Quality Assurance in the chemical analysis

All the actions we take when

executing an analytical
methodology to get the right
answer for our purpose. For
instance: sufficient accuracy and
precision to support subsequent
decisions.
Raw data: measurements; peak
areas
Treated data: concentrations
derived from raw data by use of
calibration methods; Molarity
Results: quantities reported after

Instrumentation
validation

Certification of
lab facilities

Statistical
procedures

GL
P

Accountability

QA

Certification of
analysts

Documentation
and records

Reagent/materi
al certification

Specimen/samp
le tracking

Quality Assurance in the chemical analysis

Use objectives: states the purpose for which results will be
used. Kind of data and the way it is collected depend on how
you
plan to usestates
it.
Specifications:
how good the numbers need to be and
what precautions are required in the analytical procedure.
Sampling requirements
Accuracy and precision
Rate of false results
Selectivity
Sensitivity
Acceptable blank values
Recovery of fortification
Calibration checks
Quality control samples

Sampling requirements
Representative samples and preserved after collection
Trace metal analysis: plastic or Teflon containers
Organic samples: glass containers

Rate of false results

False positive: concentration of a sample exceeds legal limit

when in fact it is below
False negative: concentration of a sample below the limit when
it is actually above

Selectivity and Sensitivity

Selectivity: being able to distinguish analyte from other species

in the sample
Sensitivity: capability of responding reliably and measurably to
changes in analyte concentration

Acceptable blank values

Method blank: sample with all components except the analyte
Reagent blank: sample with all components but not subjected to
sample preparation procedures
Field blank: sample with all component and exposed to the site of
sampling

Recovery of fortification

Spike: known amount of analyte added to a sample to test the

response of the calibration curve

alibration checks:

Instrument working properly and calibration is valid

Performance test samples or quality control samples or blind samples

Standard Operating Procedures (SOPs)

SOPs are the center of quality assurance. Stating the procedures
to follow (instructions to follow and how) in specific situations.
SOPs: Instrument maintenance and calibration
SOPs: Recording temperatures of refrigerators, calibrating
balances, replacing reagents

Assessment

Collecting to show that analytical procedures

are operating within specified limits
Verify that final results meet objectives

Quality assurance process summarized

Chemical Analysis

The Chemical analysis

Terminology: Analysis, analytes, matrix, determination,

measurement, technique, method, procedure, protocols
Analytical techniques:
total analysis techniques and concentration techniques
Parameters to select the analytical method: accuracy,
precision, sensitivity, selectivity, robustness, ruggedness,
scale of operation, equipment, time and cost
Developing the procedure: Interferences, calibration and
standardization, sampling, validation, and protocol

Terminology
Analysis: A process that provides chemical or physical
information about the constituents in the sample or the
sample itself.
Analytes: The constituents of interest in a sample
Matrix: All other constituents in a sample except for
the analytes
Determination: Analysis of a sample to find the
identity, concentration, or properties of the analyte
Measurement: An experimental determination of an
analytes chemical or physical properties
Technique: A chemical or physical principle that can be
used to analyze a sample
Method: A means for analyzing a sample for a specific
analyte in a specific matrix
Procedure: Written directions outlining how to analyze
a sample
Protocol: Set of written guidelines for analyzing a
sample specified by an agency
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Classifying analytical techniques

Total analysis techniques ( classical techniques): signal is
SA
kn A of analyte.
proportional to an absolute
amount
nA: moles of grams of analyte, SA signal due to analyte.
Mass, volume, and charge are the most common signals for total
analysis techniques Examples are: gravimetry, titrimetry and
coulometry. With a few exceptions, the signal in a total analysis
technique results from one or more chemical reactions involving
the analyte. These reactions may involve any combination of
precipitation, acidbase, complexation, or redox chemistry.

