European Pharmacopoeia Research Articles

Development of molecular sterility assay for rapid quality release of cord blood erythrocytes units for transfusion.

Umbilical cord blood (CB) units stored in banks are an important source of hematopoietic stem cells for transplantation and other cell therapies. New applications, such as their use in transfusions, require rapid quality release as cord blood red blood cells (CB-RBC) have a shorter shelf life. This project aims to investigate the most prevalent microbial contaminants in CB preparations and validate a rapid sterility testing strategy for CB-RBC based on an automated system (BACT/ALERT®) in tandem with a molecular assay (real-time PCR) capable of detecting at least 100 CFU/mL of Cutibacterium acnes in CB-RBC to accelerate the detection of the most common slow-growing bacteria. Microbial contamination incidence was assessed by reviewing 4696 CB sterility tests, revealing a positivity rate of 3.4%, with C. acnes being the most common slow-growing pathogen. The BACT/ALERT® system, which was validated according to European Pharmacopeia guidelines, was an appropriate method for sterility testing of CB-RBC, although it required up to 14 days of culture to detect C. acnes when iFAPlus and iFNPlus bottles were used to neutralize antimicrobials. Interestingly, the BACT/ALERT® method detected C. acnes at 30 CFU/mL within 14 days, while real-time PCR identified concentrations ≥65 CFU/mL by Day 4. In conclusion, we developed a rapid sterility testing strategy that combines automated culture systems and real-time PCR for early microbial contamination, enhancing CB-RBC shelf life for transfusion and emphasizing the importance of combining detection methods.

The lessons from DynHom Research Project

Initiated in 2014, the DynHom (Dynamisation & Homeopathy) project was set up to answer the question: “How are highly dynamized dilutions prepared according to the homeopathic European Pharmacopoeia work?”. This vast program was carried out in collaboration with French and Belgian university teams. Based on the work of Prof. JL Demangeat, already published in nuclear magnetic resonance [1], we confirmed, like other teams around the word, that homeopathic preparations are not simply water but structured solutions, even for dilutions beyond the Avogadro number [2]. This major observation raises a new question: how and by what are these homeopathic solutions? After 8 years of measurements on multiple series of preparations (dynamized products, simply diluted products, and dynamized solvents as controls) using sophisticated laser microscopy (Nanoparticle Tracking Analysis, NTA), the answer is clear: the presence of nanoparticles in all homeopathic preparations is definitively established (3). Homeopathic potentization cannot be considered as simple dilutions. The molecular composition of these preparations was studied in parallel after lyophilization using electron microscopy and infrared spectrometry (SEM-EDX-FTIR). A specific proportion of carbonates and silicates is observed. Sodium bicarbonate is the main component. This remains true throughout manufacturing, even for the highest potentization [4]. In parallel, systematic measurements of the pH were performed on all these serial preparations after pooling manufacturing lines. As before, we conclude that pH profiles are specific for each product. Once again, this is just as evident in high dilutions. The particulate composition seems able to stabilize specific electric fields for each homeopathic medicine (in the publication process). Another key question is whether the specific characteristics of liquid homeopathic medicines can still be detected after impregnation with globules, granules, or powders. To find out, we need to use an innovative approach, electro-photonic analysis (EPA). A high-voltage current (12,000 volts) applied to the granules excites the electrons, which, when the stimulation stops, return to their initial resting state, emitting light. An imprint of the solution impregnated into the granule remains in the form of a specific light spectrum (future publication). These years of research into the pharmacognosy of homeopathic medicines have therefore been useful, but we still have a long way to go to arrive at a pharmacological explanation of our medicines.

Invivo and ex-vivo evaluation and characterization of wound healing effect of topical calendulaofficinalis and turmeric cream in diabetic rats.

