Requirements Of European Pharmacopoeia Research Articles

A comparative analysis of compounding aqueous extracts from the medicinal plant raw material in accordance with the requirements of the regulatory legal acts of Ukraine and the European Union

In the world pharmaceutical practice, there are significant differences regarding the requirements for the technological parameters of aqueous extracts from medicinal plant raw materials.
 Aim. To conduct a comparative analysis of the requirements of regulatory documents of Ukraine, the European Union and Poland regarding the compounding of water extracts from medicinal plant raw materials.
 Materials and methods. Comparative analysis was carried out using systemic and structural-logical analysis. The object of research was normative legal acts regulating medicines compounding in Ukraine, European Pharmacopoeia (EP) XI ed., European Medicines Agency (EMA), Pharmacopoeia of Poland (PP) VI and IX ed., textbook "Farmacja stosowana, Technologia formaci leku; podręcznik dla studentów farmacji" and others.
 Results and discussion. As a result of the analysis, a few distinctive features in the technology of water extracts from medicinal plant raw materials were revealed compared to the requirements of the EU and Poland. According to the requirements of European Pharmacopoeia and the European Medicines Agency, aqueous extracts from medicinal plant raw materials, so-called herbal teas, are prepared with the methods of maceration, infusion and decoction. Macerates are prepared from mucilaginous medicinal plant raw materials with the method of cold infusion at room temperature for 30 minutes. Infusions are usually prepared from delicate parts of the plant, while medicinal plant raw materials are poured with hot water with an initial temperature of 90-100 ºС and infused by gradually cooling the extract at room temperature for 5-15 minutes. Decoctions are usually prepared from roots, rhizomes and bark by boiling medicinal plant raw materials in a boiling water bath for 15-30 minutes, while the raw materials are initially poured with an extractant at room temperature, and after boiling, the extract is immediately filtered.
 The compounding peculiarities of water extracts from medicinal plant raw materials in Poland are as follows: the ratio of medicinal plant raw materials to the extractant with weak biologically active substances is 1:10, with mucus-containing medicinal plant raw materials – 1:20; for medicinal plant raw materials with cardiac glycosides and alkaloids – 1:100; water extracts are prepared by mass, while water absorption coefficients or consumption coefficients are not taken into account; for acidification, 0.5 g of citric acid per 100 g of extractant is added to extracts from alkaloid-containing raw materials; macerates are prepared from mucilage-containing raw materials only by the cold infusion method (for example, flax seed macerate); infusions are prepared only from medicinal plant raw materials containing cardiac glycosides (Lily of the Valley herb, Adonis vernalis herb), where raw materials are poured with boiling water, infused in a water bath for 15 minutes and cooled at room temperature for 15 minutes; decoctions are always prepared from other types of medicinal plant raw materials, where the raw materials are poured with an extractant at room temperature, heated and boiled in a boiling water bath for 30 minutes, filtered without cooling.
 Conclusions. A comparative analysis of the technological parameters of water extracts compounding from medicinal plant raw materials in accordance with the requirements of regulatory documents of Ukraine, the European Union and the Republic of Poland was carried out. A number of differences have been established, particularly in the methods of recipes prescribing, features of calculations, in the ratio of medicinal plant raw materials to the extractant, temperature regimes, extraction time, medium pH, the concepts of the processes of maceration, infusion and decoction. It was found that compared to domestic regulations, the technological process in the EU and Poland is optimized in terms of time, energy and in some cases, raw material expenditure. The obtained results have a practical importance, as they are the basis for further scientific research on the substantiation of the influence of technological factors on the quantitative content of the main groups of biological active substances in the compounded water extracts.

