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Calibration

The document describes three common calibration techniques for analytical instrumentation: 1. The calibration curve method involves preparing standard solutions of a known analyte and measuring the analytical signal to generate a calibration curve. 2. The standard additions method is useful when the sample matrix is complex and no ideal blank is available. It involves adding increments of a standard to aliquots of the sample and measuring the signal. 3. The internal standard method adds a constant amount of a second species to standard and sample solutions. It corrects for variations in sample size and matrix by measuring the ratio of analyte and internal standard signals.

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155 views32 pages

Calibration

The document describes three common calibration techniques for analytical instrumentation: 1. The calibration curve method involves preparing standard solutions of a known analyte and measuring the analytical signal to generate a calibration curve. 2. The standard additions method is useful when the sample matrix is complex and no ideal blank is available. It involves adding increments of a standard to aliquots of the sample and measuring the signal. 3. The internal standard method adds a constant amount of a second species to standard and sample solutions. It corrects for variations in sample size and matrix by measuring the ratio of analyte and internal standard signals.

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joshjeth
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Calibration Techniques

1. Calibration Curve Method


2. Standard Additions Method 3. Internal Standard Method

Calibration Curve Method


1. Most convenient when a large number of similar samples are to be analyzed.
2. Most common technique.

3. Facilitates calculation of Figures of Merit.

Calibration Curve Procedure


1. Prepare a series of standard solutions (analyte solutions with known concentrations).
2. Plot [analyte] vs. Analytical Signal. 3. Use signal for unknown to find [analyte].

Example: Pb in Blood by GFAAS


[Pb] (ppb) 0.50 1.50 2.50 3.50 4.50 5.50 Signal (mAbs) 3.76 9.16 15.03 20.42 25.33 31.87
Results of linear regression: S = mC + b m = 5.56 mAbs/ppb b = 0.93 mAbs

35

30

y = 5.56x + 0.93

25

mAbs

20

15

10

0 0 1 2 3 4 5 6

Pb Concentration (ppb)

A sample containing an unknown amount of Pb gives a signal of 27.5 mAbs. Calculate the Pb concentration.
S = mC + b C = (S - b) / m C = (27.5 mAbs 0.92 mAbs) / 5.56 mAbs / ppb C = 4.78 ppb (3 significant figures)

Calculate the LOD for Pb


20 blank measurements gives an average signal 0.92 mAbs with a standard deviation of bl = 0.36 mAbs LOD = 3 bl/m = 3 x 0.36 mAbs / 5.56 mAbs/ppb LOD = 0.2 ppb (1 significant figure)

Find the LDR for Pb


Lower end = LOD = 0.2 ppb (include this point on the calibration curve) SLOD = 5.56 x 0.2 + 0.93 = 2.0 mAbs

(0.2 ppb , 2.0 mAbs)

Find the LDR for Pb

Upper end = collect points beyond the linear region and estimate the 95% point.
Suppose a standard containing 18.5 ppb gives rise to s signal of 98.52 mAbs This is approximately 5% below the expected value of 103.71 mAbs (18.50 ppb , 98.52 mAbs)

Find the LDR for Pb

LDR = 0.2 ppb to 18.50 ppb


or

LDR = log(18.5) log(0.2) = 1.97


2.0 orders of magnitude

or
2.0 decades

Find the Linearity


Calculate the slope of the log-log plot

log[Pb] -0.70 -0.30 0.18 0.40 0.54 0.65 0.74 1.27

log(S) 0.30 0.58 0.96 1.18 1.31 1.40 1.50 1.99

Not Linear??
2.50

y = 0.0865 x + 0.853
2.00

log(Signal)

1.50

1.00

0.50

0.00 -1.00

-0.50

0.00

0.50

1.00

1.50

log(Pb concentration)

Not Linear??
120

100

80

Signal (mAbs)

60

40

20

0 0 2 4 6 8 10 12 14 16 18 20

Pb Concentration (ppb)

Remember

S = mC + b
log(S) = log (mC + b)

b must be ZERO!!
log(S) = log(m) + log(C)

The original curve did not pass through the origin. We must subtract the blank signal from each point.

