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Good Manufacturing Practices

 Regulates the manufacture of pharmaceuticals.
 Includes requirement for having a QC lab to test
starting materials, intermediates and finished
products, and also the packaging materials.

 Assessment
 Collecting data to show that AP’s are operating within
specified limits
 Verifying that final results meet use objectives

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Control Charts Control Charts

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Method Validation
Method Validation
 Process of proving that an analytical method is acceptable
for its intended purpose
 Pervasive use of analytical measurements in trade,
healthcare, environment protection and monitoring,
materials and products development, legal system etc
 Use of analytical measurements as data-input for major
decision making processes
 Use of validated methods is important for an analytical
laboratory to show its qualification and competency
 Analytical measurements must be trusted by the end-user,
ergo, the analytical chemist must be able to provide
measurements that can answer the end-user’s problem with
confidence
I.Taverniers, M. De Loose & E. Van Bockstaele, Trends in Analytical Chemistry, Vol.
23, No. 8, 2004

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Key Performance Characteristics of

Method Validation (Figures of Merit) 1. Accuracy
 Accuracy  Indicates the closeness of measurement to its true or
 Precision accepted value
 Specificity / Selectivity  Measures agreement between a result and its true
 Sensitivity value.
 Absolute error
 Limit of Detection (LOD)
 %relative error
 Limit of Quantitation (LOQ)
 Linearity  Lower AE, %RE higher accuracy
 Range  Confidence Interval – allows to estimate the range
within which the true value might fall w/in a given
 Robustness
probability.

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2. Precision 2. Precision
 Agreement among data obtained from several  Instrument precision (injection precision) 
measurements reproducibility observed when same quantity of one
sample is repeatedly introduced (≥10 times) into an
instrument
 Intra-assay precision  evaluated by analyzing
aliquots of a homogeneous material several times by
one person on one day with same equipment
 Ruggedness (intermediate precision)  variation
observed when an assay is performed by different
people on different instruments on different days in
same lab

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2. Precision 2. Precision
 Interlaboratory precision  Measures of Precision
 Most general measure of  Absolute standard deviations
reproducibility observed  Variance
when aliquots of same  Relative standard deviation
sample are analyzed by
 Coefficient of variation
different people in
different labs (PT)  Standard deviation of the mean
 Becomes poorer as level  Spread or range
of analyte decreases
(Horwitz trumpet)

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3. Sensitivity 4. Limit of Detection

 Ability to discriminate small differences in analyte
concentration; determined by slope of the calibration
curve and precision
 S = ΔR / ΔC
 Higher change in response per unit change in
concentration – higher sensitivity
 The minimum concentration or amount of the analyte
that can be detected at a known confidence level; the
minimum concentration that will produce a
measurement signal which is significantly different from
the signal obtained from the background (or blank)
solution.
 Rm = Rblk + 3σblk

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4. Limit of Detection 4. Limit of Detection

 Procedure that produces detection limit with ~99%  The corrected signal, ysample - yblank is proportional to
chance of being greater than blank: sample concentration:
 After estimating the detection limit from previous  ysample – yblank = m x sample concentration
experience with method, prepare a sample whose  Minimum detectable concentration or detection limit
concentration is ~1 to 5 times detection limit.
 MDL/LOD = 3 σblank /m
 Measure the signal from n replicate samples (n ≥ 7).

 Compute the standard deviation (s) of n measurements.
 Measure the signal from n blanks (containing no analyte),  Limit of quantitation
and find the mean value, yblank.  smallest amount that can be measured with reasonable
 The minimum detectable signal, ydl is defined as accuracy
 LOQ = 10 σblank /m
 ydl = yblank + 3σblank

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4. Limit of Detection 5. Dynamic (or Linear) Range

 Instrument detection limit  Gives the range of concentration over which a linear
 Obtained by replicate concentration curve is obtained.
measurements (n≥7) of
aliquots from one sample
 Within this range, the response is linearly related to the
analyte concentration
 Method detection limit
 Obtained by preparing n≥7
individual samples and
analyzing each one once
 Reporting limit
 concentration, below which
regulatory rules say that a
given analyte is reported as
“not detected”

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5. Dynamic (or Linear) Range 6. Specificity / Selectivity

 Notes on Linear range
 Concentration interval, over which linearity, accuracy, and
precision are all acceptable
 A linear calibration range is preferred because it is easy to
determine the “best” line and carry out an interpolation to
determine the concentration corresponding to a measured
response.
 The degree of linearity is measured using the correlation
coefficient (r)
 The wider the linear range, the better  Ability of an analytical method to distinguish analyte from everything else
that might be in sample
 r = 0.9900 or better – condition for linearity
 Baseline separation
 Ex: range for a major component of a mixture providing a  The detector signal returns to its baseline before next compound reaches
correlation coefficient of R2 ≥ 0.995, spike recovery of 100 ± 2%, detector.
and interlaboratory precision of ±3%  Unresolved impurities at their maximum expected concentration should
not affect the assay.

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7. Robustness or Reliability
 Ability of an analytical method to be unaffected by
small deliberate changes in operating parameters
 For example, the chromatographic method is robust if it
gives acceptable results when small changes are
made in solvent composition, pH, buffer
concentration, temperature, injection volume, and
detector wavelength.
Questions???

jonylgarcia@gmail.com

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