Macro and microscopic analysis of Alfredia nivea KAR. & KIR herb

Kazakhstan flora is rich in promising little-studied plants that are used in folk medicine, but additional deep research is required for their introduction into medical practice with the help of modern scientific techniques. An interesting object for introduction into the official medical and pharmaceutical practice is the Alfredia Nivea Kar. & Kir, family Asteraceae, whose extracts have a neurotropic effect.
 The aim. Regarding mentioned above, in order to create new medicines based on the grass of the Alfredia nivea, it is necessary to develop methods of its macro and microscopic identification.
 Materials and methods. Macro- and microscopic studies of the grass of Alfredia nivea were carried out in accordance with the methodology of SPhU 2.8.23 "Microscopic examination of medicinal vegetable raw materials". Macroscopic studies were performed using magnifying glass and binocular microscope MBS-9, microscopic-using MS Microscopes 10 (eyepiece X5, X10, X15, lenses x10, x40), micromed XS-4130 (WF15X, Opera, lens X40/0, 65, x10/0,25) with microphoto addon (China).
 Results. The morphological and anatomical features of the Alfredia nivea grass. Diagnostic features of the structure of stems, leaves and flowers of this species were revealed.
 Conclusions. The macroscopic and anatomical identification parameters of the grass of the Alfredia of Snow. The study of the morphological and anatomical features of the stems, leaves and flowers of Alfredia nivea, considering the requirements of European pharmacopoeia. Diagnostic features of the structure of stems, leaves and flowers of the species were found, which were used as key quality indicators for standardization and identification of raw materials

SmartFilm Tablets for Improved Oral Delivery of Poorly Soluble Drugs.

(1) Background: Numerous oral drugs exhibit limited bioavailability due to their poor solubility and poor intestinal permeability. The smartFilm technology is an innovative approach that improves the drug aqueous solubility via incorporating the drug in an amorphous state into a cellulose-based matrix, i.e., paper. smartFilms can be transformed into a free-flowing physical form (i.e., paper granules) that can be compressed into tablets with optimum physico-chemical and pharmaceutical properties. The aim of this study was to investigate if smartFilm tablets are suitable for improved oral delivery of poorly water-soluble drugs. (2) Methods: Curcumin is a poorly soluble drug with low intestinal permeability and was used for the production of curcumin-loaded smartFilms. The curcumin-loaded smartFilms were transferred into smartFilm granules which were then compressed into curcumin-loaded smartFilm tablets. The tablets were characterized regarding their physico-chemical and pharmaceutical properties, and the intestinal permeability of curcumin was determined with the ex vivo porcine intestinal model. The ex vivo intestinal permeability of curcumin from the smartFilm tablets was compared to a physical mixture of curcumin and paper and to a classical and to an innovative commercial product, respectively. (3) Results: The produced curcumin-loaded smartFilm tablets fulfilled the European Pharmacopoeia requirements, incorporated curcumin in amorphous state within the cellulose matrix and exhibited an enhanced dissolution rate. The ex vivo intestinal permeation data were shown to correlate to the in vitro dissolution data. The ex vivo intestinal permeation of curcumin from the smartFilm tablets was about two-fold higher when compared to the physical mixture and the classical commercial product. No differences in the ex vivo bioavailability were found between the smartFilm tablets and the innovative commercial product. (4) Conclusions: smartFilm tablets are a cost-effective and industrially feasible formulation approach for the formulation of poorly water-soluble drugs, i.e., BCS class II and IV drugs.

Preparative Production and Purification of Recombinant Human Cyclophilin A.

In this work, we developed the method of preparative production of recombinant human cyclophilin A (rhCypA) in Escherichia coli. The full-length cDNA encoding the gene of human CypA (CYPA) was amplified by RT-PCR from the total RNA of human T cell lymphoma Jurkat. The nucleotide sequence of CYPA was optimized to provide highly effective translation in E. coli. Recombinant CYPA DNA was cloned into the pET22b(+) vector, and the resulted expression plasmid was used to transform E. coli strain BL21(DE3)Gold. The recombinant producer strain of E. coli produced soluble rhCypA in the bacterial cytoplasm. The synthesis efficiency of rhCypA was up to 50% of the total cell protein allowing to produce rhCypA in the amount of 1 g per liter of the culture. We also developed the method for rhCypA purification, consisting of a single-step tandem anion exchange chromatography on DEAE- and Q-Sepharose columns. The protein purity was 95% according to electrophoresis (SDS-PAGE), and its contamination with endotoxin did not exceed 0.05 ng per 1 mg of the protein that met the requirements of European pharmacopoeia for injectable preparations. The produced recombinant protein exhibited functional features of native CypA, i.e., isomerase activity and chemokine activity as assessed by stimulation of migration of mouse bone marrow hematopoietic stem cells in vivo. The generated producer strain of E. coli is a super-producer and could be used for large-scale experimental studies of rhCypA and in its preclinical and clinical trials as a drug.