Concentration techniques: ( Instrumental techniques):

kC A concentration.
signal is proportional S
toA analytes
CA: concentration of analyte in sample, SA signal due to analyte.
Instrumental techniques rely on measuring an optical or
electrical signal. Examples: spectroscopy, Chromatography,
potentiometry, and voltammetry
9

Common components on instrumental techniques

10

Parameters to select the analytical

method
1. Accuracy
2. Precision
3. Sensitivity
4. Selectivity
5. Robustness
6. Ruggedness
7. Scale of operation
8. Analysis time
9. Availability of equipment
10.Cost
11

Parameters to select the analytical

method
Accuracy: how closely the result of a
measurement agrees with the
expected result.

1. Relative error within1% of correct

result: Highly accurate
2. Relative error between 1% and 5%:
Moderately accurate
3. Relative errors greater than 5%: low
accuracy

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Parameters to select the analytical

method
Precision: An indication of the
reproducibility of a measurement or
result.
More precise data

Less precise data

13

Parameters to select the analytical

method
Sensitivity: A measure of a methods
ability to distinguish between two
samples; reported as the change in
signal per unit change in the amount of
analyte (k).

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Parameters to select the analytical

method
Selectivity: A measure of a methods freedom
from interferences as defined by the methods
selectivity coefficient.
kA and kI: sensitivities for
the analyte and the
interferent.
nI and CI are the moles
(or grams) and
concentration of the
interferent in the sample

Selectivity coefficient (KA,I): A measure of a

methods sensitivity for an interferent relative to
that for the analyte.
15

Parameters to select the analytical

method
Robustness: A method relatively free from
chemical interference that can be applied in a
wide variety of sample matrices
Ruggedness: A method that is insensitive to
changes in experimental conditions. For instance:
temperature, acidity, or reaction time
Scale of operation: Consider that scale of the
analysis.
1. Amount of sample available
2. Concentration of analyte in sample
3. Absolute amount of analyte needed to obtain
measurable signal
16

Parameters to select the analytical

method
Scale of operation:
Consider the scale of the
analysis.
1. Amount of sample
available
2. Concentration of
analyte in sample
3. Absolute amount of
analyte needed to obtain
measurable signal
Equipment, time, and
cost: equipment need, time
required to complete the
analysis and cost per
sample.

17

Method validation
Process of proving that an analytical methodology
is acceptable for its intended purpose
Parameters to measure include:
Specificity
Linearity
Accuracy
Precision
Range
Limit of detection and quantification
Robustness

18

Specificity
Ability of an analytical method to distinguish
analyte from everything else that might be in the
sample
Baseline requirement: detector signal return to
its baseline before next compound reaches
detector

19

Linearity
How well a calibration sample follows a straight
line, showing that response is proportional to the
quantity of the analyte
Calibration curves should span 0.5 to 1.5
expected analyte concentration
Each standard should be prepared and analyzed
three times
Blank

20

Accuracy
how closely the result of an experiment agrees with
the expected result

Analyze a certified reference material

Compare results from two or more different
analytical methods
Blank sample spiked with known amount of
analyte
Standard addition

1. Relative error within1% of correct result: Highly

accurate
2. Relative error between 1% and 5%: Moderately

21

Precision
An indication of the reproducibility of a measurement
or result. How well replicate measurement agree with
one another.
More precise data
Less precise data

Repeatability: spread of results when one person uses

one procedure to analyze the sample by the same
method multiple times
Reproducibility: spread of results when different
people in different labs using different instruments
each try to follow the same procedure

22

Precision
Instrument precision: repeatability of the same
quantity of one sample is repeatedly introduced (>
10 times) into an instrument
Intra-assay precision: evaluated by analyzing
aliquots of a homogenous material several times by
one person on one day with the same equipment
Intermediate precision (ruggedness): variation
observed when an assay is performed by different
people on different instruments on different days in
the same lab
Interlaboratory precision(reproducibility):
aliquots of the same sample are analyzed by

23

Precision

Coefficient of variation of interlaboratory

results as a function of sample
concentration (expressed as g analyte/g
sample). The shaded region has been
referred to as the Horwitz trumpet
because of the way it flares open. [From
W. Horwitz, Evaluation of Analytical
Methods Used for Regulation of Foods and
Drugs, Anal. Chem. 1982, 54, 67A.]

