T&E - Analítica



The study of Impurities of Degradation regulated by RDC No. 53 of 04/12/2000-ANVISA and Guide 04/2015 applies to the Active (raw material-IFA), Placebo (excipients of the formulation) and Final Product (Formulated: Placebo + IFA) with the objective of subjecting the three components of the drug to verify possible degradations and interactions between IFA and Placebo that could affect the user.

Here are three tests recommended by DRC 53 of December 4, 2015:


·         Notification limit: value above which a degradation product should be reported in the stability study (s);

·         Limit of identification: value above which a degradation product must have its chemical structure identified;

·         Limit of qualification: value above which a degradation product should be qualified.



Table of Limits


The degradation profile consists of a description of the results and the analytical activities used in the detection, identification, structural elucidation and quantitative determination of the degradation products present in the active pharmaceutical ingredient and in the medicament.

The drug stress test is chemically and physically treated by subjecting Placebo, IFA and Finished Product in contact with water, acid solution, alkaline solution, oxidizing solution and solution containing metals such as chemical tests and dry temperature and light as physical tests. After a certain contact time or exposure of Placebo, IFA and Finished Product, there is a comparison of these samples before stress with those resulting after stress period during the study of Limits of Notification. The method used in the reporting study should be indicative of stability and should be duly validated.

The indicative existence of a new substance, if present, also in the stability studies, is treated in order to identify the impurity, in the second step, study of Identification Limit, which after identification, should be evaluated as to the need to be submitted to toxicity tests and / or carcinogenicity.

Prediction of Impurity: it is the deep theoretical study of the molecule in front of the stresses. It involves advanced bibliographic research and chemical study on the chemical functions that make up the structure of the molecule. In this way, possible impurities can be predicted during the practical study.

To carry out the study of IMPURITIES OF DEGRADATION, the T & E Center has a diversity of analytical techniques of separation, quantification, preparation and identification, as well as experienced professionals to perform and interpret the phenomena that occurred during the experiment. Thus, the T & E Analytical Center serves the pharmaceutical industry in studies by RDC No. 53/2015 in:

·         Theoretical prediction studies;

·         Limit of notification test;

·         Identification limit test;

·         Limit of qualification test;

Note: T&E has published the only book in Brazil on Degradation Impurities and offers free mini-course to the T&E partnerships to share our experience and guide the complexity in carrying out this essay.



Identification and Quantification

For the identification and quantification the analytical techniques will be in function of the molecule and of the analytical resources employed, among the techniques most used are:

·         Gas-phase Chromatography (GC-FID-ECD-TCD-MS-Head Space);

·         Liquid phase chromatography (UPLC or HPLC-UV / Vis; Fluorescence; ELSD);

·         LC-MS / MS: Mass Spectrometry coupled to HPLC or UPLC;

·         ICP-OES (Induced-Simultaneous and Sequential Plasma);

·         AA - (Atomic Absorption) and FC (photometry of flame).

·         DSC; TG / DTG; IVFT; UV / Vis; High resolution microscopy, wet testing; among others.

·         NMR  H and C; Diffraction X.

·         Determination of peak purity using DAD


During the analytical test it may be necessary to extract or concentrate a possible contaminant signal, in this case one may use various methodologies such as:

·         Liquid-Liquid Extraction,

·         SPE (Solid Phase Extraction) extraction,

·         Solid Phase Micro Extraction (SPME) extraction,

·         SBE Extraction (Stil Bar Extraction)

·         Isolation and Extraction via Preparative Liquid Chromatography

·         Isolation by colligative properties

·         Resin insulation

·         Direct analysis by instrumental couplings

·         Among others, both for organic and for metals.


Every study begins with a protocol and ends with a Report and Certificate as standardized by ANVISA. The methodologies adopted are preferably pharmacopoeial (Brazilian Farm, American Farm, British Farm, European Farm, Japanese Farm ..), and when none of these compendia is available, we can propose the development of a methodology for the products or use of the methodology provided by the contracting company, performing prior to the conduction of the study a total or partial validation according to RDC 166 of July 24, 2017, as well as, all other items are governed by ANVISA or international standards. The documentation: is archived in the center the provision for consultation at any time by the contractor or regulatory agency.