ANALYTICAL DEVELOPMENT AND VALIDATION
ABOUT ANALYTICAL DEVELOPMENT OF A METHOD OF CONTENT
It is perhaps the most complex stage of the search for an analytical technique, since it involves materials and human resources. Material resources due to the detectability of all possible components (impurities or substances reported) present in the final product and to ensure that there are no shocking interferences, of these impurities in the quantification of the content of the active (drug). There are several techniques for developing methodologies of either active or impurities. Pharmaceutical preference falls to separative equipment, primarily by Liquid Chromatography (HPLC) if possible with detection by ultraviolet light absorption. However, a range of HPLC detectors may be required depending on the structure of the molecule. Gas-phase Chromatography (CFG) in its different detectors may be simpler to resolve the development.
DEVELOPMENT: Two definitions are important:
I. ANALYTICAL CONDITION: it is the operational condition of the equipment in which the signal of active (drug) is observed eluting of majority form. The analytical condition, by itself, is not an analytical method indicative of stability,
II. ANALYTICAL METHOD: is the Analytical Condition submitted to challenges, such as precision, accuracy, linearity, recovery, robustness, limits, among others. At this moment the partnership with the sponsor is fundamental.
There are several ways to start a development:
· Easy: to find an analytical condition or method, available in bibliographic research, for that drug in that product.
· Medium Difficulty: to find an analytical condition or method, available in the bibliographic search for that drug, or drug of similar structure in distinct pharmaceutical forms.
· Complex: after having tested the Easy or Medium Difficulty and they did not respond satisfactorily to the product under analysis.
· Difficult: to find no bibliographical references about that molecule, now inserted in the formulated under analysis.
VALIDATION: The validation of content and or impurities, follows RDC 166 of July 24, 2017.
For the analysis of the Easy and Medium Difficulty, and especially of the Complex and Difficult Analytical T&E has wide experience in these 20 years of existence. It has the material and human resources for sustainable development and indicative of stability.
Still considering the complexity, in a formulation, there may be more than one substance to be analyzed, in some cases there are 2 to 5 compounds and if we consider vitamin complexes, we can reach more than 10 compounds. In these cases the T&E Center seeks to obtain analytical condition that contains the maximum assets in order to reduce equipment, time and obviously costs.
In the case of HPLC, two are the situations to be observed, depending on the structure of the molecule besides the formulated itself to present interaction with the excipients present interfering in the separative process. In this way the search, considering the HPLC, could fall to the chromatography in Normal Phase (operationally) or Reverse Phase which allows greater operational flexibility and accepted in several detectors coupled to the HPLC. For the CFG, there will be a need for a volatile molecule, so the T & E Center has experience in analyzing the product itself or being derivatized to attribute volatility. Finally, the T&E Center will be prepared to meet the need to develop your company.
After determination of the ANALYTICAL CONDITION, the T&E Center can carry out the Validation or the company itself can carry it out and if necessary with our monitoring, orientation and or validation in the company itself.
Matrices or Formulated Products and Raw Material.
The T&E Analytical Center has experience in several types of products, such as:
· Pharmaceutical: levels in any matrix, including phytotherapics (using the QuEChERS procedure), vitamins and products at macro and micro levels;
· Veterinary: a process similar to that of drugs;
· Rations, Mix and Food: various methodologies, which are included in sample preparation, extractions, among others;
· Special: chemistry, petrochemical, aromas, cosmetics, special cases (incrustations, etc).
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:
· 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;
· RMN H and C; Diffraction X;
· Determination of peak purity using DAD;
For the inorganic analysis as:
Metals, semi-metals, amethals, organic or organic-metallic compounds (complexes or chelates) are usually destroyed either by solvation, sulfuric dehydration followed by nitric oxidation or by peroxides, ashes at different temperatures, reactions as a dry route, Schonninger, Mufla, and microwave reactions, among others, in order to obtain the element in a state of solubilization for introduction into the given technique.
In the case of metals, the instruments provided by the T&E Analytical Center are:
· ICP-OES (Induced-Simultaneous and Sequential Plasma);
· AA - (Atomic Absorption Spectrometer);
· FC (Flame Photometry);
· Analysis with hydride generation;
· Volumetric and potentiometric analyzes, complexiometric;
In the development process, in many situations it is necessary at some stage to carry out the extraction of the asset, either in the form as such or in the derivatized form. For extractions, the T & E Center seeks constant updating and it is cited the most used: extraction L-L, SPE, SPME; SBSE, Cold or Hot Extraction, "Head Space", among others. Extractions with programmed pHs, and often the "clean-up" used in the QuEChERS process, which is briefly described below:
QuEChERS (Quick, Easy, Cheap, Effective, Rugged, Safe)
I. The QuEChERS method is a modern method of sample preparation for multi-residue determination of pesticides in food by chromatographic methods coupled with mass spectrometry.
II. It is based on the extraction of the sample with acetonitrile containing 1% acetic acid and liquid-liquid partition simultaneously, formed by the addition of anhydrous magnesium sulfate and sodium acetate.
III. The extraction of the method consists in placing, in a centrifuge tube, 1 mL of acetonitrile containing 1% acetic acid and add 0.4 g of anhydrous sodium sulfate and 0.1 g of anhydrous sodium acetate per gram of sample.
IV. Then the tube should be centrifuged at 3000 rpm and one portion of the extract is transferred to another tube containing 50 mg of a secondary primary amine sorbent (PSA) and 150 mg of magnesium sulfate per ml of extract.
V. Dispersive Solid Phase Extraction (DSPE) - QuEChERS- is mainly used to remove polar components from the matrix (organic acids, some peppers and sugars)
VI. After mixing and centrifugation, the extract is analyzed.
Microbiological analyzes for sterile, non-sterile and antineoplastic products.
Determination of Content - Development and Validation, among other tests:
· Coliformes Thermo tolerant;
· Search of Salmonella;
· Countable viable cells;
· Challenge test;
· Endotoxins LAL;
· Determination of content (power);
· Vitamin analysis;
· Testing in lids and containers;
· Supports restaurant kitchens in contamination;
· Sampling and analysis of process air and packaging;
· Process validation analysis;
· Verification of industrial contamination;
Other situations, do not hesitate to contact us. The exchange of technical information may result in solving your company's problem.