Iso8466-1 - 1990 - Calibracion Lineal
Iso8466-1 - 1990 - Calibracion Lineal
STANDARD 8466-1
First edition
1990-03-01
Part 1:
Statistical evaluation of the linear calibration function
Reference number
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ISO 8466-1 : 1990 (EI
Contents Page
.. .
Foreword ............................................................ Ill
1 Scope .............................................................. 1
2 Definitions.. ........................................................ 1
3 Symbols.........................................................~ .. 2
4 Performance ........................................................ 3
5 Example ............................................................ 6
0 ISO 1990
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ISO 8466-1 : 1990 (E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of
national Standards bodies (ISO member bedies). The work of preparing International
Standards is normally carried out through ISO technical committees. Esch member
body interested in a subject for which a technical committee has been established has
the right to be represented on that committee. International organizations, govern-
mental and non-governmental, in liaison with ISO, also take patt in the work. ISO
collaborates closely with the International Electrotechnical Commission (IEC) on all
matters of electrotechnical standardization.
International Standard ISO 8466-1 was prepared by Technical Committee ISO/TC 147,
Water quaiity.
ISO 8466 consists of the following Parts, under the general title Water gua/ity -
Calibration and evaluation of analytical methods and estimation of Performance
charac teris tics :
Part 1:
Statistical evaluation of the linear calibration function
1 Scope
1 Originai Sample 1
This part of ISO 8466 describes the Steps to be taken in
evaluating the statistical characteristics of the linear calibration
_ , MeasUri!g ;X~~nstructi;
function. lt is applicable to methods requiring a calibration. Fur-
ther Parts of ttis lnternationa~ Standard will cover the deter-
mina$.i!! ok IGr%t of detection and FEmit of determination, the
effect of Ln$erferences and other performante chara,cteristics
1
method and for the calculation of Performance characteristics measuring instructions
of the calibration function.
r-YG&q
calibrating and evaluating instructions
2 Definitions
For the purposes of this part of ISO 8466, the following defini-
tions apply.
2.2 calibrating instruction : Describes the approach to 2.4 measured values : The concentration-dependent initial
determine the calibration function from information values, yi, values (e.g. extinction) of a measuring System.
obtained by measuring given Standard concentrations, xi. The
slope of the calibration function, b, as a measure of sensitivity NOTE - Information value and measured volume are synonymous.
of the analytical method and the Standard deviation of the
method, sXO, are figures of merit and characteristics which 25 residual Standard deviation, s : The residual Standard
result from the calibration experiment. deviation describes the scatter of the information values about
the calculated regression line. lt is a figure of merit, describing
The Standard deviation, sXO, allows the comparison of indepen- the precision of the calibration.
dent analytical methods.
For the purpose of this Standard, the Standard deviation of the
For the user of the method, these characteristics present method means the Standard of deviation of the calibration pro-
criteria for the internal laboratory quality control. cedure.
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ISO 8466-1 : 1990 (EI
2.6 Standard deviation of the method sXO: The ratio of Yi Information value of the Standard concentra-
the residual Standard deviation, s,,, to the sensitivity of the tion Xi calculated from the calibration function.
calibration function, b. lt is a figure of merit for the perfor-
mance of the analytical method, and is valid within the working s2
i
Variante of the information values for the
range (sec equation 13). analyses of Standard samples, having the con-
centration Xi.
For the purpose of this Standard, the Standard deviation of the
method means the Standard of deviation of the calibration pro-
fi Degrees of freedom for the calculation of the
cedure. variance (f i = tli- 1).
2.7 coefficient of Variation of the method, VXO: The ratio a Calculated blank (Ordinate intercept of the cali-
of the Standard deviation of the method sXOto the appertaining bration straight line).
mean, F, which is the centre of the working range.
See also note to 2.5 and 2.6. b Sensitivity of the method (slope of the cali-
bration line; coefficient of regression
2.8 working range (of an analytical method): The interval,
being experimentally established and statistically proved by the x Mean of the Standard concentrations
resulting from the calibration experiment.
calibration of the method, between the lowest and highest
quantity or mass concentration. The lowest possible limit of a
working range is the limit of detection of an analytical method. Y Mean of the information values Yit resulting
from the calibration experiment.
2.9 homogeneity of variances: Homogeneity of variances
Residual Standard deviation.
of pooled data, such as those resulting from replicate analyses sY
at different levels, is confirmed if these variances are not
Residual Standard deviation obtained by linear
significantly correlated to their appertaining concentrations. sY1
regression calcula tion.
2.10 sensitivity of the analytical method: The slope of
sY2 Residual Standard deviation obtained by non-
the calibration function of the complete analytical method, in-
linear regression calculation.
clusive of all procedural Steps, within the working range in
question.
