CHAPTER

Sampling, Standardization,
and Calibration
Figure 8-1
Classification of analyses by sample size.
Figure 8-2
Classification
of constituent
types by
analyte level.
 Matrix Effects
• Matrix can contain species that have chemical
properties similar to the analyte.
• Such effect can be induced not only by the
sample itself but also by the reagents and
solvents used to prepare the sample for the
determination.
 採樣真的很難
Sampling is often the most
difficult aspect of an analysis
Figure 8-4
Steps in obtaining a laboratory
sample. The laboratory sample
consists of a few grams to at most
a few hundred grams. It may
constitute as little as 1 part in 107
or 108 of the bulk material.
Both goals require obtaining a
random sample
Sampling Uncertainties
Both systematic and random errors in analytical data
can be traced to instrument, method, and personal
causes

Systematic error ~ eliminate by exercising care, by

calibration, and by the proper use of standards,
blanks, and reference materials
Random error ~ reflected in the precision of data,
which can generally kept at an acceptable level by
closely controlling the variables that influence the
measurements
Errors due to invalid sampling ~ not controllable.
Ordinarily treated separately
When Sm  Ss/3, there is no point in trying to improve the measurement precision.
If the sampling uncertainty is large and can not be improved  switch to a faster
method of analysis  more samples can be analyzed (std dev of the mean is lower
by a factor of N1/2)
10 2.23 8.60
Automated Sample Handling
轉換率
Dispersion – a band-spreading or mixing phenomenon that results
from the coupling of fluid flow with molecular diffusion

Diffusion – mass transport due to a concentration gradient

 Sampler vs. Detector

The analyte concentration profiles at the sampler and at the detector.

Normally, the height of a sample peak is related to the concentration
of the analyte.
•  syringe
Fig 2-3: 10 mL syringe

 sandwich
injection technique

Harris Fig 24-9

•  loop injectors/rotary sample valve

 sample loading  sample injection flow
syringe meter
eluent to eluent to injector detector
eluent in column eluent in column
flow
two-stage splitter recorder
pressure
regulator

flow column
controller
sample in out sample in out

carrier gas Skoog

supply column oven Fig 27-1
 injector (Harris 25-2)
 sample introduction (0.1 ~ 100 mL) w/o depressurization

•  fixed-volume sample loop

 loading  injection

sample
Skoog loops
Fig 28-7
Fig 3-4

Harris Skoog
Fig 25-16 injector Fig 28-4
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191
http://bmrc.berkeley.edu/courseware/ICMfg92/images/gif/photolith.gif
LOD: Limit of Detection 跟零點有差異的
LOQ: Limit of Quantification 儀器達到最低的濃度，可能會和LOD重疊
LOL: Limit of linearity
Comparison with Standards
Figure 1F-2
Constructing and using a
calibration curve to determine the
concentration of arsenic. The
absorbance of the solutions in the
cuvettes are measured using a
spectrophotometer. The
absorbance values are then plotted
against the concentrations of the
solutions in the cuvettes, as
illustrated in the graph. Finally, the
concentrations of the unknown
solutions are read from the plot, as
shown by the dark arrow.
The comparison method for aflatoxin
(a)
(b) (c)

Aflatoxin threshold 20 ppb

Set by US FDA

During the analysis, an enzyme

reaction causes a blue product to
form. [Aflatoxin] , blue color 

191
 External Standard Calibration
• External standard is prepared separately
from the sample
• Internal standard is added to the sample
itself
• The least squares method
Draw a best line among data points and also specifying
the uncertainties associated with – Regression analysis
 Least-square method: 2 assumptions
(1) A linear relationship actually exist between x and y.

