Environmental Data Analysis

Lecture 7

Dr. Zhi NING

Agenda

• Limit of detection
• Calibration
• Sizing experiment
Limit of detection

• General concepts
– Method limit of detection, method detection limit
(MDL);
– The ability of a measurement method to determine
an analyte in a sample matrix;
– Statistic terms rather than a chemical concept;
Limit of detection

• US. EPA definition

• “The method detection limit (MDL) is defined as the
minimum concentration of a substance that can be
measured and reported with 99% confidence that
the analyte concentration is greater than zero and is
determined from analysis of a sample in a given matrix
containing the analyte”
– A minimum of seven aliquots of the prepared
solution shall be used to calculate the MDL.
– Solution has a concentration near the expected
limit of detection.
Limit of detection

• Instrument limit of detection (IDL) is based on

the ability to detect the difference
between signal and noise.
– Purely instrumental measurement error.
• Different from MDL
– More of a statistic term
Limit of detection

Measurements are made on a minimum

of seven aliquots (n ≥ 7) of a prepared
solution that has a concentration near
the expected limit of detection. These
data are used to calculate the standard
deviation of the replicate measurements:

• The method detection

limit (MDL) is:
Limit of detection

• Analyze >7 replicate aliquots at a slightly

different concentration to “verify the
reasonableness” of the estimate of the MDL.
• Combined (pooled) variances to obtain a
single estimate for s2. The general relation for
k pooled variances is:

• v=(n1-1)+(n2-1)+…(nk-1)
Limit of detection

• Exercise
• See data example

• Practice
Calibration

• Instrumental methods of chemical analysis

– Construct a calibration curve
Will translate highly precise physical signal (light
absorption, peak height, voltage, etc.) into an
estimate of the true concentration
– By interpolation and not by extrapolation.
Calibration

• Precision of the measured concentrations

1. Does the plot of the calibration curve appear to be
well described by a straight line? If it appears to be curved,
what is the mathematical form of the curve?
2. Bearing in mind that each point on the calibration curve is
subject to error, what is the best straight line (or curved line)
through these points?
3. If the calibration curve is actually linear, what are the
estimated errors and confidence limits for the slope and
intercept of the line?
4. When the calibration curve is used for the analysis of a test
specimen, what are the error and confidence limits for the
determined concentration?
Calibration

• HPLC peak area and dye concentration

Dye conc. HPLC peak area
0.180 26.666
0.350 50.651
0.055 9.628
0.022 4.634
0.290 40.206
0.150 21.369
0.044 5.948
0.028 4.245
0.044 4.786
0.073 11.321
0.130 18.456
0.088 12.865
0.260 35.186
0.160 24.245
0.100 14.175

• If measured peak area is 22.0, then

Calibration

• Working-Hotelling confidence band

Sizing the Experiment

• How large a sample do I need?

Replication and Experimental Design

• Informative power of experiment

– Refine equipment
– Minimize variance from system errors
– Replications
• Reconsider definition of standard
error/variation
– Standard error is random variation of an estimate
from the whole experiment.
– Standard deviation refers to the intrinsic variation
of the observations within individual units.
• Note the role of n (replications)
– Increasing n doesn’t reduce standard deviation
– But reduce standard error.
• 100-fold increase reduces error by 10;
• Four-fold increase reduces error by 2;

– If error is too large, then experiment is useless;

– If error is too small than necessary, then resources
will be wasted.
Let’s review this page…

• Procedures:

1. Calculate the standard deviation for the sample by using this

formula:
Where, S = Standard deviation, = sample mean,
n = number of observations in sample
2. Calculate the value of the one sample t-test, by using this formula:
Where, t = one sample t-test value,
µ= sample mean

3. Calculate the degree of freedom by using this formula:

V=n–1 Where, V= degree of freedom

4. Hypothesis testing:
Compare the calculated value with the table value. If the calculated value is
greater than the critical value, then we will reject the null hypothesis, and accept
the alternative hypothesis.
Confidence Interval for a Mean

• The (1 – α)100% confidence interval for the

mean η has the form:

• E is
• The sample size n can be expressed as

• We can assume sample size large enough to

be normal distribution; it can be t -dist if n is
small
• To determine n, E, z and  needed

• True value of  is often unknown, but it can

be estimated based on prior data;
– We assume the system doesn’t change during
next phase of sampling;
– Data checking and revise sampling plan if
necessary.
• t decreases as n increases, but little change
once n exceeds 5;
• The gain of reducing interval comes from n.
• Example
• Example
– A monitoring study is intended to estimate the
mean concentration of a pollutant at a sewer
monitoring station. A preliminary survey consisting
of ten representative observations gave [291, 320,
140, 223, 219, 195, 248, 251, 163, and 292].
– The average=? and standard deviation is s = ?.
– The 95% confidence interval of this estimate is
calculated using t 9,0.025 = -2.262:

– What sample size is needed to estimate the true

mean with ± 20 units?