VALIDASI METODA ANALISIS

Kelompok 6

Rahmita Bimasari (1611011052)

Supina Darmala Sari (1611011054)
Nevi Syafitri (1611011056)
Isra Hasanah (1611011058)
Valdy Filando Sardi (1611011060)
Definition

Method validation is the process used to confirm that the analytical procedure employed for a specific test is suit
able for its intended use.
Results from method validation can be used to judge the quality, reliability and consistency of analytical results; it
is an integral part of any good analytical practice.

Analytical methods need to be validated or revalidated before their introduction into routine use;
whenever the conditions change for which the method has been validated (e.g., an instrument with different char
acteristics or samples with a different matrix); whenever the method is changed and the change is
outside the original scope of the method.

(Huber, 2016)
 Steps
Eight steps for validation:
Accuracy
Precision
Specificity
Limit of detection
Limit of quantitation
Linearity and range
Ruggedness
Robustness

(Huber, 2016)
 Accuracy and Recovery
Accuracy can also be described as the closeness
of agreement between the value that is
adopted, either as a conventional, true or
accepted reference value, and the value found.

How? : compare the results of the method with re

sults from an established reference method.
This approach assumes that the uncertainty of
the reference method is known.

(Huber, 2016)
Precision and Reproducibility
The precision of a method is the extent to
which the individual test results of multiple
injections of a series of standards agree

(Huber, 2016)
 Precision and Reproducibility
Precision may be considered at three levels:
1. Repeatability expresses the precision under the same o
perating conditions over a short interval of time. Repeat
ability is also termed intra-assay precision.
2. Intermediate precision expresses variations within labor
atories, such as different days, different analysts, differe
nt equipment, and so forth.
3. Reproducibility expresses the precision between labora
tories (collaborative studies usually applied to standardi
zation of methodology).
(Huber, 2016)
Specificity

the term specific generally refers to a method t

hat produces a response for a single analyte onl
y, while the term selective refers to a
method that provides responses for a number o
f chemical entities that may or may not be
distinguished from each other. If the response is
distinguished from all other responses, the met
hod is said to be selective

(Huber, 2016)
Limit of Detection
The limit of detection is the point at which a m
easured value is larger than the
uncertainty associated with it. It is the lowest c
oncentration of analyte in a sample that
can be detected but not necessarily
quantified.

(Huber, 2016)
Limit of Quantitation
The limit of quantitation is the minimum
injected amount that produces quantitative m
easurements in the target matrix with
acceptable precision in chromatography,
typically requiring peak heights 10 to 20
times higher than the baseline noise.

(Huber, 2016)
Linearity and Range
The linearity of an analytical method is its
ability to elicit test results that are directly
proportional to the concentration of analytes
in samples within a given range or
proportional by means of well-defined
mathematical transformations.

(Huber, 2016)
Linearity and Range
The range of an analytical method is the interva
l between the upper and lower levels
(including these levels) that have been
demonstrated to be determined with
precision, accuracy and linearity using the
method as written. The range is normally
expressed in the same units as the test results
(e.g., percentage, parts per million) obtained
by the analytical method.

(Huber, 2016)
Ruggedness
Ruggedness is a measure of reproducibility
of test results under normal, expected
operational conditions from laboratory to labor
atory and from analyst to analyst.
Ruggedness is determined by the analysis of a
liquots from homogeneous lots in different lab
oratories

(Huber, 2016)
Robustness
Robustness tests examine the effect that
operational parameters have on the analysis
results. For the determination of a method’
s robustness, a number of method parameters, fo
r example, pH, flow rate, column temperature, inj
ection volume, detection wavelength or
mobile phase composition, are varied within a
realistic range, and the quantitative influence of t
he variables is determined.

(Huber, 2016)
Robustness
If the influence of the parameter is within a
previously specified tolerance, the parameter i
s said to be within the method’s
robustness range.

(Huber, 2016)
Selecting Validation Parameters and Limits
The selection of validation parameters and
acceptance criteria should be based on
business, regulatory and client requirements a
nd should be justified and documented.

(Huber, 2016)
(Huber, 2016)
(Huber, 2016)
Test Excecution

The optimal sequence for a liquid chromatographic method

1. Specificity/selectivity
2. Repeatability of retention times and peak areas
3. Linearity, limit of quantitation, limit of detection, range
4. Accuracy at different concentrations
5. Intermediate precision
6. Reproducibility
Daftar Pustaka
Huber, Ludwig. (2016). Validation of Analytical Methods and Procedures. FDA

(Huber, 2016)
Thank You!