Figure 8-10
The slope-intercept form of
a straight line.

b is the y intercept and m is the slope of the line. (2) Assuming deviation of the
individual points from the straight line arises from error in the measurement; there is
no error in the x values of the points (concentration)
Summary
• Least square method: find the sum of the square of the
residuals (SSresid)
• SSresid=Σ[yi-(b+mxi)]2, N=# of points used
• Sum of square of deviation from mean for individual value of x
and y
– Sxx = Σ(xi-x)2
– Syy = Σ(yi-y)2
– Sxy = Σ(xi-x)(yi-y)
• The slope of the line, m = Sxy/Sxx [eq 8-13]
• The intercept, b = y-mx [eq 8-14]
• The std dev about regression, Sr=√ (Syy-m2Sxx)/N-2 [eq 8-15]
• The std dev of the slope, Sm = √Sr2/Sxx [eq 8-16]
• The std dev of the intercept, Sb = Sr√(Σxi2/(NΣxi2-(Σxi)2) [eq 8-17]
• The std dev about regression, Sr is the std dev
for y when the deviations are
• measured not from the mean of y but from the
straight line that results from the least-squares
prediction. The value of Sr is related to SSresid

Sr= √Σ[yi-(b+mxi)]2 / N-2 = √SSresid / N-2

The standard deviation about regression (Sr),
also called the standard error of the
estimate or just the standard error, is a
rough measure of the magnitude of a
typically deviation from the regression
line
Figure 8-13
Spreadsheet to illustrate the internal standard method
for the flame emission determination of sodium.
Please carry out a least-square analysis.
m = Sxy/Sxx
 b = y-mx

Sb = Sr√(Σxi2/(NΣxi2-(Σxi)2)
 Interpretation of the Least-Square Results
Measures the variation in the observed values of the dependent
• SSresid =variable (y values) SSresid=Σ[yi-(b+mxi)]2
• Define total sum of the square, SStot = Syy
– Total sum of the square is a measure of the total
variation in the observed values of y, because the
deviations are measured from the mean value of y.
• Coefficient of determination (R2)
– Is a measure of the fraction of the total variation in y
that can be explained by the linear relationship
between x and y.
– The closer R2 is to unity, the better the linear model
explains the y variation.
 Sensitivity
– Detection Sensitivity (Limit of Detection): The smallest concentration of analyte
that can be statistically significantly distinguished from zero for a given sample
matrix with a stated degree of confidence.
– Limit of Quantitation: The smallest concentration of analyte that can be
measured in samples and yield a predicted concentration with states relative
precision or accuracy (or both).
– Analytical sensitivity and detection limit. These terms are commonly confused.
The International Union of Pure and Applied Chemistry (IUPAC) defines
analytical sensitivity as the ability of an analytical procedure to produce a
change in signal for a defined change of the quantity This is often visualized as
the slope of the calibration curve.
• Detection limit (or limit of detection) is defined as the lowest concentration or
quantity of an analyte that can be detected with a stated reasonable uncertainty
for a given analytical procedure. The operational definition of this limit must be
supplied by the author: e.g., the concentration at a signal-to-noise ratio of 10 or the
concentration corresponding to a signal 3 SD above the mean for a calibrator that is
free of analyte. This is also called the limit of the blank.
https://goldbook.iupac.org/html/L/L03540.html
Quality Assurance of Analytical
Results

A control chart is a sequential plot of some

characteristic that is criterion of quality
UCL and LCL are three std dev on either side of the population mean and
form a range within which a measured mass is expected to lie 99.7% of the time.
Figure 8-15
A control chart for a modern analytical balance. The
results appear to fluctuate normally about the mean
except for those obtained on day 17. Investigation led to
the conclusion that the questionable value resulted from
a dirty balance pan. UCL = upper control limit; LCL = lower
control limit.
Figure 8-16
A control chart for monitoring the concentration of benzoyl peroxide
in a commercial acne preparation. The manufacturing process
became out of statistical control with sample and exhibited a
systematic change in the mean concentration. UCL = upper control
limit; LCL = lower control limit.
The chart shows that until day 83, the manufacturing
process was in statistical control, with normal
random fluctuation in the amount of benzoyl
peroxide.