Stability study of metronidazole cream, extemporaneously made in pharmacy

Abstract Background Rosacea is a chronic recurrent disease, characterized by facial redness, dilated blood vessels, inflammatory rash elements on the face. The pharmacy produces 2% metronidazole cream extemporaneously for the treatment of the disease, as there is no such cream concentration on the market. The treatment of the disease is usually long, so the shelf life of the cream is very important for the patient, especially for those who do not have the opportunity to reach the manufacturing pharmacy. The aim of this investigation was to determine the stability and shelf life of metronidazole cream. Methods Long-term stability study of metronidazole cream was applied at 25 ± 2° C, 60 ± 5% relative humidity; accelerated stability test at 40 ± 2 °C, 75 ± 5% relative humidity. The influence of stress conditions were evaluated on the stability of metronidazole cream. It was determined metronidazole cream texture and microbiological examination was applied. Results It was found that color (white), odor (odorless), integrity (homogeneous), pH (6.8) did not change during storage of metronidazole (6 months). Mechanical properties of the metronidazole cream: firmness, shear, consistency and cohesion throughout the study remained stable. High performance liquid chromatography in the full stability study showed that metronidazole concentrations did not fluctuate within ± 5%. Stress factors such as acid media, alkaline media, hydrogen peroxide, daylight and heat did not affect the stability of the cream. A microbiological examination of the metronidazole cream suited European Pharmacopoeia requirements. Conclusions The extemporaneously made metronidazole cream were stable for 6 months.

Long-term physicochemical stability of acyclovir 5 mg/mL solution stored in polypropylene bags as a simulated hospital stock preparation.

To investigate the long-term chemical and physical stability of 5-mg/mL acyclovir solution in polypropylene bags stored at 5°C ± 3°C for 2 months in order to determine the feasibility of batch production by a centralized intravenous additive service. Eight empty 100-mL polypropylene bags (bags A) and 8 empty 250-mL bags (bags B) were respectively filled with 60 mL and 200 mL of 5-mg/mL acyclovir and 0.9% sodium chloride injection (NaCl) under aseptic conditions through a semiautomated manufacturing process and vacuum packed before storage at 5°C ± 3°C. Four bags A and 4 bags B were tested for chemical stability via a stability-indicating high-performance liquid chromatography (HPLC) method immediately after preparation (time 0) and after 7, 14, 21, 28, 35, 42, and 63 days. Samples for microbiological assay were collected on days 0 and 63 from 4 bags A and 4 bags B immediately after breaking the vacuum. Osmolality, pH, and physical stability were assessed by visual examination, Subvisible particle counting was performed on 6 additional bags (3 each of bags A and B). Mean percentage loss of acyclovir relative to the mean experimental concentration at time 0 was below 5% over the 63-day study period.. No significant differences of pH, no change in color and no precipitate were observed during the study. Subvisible particle counts were compliant with European Pharmacopoeia requirements. Acyclovir solutions remained sterile over the 63 days of the study. Extemporaneously prepared acyclovir 5 mg/mL solutions in 0.9% NaCl stored in polypropylene bags were chemically and physically stable over 63 days when stored at 5°C ± 3°C.

Safety and Efficacy Profile of Live, Intermediate Plus Vaccine Against Infectious Bursal Disease Based on Strain G6.

Infectious bursal disease (IBD) is an acute, highly contagious, immunosuppressive disease of young chickens that causes considerable economic loss in the poultry industry worldwide. Vaccination with live attenuated vaccines is still the most important method used for the control and prevention of IBD in chickens. Here we present the results of in vitro characterization, as well as efficacy and safety testing of a live, intermediate plus vaccine against IBD based on strain G6. Strain characterization confirmed that G6 strain is an intermediate plus strain, showing a high degree of homology with the existing vaccine strains of the same virulence. Safety studies showed that chickens can be vaccinated from 10 days of age. Onset and duration of immunity in specific pathogen free and maternally derived antibodies (MDA) chickens was proven to be 14 and 35 days after vaccination, respectively. When immunizing MDA-positive chickens, vaccine is capable of breakthrough at a titer of ≤500 ELISA units. The field trial conducted on commercial broilers showed a 95% protection against vvIBDV challenge. Stability of the freeze-dried vaccine after reconstitution was confirmed over a period of 3 h. Overall, IBD G6 vaccine has shown good safety and efficacy profile in accordance with European Pharmacopoeia requirements.

