Applications of vibrational spectroscopy and hyperspectral imaging for the analysis of substandard and falsified medicines.

peer reviewedAccess to quality medicines is an essential right of the patients. However, in 2017, the World Health Organization estimated that 1 in 10 medical products circulating in low- and middle-income countries is either substandard or falsified. This reinforces the fact that post-marketing surveillance (PMS) of medical products by strong national regulatory authorities (NRA) is crucial. To achieve an efficient PMS, the NRA need analytical tools at the inspection, screening, confirmatory and forensics levels to control the physicochemical properties of the samples. Because of their fast, non-destructive, and relatively affordable character, vibrational spectroscopy tools are unavoidably present at each step. Handheld devices are particularly useful during inspection and screening phases since these tools can identify and/or quantify active pharmaceutical ingredients (API) even through opaque packaging in seconds. However, they generally need exhaustive and up-to-date databases for each specific product. Another limitation is the work and time needed before going into the field to develop and validate the chemometric models. Indeed, this mandatory step requires highly skilled scientists and a prior collection of certified references of the medicines to analyse. Benchtop systems and among them imaging systems are particularly useful in the confirmatory and forensic steps. Indeed, the imaging systems enable the visualization and identification of a large range of both organic and inorganic compounds used as API or excipients. In addition, thanks to the high spatial resolution, it allows the detection of trace contaminants. This information may be of particular interest during prosecutions and the clustering of criminal cases. Nevertheless, the extraction of the relevant information from the raw measurements requires once again intensive work by highly trained staff. In conclusion, vibrational spectroscopy tools have particularly interesting features for the PMS of medicines, but research is still needed to make them easier to set up and use by NRA inspectors and non-specialists

Quality control of anti-infective medicines : case of ciprofloxacin and metronidazole formulations in Butembo

The circulation of substandard or falsified medicines has increased around the world and this is one of the biggest health crimes considering the potential harm. Democratic Republic of Congo (DRC) is not spared by this scourge with an almost non-existent quality assurance and quality control system. This work aimed to assess the quality of formulations based on Ciprofloxacin and Metronidazole marketed in Butembo, specifically, by checking some parameters based on the WHO checklist for visual inspection. The pre-survey and the collection of samples were limited to 12 pharmacies known as wholesalers. 51 samples of metronidazole and ciprofloxacin formulations (tablets, oral suspensions and solutions for injection) were randomly selected, in which we found 27 different brands manufactures. Concerning the packaging and labelling, all the products presented a satisfactory result. For physical characters of the product, only two samples of tablet showed an integrity defect. Overall, 40 samples (78.4%) were out of specifications of the WHO visual inspection, in which the majority was in tablet forms (100,0 % for metronidazole and 68.2 % for Ciprofloxacin). The majority of non-compliance was due to the not-registration of samples in DRC. This study showed a great pharmaceutical regulation problem in RDC. Limited to visual inspection, it constituted the first step of the analytical approach of quality control of ciprofloxacin and metronidazole formulations found on the Butembo market

Design, Development and Optimization of Topotecan Hydrochloride Solid Lipid Nanoparticles for Oral Chemotherapy

peer reviewedThe objective of this research work was to formulate and evaluate solid lipid nanoparticles of anticancer drug topotecan hydrochloride in order to sustain its release and therefore improve its therapeutic outcomes. SLNs were prepared by hot micro emulsion method using glyceryl monostearate as lipid carrier and Tween 80 as surfactant in specific proportions. FT-IR and DSC studies revealed that there was no interaction between drug and lipid. The effect of drug: lipid ratio, surfactant concentration, speed and time of homogenizer on drug release, particle size and EE were studied. The results of particle size, encapsulation efficiency and in vitro drug release at 12hr fell between 111.9 nm and 255nm, 65.12% and 77.63%, and 72.31 and 79.79% respectively. Optimized formulation showed release of drug up to 73.47 % at 12 hrs, entrapment efficiency of 77.75 % and particle size of 158.0 nm. The analysis of 3-D graphs revealed that parameters such as drug: lipid ratio and time of homogenization have a profound effect on both encapsulation efficiency and cumulative drug release. Scanning electron microscopy studies showed particles with disuniform surface and shape. The drug release from optimized formulation was found to fit best into first order kinetic model. It also showed almost linear regression in Higuchi’s plot suggesting that diffusion is one of the mechanisms for drug release and n value of Korsmeyer-Peppas plot was found to be 0.716 indicating that the drug release did not follow fickian diffusion controlled mechanism.Nanosize orla anticance

Development of a generic near infrared (PIR) method in quality control and counterfeit medicines.