Corrected Data
[Pb] (ppb) 0.20 0.50 1.50 2.50 3.50 4.50 5.50 18.50 Signal (mAbs) 1.07 2.83 8.23 14.10 19.49 24.40 30.94 97.59

log[Pb] -0.70 -0.30 0.18 0.40 0.54 0.65 0.74 1.27

log(S) 0.03 0.45 0.92 1.15 1.29 1.39 1.49 1.99

Linear!
2.50

y = 0.9965x + 0.7419
2.00

log(signal)

1.50

1.00

0.50

0.00 -1.00

-0.50

0.00

0.50

1.00

1.50

log(Pb concentration)

Standard Addition Method


1. Most convenient when a small number of samples are to be analyzed.
2. Useful when the analyte is present in a complicated matrix and no ideal blank is available.

Standard Addition Procedure


1. Add one or more increments of a standard solution to sample aliquots of the same size. Each mixture is then diluted to the same volume.
2. Prepare a plot of Analytical Signal versus:
a) volume of standard solution added, or b) concentration of analyte added.

Standard Addition Procedure


3. The x-intercept of the standard addition plot corresponds to the amount of analyte that must have been present in the sample (after accounting for dilution).
4. The standard addition method assumes:
a) the curve is linear over the concentration range b) the y-intercept of a calibration curve would be 0

Example: Fe in Drinking Water


Sample Volume (mL) 10 10 10 10 10 Standard Volume (mL) Signal (V) 0 5 10 15 20 0.215 0.424 0.685 0.826 0.967
The concentration of the Fe standard solution is 11.1 ppm All solutions are diluted to a final volume of 50 mL

1.2

0.8

Signal (V)

0.6

-6.08 mL

0.4

0.2

0 -10 -5 -0.2 0 5 10 15 20 25

Volume of standard added (mL)

[Fe] = ?
x-intercept = -6.08 mL Therefore, 10 mL of sample diluted to 50 mL would give a signal equivalent to 6.08 mL of standard diluted to 50 mL.

Vsam x [Fe]sam = Vstd x [Fe]std

10.0 mL x [Fe] = 6.08 mL x 11.1 ppm


[Fe] = 6.75 ppm

Internal Standard Method


1. Most convenient when variations in analytical sample size, position, or matrix limit the precision of a technique.
2. May correct for certain types of noise.

Internal Standard Procedure


1. Prepare a set of standard solutions for analyte (A) as with the calibration curve method, but add a constant amount of a second species (B) to each solution.
2. Prepare a plot of SA/SB versus [A].

Notes
1. The resulting measurement will be independent of sample size and position.
2. Species A & B must not produce signals that interfere with each other. Usually they are separated by wavelength or time.

Example: Pb by ICP Emission


Each Pb solution contains 100 ppm Cu.

Signal [Pb] (ppm) 20 40 60 80 100

Pb 112 243 326 355 558

Cu 1347 1527 1383 1135 1440

Pb/Cu 0.083 0.159 0.236 0.313 0.388

No Internal Standard Correction


600

500

Pb Emission Signal

400

300

200

100

0 0 20 40 60 80 100 120

[Pb] (ppm)

Internal Standard Correction


0.450 0.400 0.350

Pb Emission Signal

0.300 0.250 0.200 0.150 0.100 0.050 0.000 0 20 40 60 80 100 120

[Pb] (ppm)

Results for an unknown sample after adding 100 ppm Cu


Run 1 2 3 4 5 mean S/N Pb 346 297 328 331 324 325 17.8 18.2 Signal Cu 1426 1229 1366 1371 1356 1350 72.7 18.6 Pb/Cu 0.243 0.242 0.240 0.241 0.239 0.241 0.00144 167

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