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See detailDevelopment and Validation of LC Method for the Diclofenac Sodium Release Determination Using Accuracy Profile Concept
Al-Kamarany, M.A.; El Karbane, M; Belomaria, M et al

in Asian Journal of Chemistry (2012), 24(10), 4393-4400

The article presents a rapid and sensitive RP-HPLC method with UV detection, using the diclofenac sodium release determination in solid pharmaceutical formulations. A novel validation strategy based on ... [more ▼]

The article presents a rapid and sensitive RP-HPLC method with UV detection, using the diclofenac sodium release determination in solid pharmaceutical formulations. A novel validation strategy based on accuracy profiles was used to select the most appropriate regression model with highest accuracy within well defined acceptance limits. Furthermore, the strategy was used to determine the limits of quantification as well as the suitable concentration range. The validation phase was completed by investigating of the risk profiles of various acceptable regression models in order to avoid obtaining measurements outside the acceptance limits fixed a priori. On the other hand, the present study shows how the LC method can be used more accurately to assess the kinetic dissolution profiles, instead the UV-visible method required by monographs of the USP. Robustness study was also performed in order to demonstrate the capability of this method to remain unaffected by a small and deliberate variation in method parameters. The LC method was validated using the total error approach, as a decision tool, guarantees that each of the future results that will be within the acceptance limits settled at ± 5 %. The UV spectrophotometric method based on the USP monograph, gives rise of impurities from diclofenac sodium in acidic condition (HCl 0.1 N). These impurities absorb at the same wave length (276 nm) as the active principal ingredient, which will yield some significant error in the per cent release during the dissolution test study. Described analytical method is a simple, sensitive, specific and more accurate indicating that this LC method is useful for manufacturing and quality control assay. [less ▲]

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See detailQuantification of rotenone in seeds of different species of yam bean (Pachyrhizus sp.) by a SPE HPLC-UV method
Lautié, E.; Rozet, Eric ULg; Hubert, Philippe ULg et al

in Food Chemistry (2012), 131(4), 1531-1538

This study describes the development of a validated method for the quantification of rotenone in yam bean. The milled seeds were submitted to a Soxhlet dichloromethane extraction which allowed extracting ... [more ▼]

This study describes the development of a validated method for the quantification of rotenone in yam bean. The milled seeds were submitted to a Soxhlet dichloromethane extraction which allowed extracting 90% of the seeds rotenone. Elimination of the lipids was obtained via solid phase extraction. Rotenone was eluted with dichloromethane/methanol and the solution dried under vacuum and solubilised directly in methanol before injection in HPLC. The whole process was realised as much as possible protected from light and at temperatures lower than 40°C which allowed high recovery rates of spiked rotenone. Total error was used as criterion for the validation process and accuracy profiles drawn. The method allows the quantification of rotenone in yam bean seeds from 0.07% up to 1.25% (w/w). This method was applied to the quantification of rotenone in the seeds of several accessions of Pachyrhizus erosus and P. ahipa. The results range from 1.13 to 2.76 mg/g dry material. [less ▲]

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See detailFlexibility and Applicability of β-expectation tolerance interval approach to assess the fitness of purpose of pharmaceutical analytical methods
Bouabidi, Abderrahim; Talbi, M.; Bourichi, H. et al

in Drug Testing and Analysis (2012), 4(12), 1014-1027

An innovative versatile strategy using Total Error has been proposed to decide about the method’s validity that controls the risk of accepting an unsuitable assay together with the ability to predict the ... [more ▼]

An innovative versatile strategy using Total Error has been proposed to decide about the method’s validity that controls the risk of accepting an unsuitable assay together with the ability to predict the reliability of future results. This strategy is based on the simultaneous combination of systematic (bias) and random (imprecision) error of analytical methods. Using validation standards both types of error are combined through the use of a prediction interval or β-expectation tolerance interval. Finally, an accuracy profile is built by connecting, on one hand all the upper tolerance limits, and on the other hand all the lower tolerance limits. This profile combined with pre-specified acceptance limits allows to evaluate the validity of any quantitative analytical method and thus their fitness for their intended purpose. In this work, the approach of accuracy profile was evaluated on several types of analytical methods encountered in pharmaceutical industrial field and also covering different pharmaceutical matrices. The four studied examples depicted the flexibility and applicability of this approach for different matrices ranging from tablets to syrups, different techniques such as liquid chromatography, or UV spectrophotometry and for different categories of assays commonly encountered in the pharmaceutical industry that are content assays, dissolution assays and quantitative impurity assays. The accuracy profile approach assesses the fitness of purpose of these methods for their future routine application,. It also allows to select the most suitable calibration curve, to evaluate adequately a potential matrix effect and propose efficient solution and to define correctly the limits of quantification of the studied analytical procedures. [less ▲]

