**3. Compendial analytical procedures**

*Analytical Chemistry - Advancement, Perspectives and Applications*

analytical activity must follow Good Analytical Practices.

*Flow chart showing essential requirements of registration of pharmaceutical product.*

drug products (**Figure 1**).

development.

ensure compliance with standards and potency should be part of Biologics license application (BLA). It is must to meet all standards of guidelines provided for analytical procedures. All these parameters should be suitable for their purpose wherever applicable. Detail Analytical procedures including detail validation parameters are the important part of Electronic Common Technical Document Specification as per International conference of Harmonization (ICH). The analytical procedure is Food and Drug Approved (FDA) if it a part of Approved NDA, ANDA or BLA. These methods can be generated from FDA recognized sources like U. S. Pharmacopeia/National Formulary (USP/NF) or if anyone submits validated procedure that will be accepted by FDA. The only validation or verification data of FDA approved methods of new products are considered for applications to various

Every manufacturer must generate large amount of corrected data for safety and efficacy of drug for commercial viewpoint. As it is mandatory to follow Current Good Manufacturing practices (cGMPs) for manufacturing purpose, likewise each

Method Validation, calibrated instrument, and training are three important tasks of Good analytical practices (GAPs). Commercially available dosage form is an outcome of several steps which are systematically carried out during product development. It is very important that all steps should be carried in systematic manner to ensure complete drug development stage. In recent years there is special focus on efficiency and efficacy of drug product and for this clinical study is most important task but apart from this there are various behind the scene activities are associated with drug development process without which pharmaceutical drug development is not possible. Among these behind the scene activities Method Development and Validation has its own uniqueness to ensure the drug

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**Figure 1.**

The analytical procedure official in pharmacopeias, are cross checked for implementation stage and its suitability should be checked. The verification protocol should include details of data which explains suitable analytical procedure official in USP/NF for drug product or drug substance.

The following points are to be included in the verification protocol


#### **3.1 Statistical analysis**

The statistical analysis is important work after finishing the method development and validation. The statistical values of validation are compared with the predetermined acceptance criteria.

The statistical parameters used are based on proper principle and required for evaluation of parameter. The methods like analysis of variance (ANOVA) for analysis of regression analysis, (R2 -Correlation coefficient) to measure the linearity are applied for studying validation characteristics. In case of observed data is not distributed then it is transformed normal distribution or distribution free approach. By using validates software or independent verification for correctness the data can be analyzed.

#### **3.2 System suitability requirement for potency assay**

Before starting actual analysis of standard sample, it is necessary to check whether system is working properly or not. This important task can be completed by analyzing system suitability. In this all integral system that is equipment, electronics, analytical operations and samples are evaluated. These system suitability parameters are depending on method or procedure under validation.

System suitability can be evaluated according to following points:

1.The system suitability measures the performance of given system of samples on a given day. 2. The variable parameters like chromatographic columns, column aging, mobile-phase variations, changes in instrumentation are checked whether they are working properly or not.


The important aspects of pharmaceutical development program are analytical method development, validation method transfer but it is fact that they are less considered in sense of total contribution in development process, time and economy. At the time of drug development phases, all the analytical method related activities are interrelated. In early development stages they are related and occur one after another in coming phases of development. During drug development process the changes may require to be performed in current methods and these changes in methods again requires validation or method transfer treatment.

If one's objective of method development and validation is achieved, then it can prove that the laboratory facilities are accurate and fit for further development process that is one can say optimized. Method validation is the "process of demonstrating that analytical procedures are suitable for their intended use." Both method validation and methods transfer have important share in drug development and further changes in methods. To generate supportive data during manufacturing and quality control, these methods provides a valuable data by comparing with specifications including all types stability study, Safety, characterization and drug performance can obtain with these supportive methods.

Method development is the simultaneous process as the gradual development of drug product continues. The system suitability parameters are set of tests to checks the proper working of the system. After performing robustness with proper statistical data collection one can set the criteria for final system suitability of the method.

