1

1. Title

The title, "Development and Validation of a Stability-Indicating Reverse Phase

High-Performance Liquid Chromatography (RP-HPLC) Method for the
Determination of Brivaracetam," describes a crucial analytical project focused on
creating and rigorously testing a specialized procedure for pharmaceutical quality
control.

 Development and Validation: This signifies the two essential and

interconnected phases of the research project. Development involves creating
the chromatographic procedure, optimizing parameters (such as mobile phase
composition, column, flow rate, and temperature), and ensuring optimal
separation. Validation involves demonstrating, through a set of defined
experiments (following guidelines like ICH Q2(R1)), that the developed
method is suitable for its intended purpose that it is accurate, precise, linear,
and robust.
 Stability-Indicating: This is the most critical feature of the method. It means
the method is designed and proven to accurately and selectively quantify the
intact drug substance (Brivaracetam) even when degradation products,
impurities, and excipients are present in the sample. This capability is
required by regulatory bodies (ICH Q1A(R2)) to monitor the quality and
safety of a drug product throughout its shelf life.
 High-Performance Liquid Chromatography (HPLC) Method: This
specifies the analytical technique used. HPLC is the gold standard separation
technique in pharmaceutical analysis, known for its high resolving power,
accuracy, and sensitivity. The method is used to separate the drug from all
other components in a mixture based on their differential interaction with the
stationary phase (column) and the mobile phase.
 Determination of Brivaracetam: This states the target analyte.
Brivaracetam (BRV) is the specific active pharmaceutical ingredient (API),
an antiepileptic drug (AED). The goal of the method is the quantitative
measurement (determination) of BRV concentration in both pure (bulk) form
and in its finished pharmaceutical dosage form.

In brief, the research is focused on creating and validating a highly selective and
robust HPLC test that can accurately measure the amount of the antiepileptic drug,
Brivaracetam, specifically ensuring that its concentration measurements are not
compromised by any breakdown products that may form during storage or
manufacturing. This is essential for regulatory compliance and drug safety.
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2. Introduction

2.1. Brivaracetam (BRV) and its therapeutic importance

Brivaracetam (BRV), chemically designated as (2S)-2-[(4R)-2-oxo-4-

propylpyrrolidin-1-yl]butanamide, represents a third-generation antiepileptic drug
(AED). It is the successor to Levetiracetam, designed to treat partial-onset seizures in
adults and children.

The drug's therapeutic efficacy stems from its highly selective and high-affinity
binding to the Synaptic Vesicle Protein 2A (SV2A), an integral membrane
glycoprotein found in presynaptic vesicles. This mechanism modulates
neurotransmitter release, providing robust anticonvulsant activity. The improved
affinity of BRV for SV2A and its rapid brain penetration relative to older AEDs
contribute to its clinical advantage.

2.2. Physicochemical Properties and Stability Concerns

BRV is a small molecule with a molecular weight of 212.26 g/mol. Its structure
contains two key functional groups that pose potential analytical challenges:

1. Chiral Center: The molecule possesses a chiral carbon at the C2 position of

the butanamide moiety. While the drug is marketed as the S-enantiomer, any
trace amount of the R-enantiomer formed during synthesis or racemization
during processing must be controlled as a critical impurity.
2. Amide Bond: The core structure contains an amide linkage, which is
notoriously susceptible to hydrolytic degradation under extreme pH
conditions (acidic or basic). This chemical liability means that degradation
products (degradants) are likely to form under stress, demanding a specialized
analytical approach.

2.3. Regulatory Framework and the Need for Stability-Indicating Methods

(SIAM)

The quality and safety of any pharmaceutical product are governed by stringent
international guidelines. A cornerstone of this regulation is the demonstration of
stability, which confirms that the drug remains safe and effective throughout its
proposed shelf life.

 ICH Q1A (R2) – Stability Testing: This guideline explicitly requires that all
analytical procedures used for the assay of the API in samples subjected to
stability testing must be Stability-Indicating (SI). This means the method
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must possess the power to resolve the intact drug from its breakdown
products and impurities.
 ICH Q2 (R1) – Analytical Validation: This mandates that the method's
specificity is the ability to measure the analyte response in the presence of all
potential interferents (degradants, impurities, excipients) must be proven and
documented.
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3. Research Rationale:

The development of this specific Stability-Indicating High-Performance Liquid

Chromatography (SI-HPLC) method for Brivaracetam (BRV) is driven by three core
objectives: Regulatory Compliance, Analytical Efficiency, and Strategic Future-
Proofing.