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See detailSolid -state characterization and impurities determination of Fluconazol generic products
Bourichi, Houda; Brik, Youness; Hubert, Philippe ULg et al

in Journal of Pharmaceutical Analysis (2012)

In this paper, we report the results of quality control based in physicochemical characterization and impurities determination of three samples of fluconazole drug substances marketed in Morocco. These ... [more ▼]

In this paper, we report the results of quality control based in physicochemical characterization and impurities determination of three samples of fluconazole drug substances marketed in Morocco. These samples were supplied by different pharmaceuticals companies. The sample A, as the discovered product, was supplied by Pfizer, while samples B and C (generics), were manufactured by two different Indian industries. Solid- state characterization of the three samples was realized with different physicochemical methods as: X- ray powder diffraction, Fourier- transformation infrared spectroscopy, differential scanning calorimetry. High performance liquid chromatography was used to quantify the impurities in the different samples. The results from the physicochemical methods cited above, showed difference in polymorph structure of the three drug substances. Sample A consisted in pure polymorph III, sample B consisted in pure polymorph II, sample C consisted in a mixture of fluconazole Form III, form II and the monohydrate. This result was confirmed by differential scanning calorimetry. Also it was demonstrated that solvents used during the recrystallization step were among the origins of these differences in the structure form. On the other hand, the result of the stability study under humidity and temperature showed that fluconazole polymorphic transformation could be owed to the no compliance with the conditions of storage. The HPLC analysis of these compounds showed the presence of specific impurities for each polymorphic form, and a possible relationship could be exist between impurities and crystalline form of fluconazole. [less ▲]

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See detailReply to the responses on the comments on “Uncertainty profiles for the validation of analytical methods” by Saffaj and Ihssane
Rozet, Eric ULg; Ziemons, Eric ULg; Marini Djang'Eing'A, Roland ULg et al

in Talanta (2012), 100

Saffaj et al., recently proposed an uncertainty profile for evaluating the validity of analytical methods using the statistical methodology of γ-confidence β-content tolerance intervals. This profile ... [more ▼]

Saffaj et al., recently proposed an uncertainty profile for evaluating the validity of analytical methods using the statistical methodology of γ-confidence β-content tolerance intervals. This profile assesses the validity of the method by comparing the method measurement uncertainty to a pre defined acceptance limit stating the maximum uncertainty suitable for the method under study. In this letter we comment on the response (T. Saffaj, B. Ihssane, Talanta 94 (2012) 361-362) these authors have made to our previous letter (E. Rozet, E. Ziemons, R.D. Marini, B. Boulanger, Ph. Hubert, Talanta 88 (2012) 769–771). In particular, we demonstrate that β-expectation tolerance intervals are prediction intervals, we show that β-expectation tolerance intervals are highly usefull for assessing analytical methods validation and for estimating measurement uncertainty and finally we show what are the differences and implications for these two topics (validation and uncertainty) when using either the methodology of β-expectation tolerance intervals or the γ-confidence β-content tolerance tolerance interval one. [less ▲]

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See detailA RAPID VALIDATED UHPLC-PDA METHOD FOR ANTHOCYANINS QUANTIFICATION FROM EUTERPE OLERACEA FRUITS
Dias, A.L.S.; Rozet, Eric ULg; Chataigne, G et al

Poster (2012)

Commercialization of Euterpe oleracea fruit has increased because of its abundance in anthocyanins [1]. Characterizations of these compounds are important for the food industry. The aim is to validate an ... [more ▼]

