50%relative humidity, 25°C, and 1 L/min flow rate for 30 min...

Figure 3 shows calibration plots of log (particle diameter) vs. elution voliame difference (AV) between marker and particle using three different monodisperse latexes at a low eluant ionic strength of 1.29 mM SLS. These results illustrate the featiire of universal calibration behavior predicted by the capillary bed model as mentioned earlier. Of note also is the fact that the curve deviates from linearity for the 38 nm particle and begins to approach the origin as also indicated by the model calculations. 

Upon calibrating the columns with narrow MWD polystyrene standards, it was noted that the polydichlorophosphazenes eluted over a relatively straight region of the calibration plot. Therefore a two parameter equation. 

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... 

Quantitation was performed in all cases using the external calibration method. A series of standards were injected and the responses plotted against their known concentrations. Peak responses in samples were compared with the calibration plots to obtain the amount found (nanograms). A fresh calibration plot was generated with each analytical set of samples. 

FIG. 14 Time-course changes of sensitivity (slope of calibration plots) for Na -ISFETs based on sol-gel-derived membranes incorporating bis(12-crown-4) by covalent bonding using (11) ( ) and by encapsulation of bis[(12-crown-4-)ylmethyl] 2-dodecyl-2-methylmalonate (O)- (From Ref 49.)... 

Applicability in biological ion assay is an important factor for biocompatible potentio-metric ion sensors. Attempts were made to determine Na" " concentrations in human blood sera by using silicone-rubber membrane Na+-ISFETs based on (5) [Fig. 17(a)] [29]. The found values for Na concentration in undiluted, 10-fold diluted, and 100-fold diluted serum samples are in good agreement with the Na" " calibration plots. Even in the undiluted serum samples, only a slight potential shift was observed from the calibration. This indicates that the calixarene-based silicone-rubber-membrane Na+-ISFETs are reliable on serum Na assay. For comparison with the silicone-rubber membrane, Na -ISFETs with corresponding plasticized-PVC membrane containing (2) or (5) were also tested for the Na assay. The found values of Na" " concentration... 

In this case, the output from a group of adjacent diodes is electronically suauMd or averaged to produce one signal output. Practically, this is accomplished by defining a central wavelength for detection and an associated effective bandwidth. These parameters, however, must be set carefully to avoid nonlinearity in the calibration plots [80]. 

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