3.1. Regulatory Imperative: Addressing the Stability-Indicating Gap

Pharmaceutical regulatory guidelines, primarily ICH Q1A (R2) and Q2(R1), mandate
that all methods used for quantifying an Active Pharmaceutical Ingredient (API)
during stability testing must be stability-indicating. While several analytical methods
exist for BRV, including UPLC and RP-HPLC methods also reported. There is a
continuous need to publish, optimize, and validate methods tailored for specific
laboratory constraints (e.g., cost, equipment).

The structural features of BRV, particularly its amide bond and chiral center, make it
inherently susceptible to hydrolysis (acid/base degradation) and potential
racemization, which must be resolved chromatographically. This research ensures the
development of a method that is explicitly proven to separate BRV from all potential
degradation products formed under forced stress conditions, thereby guaranteeing the
accuracy of the assay.

3.2. Economic and Environmental Efficiency

The choice of mobile phase solvents is a strategic cost-saving and green chemistry
measure:

 Methanol (MeOH) over Acetonitrile (ACN): MeOH is generally a more

economical solvent and often preferred in resource-constrained environments.
While ACN can offer slightly higher chromatographic efficiency in some
cases, the aim is to demonstrate that MeOH can be optimized to achieve
regulatory-compliant separation (Resolution Rs ≥ 1.5) for BRV and its
degradants. This aligns with the principles of sustainable analytical method
development being discussed in recent literature.
 Volatile Additives: The aqueous phase utilizes 0.1% Formic acid (FA) and
0.1% Acetic acid (AA), instead of non-volatile inorganic salts (like potassium
phosphate buffer) typically used in conventional RP-HPLC. The use of these
weak organic acids is an economical alternative to purchasing expensive, pre-
mixed buffers.
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3.3. Strategic Future-Proofing: LC-MS Compatibility

This is the central strategic rationale for the chosen mobile phase system:

 Degradant Identification: A key deficiency in many initial SI-HPLC studies

is the lack of structural information for unknown degradation products. To
achieve definitive identification, the method must be transferable to an
advanced technique like LC-MS/MS.
 Ionization Enhancement: Formic acid and Acetic acid are ideal mobile
phase components for Electrospray Ionization (ESI) in MS. Their volatility
ensures they evaporate cleanly in the MS interface, preventing signal
suppression and instrument contamination. The acidic nature also promotes
protonation of BRV and potential degradants, significantly enhancing the
sensitivity and quality of the mass spectrometric data.

In simple way, the research is not merely developing another HPLC method; it is
designing a cost-effective, robust SI-HPLC platform that simultaneously satisfies all
immediate regulatory requirements for quality control while being structurally and
functionally optimized for advanced structural characterization in future research
phases. This duality significantly increases the method's value and applicability.
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4. Review of Literature:

4.1. Gillard, M., Fuks, B., et al (2011)

Early research by this group established the pharmacological profile and reasoning
behind developing Brivaracetam (BRV) as a third-generation antiepileptic drug.
They provided key data on how BRV binds to the Synaptic Vesicle Protein 2A
(SV2A), which is an integral membrane glycoprotein in presynaptic vesicles. The
study showed that BRV has a highly selective and strong binding mechanism. This
mechanism controls neurotransmitter release, leading to significant anticonvulsant
activity with fewer side effects than earlier treatments. This binding site is seen as the
main target for its effectiveness in managing partial-onset seizures.

4.2. Nicolas, G.D., Kenda, B.M., et al (2016)

This work summarized the clinical role, mechanism of action, and improved
therapeutic index of Brivaracetam compared to its predecessor, Levetiracetam. It is
designed to treat partial-onset seizures in adults and children, the drug's rapid brain
penetration and higher affinity for SV2A contribute to its significant clinical
advantages. This research established the drug's regulatory and commercial
importance, reinforcing the critical need for robust analytical control systems to
ensure patient safety and dose uniformity. The study also highlighted the drug's
metabolic pathway, which became a baseline for understanding its chemical stability.

