LIQUID CHROMATOGRAPHY TANDEM MASS SPECTROMETRY DETERMINATION METHOD OF BENCYCLOQUIDIUM BROMIDE: APPLICATION TO DRUG INTERACTION STUDY IN HUMAN

Objective: This study was conducted to develop a sensitive and effective LC-MS/MS method for the determination of bencycloquidium bromide (BCQB) and its application in pharmacokinetic drug interaction study between BCQB and paroxetine. Methods: The chromatographic separation was performed on Hedera ODS-2 C18 column with a mobile phase consisted of acetonitrile-10 mmol/l ammonium acetate containing 0.2% acetic acid (33:67, v/v) at 550 μl/min, and the plasma samples were processed using solid-phase extraction. The MS/MS transitions were m/z 330.2 → 142.0 for BCQB and m/z 344.2 → 156.1 for the I. S in positive ESI mode. Results: The validated method was linear over the concentration range of 2-1200 pg/ml with the correlation coefficient r2>0.998. The intra-and inter-batch precisions of the assay were lower than 8.2% and 9.1%, respectively. The lower limit of quantification (LLOQ) was 2 pg/ml. The stability data at different storage conditions of BCQB were within±5% RE. The mean AUC0-36 of BCQB was increased by approximately 33%, after the administration of BCQB alone and upon co-administration with paroxetine during the drug interaction study. Conclusion: The LC-MS/MS method validated in this study was robust, reproducible, accurate, precise and reliable and was successfully applied in the pharmacokinetic drug interaction studies

IDENTIFICATION AND QUANTIFICATION OF PHOSPHODIESTERASE-5 INHIBITOR AS FALSIFIED IN ‘‘NATURAL’’ MALAYSIAN HERBAL APHRODISIACS SOLD IN SOME BENINESE MARKETS

Objective: Nowadays, there have been several reports of herbal products falsified with well-known synthetic molecules, leading to harmful health consequences for the consumer. The aim of this study was to assess the profile of ‘natural' herbal aphrodisiacs in the local markets of the municipalities of Cotonou and Abomey-Calavi in Benin and to screen some of them for the presence of additives such as sildenafil and tadalafil. Methods: A non-probability survey was conducted to identify the available aphrodisiacs and their characteristics. Some of them were then selected for analysis. Thin Layer Chromatography (TLC) was adopted for qualitative detection. The TLC positive extracts were then analyzed by HPLC on a C18 column with a mobile phase consisting of a mixture of 0.05M phosphate buffer (pH 5.8), acetonitrile and methanol (30:50:20). The Detection was performed at 290 nm. Results: Seventy-seven aphrodisiacs were identified and from these, 18 were selected for analysis. Six of them were adulterated with tadalafil. The concentration of tadalafil in the samples was 1.7 to 4.6 times higher than the recommended dose of 20 mg. Conclusion: This work opens the door to the need to control ‘‘natural’’ labeled products in order to ensure their quality

Evaluation of 90-day subchronic oral toxicity of aqueous extract of Gmelina arborea Roxb (Verbenaceae) leaves in Wistar rats

Purpose: To evaluate the 90 day sub-chronic toxicity of aqueous extract of Gmelina arborea leaves in Wistar rats. Methods: Rats were submitted to repeated daily oral administration of extract (250, 62.5 and 15.62 mg/kg) of Gmelina arborea leaves. The control groups were given distilled water and the rats were monitored for any toxicity symptoms as well as body and organs weights, water and food intake changes. The biochemical, haematological and histolopathological parameters were analysed. Results: The 90 days administration of the aqueous extract did not produce any toxicity signs or mortality. In addition, no significant alteration in water or food intake by the rats was observed. Although there were no changes in the body weights, significant decrease in the weight of the kidneys of the rats was observed at 250 mg/kg. Biological parameters as well as the histopathology of liver and kidneys were not significantly affected. Significant decreases were noted in glucose level at the three dose levels. In addition, significant difference in the levels of transaminases, glucose and platelets were observed. Conclusion: The 90-days subchronic toxicity test on Gmelina arborea did not produce any toxic effects. This confirms the safety of the plant leaves by traditional medicine practitioners. Keywords: Gmelina arborea, Subchronic toxicity, Wistars rats, Biological parameter

C43 - Essai de formulation d’un collutoire à base d’huile essentielle de Cymbopogon giganteus, chiov (poaceae) pour traiter l’angine de gorge