Commercialization of Euterpe oleracea fruit has increased because of its abundance in anthocyanins [1]. Characterizations of these compounds are important for the food industry. The aim is to validate an UHPLC-PDA method for major anthocyanins quantification in this fruit after fast extraction procedures and samples preparation. Fruits were harvested in Abaetetuba (Brazil) and extracted sequencially by EtOAc, MeOH and MeOH 50% all at 0.1% HCl. A HSS C18 column (1.8µm) was used with a gradient elution of ACN and 5% HCOOH. Total error and accuracy profiles were used as validation criteria. A first EtOAc extraction removes the lipophilic compounds and allows an easier extraction by MeOH and quantification of anthocyanins in this extract. It was found to be faster (17 min) that HPLC-UV methods [2]. Calibration in the matrix was found to be more accurate than calibration without matrix. Trueness (<6.76% relative bias), repeatability (<4.6% RSD), intermediate precision (<5.3% RSD), selectivity (by UHPLC-ESI+-HRMS), response function and linearity for cyanidin-3-glucoside and cyanidin-3-rutinoside were evaluated. The concentration range validated was 1 to 48 µg/mL for both compounds. [less ▲]

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See detailValidation of analytical methods involved in dissolution assays: Acceptance limits and decision methodologies
Rozet, Eric ULg; Ziemons, Eric ULg; Marini Djang'Eing'A, Roland ULg et al

in Analytica Chimica Acta (2012), 751

Dissolution tests are key elements to ensure continuing product quality and performance. The ultimate goal of these tests is to assure consistent product quality within a defined set of specification ... [more ▼]

Dissolution tests are key elements to ensure continuing product quality and performance. The ultimate goal of these tests is to assure consistent product quality within a defined set of specification criteria. Validation of an analytical method aimed at assessing the dissolution profile of products or at verifying pharmacopoeias compliance should demonstrate that this analytical method is able to correctly declare two dissolution profiles as similar or drug products as compliant with respect to their specifications. It is essential to ensure that these analytical methods are fit for their purpose. Method validation is aimed at providing this guarantee. However, even in the ICHQ2 guideline there is no information explaining how to decide whether the method under validation is valid for its final purpose or not. Are the entire validation criterion needed to ensure that a Quality Control (QC) analytical method for dissolution test is valid? What acceptance limits should be set on these criteria? How to decide about method’s validity? These are the questions that this work aims at answering. Focus is made to comply with the current implementation of the Quality by Design (QbD) principles in the pharmaceutical industry in order to allow to correctly defining the Analytical Target Profile (ATP) of analytical methods involved in dissolution tests. Analytical method validation is then the natural demonstration that the developed methods are fit for their intended purpose and is not any more the inconsiderate checklist validation approach still generally performed to complete the filing required to obtain product marketing authorization. [less ▲]

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See detailA rapid validated UHPLC–PDA method for anthocyanins quantification from Euterpe oleracea fruits
Dias, A.L.S.; Rozet, Eric ULg; Chataigné, G. et al

in Journal of Chromatography. B : Analytical Technologies in the Biomedical & Life Sciences (2012), 907

The aim of this work is to develop the first validated UHPLC–PDA method for major anthocyanins quantification in Euterpe oleracea fruits after fast extraction procedures and samples preparation. The ... [more ▼]

The aim of this work is to develop the first validated UHPLC–PDA method for major anthocyanins quantification in Euterpe oleracea fruits after fast extraction procedures and samples preparation. The separation was performed on HSS C18 column (1.8 m) using a gradient elution with acetonitrile and 5% formic acid in a total run time of only 17 min. Total error and accuracy profiles were used as criteria for the validation process. Calibration in the matrix was found to be more accurate than calibration without matrix. Trueness (<6.76% relative bias), repeatability (<4.6% RSD), intermediate precision (<5.3% RSD), selectivity, response function and linearity for major anthocyanins, cyanidin-3-glucoside and cyanidin-3-rutinoside, were evaluated. The concentration range validated was 1–48 g/mL for both compounds. In addition two cyanidin-di-O-glycosides were detected for the fist time in this fruit. We also showed that a first extraction of the fruits with ethyl acetate removes the lipophilic compounds and allows an easier extraction by methanol and quantification of anthocyanins in this extract. [less ▲]