DS2 Differente of variances.
values 7.
N Number of concentration levels (for this part of
ISO 8466, N = IO). tV;, l-a) Tabled value of the t-distribution with fl =
N - 2 degrees of freedom and a confidence
Xl Concen tration of the Standard Sample at the lower level of (1 - a) (f-factor of Student’s distribu-
level of the working range ( 1st Standard Sample). tion).
% Number of replicates per level Xi. VB (x, Confidence interval for the concentration .?.
Mean of the information values Yi,j of Standard Confidence interval of the mean Z?of the con-
samples, having the concentration Xi. centration.
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ISO 8466-1 :, 1990 IE)
4.1 Choice of working range Both data sets of the concentrations x1 and xlo are used t~
calculate the variances sf and s$, as given in equation (1) :
Esch calibration experiment is started with the choice of a 10
preliminary working rangeL3].
t: (Y i t j - El*
j=l
The working range depends on S*i = . . . (11
Ili -1
a) the practice-related objective of the calibration.
with the mean
The working range shall cover, as far as possible, the 10
application range for water, waste water, and sludge analy-
sis. The most frequently expected Sample concentration c Y-‘,J *
.- j=l
should lie in the centre of the working range. Yj = fori = 1 ori = 10 . . . (2)
yli
b) feasibilities of technical realizability.
The variances are tested (F-test) for significant differentes at
The measured values obtained must be linearly correlated to the limits of the working rangeL5t 6].
the concentrations. This requires that the measured values
obtained near the lower limit of the working range tan be The test value PG is determined for the F-test from
distinguished from the blanks of the method. The lower equation (3).
limit of the working range should therefore be equal to or
greater than the limit of detection of the method. Dilution
and concentrating Steps should be feasible without the risk PG = 2 forsyo > sf . . . (3)
of bias. 1
range. In Order to check for the homogeneity of the variances, variances sf and s$ is significant.
ten replicates of each of the lowest and the highest concentra-
tions (x, and x,~) of the working range are determined. Ten in- If the differente between the variances is significant, the
formation values, )pi jl result from these series of measurements preliminary working range should be made smaller until the dif-
(see table 1). ’ ference between the variances is found to be random only.
4.13 Test for linearity[*J ‘# *] The measurement against a blank is not allowed, since thereby
valuable information on the magnitude of the blank will be lost.
The easiest test for the linearity is the graphical representation The comparison medium for zeroing the instrument is always, if
of the calibration data with the calculated regression line. Any possible, a pure solvent (e.g. pure water).
unlinearity is evident (see figure 2).
051 I
z-
+E
8
04J
--y-----------l-----
1
g:ppIIj
3
03 J
-- ---- --L------
4 -t- , t ----t---l----l
5 .-----j----~----~--q-----j
\ , , , ., , ; . ,)c
~--
6
7
.--~~ --L-----
1 t------t-m
I
I
sv, which is a measure of the scatter of the information values Fromsthe law of error propagation it follows that, for each value
about the calibration line and is given by equation (9). X, a confidence interval for the true value y exists whose
limiting Points are on two hyperbolic paths bracketing the
calibration line. Between these paths the true calibration func-
N
tion tan be expected with a significance level of a (fl : N - 2,
[Y j - (a + bx,)l*
c i=l
confidence level = 1 - a), determined by Student’s t-factor.
(9)
N-2 The confidence intervals for analytical results, calculated from
the calibration function, are given by the intersections with the
respective hyperbolic paths in figure 3. The estimation of the
4.3 Assessment
confidence intervals are given by equation (12) [71
The concentration of an analyzed Sample is obtained
7,,* = % + VB (.%,
a) from the measured valuey, to give ?
.a
Y -a Y -a
x =- . . . (IO) .2,,2 = b
b
or
As to the uncertainty of an analytical result, keep in mind that Equation (12) indicates that the confidence interval VB(?)
the analytical error is a combination of the uncertainty of the brackets the true analytical value with a range governed by the
determination of the measured value, and the uncertainty of statistical security of Student’s distribution. The magnitude of
the estimation of the regression coeff icients[*! VB(g) is mainly determined by the number of replicates n^ and
0
7
0 I
I
I
I
/ 1 I
a /’ I I
- vl3m
I I
I I
I
I
- 4-
/ 4 - X 10
Working range
Figure 3 - Working range x, to xlo, calibration line with confidence band and a Single analytical result with its
appertaining confidence interval
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ISO 8466-1 : 1990 (E)
their results, the mean of the information values 7, as weil as 5.1 Choice of working range
the characteristics of the method, the residual Standard devi-
ation s,,, and the sensitivity b. For the analysis of drinking water and surface water, a working
range of 0,05 mg to 0,5 mg (NO;)/1 is appropriate.