Les médicaments antirétroviraux de mauvaise qualité sont l'un des principaux problèmes de santé publique en Afrique. Ils constituent une menace réelle pour les malades vivant avec les VIH/sida. Pour aborder une partie de ce problème, une méthode analytique permettant de détecter et de quantifier les antirétroviraux ; la spectroscopie Proche Infrarouge (PIR) a été sélectionnée pour l'analyse des médicaments ARV en raison de son faible coût, de sa rapidité et de son caractère non destructif ne nécessitant pas d’étape de préparation des échantillons. Compte tenu de ces avantages, le PIR pourrait être utilisé en première ligne dans la lutte contre la contrefaçon des médicaments ARV. Les spectres proches infrarouges des différents échantillons des médicaments ARV contenant Zidovudine ont été prélevés et comparés avec le spectre proche infrarouge de Zidovudine SCR. L’analyse de ces différents spectres a permis de sélectionner une gamme spectrale comprise entre 4500 et 8000 cm-1 dans laquelle le principe actif et les excipients absorbent modérément. La présente étude a permis de démontrer le potentiel ainsi que l’intérêt de la spectrophotométrie proche infrarouge dans la détection rapide des médicaments contrefaitscréation d'un centre d'excellence en Assurance et Contrôle de Qualité des médicaments et produits de sant

Toxicological risks associated with poor quality medicines

Objective: Poor quality medicines regrouping counterfeit/falsified, sub-standards and degraded are a scourge for developing countries (1). Their public health consequences are often observed for cases of low dosage of active ingredients and sometimes for the absence of these. On the basis of the observed facts as well as suspected reported cases, we were interested in deepening the information through laboratory tests. Methods: Liquid chromatography (LC), thin layer chromatography (TLC), nuclear magnetic resonance (NMR), mass spectrometry (MS), Raman imagery (Rim) and near infrared spectroscopy (NIR) were used as targeted analytical techniques in this study, associated with the principal component analysis (PCA), while the suspect samples were obtained via the public health authorities of the DR Congo, Benin and Rwanda after their seizure. Results: The observed cases are presented by therapeutic class, namely: - analgesics: (1) cases of tablets supposed to contain paracetamol and which have caused abnormal adverse effects in patients who have consumed it, namely sedative effects, polyuria and hypotension. Using LC and complementary surveys by people interview, we were able to demonstrate the presence of a benzodiazepine and at very high doses. (2) Cases of paracetamol syrup whose excipient (glycerol) was substituted by diethylene glycol and which had caused the death of a hundred babies. Using the NIR and the PCA, we were able to elucidate this substitution. - antimalarials: (case 1) cases of arthemether-lumefantrine tablets which had no therapeutic effect. By means of TLC and LC, the absence of these two active ingredients was clearly demonstrated, but the presence of starch was revealed by Rim. (Case 2) case of quinine tablets that had no expected pharmacological effect. The use of several combined techniques (TLC, LC-MS, NMR and Rim) made it possible to demonstrate the presence of a substance with an imidazole structure (antamoeba). - antibiotics: (1) amoxicillin powder which after reconstitution in hospital caused poisoning of the babies until death for some. The reconstitution of the suspension had revealed the misuse of hydrogen peroxide instead of distilled water. (2) Finally a large-scale study (80 samples of amoxicillin powder) showed that 8% of the samples were above the claimed dosage. Conclusion: The cases presented indicate that there are effectively poor quality medicines and that they are responsible of public health problems and in particular of toxicity. Appropriate measures should be taken to protect users.Lutte contre la falsification des médicament

Design, development and optimization of Solid Lipid Nanoparticles of Temozolomide as anticancer drug