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See detailCritical Review of Near-Infrared Spectroscopic Methods Validations in Pharmaceutical Applications
De Bleye, Charlotte ULg; Chavez, Pierre-François ULg; Mantanus, Jérôme ULg et al

in Journal of Pharmaceutical & Biomedical Analysis (2012), 69

Based on the large number of publications reported over the past five years, near-infrared spectroscopy (NIRS) is more and more considered an attractive and promising analytical tool regarding Process ... [more ▼]

Based on the large number of publications reported over the past five years, near-infrared spectroscopy (NIRS) is more and more considered an attractive and promising analytical tool regarding Process Analytical Technology and Green Chemistry. From the reviewed literature, few of these publications present a thoroughly validated NIRS method even if some guidelines have been published by different groups and regulatory authorities. However, as any analytical method, the validation of NIRS method is a mandatory step at the end of the development in order to give enough guarantees that each of the future results during routine use will be close enough to the true value. Besides the introduction of PAT concepts in the revised document of the European Pharmacopoeia (2.2.40) dealing with near-infrared spectroscopy recently published in Pharmeuropa, it agrees very well with this mandatory step. Indeed, the latter suggests to use similar analytical performance characteristics than those required for any analytical procedure based on acceptance criteria consistent with the intended use of the method. In this context, this review gives a comprehensive and critical overview of the methodologies applied to assess the validity of quantitative NIRS methods used in pharmaceutical applications. [less ▲]

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See detailChapter 3 Method Transfer Between Conventional HPLC and UHPLC
Debrus, Benjamin ULg; Rozet, Eric ULg; Hubert, Philippe ULg et al

in Guillarme, Davy; Veuthey, jean-Luc (Eds.) UHPLC in Life Sciences (2012)

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See detailAPPLICATION OF DESIGN OF EXPERIMENTS AND DESIGN SPACE METHODOLOGY FOR THE HPLC-UV SEPARATION OPTIMIZATION OF APORPHINE ALKALOIDS FROM LEAVES OF Spirospermum penduliflorum THOUARS
Rafamantanana, Mamy; Debrus, Benjamin ULg; Raoelison, Guy et al

in Journal of Pharmaceutical & Biomedical Analysis (2012), 62

Spirospermum penduliflorum Thouars (Menispermaceae) is an endemic species of Madagascar traditionally used as vasorelaxant. Recently, two aporphine alkaloids known to possess antihypertensive activity ... [more ▼]

Spirospermum penduliflorum Thouars (Menispermaceae) is an endemic species of Madagascar traditionally used as vasorelaxant. Recently, two aporphine alkaloids known to possess antihypertensive activity (dicentrine and neolitsine) were isolated and identified from the leaves of this plant. In the present study, a HPLC-UV method allowing the separation of all alkaloids and the quantification of dicentrine in the alkaloidic extract of leaves was developed using design of experiments and design space methodology. Three common chromatographic parameters (i.e. the mobile phase pH, the initial proportion of methanol and the gradient slope) were selected to construct a full factorial design of 36 experimental conditions. The times at the beginning, the apex (i.e. the retention time) and the end of each peak were recorded and modelled by multiple linear equations. The corresponding residuals were normally distributed which confirmed that the models can be used for the prediction of the retention times and to optimize the separation. The optimal separation was predicted at pH 3, with a gradient starting at 32% of methanol and a gradient slope of 0.42%/min. Good agreement was obtained between predicted and experimental chromatograms. The method was also validated using total error concept. Using the accuracy profile approach, validation results gave a LOD and LOQ for dicentrine of 3 µg/ml and 10 µg/ml, respectively. A relative standard deviation for intermediate precision lower than 10% was obtained. This method was found to provide accurate results in the concentration range of 10 µg/ml to 75 µg/ml of dicentrine and is suitable for routine analysis. [less ▲]

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See detailDesign space approach in the optimization of the spray-drying process
Lebrun, Pierre ULg; Krier, Fabrice ULg; Mantanus, Jérôme ULg et al

in European Journal of Pharmaceutics & Biopharmaceutics (2012), 80(1), 226-234

From a quality by design perspective, the aim of the present study was to demonstrate the applicability of a Bayesian statistical methodology to identify the design space (DS) of a spray-drying process ... [more ▼]

