Validasi Metode Analisis

Dr. rer. nat. Tanti Tatang Irianti, M.Sc., Apt.

intanti@ugm.ac.id
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 Validasi Metode Analisis
❖Kemanfaatan metode harus konsisten dengan tujuan utama
validasi metode diusulkan, yaitu,
Prosedur analisis harus divalidasi:

a.secara menyeluruh, termasuk penyiapan sampel sebelum

analisis.
b.mencakup rentang konsentrasi analit yang dinyatakan
dalam lingkup metode.
c.untuk masing-masing jenis matrik dimana metode tersebut
akan digunakan.

Gonzales and Herrador, 2007, Trend in Analytical Chemistry, 26 (3): 227-238

4
The Manager ’s Guide to VAM, UK Department of Trade and Industry,
Valid Analytical Measurement Programme [ Eurachem Guide, 1998 ].

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 Eurachem Guide, 1998

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 Definisi

Eurachem Guide, 1998

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 Kapan Suatu Metode Analisis Perlu
divalidasi atau revalidasi ?
[ Huber, in: Nash and Wachter, 2003 ]

❖Sebelum metode diterapkan atau diberlakukan untuk

penggunaan rutin.

❖Bila kondisi berubah dilihat dari tujuan metode tersebut

divalidasi [misal: instrumen dengan karakteristik yang berbeda].

❖Bila metode tersebut berubah dan perubahan tersebut diluar

lingkup awal metode tersebut saat divalidasi.

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 lanjutan….

❖Bila kontrol kualitas mengindikasikan bahwa metode baku

tersebut berubah dengan berubahnya waktu.

❖Ada maksud untuk membuktikan kesetaraan antara dua metode

(metode baru dan metode baku).

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Metode Compendial/Farmakope

Metode yang termuat di dalam farmakope

sudah divalidasi

Jadi tidak perlu dilakukan validasi lagi

namun

diperlukan verifikasi

Kazakevich, Y. and LoBrutto, R., 2007, HPLC for Pharmaceutical Scientists,

Chpt. 9
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 Assignment of Validation Parameters

Kazakevich, Y. and LoBrutto, R., 2007, HPLC for Pharmaceutical Scientists,

Chpt. 9, p.459.
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Bukti Bahwa Metode Analisis yang ada
di Farmakope/Compendia
Sudah di Validasi
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 Method
Adjustment or Modification
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 Verification
Verification of an analytical procedure is:

The demonstration that a laboratory is capable of replicating

with an acceptable level of performance a standard method.

Verification under conditions of use is demonstrated by

meeting system suitability specifications establihed for the
method, as well as a demonstration of
accuracy and precision
or
other method parameters for the type of method
Ora Laboratory Procedure, FDA: Ora-Lab.5.4.5
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 Typical Analytical Characteristics Used
In Method Validation

Specificity/Selectivity

Precision

Accuracy

Limit of Detection

Limit of Quantitation

Linearity/Range

Robustness

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 Table 2: Data Elements Required for Validation
[ USP-35, p.881 ]
Analytical Category Category II Category Category
Performance I Quanti- Limit III IV
Characteristics
tative Tests
Accuracy Yes Yes * * No
Precision Yes Yes No Yes No
Specificity Yes Yes Yes * Yes
Detection Limit No No Yes * No

Quantitation No Yes No * No
Limit
Linearity Yes Yes No * No
Range Yes Yes * * No

* May be required, depending on the nature of the specific test

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 United State Pharmcopeia, No.31-NF 26, (2008), ch. 621

2022 Snyder dkk., 2010, Introduction to Modern Liquid Chromatography, Chpt. 12 23

Categories of assays for which validation data
[ USP -35, p.881 ]
◼ Category I :
Analytical procedures for quantitation of major
components of bulk drug substances or active ingredients
(including preservatives) in finished pharmaceutical
products.

◼ Category II :
Analytical procedures for determination of impurities in
bulk drug substances or degradation compounds in finished
pharmaceutical products. These procedures include
quantitative assays and limit tests.

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Categories of assays for which validation data
[ USP -35, p.881 ]

◼ Category III :

Analytical procedures for determination of performance

characteristics (e.g., dissolution, drug release, etc.).

◼ Category IV : Identification tests

For each assay category, different analytical information
is needed. Listed in Table 2 are data elements that are
normally required for each of these categories.

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 Guidance for Method Validation

Ahuja dan Dong, 2005.

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 2011

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 Identitas Analit, p.182

Confirmation of the identity of an analyte’s spot can be

also determined by two- dimensional TLC with full scan
UV/VIS and cochromatography.
The analyte’s spot should be intensified by doing a
cochromatography, and new spot should not appear and the
visual appearance shall not change; Rf value of the analyte
shall agree with the reference standard within +/- 5%.

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 Peak purity, p.182

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 Clarke’s, 2011, 4th Ed., p.607

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 Moffat, A.C., 1986, [ Editor ]
Clarke’s Isolation and Identification of Drugs Second
Edition, p.308

The A1%
1cm value are divided into 3 categories:
Category 1:

The letter a after figure A indicates that the value is a

1%
1cm

mean value based on several reported figures all of which

lie within a range of
+ 10%
of the mean

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 Moffat, A.C., 1986, [ Editor ]
Clarke’s Isolation and Identification of Drugs Second
Edition, p.308

Category 2:

The letter b after a figure A indicates that the value is a

1%
1cm

single reported value of

unknown reliability

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 Moffat, A.C., 1986, [ Editor ]
Clarke’s Isolation and Identification of Drugs Second
Edition, p.308

Category 3:

The letter c after a figure A1cm indicates that the value is a

1%

mean value based on several reported figures some of

which lie outside
+ 10%
of the mean

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 Hub. pKa vs pH vs Ionisasi Anait

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 BUFER

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Metode H P L C
 Spesifisitas

❖ Metode HPLC

Puncak-puncak kromatogram terpisah sempurna dilihat dari

Resolusi yang dihasilkan

Juga harus diperhatikan baseline resolutionnya

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 Selektivitas

Ahuja dan Dong, 2005.

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 Resolusi

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 HPLC

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 Ampisilin, Amoksisilin, Pensilin G-Na,
Penicilin V.

Amoksisilin Ampisilin

Penisilin G Na Penisilin V

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 Ampisilin, Penisilin G, dan Penisilin V

Pertanyaan:

Apakah ketiga obat tersebut dapat di analisis:

Secara Spektrofotometri Ultraviolet ?

Secara Titrimetri (Iodometri) ?

Mengapa Demikian ?

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 Antibiotik

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 Contoh
forced degradation
study
 Forced Degradation Studies

Ahuja dan Dong, 2005.

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 Forced Degradation Study (Ahuja &Dong, 2005, p.366)

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 FDS

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 Contoh
Forced Degradation
Study

Dr. rer. nat. Tanti Tatang Irianti, M.Sc., Apt.

intanti@ugm.ac.id
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 Forced Degradation Studies

Ahuja dan Dong, 2005.

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 Carbamazepin

Carbamazepin

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 Oksidatif-hidrolisis

Carbamazepin

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 Carbamazepin

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 [ minute ]

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Terimakasih