These methods focus on active pharmaceutical ingredient (API) behavior. As the knowledge about API and drug product goes on progress the analytical methods become more refined. The important aspects of analytical method are that should be robust, simple and meeting the regulatory guidelines. Various trial an error experiments are to be carried out to develop the method. The performance criterion's to be finalized before the final validation of method. Forced degradation study which is integral part of stability-indicating method and system suitability tests are one of the key points of method development and final validation [3].

#### **3.3 Impurity profiling**

The International Conference for Harmonization (ICH) guidances are available that are related to the qualification of impurities in new drug substances that are produced by chemical synthesis. These impurities can be addressed in two perspectives that are chemistry aspects and safety aspects. The chemistry aspects explain the identification and classification of impurities, the various analytical procedures and setting of specifications. In the safety aspects explains the qualification of impurities which are not addressed in clinical trials. In this aspect the threshold limits are defined. This ICH guidance classifies impurities in three classes as Organic, Inorganic and Solvent. Each class of impurities should be properly reported, with all aspects, developed during synthesis, storage of the new drug product. The

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*Analytical, Bioanalytical, Stability-Indicating Methods: Key Part of Regulatory Submissions*

analytical procedure including validation reports related to impurities should be properly reported [4]. In concern with the above discussed guidelines related to qualification of impurities in new drug substances produced by chemical synthesis, impurities which are classified as degradation product developed with the reaction with excipients or container closure system may termed as reaction products. All the observed degradants during manufacturing and stability study should be reported. All the data related to their identification, specification, analytical procedures for quantification, their limits of detection and quantification should be

To ensure safety, efficacy and quality of drug product there are need of stability indicating methods. The Food and Drug Administration (FDA) defines the stability indicating method as a validated analytical procedure that accurate and precisely measure active ingredients (drug substance or drug product) free from process

To obtain forced degraded samples for assessing selectivity of method, method development and method validation are three important steps of stability indicat-

As per guidance document available for stability indicating method does not contain any explanation about extent up to which the degradation should be carried out. There are no certain guidelines regarding stress degradation. Therefore, it is always necessary to keep all experimental conditions of degradation with more

The main purpose of forced degradation is to obtain stability of drug. It should provide information about route of degradation and utility towards the stability

Forced degradation can be able to judge excipients or non-drug substances. It also provides information useful for structure elucidation of degradation products. Importantly it gives data related to thermal, hydrolysis, and oxidation and photo degradation behavior of drug substance and drug product. It is very important to know about chemical behavior of drug product and drug substance in formulation development, manufacturing and packaging. The data helps in quality improvement [7].

There are vast discussions among the various scientists that what should be the limits of stress testing? Generally, values in between 5–20% are proper and acceptable for chromatographic assays. The acceptable stability limit for small molecules are 90% of label claim and generally employed by pharmaceutical scientist that is 10% degradation is optimum for use in analytical validation. There are some experiments in which very little or no degradants are obtained due to exceptional stability

be carried out. If positive result is obtained, then the stability of drug is noted. But

According to the recent recommendations of Food and Drug administration (FDA) and ICH guidelines, the stability indicating property of analytical method can be obtained by carrying out forced degradation study. The pathway

c for 6 months should

*DOI: http://dx.doi.org/10.5772/intechopen.93566*

**4. Stability-indicating method**

**4.1 Importance of forced degradation**

reality and deliberate degradation.

**4.2 Acceptance criteria for forced degradation**

of molecule under study in such case accelerated storage 400

overstressing the drug substance may produce false results [7].

impurities, excipients and degradation products [6].

reported [5].

ing methods.

indicating [6].

*Analytical, Bioanalytical, Stability-Indicating Methods: Key Part of Regulatory Submissions DOI: http://dx.doi.org/10.5772/intechopen.93566*

analytical procedure including validation reports related to impurities should be properly reported [4]. In concern with the above discussed guidelines related to qualification of impurities in new drug substances produced by chemical synthesis, impurities which are classified as degradation product developed with the reaction with excipients or container closure system may termed as reaction products. All the observed degradants during manufacturing and stability study should be reported. All the data related to their identification, specification, analytical procedures for quantification, their limits of detection and quantification should be reported [5].