Objective: To evaluate wound healing by topically administration of calendulaofficinalis and tumeric cream to diabetic rats. Study Design: Observational Cross-sectional study Setting: Department of Pharmaceutics, Faculty of Pharmacy, BahaUddin Zakariya University, Multan, Pakistan Period: October 2020-march 2021. Method: Different batches of hydrophilic cream bases (C1-C6) were developed and characterized. The optimized base composition (C6) was loaded with calendula Officinal is extract and turmeric powder as an active ingredient (C7) and evaluated the rate at which wounds heal in diabetic rats. Calendula officinal is and turmeric were combined in a cream, as different stability parameters of the creams were analysed at different temperature. Microbiological action of the cream was also examined. Results: The formulation was homogenous, stable with pH 6.29±0.03, viscosity 381± 6.96 Cps and the microbiological quality met the requirements of European pharmacopeia. The total flavonoid content was less than 1. The formulation showed accelerated wound healing with more than 95% wound contraction after tenth day. Conclusion: The result were favorable and suggested that topical administration of Calendula officinal is and turmeric powder combination would enhance wound healing in diabetic rats with potential for diabetic foot ulcer treatment. Moreover, the designed formulation poses good quality emulsion system stabilizing the active ingredient as well as appropriate release of medicinal agents.

Findings on the heavy metal content in herbal drugs and essential oils – An updated database evaluation

In order to guarantee the safety of patients and consumers, the European legal framework restricts the content of heavy metals and further elements in plant material used for the production of medicinal and food products. This data evaluation comprises findings on seven elements between 2016 and 2021 which were assessed against the requirements of the European Pharmacopoeia (Ph.Eur.) and the ICH Q3D guideline on elemental impurities. !5.328 herbal drug samples from 427 products and 537 essential oil samples from 25 products were included in this evaluation which demonstrates that the Ph. Eur. limits set for cadmium, lead and mercury are considered appropriate. Samples from organic production revealed a lower occurrence of cadmium, lead and mercury as compared to samples from conventional production. With regard to the ICH Q3D requirements, none of the herbal drug samples exceeded the respective limit for cobalt, nickel and vanadium, while for arsenic higher values were observed in individual cases. A comparison of the findings with a former period of evaluation showed an increase in positive findings and also a few more exceedances of the Ph.Eur. limits. For essential oils, it can be confirmed that independent from the production process, the probability of heavy metal contamination is low.

Phytochemical variability in essential oils of Juniperus communis var. saxatilis populations grow wild in western Himalaya, India

Needle and berry samples of Juniperus communis L. var. saxatilis Pall. were sampled separately during post rainy season to comparatively investigate the essential oil (EO) yield and phytochemical variability in eight populations grow wild in western Himalaya, India. The EOs obtained from air-dried needle and berry samples separately by hydrodistillation method using Clevenger type apparatus varied in yield with geographic origin of populations, ranged between 2.5-3.4% and 1.8-2.8% (v/w, dried weight basis), respectively. GC-MS analysis of EO profiles resulted in identification of a total ninety-four phytochemicals, representing over 95% of total oil content. Both, needle as well as berry EO samples were dominated by monoterpene hydrocarbons α-pinene (18.52-30.65% and 14.31-27.37%), D-limonene (17.01-28.96% and 15.80-29.70%), β-myrcene (3.74-5.59% and 4.12-14.23%), followed by sesquiterpene hydrocarbons β-elemene (0.86-4.13% and 0.77-3.65%) and germacrene D (0.27-2.57% and 0.20-6.15%), respectively. The oxygenated sesquiterpenes α-cadinol (0.34-5.90% and to 5.63%) and α-bisabolol (to 3.39 and 0.95-6.71%) constituted the third most abundant group of terpenes while oxidized monoterpenes with α-terpineol (0.73-2.17% and 0.10-3.51%) and α-terpinyl acetate (2.10-4.76% and 0.05-3.54%) were the least dominant group. The principal component and hierarchical cluster analysis indicated that selective local environmental forces driving the microhabitat conditions appeared to be linked to high variability in essential oil composition of different common Juniper populations instead of their geographical proximity. The present study revealed that common Juniper essential oils from western Himalaya, India fulfil minimum requirements of European Pharmacopoeia and warrants its further exploitation as a natural resource of commercial significance for dietary and therapeutic applications.

The development of a formulation, the selection and harmonization of the nutrient medium for determining the activity of aminoglycoside antibiotics