4.3. Mhaske, D.V., & Mali, N.V. (2016)

These study conducted early HPLC studies focused on the degradation behavior of
Brivaracetam to develop a validated stability-indicating assay method. This work
served as an essential precursor in the analytical field, identifying fundamental
challenges such as the selection of an appropriate UV detection wavelength and
evaluating the solubility of the drug in various solvent systems. Their findings
provided the initial framework for understanding how BRV reacts under various
environmental stressors, emphasizing that the amide group in the pyrrolidine ring is a
potential site for hydrolytic cleavage.

4.4. Pasha, S. I., Varanasi, M. B., et al (2017)

By using chromogenic spectrophotometric estimation for Brivaracetam in both bulk

drug and formulation, the authors investigated an alternative analytical route. They
demonstrated a quantitative measurement method that differed from
chromatographic techniques by using Folin's phenol reagent. Since
spectrophotometry lacks the separation power needed for contemporary stability-
 7

indicating protocols, this study demonstrated the continued need for more selective
methods capable of differentiating the drug from complex impurities and excipients,
even though it is useful for basic estimation in low-resource settings.

4.5. Vasanth, D. A., & Rajkamal, B. A. (2017)

This study developed and validated a highly sensitive LC-MS/MS method

specifically for pharmacokinetic studies of Brivaracetam in healthy rabbits. By
successfully measuring drug concentrations in biological matrices with high
precision and low limits of detection, the research confirmed that mass spectrometry
is a superior tool for BRV analysis. This work laid the groundwork for future
researchers to consider LC-MS compatibility when designing new HPLC methods,
particularly regarding the use of volatile buffers instead of non-volatile phosphate
buffers.

4.6. Vishweshwar, V., Babu, J. M., et al (2018)

In a significant advancement for quality control, these researchers developed and

validated a Stability-Indicating UPLC method for the determination of Brivaracetam
and its related impurities. The method utilized sub-2-micron particle columns to
resolve the intact drug from its various degradation products with high precision and
significantly reduced run times. This work set a rigorous standard for resolution
parameters and validation criteria required by regulatory bodies like the ICH,
demonstrating that UPLC is the preferred choice for high-throughput pharmaceutical
analysis.

4.7. Jain, P. S., & Jain, N. K. (2019)

This study reported the development and validation of a stability-indicating RP-

HPLC method specifically for the estimation of Brivaracetam. Their research was
pivotal in documenting BRV’s susceptibility to hydrolytic and oxidative stress
conditions. By performing forced degradation studies according to ICH guidelines,
the authors proved that specialized analytical procedures are required to ensure that
concentration measurements are not compromised by breakdown products formed
during long-term storage or under accidental exposure to heat and moisture.

4.8. Devarajan, V., & Ganesan, V. (2019)

The authors given a validated stability-indicating RP-HPLC method for determining

Brivaracetam in both bulk drug and pharmaceutical dosage forms. Their work
focused on ensuring that the method was sensitive enough to detect low levels of
impurities that might impact the drug's safety profile. This research consistently
confirmed the industry-wide requirement for methods that can effectively separate
 8

the API from its various chemical liabilities, ensuring that the therapeutic efficacy of
the tablets remains consistent across different batches.

4.9. Baksam, V. N., Pocha, V. R., et al (2020)

This research used the critical and complex issue of stereoisomeric impurities in
Brivaracetam using chiral HPLC. Because BRV is marketed as the S-enantiomer,
any formation of the R-enantiomer during synthesis or processing is considered a
critical impurity. The authors developed a novel method to resolve these
stereoisomers, highlighting a specific but vital aspect of stability-indicating analysis.

4.10. Shanbhag, V. A., et al (2020)

This research studied on high-throughput analysis by developing and validating a

UPLC method for the determination of Brivaracetam in dosage forms. Their research
emphasized the use of modern instrumentation to achieve faster analysis times
without sacrificing the accuracy or precision required for regulatory compliance.
This study contributed to the trend of evolving analytical methods toward greater
laboratory efficiency.

4.11. Ilayaraja, P., Maniavannan, M., et al (2022)

This study focused on the simultaneous determination of Brivaracetam and its

various isomers using RP-HPLC. By demonstrating the difficulty of separating
closely related chemical structures, the authors highlighted the need for high
chromatographic selectivity. This research is particularly relevant for ensuring the
high purity of the active pharmaceutical ingredient in commercial products.