Introduction : Sur la base d’études antérieures, ayant démontré une efficacité in vitro de l’huile essentielle de Cymbopogon giganteus sur les germes impliqués dans l’angine de gorge, l’objectif de cette étude était de développer une formulation adaptée pour la prise en charge de cette pathologie. Matériel et méthodes : L’huile d’olive vierge, le TWEEN 60 et le SPAN 60 ont été utilisés pour le développement du collutoire à 0,5% v/v d’huile essentielle de Cymbopogon giganteus. La caractérisation du collutoire a été effectuée au moyen de tests organoleptiques et microscopiques (Balla et al 2022). Le sens de l’émulsion est déterminé par la méthode des colorants et confirmé par le test de conductivité électrique. L’absence d’instabilité du collutoire vérifiée au moyen du test à la centrifugeuse. En prélude à des futurs essais cliniques, la stérilité du collutoire a été recherchée par ensemencement sur différents milieux gélosés. Résultats et Discussion : Après plusieurs essais, seule la préparation à 40% d’huile d’olive à l’aspect blanc laiteux et au toucher gras était stable avec une absence de floculation et de coalescence et sans observation de phénomènes d’instabilités à type de crémage ou de sédimentation. Les tests de conductivité (49,2 µS/cm) ont bien confirmé le sens de l’émulsion H/E. Les tests de stérilité ont été concluants en l’absence de prolifération de microorganisme après 5 jours d’incubation. Conclusion : Cette étude a permis la formulation d’un collutoire à base d’huile essentielle de Cymbopogon giganteus présentant un bon profil de stabilité et de qualité

C4 - Formulation d’un émulgel à base d’un mélange d’huile essentielle d’Ocimum gratissimum L. (Lamiaceae) et du gel d’Aloe vera (L.) Burm. F. (Asphodelaceae) contre les acnés vulgaires

Introduction : L’acné due à une production excessive de sébum associée ou non à une prolifération bactérienne est habituellement traitée avec des topiques. L’Aloe vera (AV) et l’Ocimum gratissimum (OG) sont deux plantes traditionnellement utilisées dans le traitement de l’acné. Ainsi, notre étude a consisté en des essais de formulation et d’évaluation de l’efficacité antibactérienne d’un émulgel combinant ces deux plantes. Méthodologie : Les essais ont été faits à partir du gel d’AV à la proportion de 10% et 20% respectivement associée à l’huile essentielle (HE) d’OG à 5% et 10% permettant d’élaborer un émulgel à base de l’HE gélifiée avec du carbomère 980 à 1%. La stabilité de nos formulations a été évaluée par des tests pharmaco-techniques que sont la détermination des caractères organoleptiques, l’examen microscopique, le test de stérilité et la mesure de quelques paramètres physico-chimiques (conductivité, pH, viscosité et aptitude à l’étalement) et l’activité antibactérienne a été déterminée sur les germes du genre Staphylococcus. Résultats : La formule à 10% d’AV et 5% d’OG a été retenue au regard de sa plus grande stabilité. Au plan organoleptique, elle présentait un aspect homogène, une uniformité de couleur, une bonne consistance et une odeur caractéristique de l’OG. Sa conductivité évaluée à 1935µS/cm a confirmé le sens de l’émulsion Huile/Eau. Quant à la viscosité, elle était de 344400 mPa.s suggérant une bonne aptitude de l’émulgel à l’étalement. Enfin le pH (7,38) indiquait une bonne compatibilité avec celui de la peau, confirmée par les tests d’irritabilité sur le lapin albinos (Indice d’irritabilité IIP ˂ 0.5). L’activité antibactérienne sur les germes du genre Staphylococcus méticillino-résistante a montré une bonne sensibilité à cette formulation avec des zones d’inhibition ayant des diamètres > 8mm. Conclusion : Les résultats, concluant sur la stabilité et l’efficacité in vitro de l’émulgel à 5% d’HE d’OG, ouvrent la voie à la réalisation d’une étude clinique avant mise sur le marché

Evaluation of the knowledge and attitude of pharmacists about the national malaria control policy in southern Benin

Abstract Background The national strategy against malaria in an endemic country should involve all the health stakeholders. In Benin, the private sector is rarely present in the activities of the National Malaria Control Programme (NMCP), and its surveillance system does not cover private sector outlets that are a non-negligible part of the healthcare system. Objective The aim of this study was to describe the drug delivery practices within private pharmacies of Cotonou and Porto-Novo and the awareness of medicine providers concerning the national policy of malaria treatment. Methods A survey was performed among pharmacy staff members responsible for dispensing medicines and providing advice to patients within pharmacies of Cotonou and Porto-Novo. Dispensing/pharmacy assistants (‘dispensators’) from 82 pharmacies in Cotonou and 19 in Porto-Novo were surveyed. Data entry was performed using Epidata 3.1 software and data analysis was carried out using SPSS software version 21.1. Chi square test was used to compare proportions. A significance threshold of 0.05 was defined for the p value. Results 46% of providers did not know the artemisinin-based combination therapy recommended by the NMCP for treating uncomplicated malaria. 58.7% were not able to recognize the gravity signs of malaria. 89.8% of dispensators were used to deliver an anti-malarial upon patient request, without prior biological confirmation as requested by the NMCP policy. Conclusions Dispensing practices within the studied pharmacies from Cotonou and Porto-Novo were not in adequacy with the NMCP guidelines for uncomplicated malaria, which is a striking weakness in the training of drug providers on key elements of the guidelines for managing malaria. The NMCP needs to help dispensator from private pharmacies sector to standardize drug delivery practices according to its guidelines