The work is devoted to research on the selection of peptone for the optimization of the nutrient medium for the antimicrobial activity by the agar diffusion method of neomycin (framycetin), which have a similar chemical structure. The paper substantiates the relevance and importance of developing a formulation, selecting the optimal composition (ingredients), the quantitative ratio of ingredients, the pH of the medium, compliance of the proposed nutrient media with the State Pharmacopoeia of the Russian Federation and the European Pharmacopoeia for the standardization of antibiotics. The aim of the work was to study the components of the nutrient medium in comparison with the pharmacopoeial standards recommended for determining the effectiveness of antibiotics. It is shown that for carrying out microbiological tests in the standardization of antimicrobial drugs, the nutrient medium is the main one, and the correctness and accuracy of the interpretation of the research results depend on their quality. The experiment studied four brands of dry peptone and broth from different manufacturers – LLC NITSF and FBUN GNTs PMB (Obolensk) when working with the test strain of the spore culture Bacillus subtilis ATCC 6633. An optimal nutrient medium is proposed, the composition and conditions of its preparation are selected. To determine the antimicrobial activity of framycetin by the agar diffusion method, a developed medium for determining the effectiveness of neomycin when working with the spore-forming form of the test strain Baccillus subtillis ATCC 6633 is proposed.

Analysis of Current Approaches to Monitoring the Content of Depressor Substances

INTRODUCTION. Raw materials of biological origin can contaminate medicines with depressor substances, such as histamine, acetylcholine, bradykinin, serotonin, and prostaglandins. In the Russian Federation, tests for the content of these impurities are conducted in anaesthetised healthy cats in accordance with the General Chapter Test for depressor substances of the State Pharmacopoeia of the Russian Federation. The global trend towards ending animal testing and the search for ways to eliminate animal tests for depressor substances motivate researchers to study international approaches to the determination of histamine-like substances.AIM. This study aimed to analyse Russian and international approaches to the control of biologicals for depressor substances and to consider the applicability of alternative instrument-based physicochemical methods for the determination of histamine-like impurities, with a view to reducing animal testing.DISCUSSION. The rationale for reducing the use of animals as test systems is set forth in Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. In 2023, the European Pharmacopoeia Commission initiated the removal of in vivo tests for histamine-like substances from the European Pharmacopoeia. However, China and India use animal testing as the basic method for assessing the safety of medicines. In the Russian Federation, the control of depressor substances is regulated under General Chapter 1.2.4.0008.18 of the State Pharmacopoeia of the Russian Federation. Since the publication of General Chapter 42-0063-07 Test for histamine in edition XII in 2007, the State Pharmacopoeia of the Russian Federation has included a compendial text providing for a reduction in the number of cats used for testing. At present, Russian scientists are searching for targeted physicochemical and immunochemical methods for the determination of depressor substances. It has been demonstrated that a reasonable approach is to assess the need for testing a particular medicinal product in animals on a case-by-case basis. This approach will help minimise the number of in vivo tests and maintain an adequate quality of medicines at the same time.CONCLUSIONS. Despite the global trend towards ending animal testing, it is still necessary and required to control medicines for the content of depressor substances. Nevertheless, there is no global consensus on the need to use animals for this purpose. The absence of a separate general chapter/monograph, or references to it, will not change the fact that histamine-like substances are controlled worldwide. The decision to exclude in vivo methods from the European Pharmacopoeia necessitates the search for alternative test methods and the implementation of additional manufacturing controls to minimise the contamination risk for finished medicinal products. Studies have shown that high-performance liquid chromatography can determine the content of histamine in a limited amount of a medicinal product.

Simplified process for preparing native and depolymerized capsular polysaccharides of Streptococcus pneumoniae

Streptococcus pneumoniae is a major pathogen of bacterial pneumonia, meningitis, sepsis, and otitis media. The pathogenicity of this bacterium is largely attributed to its polysaccharide capsule, a protective layer around bacterial cell that enables bacteria to resist against host defense. Capsular polysaccharides (CPSs) of S. pneumoniae have been used as antigens to develop a variety of pneumococcal vaccines against invasive pneumococcal disease (IPD). These vaccines have been proven to be effective in reducing the incidence of IPD cases that are caused by vaccine-covered serotypes at the global scale. A crucial step in the manufacture of pneumococcal polysaccharide and conjugate vaccines is to purify native and depolymerized CPSs to meet strict quality standards in purity and structural integrity. The major impurities comprise proteins, nucleic acids and cell wall polysaccharides (CWPS). Traditionally, the removal of impurities to obtain purified native CPSs involves a complex process of purification, after which purified CPSs need to be further size-reduced to obtain depolymerized CPSs by multi-step approaches. In this study, we streamlined the process of CPS purification, which involves firstly ultrafiltration, followed by one-step acid precipitation, and finally diafiltration to obtain pure native CPSs. Furthermore, hydrolysis using trifluoroacetic acid (TFA) was integrated into the process to obtain purified depolymerized CPSs. The native and depolymerized CPSs produced by this optimized process were comparable to the materials obtained by the traditional approaches in purity and structural integrity, which would meet the quality standards of CPSs for vaccine production in the current edition of the European Pharmacopeia.

