Analytical Method and Analytical Method Development Approaches

Table of Contents

• Introduction and Outcome
• Analytical method development and Analytical Method
• Types of Analytical Method Development
• Techniques for Analytical Method Development
• Strategy for Analytical Method Development
• Essential components of any new method
• Applications of a new Analytical Method
• Difference between Analytical method and Analytical method development
• Conclusion

The Analytical Method Development Approach determines the reliability of the Analytical Method. Be it reaction monitoring, impurity profile, content test, residual solvents test, assay or characterisation, all are conducted through Analytical methods. Therefore, the Analytical method plays a crucial role in ensuring the quality, efficacy and safety of any drug substances or their products. In this article; I will share skill-based knowledge on Analytical method development so that you can apply it during Analytical method development. This article will also enable you to answer several questions:

• What is the Analytical method?
• What is the Analytical method development and how it is performed?
• What are the different methods?
• What are the different techniques used for Analytical method development?
• What is the difference between Analytical method and Analytical method development?

Related topic: Analytical Method Development and Validation in Pharma

What is Analytical Method?

The Analytical method is the pre-established procedure for measuring the concentration of an active moiety or impurities in the drug substance or its stages and in the drug products.

What is analytical development Approach?

Analytical method development is the process of establishing a set of analytical parameters/conditions that when executed give reliable results.

Examples:

• In the HPLC method, various analytical parameters like column, mobile phase, diluent, sample concentration, wavelength, flow rate, column temperature, injection volume are established.
• In assay by chemical method: Titration condition parameters like sample weight, volumetric solution, solvent, and indicator are established
• In UV; Parameters like wavelength or wavelength range, sample concentration, and diluent are established
• In IR; Parameters like wavenumber range, number of scans, and sample preparation are established

Qualitative and Quantitative Analysis or Types of Analytical Analysis

• Qualitative method: It focuses on identifying the pharmaceuticals or presence of specific substances within a sample.
• Quantitative: It focuses on the exact amount or concentration of a substance in a sample

Difference between Qualitative and Quantitative analysis

Both qualitative and quantitative analysis play a vital role in pharmaceutical development. The following is the difference between qualitative and quantitative analysis:

Qualitative Pharmaceutical Analysis:

• Purpose: It focuses on identifying the chemical composition or presence of specific substances within a sample.
• Process: Involves methods to detect or confirm the presence of active pharmaceutical ingredients (APIs), impurities, or contaminants, but does not quantify their amount.
• Techniques:
  • Chromatography (e.g., TLC, HPLC for identification): To confirm the identity of a compound.
  • Spectroscopy (e.g., UV-Vis, IR): To detect and confirm the presence of specific functional groups or molecules.
  • Chemical Reactions: Specific chemical tests that indicate the presence of certain compounds.
• Outcome: The result is usually a yes/no determination or a qualitative statement (e.g., “This substance is present”).

Quantitative Pharmaceutical Analysis:

• Purpose: It determines the exact amount or concentration of a substance in a sample.
• Process: Focuses on measuring the concentration of APIs, excipients, or contaminants to ensure they meet specific standards and regulatory requirements.
• Techniques:
  • Titration: To quantify the amount of a substance by reacting it with a reagent of known concentration.
  • Chromatography (e.g., HPLC, GC): Quantification of substances based on their peak areas or retention times.
  • Spectrophotometry: Measures the absorbance or emission of light to quantify a substance.
  • LCMS and GCMS: Quantifies the amount of each molecule based on its mass and intensity.
• Outcome: The result is a numeric value indicating concentration or amount (e.g., %, Each 5 ml contains 100mg of paracetamol).

Note:

• Qualitative analysis identifies the what is in a sample.
• Quantitative analysis measures the how much of a substance is in a sample.

Analytical Tests and Analytical Methods

The following Analytical Tests are widely used to ensure the quality, efficacy and safety of any pharmaceuticals:

• Method for Identification test
• Method for Related Substances/Impurity profile test
• Assay
• Method for Characterization
• Content test

Method for Identification test

Chromatographic techniques (e.g.HPLC, GC and TLC), spectroscopic techniques (e.g. FTIR, UV) and chemical techniques (e.g. colour development) are used for identification test

Method for Related Substances/Impurity profile test

HPLC and GC are widely used for this test in the Pharmaceutical industries

Assay

Chromatographic techniques (e.g.HPLC, GC), spectroscopic techniques (e.g. UV) and chemical techniques (e.g. titration) are used for Assay test

Content test

Chromatographic techniques(e.g.HPLC, GC), spectroscopic techniques (e.g. UV, AAS), mass spectroscopy techniques (e.g. LC-MS, GC-MS, ICP-MS) and chemical techniques (e.g. colour development) are used for identification test

Method for Characterization

NMR, chromatographic techniques(e.g.HPLC, GC), spectroscopic techniques (e.g. UV, AAS), mass spectroscopy techniques (e.g. LC-MS, GC-MS, ICP-MS) and chemical techniques (e.g. color development) are used for characterisation of Drug substances and their stages