P30 Etude clinique d’un collutoire à base d’huile essentielle de Cymbopogon giganteus, chiov (poaceae) chez les adultes souffrant de l’angine de gorge au CHU de SURU-LERE de Cotonou

Introduction : L’angine de gorge est une inflammation de la gorge et en particulier des amygdales, d’origine infectieuse, induisant souvent un recours inapproprié aux antibiotiques. La prise de phyto médicaments peut être envisagée afin de contourner le risque d’antibiorésistance. D’où l’intérêt de cette étude à évaluer l’efficacité antimicrobienne d’un collutoire à base de C. giganteus en clinique.   Matériel et méthodes : L’étude clinique phase 2 à un bras, réalisée a porté sur des patients adultes (18 ans à 65 ans) dont le diagnostic positif a été posé par un médecin et qui ont été traités en ambulatoire durant 07 jours à raison de deux pulvérisations dans la gorge chaque 4h en association avec 1g de paracétamol pris matin, midi et soir pendant 48 h puis si douleur. L’efficacité thérapeutique a été évaluée à l’aide des principaux signes cliniques spécifiques relevés à J0, J3 et J7 à travers un même score préétabli. Résultats et Discussion : Sur 36 patients inclus, 34 efficacités thérapeutiques ont été observées contre 02 échecs. Aucun évènement indésirable majeur signalé, excepté une hypersalivation et une sécheresse de la gorge chez 4 patients, rapidement résorbés par la prise d’eau. Conclusion : Les données recueillies au cours de l’étude suggèrent que le collutoire à base C. giganteus associés au paracétamol soulage l’angine de gorge et les symptômes associés et s’avère un candidat intéressant pour une demande de mise sur le marché

Overview of regulations on medicines derived from traditional Pharmacopoeia in Benin and Burkina Faso

The success of the promotion policies of the Medicines Derived from Traditional Pharmacopoeia (MDTPs) requires a regulation that is adapted to realities and guarantees their quality, efficiency and safeness. This study aims to analyze the legal texts and the current guiding principles obtained from the heads of the departments in charge of traditional medicine in Burkina Faso and Benin. The documents collected from the two countries have been analyzed comparatively and also in relation to WAEMU regulations and the WHO recommendations. Several texts, dealing with the activity, products, facilities and advertisement related to traditional medicine, have been recorded in both countries. The regulation battery of Burkina Faso is more extensive than that of Benin, especially on traditional medicine and pharmacopoeia facilities. In addition, unlike biomedicines, the West African Economic and Monetary Union (WAEMU) and the West African Health Organization (WAHO) have not yet passed community laws on MDTPs. To limit disparities in legal frameworks between the countries of the same sub-region, it is important that the WAEMU or WAHO be involved in the harmonization of pharmaceutical regulations by setting Community rules in the domain

In vitro antiplasmodial activity of plants used in Benin in traditional medicine to treat malaria.

AIM OF THE STUDY: The aim of the study was to evaluate the in vitro antiplasmodial activity of crude extracts of 12 plant species traditionally used in Benin for the treatment of malaria in order to validate their use. MATERIALS AND METHODS: For each species, dichloromethane, methanol and total aqueous extracts were tested. The antiplasmodial activity of extracts was evaluated using the measurement of the plasmodial lactate dehydrogenase activity on chloroquine-sensitive (3D7) and resistant (W2) strains of Plasmodium falciparum. The selectivity of the different extracts was evaluated using the MTT test on J774 macrophage-like murine cells and WI38 human normal fibroblasts. RESULTS: The best growth inhibition of both strains of Plasmodium falciparum was observed with the dichloromethane extracts of Acanthospermum hispidum DC. (Asteraceae) (IC(50)=7.5 microg/ml on 3D7 and 4.8 microg/ml on W2), Keetia leucantha (K. Krause) Bridson (syn. Plectronia leucantha Krause) (Rubiaceae) leaves and twigs (IC(50)=13.8 and 11.3 microg/ml on 3D7 and IC(50)=26.5 and 15.8 microg/ml on W2, respectively), Carpolobia lutea G.Don. (Polygalaceae) (IC(50)=19.4 microg/ml on 3D7 and 8.1 microg/ml on W2) and Strychnos spinosa Lam. (Loganiaceae) leaves (IC(50)=15.6 microg/ml on 3D7 and 8.9 microg/ml on W2). All these extracts had a low cytotoxicity. CONCLUSION: Our study gives some justifications for the traditional uses of some investigated plants

A comparative study of Caesalpinia bonduc (L.) Roxb. root extracts on sexual behaviour in male Wistar rats

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