The combination of infectious bronchitis virus BR1 and Mass vaccines provides broad protection

ABSTRACT Two vaccination-challenge trials were performed using a commercial infectious bronchitis virus (IBV) BR1 vaccine, given alone or combined with a commercial IBV Mass vaccine against challenges with IBV M41, 793B, D388 (QX), Q1, Brasil-1 or Variant 2 challenge viruses, which includes the IB viruses that are dominant in South America. The efficacy of the vaccines against the challenge viruses was investigated by determination of the ciliary activity of the tracheal epithelium after challenge. The level of protection induced by the IBV BR1 vaccine alone against the six IBV challenge strains, of which five were of heterologous genotypes, varied from 50% to 100% with an average of 80%. The level of protection induced by the combination of the IBV BR1 and IBV Mass vaccines against the six IBV challenge strains, of which four were of heterologous genotypes, varied from 80% to 100% with an average of 92%. Vaccination with IBV BR1 alone provided a high level of protection against most tested challenge viruses, though the combination of IBV BR1 and IBV Mass was more consistent, showing less variation and compliance with the criterium mentioned in the European Pharmacopoeia 10th edition (at least 80% protection) for all tested challenge viruses. These trials show that vaccination with a combination of IBV BR1 and IBV Mass vaccines provides high levels of protection against the circulating IBV strains in South America.

Optimization of manufacturing process for serotype 14 pneumococcal capsular polysaccharide

Streptococcus pneumoniae is a pathogenic bacterium that causes infections such as pneumonia, meningitis, otitis media, and bacteremia. The prevention of pneumococcal disease by vaccination has become more urgent due to increased antibiotic resistance. Pneumococcal capsular polysaccharides (CPS) are effective vaccine antigens that stimulate the host to produce protective antibodies. S. pneumoniae serotype 14 is one of most prevalent types in Latin America and across the world. However, the yield of S. pneumoniae serotype 14 CPS from existing fermentation processes remains low and requires improvement. In this study, various aspects of the fermentation process were optimized to improve pneumococcal growth and polysaccharide productivity, including feed medium, cultivation gas environment, fermentation pH, and temperature. A simplified purification method was also developed to obtain pure CPS, including ultrafiltration, acid and ethanol precipitation, diafiltration, and lyophilization. These fermentation optimizations significantly enhanced the optical density of pneumococcal bacterial cultures and increased fermentation yields to 2.4–2.6 g/L—significantly higher than previously achieved. Furthermore, the test results of pure CPS could meet the requirements in the European Pharmacopoeia (11th edition). These optimizations provide valuable insights into the nutritional requirements and impact of varying fermentation process parameters on pneumococcal growth and CPS productivity, thus contributing to the development of a more efficient and cost-effective method for the production of pneumococcal CPS—essential for manufacturing vaccines against pneumococcal infections.

The Design of Novel 3D-Printed, Moulded, and Oral Viscous Budesonide Formulations for Paediatrics: A Comparative Evaluation of Their Mucoadhesive Properties.

Paediatric eosinophilic oesophagitis (EoE) treatment is challenging due to the limited number of age-appropriate formulations. This study aims to develop and evaluate oral viscous suspensions and solid formulations of budesonide (BUD), focusing on their in vitro mucoadhesive properties, to enhance drug delivery and therapeutic outcomes in paediatric EoE. This study encompasses the development of oral viscous suspensions and orodispersible solid formulations (moulded tablets and 3D-printed dosage forms) containing BUD. The formulations underwent quality control tests as per the European Pharmacopoeia, chemical stability assessments, and an in vitro evaluation of their mucoadhesiveness properties. A validated analytical method enabled accurate BUD quantification and efficient extraction, and all developed formulations demonstrated chemical stability for 30 days, meeting Ph. Eur. quality standards. Three-dimensional printing using SSE successfully produced 1 mg and 0.5 mg BUD printlets, complying with quality tests for conventional tablets. Formulations containing xanthan gum (L2-XG and P1-0.5-XG) exhibited superior mucoadhesive properties. L2-XG showed significantly higher mucoadhesion than L1-MC. Among the solid formulations, P1-0.5-XG demonstrated the highest mucoadhesive properties. This is the first study to develop solid oral dosage forms of BUD at a very low dose, specifically for paediatric use. The results highlight the potential of 3D printing for developing individualised orodispersible BUD formulations with improved bioadhesion for paediatric EoE treatment. The L2-XG formulation and the XG-containing printlets are the most promising formulations in terms of increasing contact time with the oesophageal mucosa, which could translate into improved therapeutic efficacy in this patient population.