4.12. Saini N., Sharma M., et al (2023)

The authors developed a novel stability-indicating RP-HPLC method, reflecting the

continuous effort to optimize Brivaracetam analysis for better robustness. Their
research prioritized the use of accessible and cost-effective mobile phases, such as
mixtures of methanol and phosphate buffers, while maintaining the strict resolution
standards required for impurity profiling. This work serves as a modern example of
balancing analytical performance with laboratory economic constraints without
compromising data quality.

4.13. Mounika, K., Kuchana, V., et al (2023)

This work focused on the estimation of Brivaracetam in marketed tablet formulations

using a newly developed RP-HPLC method. The study emphasized the practical
application of analytical methods in a commercial setting, ensuring that the presence
 9

of excipients does not interfere with the accurate measurement of the drug. Their
validation data confirmed the method's reliability for routine pharmaceutical
analysis.

4.14. Xu, K., et al (2024)

This research utilized LC-QTOF-MS for the identification and pathway elucidation
of Brivaracetam degradation products. By providing structural information and mass-
to-charge ratios for previously unknown degradants, the study highlighted a major
deficiency in traditional HPLC methods that only provide retention times. Their
findings strongly support the strategic design of HPLC methods that are compatible
with mass spectrometry, such as those using ammonium acetate or formic acid, to
allow for deeper chemical characterization.

4.15. Sawant, S. D., & Godse, S. D. (2025)

This research work directly addresses the integration of LC-ESI-MS characterization

with the development of stability-indicating RP-HPLC methods. By aligning the
chromatographic separation with mass spectrometric detection, this research creates
a comprehensive platform for drug control. It represents the future direction of the
field, where routine quality control and advanced structural analysis are combined
into a single, efficient workflow, allowing researchers to monitor not just the
quantity of the drug, but also the identity of every peak in the chromatogram.
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5. Aims and Objectives, Hypothesis, Methodology, Tools/Techniques

5.1. Aims and Objectives

The primary Aim of this research is the development and rigorous validation of a
new stability-indicating Reversed-Phase HPLC (RP-HPLC) method for the accurate
determination of Brivaracetam in its pharmaceutical dosage forms. To achieve this,
the research pursues several key Objectives. The specific objectives are:
 To develop a simple, selective, economical, and LC-MS-compatible
Reversed-Phase HPLC (RP-HPLC) method for the quantitative determination
of Brivaracetam (BRV) in bulk and pharmaceutical formulations.
 To subject the BRV drug substance to forced degradation studies under
various stress conditions (acidic, basic, oxidative, thermal, and photolytic) as
per ICH Q1A (R2) guidelines.
 To establish the stability-indicating capability of the developed method by
ensuring complete chromatographic resolution ( Rs ≥ 1.5) of the Brivaracetam
peak from all degradation products and impurities.
 To validate the developed stability-indicating method rigorously for all
essential parameters including Specificity, Linearity, Accuracy, Precision,
Limit of Detection (LOD), Limit of Quantitation (LOQ), and Robustness in
compliance with ICH Q2 (R1) guidelines.
 To apply the fully validated method for the routine assay and stability
analysis of Brivaracetam in its commercially available pharmaceutical dosage
forms.

5.2 Hypothesis

A Reversed-Phase HPLC (RP-HPLC) method optimized using a volatile, acidic

mobile phase of Methanol (MeOH) and 0.1% Formic acid (FA) + 0.1% Acetic acid
(AA) can achieve the necessary chromatographic selectivity and resolution to serve
as a Stability-Indicating Assay Method (SIAM) for Brivaracetam, while
simultaneously providing an adaptable platform for future LC-MS based degradation
product identification.

5.3. Methodology

The research will proceed in a three-phase structure:

a. Phase I: Optimization and Method Development

1. Selection of Chromatographic System: Use of a C18 column 250 mm times

4.6 mm, 5 µm) as the stationary phase, which is standard for RP-HPLC of
moderate-polarity compounds like BRV.
 11

2. Mobile Phase Optimization: Initial screening will test varying ratios of

MeOH vs. Aqueous Phase (0.1% FA + 0.1% AA). Gradient elution may be
explored if isocratic separation proves insufficient for all degradants.
3. Chromatographic Conditions:

 Flow Rate: Optimized between 0.8 to 1.2 mL/min.