The quality of the analytical procedure increases therefore with
increasing sensitivity and decreasing residual Standard deviation. 5.1.1 Testing the hornogeneity sf the variancesl)
The Standard deviation of the method sXO[see equation (1311 is
the characteristic which allows the analyst to check the quality According to the approach outlined in 4.1 .l, the variances ~2
of his own work. of the information values obtained from the Standard concen:
trations at the lower or upper limit of the working range respec-
sXO =2 . . . (13) tively, were determined (sec table 4). ’
b
The test value PG for the F-test is calculated from equation (3)
For the comparison of different standardized analytical
methods, the coefficient of Variation of the method, expressed S2 13,54 x IO-6
as a percentage, is given by equation (14) = 2,9
PG=s= 1 4,67 x 10-6
The photometric determination of nitrite is used to demonstrate The comparison of the calculated value PC with the tabled one
the calibration and the subsequent estimation of the statistical indicates a random differente between the variances under
characteristics of the method and their influence on the final examination. As the variances are homogeneous a simple
results of the evaluation. regression analysis may be petformed.
i xi
mgll Y*l,l Yi,2 Yi,3 Y-1,4 Y-lt5 ’ Yfj
i, Y-1,7 i, R,a , &,!3 j Yi, 10
1 0,05 0,140 0,143 0,143 0,146 0,144 0,145 0,344 ' 0,146 0,145 0,148
2 0,lO 0,281
1
3 0,15 0,405 j
I
4 0,20 0,535 /
I
5 0,25 0,662 )
I
6 0,30 0,789 1
I
7 0,35 0,916 1
1
8 0,40 1,058
9 0,45 1,173
10 =N 0,50 1,303 1,302 1,300 1,304 1,300 1,296 1,295 1,301 1,296 1,306
1) Forthe sake oftransparency, alldimensions have intentionally been omitted in allequations, withoutambiguity. The dimensions are finallyadded
to the result.
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5.1.2 Testing linearity The Standard deviation of the method is calculated from
equation ( 13)
A non-linear regression function may be derived [*l as
The residual Standard deviations of the linear and the non-linear The coefficient of Variation of the method, expressed as a
calibration function, sY, and su2, are compared: percentage, is given by equation (14)
= 0,005 2 mg/1 (see 5.2 for calculation procedure) Sxo 0,002o x 100
sYl V xo = x x loo= = 0,73
0,275
sY*
= 0,005 2 mg/l.
As both residual Standard deviations are equal, the differente 5.3 Evaluation
of the variances DS* [see equation (411 does not need to be
5.3.1 Single determination
calculated. The non-linear calibration function does not lead to
a significantly better adjustment, e.g. the calibration function is The analysis of an unknown Sample, performed in the same
linear. way as the analysis of the Standards, gave an information value
jk 0,641 (extinction). The analytical result was obtained using
5.2 Calibration and characteristics of the method equation (12) with a confidence interva195 %, t(8; 0,951 = 2,31
Since the prerequisites for the Performance of a simple linear 0,641 - 0,018
regression are fulfilled, the calibration function and the %,2 = +
2,575
characteristics of the method tan be calculated using equations
(71, (8) and (13). The results are shown in table 5.
(0,641 - 0,726 2)*
+ 0,002 0 x 2,31 - + - +
The slope, b, as a measure for the sensitivity, is calculated from (2,575)* x 0,206 25
equation (7)
= (0,242 + 0,005) mg/1
N
Thus the true value of the concentration tan be expected
(Xi - 33 ’ (yi - jd
c within the range 0,237 < x < 0,247 mg/l, with a confidence
i=l
b = = 2,575 level of 0,95.
N
(Y i - yi)*
- +
(0,635 - 0,726 2)*
(2,575)* x 0,206 25
sy = = 0,005 2 [Ext.l
N-2 = (0,240 + 0,003) mg/l
The equation for the straight line is given by equation (4) Thus the true value of the concentration tan be expected
within the range 0,237 < x < 0,243 mg/I, with a confidence
Y = 0,018 + 2,575 2 x level of 0,95.
1 1 1 $i 1 ,66, i
mg0
xi
dV*1
4 I 0,20 1 0,535
I 6
8
0,30
0,35
0,40
0,789
0,916
1,058
-
T--t--
--
r / 10
9
c I 2f75l
i=l I
0,45
0,50
I
1,173
1,303
7,262
I
l
Annex A
(informative)
Bibliography
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