Pharmacodynamic and Clinical Effects of Ginkgo Biloba Extract EGb 761 and Its Phytochemical Components in Alzheimer's Disease.

Extracts made from plants are complex mixtures of substances with varying compositions depending on the plant material and method of manufacture. This complexity makes it difficult for scientists and clinicians to interpret findings from pharmacological and clinical research. We performed a narrative review summarizing information on ginkgo biloba leaf extract, its composition, pharmacological data and clinical evidence supporting its administration for the treatment of Alzheimer's disease (AD). Medicinal products containing ginkgo biloba leaf extract which are manufactured in compliance with the requirements of the European Pharmacopoeia are approved as medicinal products for the treatment of dementia and related conditions by drug regulatory agencies in Europe, Asia and South America. As multicomponent mixtures, they may affect various targets in the pathogenesis of AD, the most common form of dementia. Pharmacodynamic studies demonstrate the effects of EGb 761 and individual constituents on various pathophysiological features of experimentally induced cognitive impairment and neurodegeneration that could contribute to its clinical efficacy. The safety and efficacy in the treatment of AD and cognitive decline has been studied in randomized, placebo-controlled clinical trials. Most of the studies that investigate the effects of ginkgo biloba extract (GbE) used the special extract EGb 761, which makes it the best-researched plant preparation worldwide. It is therefore the only herbal alternative to standard-of-care anti-dementia drugs. However, the mechanism of action has not been fully elucidated yet, and the clinical studies in AD show heterogeneity.

Synthesis, Isolation and Characterization of Nine Pharmacopeial Impurities Related to the Active Pharmaceutical Ingredient Olsalazine Sodium

AbstractOlsalazine sodium is an anti-inflammatory agent used to treat ulcerative colitis. Both British and European Pharmacopoeia list nine process-related impurities in their monographs. Quantification of the impurities in the final synthesized active pharmaceutical ingredient (API) is required to meet the quality standards set by the International Council for Harmonisation (ICH). Both literature reports and commercial vendors for these impurities are few and far between. The present report describes the synthesis, isolation, purification and characterization of these nine pharmacopeial impurities.

Validation of a radiosynthesis method and a novel quality control system for [68 Ga]Ga-MAA: is TLC enough to assess radiopharmaceutical quality?

BackgroundTechnetium-99 m-labelled macroaggregated human serum albumin ([99mTc]Tc-MAA) is commonly used for lung perfusion scintigraphy. The European Pharmacopoeia (Eu.Ph.) specifies thin-layer chromatography (TLC) as the only method to assess its radiochemical purity (RCP). Similarly, TLC is the sole method reported in the literature to evaluate the RCP of Gallium-68-labelled MAA [68 Ga]Ga-MAA, recently introduced for lung perfusion PET/CT imaging. Since [68 Ga]Ga-MAA is prepared from commercial kits originally designed for the preparation of [99mTc]Tc-MAA, it is essential to optimize and validate the preparation methods for [68 Ga]Ga-MAA.ResultsWe tested a novel, simplified method for the preparation of [68 Ga]Ga-MAA that does not require organic solvents, prewash or final purification steps to remove radioactive impurities. We assessed the final product using radio-TLC, radio-UV-HPLC, and radio SDS-PAGE. Overall, our quality control (QC) method successfully detected [68 Ga]Ga-MAA along with all potential impurities, including free Ga-68, [68 Ga]Ga-HSA, unlabeled HSA, which may occur during labelling process and HEPES residual, a non-toxic but non-human-approved contaminant, used as buffer solution. We then applied our QC system to [68 Ga]Ga-MAA prepared under different conditions (25°–40°–75°–95 °C), thus defining the optimal temperature for labelling. Scanning Electron Microscopy (SEM) analysis of the products obtained through our novel method confirmed that most [68 Ga]Ga-MAA particles preserved the morphological structure and size distribution of unlabeled MAA, with a particle diameter range of 25–50 μm, assuring diagnostic efficacy.ConclusionsWe optimized a novel method to prepare [68 Ga]Ga-MAA through a QC system capable of monitoring all impurities of the final products.