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Range
Concentration interval over which linearity,
accuracy, and precision are all acceptable.
R2 0.995, spike recovery: 100 2%, precision: 3%
Linear range: concentration over which calibration curve
is linear
Dynamic range: concentration over which there is a
measurable response

25

Limit of Detection
Smallest quantity of analyte that is
significantly different from the blank.

1. After estimating the detection limit from previous

experience with the method, prepare a sample whose
concentration is to 5 times the detection limit.
2. Measure the signal from n replicate samples (n 7).
3. Compute the standard deviation (s) of the n
measurements.
4. Measure the signal from n blanks (containing no analyte)
and find the mean value, yblank.
5. The minimum detectable signal, ydl, is defined as Signal
detection limit:
6. The corrected signal, ysample - yblank, is proportional
to sample concentration:
Calibration line: ysample - yblank = m sample
concentration
Detection limit: Minimum detectable concentration =

26

Detection Limit (DL)

Limit of Quantitation
A signal that is 10 times greater than the
noise in the blank or a small signal.
Smallest quantity that can be measured with
reasonable accuracy
LOQ = 10 s/m

28

METHODS OF CALIBRATION
Calibration: process of establishing the relationship between the
signal we measure (such as absorbance) and known
S A kC A
concentrations of analyte
k is determined by measuring the signal for one or more standards, each
containing a known concentration of analyte and ploting Smeas vs. concentration
(Calibration curve)

External Calibration

Unknown sample with absorbance of 1.30 ( just like our 60.0

mg/100 mL standard) and used our original calibration curve
extrapolated to higher absorbances as shown by the black squares,
29
we would calculate a concentration of 51.9 mg/100 mL for the

External Calibration
A calibration curve shows the response of a chemical analysis
to known quantities (standard solutions) of analyte.
When there is a linear response, the corrected
analytical signal (signal from sample - signal
from blank) is proportional to the quantity of
analyte.
The linear range of an analytical method
is the range over which response is
proportional to concentration.
The dynamic range is the range over
which there is a measurable response to
analyte, even if the response is not linear.

Standard Addition
A standard addition is a known quantity of analyte
added to an unknown to increase the concentration of
analyte.
Standard additions are especially useful when matrix
effects are important.
Use Equation below to compute the quantity of analyte
[X]i after a single standard addition.
The initial concentration [X]i, increases to [X]f while the
initial signal, Ix increase to Ix+s. [s]f is the concentration of
standard added.

Ix

Xi

I x s X f S f

Standard Addition

Multiple Standard Additions and

Uncertainty
For multiple standard additions to a single solution,
use Equation 5-9 to construct the graph in Figure 5-6, in
which the x-intercept gives us the concentration of
analyte.
For multiple solutions made up to the same final
volume, the slightly different graph in Figure 5-7 is
used.
Equation 5-10 gives the x-intercept uncertainty in
either graph.

Multiple Solutions Made up to the Same Final Volume

For multiple solutions made up to the same final

volume, the slightly different graph in Figure 5-7 is used.

Multiple Standard Addition to a Constant Volume

Uncertainty in the x-Intercept (ux)

ux

sy
m

1 1
( y)
2
2
k n m ( xi x )

m = slope
k = number of replicate measurements for unknown
n = number of data points for calibration line
= mean value of measured y for unknown x
Sy = error of the regression

Internal Standard
An internal standard is a known amount of a compound,
different from analyte, that is added to the unknown.
Signal from analyte is compared with signal from the
internal standard to find out how much analyte is present.
Internal standards are useful when the quantity of sample
analyzed is not reproducible, when instrument response
varies from run to run, or when sample losses occur in
sample preparation.
The response factor F is the relative response to analyte
and standard.

Internal Standard
Compensates for changes in experimental conditions,
when Ix varies for same concentration [X]
Internal Standard: Added standard is different from
analyte
A
A
x

[X]

IS

[IS]

Internal Standards Calibration

Internal Standard
For the sake of accuracy, it is best to prepare a series of
standard mixtures of analyte plus internal standard and
prepare a graph such as Figure 5-10.
The response factor is the slope of the graph, and the
intercept should be within statistical limits of zero.
The graph should confirm linear response over the
desired analytical range.

Internal Standard

Calibration Curve