Chapter XI. Instrumental Methods For Quantitative Analysis (2011)

JUTTI LEVITA, 2011
JUTTI LEVITA-UNIVERSITAS
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When the instrument readout is totally free of

matrix effects, sample loss effects, or any other
potential error-causing effects, this method
works well.
The usual procedure is to accurately prepare a
stock standard solution of the analyte and then
to prepare a series of standards by diluting this
stock.
PADJADJARAN
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120
100
Signal (mAbs)
80
60
40
20
0
0
10
12
JUTTI
PbLEVITA-UNIVERSITAS
Concentration (ppb)
PADJADJARAN
14
16
18
20
The standard additions method (often referred

to as "spiking method") is commonly used to
determine the concentration of an analyte that
is in a complex matrix, such as biological fluids,
soil samples, etc.
The reason for using the standard additions

method is that the matrix may contain other
components that interfere with the analyte
signal, causing inaccuracy in the determined
concentration.
PADJADJARAN
The idea is to add analyte to the sample ("spike"

the sample) and monitor the change in instrument
response.
The change in instrument response between the

sample and the spiked samples is assumed to be
due only to change in analyte concentration.
The procedure for standard additions is to split the

sample into several even aliquots in separate
volumetric flasks of the same volume.
The first flask is then diluted to volume with

the selected diluent.
PADJADJARAN
A standard containing the analyte is then added in

increasing volumes to the subsequent flasks and
each flask is then diluted to volume with the
selected diluent.
The instrument response is then measured for all

of the diluted solutions and the data is plotted with
volume standard added in the x-axis and
instrument response in the y-axis.
Linear regression is performed and the slope (m)

and y-intercept (b) of the calibration curve
are used to calculate the concentration of analyte
in the sample.
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Vx = volume of the sample aliquot

cx = concentration of the sample
cs = concentration of the standard
PADJADJARAN
m = slope
An internal standard is a known amount of a

compound, different from the analyte, added to the
unknown.
The signal from the analyte is compared (ratio) with the

signal from the internal standard to determine analyte
concentration.
Internal standards are used when the quantity of

sample analyzed or the instrument response varies
from run to run and difficult to control. Also used when
sample loss is unavoidable during sample preparation
steps prior to analysis.
PADJADJARAN
If you are injecting or pipetting very small

volumes, any variation from run to run can lead
to very large relative errors.
Instruments or techniques that are fully or

partially automated tend to reduce these
variations and thus improve precision
(decrease the standard deviation).
The curve is made by preparing a set of

standard solutions for analyte (A) with the
addition of a constant amount of a second
species (B) to each solution.
PADJADJARAN
Run 1:
Run 2: You injected a

slightly smaller volume.
Both signals, analyte and internal standard, are affected the

same way. Even though the analyte signal is smaller in run 2,
the ratio to the standard will be the same as run 1.
PADJADJARAN
To use an internal standard, we prepare a

known mixture of standard and analyte to
measure the relative response of the detector
to the two species.
An instrument may not respond exactly the

same way to both compounds.
F is the response factor.

Thus, if F is 1.56, then we would say that the instrument responds
to the analyte 1.56 times greater than the standard.
Use F to determine an unknown analyte concentration with a
known amount of internal
standard.
PADJADJARAN
Each Pb solution contains 100 ppm Cu.
Signal
[Pb]
(ppm)
20
40
60
80
100
Pb
Cu
112
1347
243
1527
326
1383
355
1135
558 PADJADJARAN1440
Pb/Cu
0.083
0.159
0.236
0.313
0.388
PADJADJARAN
No Internal Standard Correction

600
Pb Emission Signal
500
400
300
200
100
0
0
20
40
60
[Pb] (ppm)
PADJADJARAN
80
100
120
Internal Standard Correction
PADJADJARAN
A series of Standard Solutions has been prepared, each having a different amount of "A"
in it. The volume% of "B" and of the
Internal
Standard "IS" is held constant in each
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LEVITA-UNIVERSITAS
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sample.
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Chapter XI. Instrumental Methods For Quantitative Analysis (2011)

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Chapter XI. Instrumental Methods For Quantitative Analysis (2011)

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JUTTI LEVITA, 2011

When the instrument readout is totally free of

The standard additions method (often referred

The reason for using the standard additions

The idea is to add analyte to the sample ("spike"

The change in instrument response between the

The procedure for standard additions is to split the

The first flask is then diluted to volume with

A standard containing the analyte is then added in

The instrument response is then measured for all

Linear regression is performed and the slope (m)

Vx = volume of the sample aliquot

An internal standard is a known amount of a

The signal from the analyte is compared (ratio) with the

Internal standards are used when the quantity of

If you are injecting or pipetting very small

Instruments or techniques that are fully or

The curve is made by preparing a set of

Run 2: You injected a

Both signals, analyte and internal standard, are affected the

To use an internal standard, we prepare a

An instrument may not respond exactly the

F is the response factor.

Each Pb solution contains 100 ppm Cu.

No Internal Standard Correction

Internal Standard Correction

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