From a quality by design perspective, the aim of the present study was to demonstrate the applicability of a Bayesian statistical methodology to identify the design space (DS) of a spray-drying process. Following the ICH Q8 guideline, the DS is defined as the “multidimensional combination and interaction of input variables (e.g., materials attributes) and process parameters that have been demonstrated to provide assurance of quality”. Thus, a predictive risk-based approach was set up in order to account for the uncertainties and correlations found in the process and in the derived critical quality attributes such as the yield, the moisture content, the inhalable fraction of powder, the compressibility index and the Hausner ratio. This allowed quantifying the guarantees and the risks to observe whether the process shall run according to specifications. These specifications describe satisfactory quality outputs and were defined a priori given safety, efficiency and economical reasons. Within the identified DS, validation of the optimal condition was effectuated. The optimized process was shown to perform as expected, providing a product for which the quality is built in by the design and controlled set-up of the equipment, regarding identified critical process parameters: the inlet temperature, the feed rate and the spray flow rate. [less ▲]

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See detailUsefulness of capability indices in the framework of analytical methods validation
Bouabidi, Abderrahim ULg; Ziemons, Eric ULg; Marini Djang'Eing'A, Roland ULg et al

in Analytica Chimica Acta (2012), 714

Analytical methods capability evaluation can be a useful methodology to assess the fitness of purpose of these methods for their future routine application. However, care on how to compute the capability ... [more ▼]

Analytical methods capability evaluation can be a useful methodology to assess the fitness of purpose of these methods for their future routine application. However, care on how to compute the capability indices has to be made. Indeed, the commonly used formulas to compute capability indices such as Cpk, will highly overestimate the true capability of the methods. Especially during methods validation or transfer, there are only few experiments performed and, using in these situations the commonly applied capability indices to declare a method as valid or as transferable to a receiving laboratory will conduct to inadequate decisions. In this work, an improved capability index, namely Cpk-tol and the corresponding estimator of proportion of non conforming results ( ) has been proposed. Through Monte-Carlo simulations, they have been shown to greatly increase the estimation of analytical methods capability in particular in low sample size situations as encountered during methods validation or transfer. Additionally, the usefulness of this capability index has been illustrated through several case studies covering applications commonly encountered in the pharmaceutical industry. Finally a methodology to determine the optimal sample size required to validate analytical methods is also given using the proposed capability metric. [less ▲]

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See detailQuality by design compliant analytical method validation
Rozet, Eric ULg; Ziemons, Eric ULg; Marini Djang'Eing'A, Roland ULg et al

in Analytical Chemistry (2012), 84

The concept of quality by design (QbD) has recently been adopted for the development of pharmaceutical processes to ensure a predefined product quality. Focus on applying the QbD concept to analytical ... [more ▼]

The concept of quality by design (QbD) has recently been adopted for the development of pharmaceutical processes to ensure a predefined product quality. Focus on applying the QbD concept to analytical methods has increased as it is fully integrated within pharmaceutical processes and especially in the process control strategy. In addition, there is the need to switch from the traditional checklist implementation of method validation requirements to a method validation approach that should provide a high level of assurance of method reliability in order to adequately measure the Critical Quality Attributes (CQAs) of the drug product. The intended purpose of analytical methods is directly related to the final decision that will be made with the results generated by these methods under study. The final aim for quantitative impurity assays is to correctly declare a substance or a product as compliant with respect to the corresponding product specifications. For content assays, the aim is similar: making the correct decision about product compliance with respect to their specification limits. It is for these reasons that the fitness of these methods should be defined, as they are key elements of the Analytical Target Profile (ATP). Therefore, validation criteria, corresponding acceptance limits and method validation decision approaches should be settled in accordance with the final use of these analytical procedures. This work proposes a general methodology to achieve this in order to align method validation within the QbD framework and philosophy. β-expectation tolerance intervals are implemented to decide about the validity of analytical methods. The proposed methodology is also applied to the validation of analytical procedures dedicated to the quantification of impurities or active product ingredients (API) in drug substances or drug products and its applicability is illustrated with two case studies. [less ▲]

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See detailContribution au développement des capacités d’enseignement et de formation pour l’amélioration de la qualité du médicament (acronyme : DEV-AQM)
Marini Djang'Eing'A, Roland ULg; Hubert, Philippe ULg

Report (2011)

Contribution to the development of teaching capacity and training for the improvement of quality of the medicines. Contextualisation: The quality of medicines is a major problem of public health in the ... [more ▼]