Evaluation of the protective effect of Nigella sativa oil’s unsaponifiable fraction in carbon tetrachloride-kidney damage model

Background/Aim: Kidney diseases is a global public health problem. This study was conducted to evaluate the renoprotective effect of the total oil’s unsaponifiable fraction (UF) of a medicinal plant called Nigella sativa. Methods: The extraction of unsaponifiables from the total seed oil of the medicinal plant Nigella sativa was carried out according to the standardized methods of the European Pharmacopoeia (Edition 5.0). Carbon tetrachloride (CCl4) is known to induce kidney toxicity. Acute poisoning of Male albino rats with 1:1 (v/v) mixture of CCl4 and olive oil (3 ml/Kg b.w. of rat by subcutaneous injection) induces considerable kidney pain. Results: Nigella sativa UF is found to be rich in tocopherols, its administration as a curative or preventive treatment to poisoned animals results in a very significant reduction in renal markers (CREA, URE). Histological study shows that kidneys are in better condition than those of intoxicated rats. Conclusion: Measurable biological evidence has been given to the use of such extract as effective substance against kidney diseases. Keywords: CCl4, Renoprotective activity, Nigella sativa, total oil, Unsaponifiable fraction

Development, Validation and Application of a Fast Sequential Method for Na, K, Ca and Mg Determination in Hemodialysis Solutions by HR-CS F AAS.

Hemodialysis solutions are liquid concentrates used during hemodialysis sessions. They are commonly used as a renal replacement therapy. This study aimed to develop a method to determine Na, K, Ca and Mg in hemodialysis solutions by high-resolution continuum source flame atomic absorption spectrometry (HR-CS F AAS) using Design of Experiments (DOE). The combination of the Doehlert Matrix with the desirability function was used to establish a compromise condition in the optimization of the significant variables studied: 60 L/h for acetylene flow rate, 8 mm for burner height, 0.25% (w/v) and 0.60% (w/v) for Cs and La concentrations. The method was validated by following the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and parameters such as the selectivity, linearity, precision, accuracy and robustness were evaluated. Calibration curves with the ranges of 25-130 mg/L for Na, 1.0-5.0 mg/L for K and Ca, and 0.30-0.70 mg/L for Mg were employed. The method proved to be precise (RSD lower than 2.7%) and accurate (mean recovery data, contemplating the three levels added were: 103.0% for Na, 100.5% for K, 101.3% for Ca and 101.5% for Mg). The developed method enables the sequential multielement determination of Na, K, Ca and Mg in a single run. Requiring only one dilution step, this method significantly reduces analysis time for both sample and standard solutions preparation and measurement. This study presents the first method reported in the literature for multielement determination in hemodialysis solutions, offering an alternative approach to the current European Pharmacopeia method.

Evaluation of different techniques for wet granulation and pelletization processes using milk as innovative pharmaceutical excipient for pediatric use

The present study focused on the use of milk as a novel excipient for the manufacture of pharmaceutical dosage forms specifically designed for the pediatric population. Dairy milks with different fat contents were studied to deliver paracetamol orally. The World Health Organization included milk in the list of GRAS (generally recognized as safe) substances, which together with its taste-masking ability and solubility solving properties, makes it a good candidate as an excipient in formulations containing paracetamol for pediatrics. The influence of the fat content in the milk, the fraction of paracetamol, the type of diluent and drying temperature (considered independent variables) were systematically investigated using a Design of Experiments (DoE) approach for the preparation of granules for oral administration, by wet agglomeration using different processing techniques, enabled the construction of mathematical models reflecting the correlation between the variables. Four different techniques were evaluated: wet granulation by low shear mixer, wet granulation by high shear mixer, wet granulation by fluidized bed, and extrusion and spheronization. The granules and pellets obtained were characterized for size, and size distribution of agglomerates, and complete release of the drug (dependent variables), according to the European Pharmacopoeia. The fraction of fat content in the milk promoted an increase on the dissolution rate of paracetamol. The key finding of the first two process techniques was a migration of paracetamol from powdered agglomerates towards the larger particles, probably due to friction and attrition events, which created a fraction of smaller size granules due to the fragmentation and loss of powder from the larger granules. The study has confirmed the potential of milk to be a novel and efficient excipient that can be used as a liquid binder in various agglomeration techniques to deliver drugs orally.