Solid Lipid Nanoparticles loaded Temozolomide were prepared by simple hot nanoemulsion method and evaluated for their in vitro drug release profile. Glyceryl Monostearate was used as lipid core; Tween 80 and Pluronic F-68 as surfactant and cosurfactant. The preformulation studies (FT-IR and DSC) revealed that the excipients used are compatibles to the drug. Four formulation parameters were optimized to obtain high quality nanoparticles. The physicochemical properties of the TMZD-SLNs were investigated by particle size analysis, zeta potential measurement, drug entrapment efficiency (EE), stability and in vitro release behavior. There was no significant change in term of physicochemical properties and drug release profile between predicted and actual values. The TMZD-SLNs showed stable size distribution at 151.4 nm with Z-potential of -24.2 mV, ideal drug EE (58.14±0.44%) and relative long term physical stability after being stored for 3 months. The drug release of TMZD-SLNs up to 24 hours exhibited a sustained profile compared to its release from Temozolomide aqueous solution which made it a promising carrier for oral sustained release drug delivery systems. Kinetic of drug release from optimized formulation was found to be zero order (time independent). It also showed almost linear regression in Higuchi’s plot which confirms that diffusion is one of the mechanisms for drug release. In conclusion, this study showed that it was possible to obtain by this method Temozolomide solid lipid nanoparticles with small particle size and ideal EE and finally obtain the sustained release of the drug, meanwhile avoiding adverse side effects by the conventional formulatio

Identification of pharmaceutical products using Handheld NIR and Raman spectrophotometers.

Abstract: Context: Over the last decade, the growth of the global pharmaceutical market has led to an overall increase of substandard and falsified drugs especially on the African market (or emerging countries). Recently, several methods using vibrational spectroscopy have been developed for rapid and on-field drug analysis. Objective: The objective of this work was to evaluate the identification performances of handheld near infrared (NIR) and Raman systems. Methods: The formulations of artemether-lumefantrine were used in tablet dosage forms. In order to perform a critical comparison, the analytical performances of the two analytical systems were compared statistically using the method of data-driven soft independent modeling of class analogy (DD-SIMCA). Results: The overall results show good detection abilities for NIR systems compared to Raman systems based on Matthews’s correlation coefficients, generally close to one Raman systems are less sensitive to the physical state of the samples than the NIR systems, because of the strong influence of the auto-fluorescence phenomenon, the signal of highly-dosed active pharmaceutical ingredient (lumefantrine) masking the signal of low-dose and weak Raman scatterers API (e.g. artemether). Conclusion: Hence, Raman systems are less effective for authentication purposes but are interesting in the context of falsification because they allow a visual interpretation of the spectral signature (presence or absence of API)

DD-SIMCA and PLS regression models applied to NIR spectroscopy for identification and assay of ciprofloxacin tablets

peer reviewedPoor-quality medicines pose a threat to the health of populations but also constitute a hindrance to the socio-economic development of governments. Developing countries are the most affected, as they face enormous difficulties in ensuring the quality of medicines present in their markets [1]. Among other difficulties, the methods commonly advocated by pharmacopoeias using sophisticated techniques such as High-Performance Liquid Chromatography (HPLC) are not always obvious to apply. These methods are generally expensive, difficult to implement because of power shortages and logistic problems leading generally to significant results-reporting deadlines [2]. Apart from these laboratory techniques, vibrational spectroscopy is of particular interest because they are fast, easy to use, cheap and less polluting. This interest is also increased by the development of low-cost handheld devices. Nevertheless, the major difficulty lies in the interpretation of the results, which necessitates chemometric methods. Considering all these points, it has been undertaken to develop and validate methods for the identification and assay of ciprofloxacin using near-infrared (NIR) spectrophotometry coupled with chemometric methods such as Data-Driven Soft Independent Modelling of Class Analogy (DD SIMCA) and Partial Least Squares (PLS) regression. For this purpose, two types of low-cost handheld NIR devices were used, one in reflection mode (NIR-S-G1) and the other in transmission mode (NIR M-T1) both from Innospectra Corporation (Taiwan). On the one hand, a qualitative DD-SIMCA model has been developed to confirm the presence of the active ingredient ciprofloxacin tablets regardless of the brand or formulation. This first step is performed on intact tablets with the reflective module and is envisaged as a screening phase applicable during an on-field inspection. On the other hand, a quantitative PLS-R model was built for the determination of ciprofloxacin content in tablets of different brands with the announced content 500mg. The PLS model was then validated using the total error approach with the accuracy profile as a decision tool with 90.0%-110.0% specifications and a risk β set at 95%. This second step is performed on dissolved tablets with the transmissive module and is envisaged as a pre-confirmatory technique. The main advantage is that it may be performed anywhere close to the inspection site. These validated methods were then applied on 23 ciprofloxacin 500mg tablet samples collected in the Cameroonian legal and illegal market. Ciprofloxacin was correctly identified in all samples using the DD-SIMCA model. The latter allowed ciprofloxacin to be discriminated against other molecules belonging to the fluoroquinolone family such as norfloxacin, ofloxacin, moxifloxacin and levofloxacin. In addition, four substandard samples were detected using the PLS regression model. All spectroscopic results were confirmed with a validated HPLC method. The applicability of the qualitative and quantitative methods developed and validated led to encouraging results as they were confirmed by HPLC used as a reference method. Because of their portability, these methods could be easily transported to areas where laboratory facilities are not present to assess the quality of medicines. Moreover, they can be used upstream of laboratory confirmation methods with a screening objective in resource-limited countries thanks to their low analysis cost