 Detection Wavelength: UV detection optimized at the maximum
absorption (λmax) of BRV (expected around 200-210 nm).
 Column Temperature: Maintaining temperature (e.g.,30°C) for
consistent retention times.

b. Phase II: Forced Degradation Studies (Stress Testing)

BRV bulk drug substance will be exposed to severe stress conditions (as per ICH
Q1A R2), aiming for 10-20% degradation:

Stress Expected
Reagent/Parameter
Condition Degradation

Acid Amide bond cleavage

0.1 N HCl at 60°C for 6-12 hours
Hydrolysis (Hydrolysis)

Base Amide bond cleavage

0.1 N NaOH at 60°C for 6-12 hours
Hydrolysis (Hydrolysis)

3-10% H2O2 at ambient temperature Oxidation of

Oxidation
for 24 hours functional groups

Thermal
Thermal
80°C in a Hot Air Oven for 7 days decomposition,
Degradation
typically minor

Exposure to UV light (200 Wh/m2)

Photolytic Photolytic cleavage or
and visible light (1.2 million lux
Degradation rearrangement
hours)

c. Phase III: Method Validation

Validation will strictly follow ICH Q2(R1):

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1. Specificity/Selectivity: Demonstrated by baseline separation (Rs ≥ 1.5) of

BRV from its degradation products, placebo, and known impurities.
2. Linearity: Establish a working range (e.g.,50% to 150%} of assay
concentration) with a correlation coefficient (R2) greater than 0.999.
3. Accuracy (Recovery): Assessed by the standard addition method at three
levels (50%, 100%, 150%).
4. Precision (Repeatability & Intermediate Precision): Measured as the Relative
Standard Deviation (RSD), required to be ≤ 2.0%.
5. Limits of Detection (LOD) and Quantitation (LOQ): Calculated based on the
standard deviation of the response (σ) and the slope of the calibration curve
(S), using LOD = 3.3 σ/S and LOQ = 10 σ/S.
6. Robustness: Evaluation of the method's reliability under small, deliberate
variations (e.g., ±5% change in MeOH ratio, ±0.1 mL/min change in flow
rate, ±2°C change in column temperature).

5.4. Tools and Techniques

 HPLC System: High-Performance Liquid Chromatograph equipped with a

Quaternary Pump, UV PDA Detector, and Autosampler.
 Column: C18 (e.g., L1 USP classification).
 Software: Chromatographic Data System (CDS) for data acquisition and
processing.
 Reagents: HPLC-grade Methanol, Formic acid (0.1%), Acetic acid (0.1%),
HCl, NaOH, H2O2.
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6. Proposed Plan of Work, Expected Outcomes / Results and

Discussions

6.1. Proposed Plan of Work

Key
Duration
Phase Activity Detailed Activities Deliverab
(Months)
les

Detailed
chronological review
of literature (HPLC,
SI-HPLC, BRV, ICH
Complete
Q1A (R2), Q2 (R1).
Literature, Literature
Procurement of BRV
Procurement Review.
1 Month standard, HPLC-
1 & Standardiz
(M1) grade solvents, and
Standardizat ed
reagents (MeOH, FA,
ion Working
AA, HCl, NaOH,
Solutions.
H2O2).
Standardization of
primary and working
stock solutions.

Mobile Phase
Screening: Initial
Optimal
experiments using
Column
isocratic and gradient
and
elution with varying
Mobile
ratios of MeOH and
Phase
Method FA, AA mixture.
Ratio
Screening & 2 Months Column Selection:
2 establishe
Optimizatio (M2-M3) Testing C18 columns
d. Initial
n (Initial) with different
Chromato
dimensions/particle
gram with
sizes. Optimization of
Good
flow rate (0.8-1.2
Peak
mL/min) and
Shape.
detection wavelength
(λmax of BRV).
 14

Key
Duration
Phase Activity Detailed Activities Deliverab
(Months)
les

Stress Sample Forced

Generation: Degradati
Subjecting BRV to on
Acid, Base, Profiles
Oxidative, Thermal, completed
and Photolytic .
Forced
conditions. Sample Demonstr
Degradation
2 Months Analysis: Running all ation of
3 Studies
(M4-M5) stressed samples on Stability-
(Stress
the optimized Indicating
Testing)
method. Identifying Capability
the number of (Separatio
degradation products n of BRV
formed and the from all
percentage of drug degradant
degradation. s, Rs ≥ 1.5).