Development and validation of an LC-MS/MS method for highly concentrated tacrolimus and cyclosporine samples prepared from pharmaceutical products to assess drug loss from feeding tubes

IntroductionTacrolimus and cyclosporine are common immunosuppressants utilized post-organ transplantation to manage allograft rejection. Both have narrow therapeutic indices and are frequently measured to support dose adjustments. Although nasogastric tubes are commonly used to provide nutritional support and serve as a route for immunosuppressant administration, they were never validated for such purposes. ObjectiveTo develop and validate a liquid chromatography – tandem mass spectrometry (LC-MS/MS) method for highly concentrated tacrolimus and cyclosporine samples prepared from pharmaceutical products to support the validation of feeding tube administration of these immunosuppressants. MethodsThe method involved stepwise dilutions with dimethyl sulfoxide before analysis using online sample preparation and LC-MS/MS. It was validated in a CLIA-certified clinical laboratory that measures immunosuppressants by LC-MS/MS and is designed to support clinical studies evaluating drug loss from feeding tubes. ResultsThe method was linear between 6.8 µg/mL and 75 µg/mL for tacrolimus, and between 0.9 mg/mL and 10 mg/mL for cyclosporine, with r2 > 0.99 and total precision <5 % at all QC levels. The method demonstrated good recovery using cyclosporine Certified Reference Material, tacrolimus European Pharmacopeia Reference Standard, and prepared pharmaceutical products. Minimal matrix effects were observed. ConclusionAn analytical method was developed and validated for in vitro studies with simulated administration of tacrolimus or cyclosporine to assess loss during drug administration using feeding tubes.

Qualitative and Quantitative Analysis of Stigmasterol from Iraqi Calendula officinalis

Calendula officinalis L. (Asteraceae) plant origin from Mediterranean countries, and in addition to the beauty of its flowers they also have important medicinal properties in the healing of many illnesses particularly skin issues. C. officinalis in the world was reported and assessed as a medicinal plant, like British pharmacopoeia (BP), European pharmacopoeia (EU), WHO and PDR. Calendula officinalis Linn. Contains 0.4% of flavonoids as minimum, calculated as Hyperoside of dried herbal substance according to British pharmacopeia (2). The Chemical Constituents are: Triterpene saponins, Triterpene (Stigmasterol), Flavonoids: isorhamnetin glycosides and quercetin glycosides, Coumarins: umbelliferon, scopoletin and aesculetin, Carotenoids: mainly zeaxanthin and Lutein (5). Stigmasterol is an essential ingredient that has been isolated from plants. Stigmasterol considered a progesterone precursor and serves as an intermediate in corticoids, androgens estrogens biosynthesis (10) and vitamin D3 synthesis (3). Stigmasterol is a triterpenes C30-derive phytosterol, naturally happening C28 and C29 carbon steroid alcohols that is similar to cholesterol in structure as well as biological activity (4).in this study Stigmasterol was identified by TLC, Mass analysis. Structure elucidation was performed using, FTIR and Mass analysis. The quantity of Stigmasterol was measured in the Iraqi plant.

Improved Automated Radiosynthesis of [18F]Dolutegravir: Toward Clinical Applications.

Positron emission tomography imaging using radiolabeled dolutegravir (DTG) is an interesting approach to understand the biodistribution of this antiretroviral drug at HIV-1 sanctuary sites. In the course of clinical translation, we depict herein an improved and pharmaceutically compliant radiosynthesis of [18F]DTG from an original tin precursor. The radiosynthesis was achieved in two steps by copper-mediated radiofluorination, followed by enol ether deprotection using a kit-based AllInOne module. Ready-to-inject [18F]DTG was obtained in 20 ± 5% (n = 12) decay-corrected radiochemical yield within 90 min, representing a 4-fold increase compared to the previously published three-step radiosynthesis. Quality control was carried out with three consecutive [18F]DTG productions according to the current European Pharmacopoeia guidelines, which include pH determination, identity and purity (chemical, radiochemical, and radionuclide) assessments, residual solvent quantification, dosage of lithium, copper, and tin traces, sterility and bacterial endotoxin tests. [18F]DTG (∼2 GBq) was obtained with a molar activity of 59 ± 2 GBq/μmol at the time of injection and was suitable for human applications.