Field survey to evaluate the prevalence of poor quality anti-infective medicines in Cameroon

Poor quality medicines pose a threat to all health systems. It is obvious that they have harmful consequences not only from the point of view of public health, but also from the economic and socio-economic point of view [1]. Over the past 15 years, substandards and falsified drugs have received increasing attention in scientific publications. However, there is little reliable data determining their prevalence with accuracy due to the scarcity of well-designed studies identified as having good methodological quality as well as representative sampling strategy [2-4]. In this context, we have decided to evaluate the prevalence of poor quality anti-infective medicines in two Cameroon areas (Yaoundé and Douala), inspired by the Medicines Quality Assessment Reporting Guidelines (MEDGUARG) [3] and WHO recommendations [4] for the sampling strategy and the methodology. Our study will focus on the formal private sector. Pharmacies as well as drugs products will be sampled by a stratified random sampling strategy. The study will focus on two anti-infective medicines (ciprofloxacin and metronidazole 500mg tablets) in tablets, from 96 outlets in the cities of Yaoundé and Douala that are the two main cities (Yaoundé and Douala) of Cameroon representing almost 70% of private outlets of the country. Mystery shoppers will collect samples using a specific scenario. As a prelude to our field study, screening and dosage methods have to be developed and validated in Liège University. They se methods consist of vibrational spectroscopy (near infrared and Raman spectroscopy) as first screening techniques and HPLC for identification and assay. For vibrational spectroscopy, qualitative models will be developed for identification using chemometric tools. HPLC methods will be validated following the total error approach using accuracy profile as decision tool. The medicines collected will be first analysed visually (physical appearance tests), then field methods will be implemented (screening methods: Paper Analytical Devices (PADs), handled NIR device). Finally laboratory testing (assay and confirmation methods: HPLC reference method and pharmacotechnical tests) will be performed at LANACOME (Yaoundé, Cameroon). Suspect and unusual samples will be transported to Liège University for further analyses. All these methods will be applied according to a decision tree based on observed facts. The study will be submitted to the ethics committee of the Ministry of Health in Cameroon. An accurate and fast HPLC method for identification and quantification of both metronidazole and ciprofloxacin has been developed. Identification models for some brands of ciprofloxacin and metronidazole using handled NIR and Raman devices has been developed before implementation on field. This study will allow us to evaluate not only the prevalence of poor quality anti-infective medicines marketed in Cameroon but also outlets dispensing substandard and falsified medicines. They will be distinguished into sub-standard, degraded or falsified and classified according to their country of origin, manufacturer and city of sampling. The results will be notified to the drug regulatory authority in Cameroon and if poor quality medicines are detected, we will proceed with an alert to the WHO Global Surveillance System. The estimation of the prevalence of counterfeit and falsified anti-infective medicines would be extrapolated to the entire population and depending on the information obtained, evaluate the patient health risk exposed to substandard and falsified anti-infective medicines and develop capacity-building interventions in the fight against poor quality medicines