Contribution to the development of teaching capacity and training for the improvement of quality of the medicines. Contextualisation: The quality of medicines is a major problem of public health in the development countries. Since 1990, this situation has deteriorated becoming worried mainly in the Central African region due to the degradation of social, economic and politic life, consequence of a long period of conflict and war. The resurgence of non-controlled drugs, the sale of illicit, deteriorated and even falsified drugs are real examples in the current practices, that is making difficult and even practically impossible an access to a safe, reliable and efficient medical treatment. It is known that even if a diagnosis is correctly made and a medical treatment is correctly prescribed, this treatment is doomed to failure if the medicine is not of a good quality. Democratic Republic of Congo (DRC) and Rwanda are among the countries that are facing such situations. Description of project purpose: The principal objective of this project is to contribute to the improvement of the quality of medicines and thus, of the public health in DRC and in Rwanda. More precisely, the project aims to strengthen the local capacity in order to respond to the need in the quality of medicines and to develop a platform of people in the pharmaceutical sector in the field of quality assurance and control. According to this main objective, the project aims in one hand to train people working in the pharmaceutical sector including the academic, the legal and the industrial, and in another hand to develop the tools to contribute to the improvement of the quality of medicines. Training and qualification of people, improvement of the teaching and making available the control documentation on quality are the sub-objectives pursued in the framework of this project. Chapter 2 : Six main activities are undergone in this project: The first activity is the seminar that is organised in order to promote the project and to initiate the importance of knowledge of the quality of medicine by awareness of the different authorities from government, from the churches and from the health sectors. The seminar is the preliminary step of this project that is done to select the different candidates. The second activity concerns the theoretical training, focused on the basis of the quality of drugs, the drug manufacturing and drug control / analysis taking into account the activity sector of each candidate. This activity is important since most of the candidates have been graduated a long time ago while working for a long time. This activity as well as the seminar is held in the beneficiary countries for one month. The trainers are among the Professors and Researchers from the “University of Liège”, the “University Libre de Bruxelles” and the “University Catholique de Louvain”, in Belgium. They are selected on basis of their expertise. The topics considered are the Quality Assurance, the Regulatory, the Statistical applied in the pharmaceutical industry, The Manufacturing and The Quality control of medicines, the Management in the Pharmaceutical sector. The theoretical training by e-learning using internet occurs in the third place. It is done as a complement to the second activity since it allows the candidate to have access to different and more documents available through the web site platform created for this purpose. This mode of teaching allows also the candidates to interact with other trainers concerning a particular subject of their working sector. The fourth activity is the practices that are done in Belgium in the different facilities of the laboratories associated to this project. This allows the different candidates to materialize the knowledge acquired during the theoretical teaching while considering their working sector, and to familiarize with the procedures dealing with their sector. The fifth activity is related to a specific training for auditor or evaluator. Indeed, this training is a specific requirement namely the creation of Federal Agency of Drug in DRC. The need is to improve the capacity of such organism to carry out efficiently the audits and evaluations of drug registration files before their commercialization. Finally, the sixth activity is the reintegration of the candidates within their professional environment. Considering the importance of this aspect, an accompanying is necessary to ensure that the acquired knowledge is valued in the professional environment. Chapter 3 : The expected impact At the end of this project, we expect that the different Authorities are aware with regard to the quality of drugs, the activities in the pharmaceutical sectors namely, the legal and industrial are improved since the actors are trained, qualified and gained competence. We expect also the improvement of teaching capacity in Pharmacy Schools taking into account the need of the pharmaceutical market, with the possibility of starting the post-university teaching programs. Finally, we expect the efficiency of activities in the pharmaceutical sector for the benefit of the DRC and Rwanda populations. Contact person : Philippe Hubert (ph.hubert@ulg.ac.be) / Roland Marini Djang’eing’a (rmarini@ulg.ac.be) Address : Service de Chimie Analytique, Département de Pharmacie, Bât. B36, Avenue de l'Hôpital, 1, 4000 Liège 1, Belgium. Tel. + 00 32 4 366 43 15 [less ▲]

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See detailDesign Space ou Espace de Conception
Boulanger, B.; Lebrun, Pierre ULg; Rozet, Eric ULg et al

Scientific conference (2011, November 29)

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