Specificity: Testing
placebo and mixed
degradants to confirm
purity (using PDA).
Linearity & Range:
Establishing the Validated
calibration curve Specificit
Method
(R2>0.999). y,
Validation 2 Months
4 LOD & LOQ: Linearity,
(Core (M6-M7)
Determining the Accuracy,
Parameters)
detection and LOD, and
quantitation limits. LOQ.
Accuracy:
Performing spike-
recovery experiments
at three levels (50%,
100%, 150%).

5 Robustness 1 Month Robustness: Testing Final

 15

Key
Duration
Phase Activity Detailed Activities Deliverab
(Months)
les

& (M8) small, deliberate Robustnes

Preliminary variations in method s results.
Application parameters (e.g., ±5% Preliminar
MeOH ratio, ± 1 y Assay of
mL/min flow rate). Dosage
Preliminary Form.
Application: Data
Applying the Ready for
validated method to a Discussio
commercial BRV n/Thesis
tablet sample. Drafting.
Data Consolidation:
Interpretation and
statistical analysis of
all results from M1-
M8.

6.2. Expected Outcomes / Results

 A validated, cost-effective, and robust stability-indicating RP-HPLC method

will be successfully developed, utilizing the LC-MS friendly mobile phase
(MeOH + 0.1% FA + 0.1% AA).
 The method will achieve baseline resolution (Rs ≥ 1.5).between BRV and its
major degradation products, particularly the hydrolytic products expected
from the amide bond.
 Validation data will show high Linearity (R2>0.999) and Accuracy (Recovery
98-102%).
 The degradation behavior of BRV under all stress conditions will be
systematically documented, confirming the compound's inherent stability
profile.

6.3. Discussions

The discussion will center on the novelty and advantages of the developed method:

1. LC-MS Compatibility: The inclusion of volatile modifiers (FA and AA)

will be highlighted as a superior feature, contrasting it with non-volatile
 16

buffer systems (Phosphate or Sulphate salts) often used in traditional HPLC

methods. This prepares the groundwork for future degradation product
identification.
2. Selectivity and Degradation Pathway: The chromatographic separation will
be discussed in the context of the anticipated chemical degradation (e.g.,
hydrolytic cleavage of the amide bond). Any unusual degradation products
will be noted, contributing new information to the BRV stability profile.
3. Robustness Analysis: Detailed discussion of the method's resilience against
small
 17

7. Conclusions and Suggestions

7.1. Conclusions

The proposed research will successfully establish a stability-indicating RP-HPLC

method for Brivaracetam that is economical, specific, and fully validated according
to international guidelines. The strategic choice of the mobile phase (MeOH/ FA/
AA) is a key achievement, creating a platform that is not only suitable for routine QC
but is also optimized for subsequent structure elucidation studies using LC-MS/MS,
thereby ensuring a comprehensive analytical control strategy for Brivaracetam.

7.2. Suggestions for Future Work

1. Structure Elucidation: Utilize the developed method and couple it with LC-
MS/MS to definitively identify the exact chemical structures of the major
degradation products.
2. Chiral Purity: Extend the analysis to develop a chiral HPLC method for
separating and quantifying the R-enantiomer impurity of Brivaracetam.
3. Dissolution Testing: Adapt the final robust method for use in dissolution
testing of BRV tablets, optimizing it for high-throughput analysis of
dissolution media.
 18

8. References

1. App, D. et al. (2024). A Brief Review of Analytical Methods for the

Estimation of Brivaracetam in Pharmaceutical Formulation and Biological
Matrices. Asian Journal of Pharmaceutical Research and Development, Vol.
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2. Baksam, V. N., Pocha, V. R., Chakka, V. B., Ummadi, R. R., & Kumar, P.
(2020). Development of an effective novel validated stability‐indicating
HPLC method for the resolution of Brivaracetam stereoisomeric impurities.
Chirality, Vol. 32(9), pp. 1208-1219.
3. Bhamare, P., Dubey, R., Upmanyu, N., & Umadoss, P. (2021). A simple
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DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF
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 19

11. ICH Q2(R1) Expert Working Group. (2005). Validation of Analytical

Procedures: Text and Methodology. ICH Harmonised Tripartite Guideline,
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