Analytical Method and Different Analytical Techniques

The following Analytical techniques are used for Analytical method development:

• HPLC: HPLC is one of the widely used analytical technique in the pharmaceutical development. It gives specific accurate and precise results.
• GC: GC is the second widely used analytical technique in the pharmaceutical development. It gives specific accurate and precise result. It is used for volatile molecules.
• FTIR: It is one of the most used instruments for the identification test. It gives fast result.
• TLC: TLC is a widely used traditional analytical technique in pharmaceutical development. It gives specific results. It is mainly used for qualitative analysis like identification test and monitoring the reaction
• LC-MS: It is widely used for unknown impurity identification and structure characterisation
• GC-MS: It is widely used for unknown impurity identification and structure characterisation for volatile molecules
• UV: It is used for both identification test and assay test.
• KF apparatus: It is used for water determination test.
• pH meter: It is one of the most used instruments in chromatographic method development as well as traditional method development.
• Autotitrator: It is one of the most widely techniques for assay test by titration method.
• XRD: It is used for polymorphic study.
• NMR: It is mainly used for structure characterisation
• LOD oven: It is used for loss and drying test
• Balance: It is used for weight measurement test

Analytical Method Development Approaches

The following 7-steps process are recommended during method development

Step-1: Define the aim of the method: In this step type of method e.g. related substances. assay, content test, identification test and their corresponding instruments should be proposed

Step-2: Literature search: A Literature search should be performed and A Literature report containing all available informations should be prepared. It is very helpful for Analytical method Development ( in deciding the method conditions)

Step-3: Design the Analytical Method Development: This step is very helpful in performing the experiment as well as concluding the result for the next experiment.

Step-4: Optimize the Analytical method: In this step optimization of the method should be done based on the conclusion in step-3 e.g. optimisation in mobile phase ratio, column temperature, flow rate, column chemistry ( In case of HPLC method development)

Step-5: Verification of the Analytical method: In this step Verification of the Analytical method should be performed. e.g. precision, linearity, recovery, and three samples analysis by two analysts using different instruments.

Step-6: Analytical method development report and STP preparation: In this step Analytical method development report and STP should be prepared and signed by the concerned persons. An Analytical method development report is a very important document since it is required by various regulatory agencies during audits. STP is used is required by QC to prepare monograph.

Step-7: Analytical method Transfer: In this step, Analytical method/STP should be provided to QC and Analytical method Transfer should be performed

Essential components of any new method

The following are the essential components of any new method:

• Chemical and reagents
• Mobile phase perpetration details
• Chromatographic conditions
• System suitability (SST)criteria
• RT and RRT Table
• Detection limit
• Quantitation limit
• Calculation
• Procedure
• Typical chromatogram (of blank, SST, QL, standard and sample)

Applications

• New Analytical method is required by QC to prepare the monograph
• Analytical method development report is required by Regulatory agencies during audit

Difference between Analytical method and Analytical method development

Analytical method is a defined and approved document required for intended analysis whereas Analytical method development is the process of establishing the analytical condition to give reliable results. Analytical method is prepared from the Analytical method development report.

Conclusion

Analytical method development is one of the major and challenging job of Analytical research department. It needs an innovative analytical skills. Hope this post has taken you to the next level and now you can develop the analytical method effectively. Write your learnings/questions related to this post in the comment section.

FAQs

What is R&D analytical development?

R&D analytical development develops the method to control the quality of medicine at different stages during the development stage. It also performs other activities like method validation, analytical technology transfer, literature search etc.

What are the steps involved in method development?

Analytical method development is the 7 steps process which includes Define the aim of the method, Literature search, Design the Analytical Method Development, Optimize the Analytical method, Verification of the Analytical method, Analytical method development report and STP preparation and Analytical method Transfer

What is AMD in pharma?

AMD is Analytical method development

What is the method development of HPLC?

HPLC method developments include several steps like selection of column, selection of mobile phase, selection of diluent, selection of wavelength, method optimisation etc.

Why is method development important?

The efficacy of the medicine depends upon its quality and the quality of a medicine is judge by the analytical method. That is why method development is important.

Why is the analytical method development required?

The analytical method development is required to develop the method to check quality of the medicine.

What is spiking in analytical method development?

In spiking method development known impurity is spiked with pharmaceutical containing unknown impurity to identify the unknown impurity.

References

• https://pharmaknowledgeforum.com/need-of-analytical-method-mini-validation/
• https://pharmaknowledgeforum.com/pharmaceutical-monograph/
• ICH guideline

Abbreviation

• ICP-MS: Inductively coupled plasma mass spectrometry
• AAS: Atomic absorption spectrometer
• STP: Standard test procedure
• QC: Quality control
• RT: Retention time
• RRT: Relative retention time
• KF: Karl Fischer
• FTIR:Fourier transform infrared spectrscopy
• UV: Ultraviolet spectrophotometer
• GC: Gas chromatography
• MS: mass spectrophotometer
• HPLC: High-performance liquid chromatography