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27 views17 pages

5 Validasyon

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pixeldenresme
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© © All Rights Reserved
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Calibration curve, linearity, measuring range

Linearity is expressed as the relationship between the concentration of


the analyte in the sample and the instrument signal in a given area
of the method. The graph shows the y-axis instrument signal and the x-
axis the corresponding concentration.
Linearity is determined using 3-6 replicates of 5 or more standards with
concentrations of 70%-120% of the expected concentration range.
The equation of a linear calibration graph is expressed as:
y= bx + a
b = slope of the line
a = y-axis intercept point
• To determine the fitness of experimental points to a linear
line the correlation coefficient, r, is calculated.

• A r value of +1 indicates perfect positive correlation, that is,


the slope of all points +
• is on a line, -1 is similarly perfect negative
• It shows that all points are on a line with a slope - correlation.
• A zero for r indicates that there is no correlation between x
and y. in analytical studies
• Generally, it is desired that the r value be 0.99 and above.
küçük
karder yöntem!
en
!l Yapmak
zortagen
• Example
• The standard aqueous solution of paracetamol was measured by UV-Vis
absorbance spectrometer and its absorbances were found as follows.

• Absorbance : 2,1 5,0 9,0 12,6 17,3 21,0 24,7


• Conc, mg/L : 0 2 4 6 8 10 12

Find the correlation coefficient, r.


Solution:
• Regression line of x versus y

• Assuming all errors come from the y-


values, passing through the calibration
points
• The best fit line equation is calculated
using the least squares method. Our
goal is to find the right equation to
minimize the deviations in the y-line
between the experimental data and the
line found.These deviations are known
as y residues.
• Since some of the deviations will be
negative and some will be positive, to
minimize the sum of squares of the
residuals is the basis of the method.
y= bx + a
Slope of the calibration curve (b) Intercept of the calibration curve (a)

Example :
• The standard aqueous solution of paracetamol was measured
by UV-Vis absorbance spectrometer and its absorbances were
found as follows.

• Absorbance : 2,1
-Dublaer
Y 5,0 9,0 12,6 17,3 21,0 24,7
X
• Conc, mg/L : 0 2 4 6 8 10 12

• Find the regression equation.


Error in the slope (Sb) and

Error in the intercept (Sa) of the regression line:

Random errors in the y direction (Sy/x) is calculated as follows.

Here, y^ values are the appropriate y values


found for each x value in the calculated regression
equation. The value of the degrees of freedom in
linear regression calculations is (n-2).
Cale ben
Example :
bu form!l Kağıdı.
Vereceden Sharda algı Vern!gen
and

hesaplar!n latm
Working Range – Linear Range

The range of minimum and maximum concentrations of the analyte in the sample at
which the method achieves an acceptable level of precision, accuracy and linearity.

It is usually determined in linearity studies.

The minimum range should be 80-120% of the test concentration for raw material and
finished product quantification; 70-130% of the test concentration for content
quantification; ±20% of the range specified in the specifications for the dissolution test
method; 120% of the impurity specification for impurity quantification.

- The lowest value of the working range is the detection limit and or measurement
limit.

- The maximum value of the working range depends on the response range of the
instrument or method.

- The working range of the method must be definable by a linear relationship.


Method 3

This method calculates the signal to noise ratio (S/N) of the lowest concentration 'spike
sample' prepared for this purpose. The 'spike concentration' used (Cspk) is divided by the
S/N ratio and 3 times the result is calculated as the LOD value.

Sensitivity:
The ability of the method or instrument to detect different concentrations.

Where the response is linear with concentration, the sensitivity of the method is
equal to the slope of the calibration line.

The greater the slope, the greater the sensitivity.

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