Effects of Different Component Contents of Colistin Methanesulfonate on the Pharmacokinetics of Prodrug and Formed Colistin in Human

To evaluate the effects of component contents in different colistin methanesulfonate (CMS) formulas on their clinical pharmacokinetics of the prodrug CMS and the formed colistin. Two CMS formulas (CTTQ and Parkedale) were investigated in a single dose, randomized, open-label, crossover study conducted in 18 healthy Chinese subjects. Both CMS formulas met the requirements of European Pharmacopoeia 9.2 with 12.1% difference in the two major active components (CMS A and CMS B). The PK parameters after a single intravenous infusion of CMS at 2.5mg/kg were calculated and the steady-state plasma colistin concentrations (Css,avg) following multiple dosing, once every 12h for 7days, were simulated with the non-compartment model. The systemic exposure (AUC0-inf) of CMS were 59.49 ± 5.90h·μg/mL and 51.09 ± 4.70h·μg/mL, and the AUC0-inf of colistin were 15.39 ±2.63h·μg/mL and 12.36 ±2.10h·μg/mL for CTTQ and Parkedale, respectively. The ratios (90% CI) of geometric mean of AUC0-inf of CTTQ to Parkedale were 116.38% (112.95%, 119.91%) and 124.49% (120.76%, 128.35%) for CMS and colistin, respectively. The predicted Css,avg (95% CI) were 0.92 (0.85, 0.99) μg/mL and 0.74 (0.69, 0.79) μg/mL for CTTQ and Parkedale, respectively. The difference in component content in the two CMS formulas had a significant (P< 0.001) impact on the systemic exposure of colistin in human, thus, warranted essential considerations in clinical applications.

3D printing of patient-tailored SNEDDS-based suppositories of lidocaine

Personalized drug delivery systems are considered an interesting perspective of the 3D printing technology, with the potential to counterbalance the pharmacological and adverse effects of pharmaceutical products. Our study was focused on the utilization of the pressure-assisted microsyringes technology to fabricate patient-tailored lidocaine-loaded suppositories. Printable inks were fabricated using a combination of an oily compound with a mixture of surfactants and evaluated for the physicochemical properties, as well as for their resemblance to the performance of self-nanoemulsifying drug delivery systems (SNEDDS). The selected ink composition was loaded with lidocaine (LID) in two concentrations (2 w/w % and 4 w/w %), showing droplet sizes in the range between 31.0 ± 8.4 nm and 42.7 ± 13.5 nm, respectively, and negative surface charge. The physicochemical characterization revealed the presence of LID in the amorphous state. The mechanical properties and in vitro performance of the SNEDDS-like suppositories complied with the European Pharmacopoeia requirements. The in vitro dissolution profiles evidenced the delayed LID released in the case of 2 w/w % LID-loaded suppositories. The work highlighted the potential of 3D printing as an alternative pathway to formulate personalized LID-loaded suppositories for local anesthesia.

Quality evaluation and comparison of immunoglobulin prepared by two various methods of human plasma fractionation

Based on European Pharmacopeia, there are some features which should be measured for any intravenous immunoglobulins prior to final release of the product. The most critical ones are the level of prekallikrein and anti-complementary activity in final formulation. For all commercial products, the national reference laboratory is prone to conduct such tests and there is no local report on quality control tests done on the products derived from Iranian human plasma. The study is to measure and control the international requirements such as prekallikrein count and anti-complementary activity for human intravenous immunoglobulins manufactured by local developed process in Iran in pilot scale. IgG-rich fraction was obtained by two fractionation methods. Cryoprecipitate was separated from tested fresh frozen plasma in both methods. In method I, for the next steps, fraction I paste, fraction II+III paste, and at the end, the fraction II paste were precipitated. In method II, the fraction I+II+III paste was simultaneously precipitated followed by deriving the fraction II paste. The paste obtained by both methods was separately subjected to the purification processes using anion and cation exchange chromatography followed by gel filtration and activity level of Prekallikrein in addition to anti-complement activity were compared with other laboratory evaluations. No difference was illustrated between protein and albumin content, pH, and conductivity of the two products. The fraction II paste obtained from both methods, was measured and compared with each other. The IgG yield compared to the primary plasma was calculated as 4.6 and 4.3 g for the aforementioned methods respectively. The absence of impurities was determined by a strong IgG bond in electrophoresis while by HPLC, the dimer/ monomer content was measured more than 99% and the polymer/ aggregate was less than 1%. The amount of prekallikrein and total anti-coagulant activity met the European Pharmacopoeia requirements for both methods.

FORMULATION AND EVALUATION OF SOMATOSTATIN ANALOGUE TABLETS

Objective: This study was undertaken with the aim of the formulation and evaluation of hypothalamic hormone somatostatin analogue tablets, which are intended for neuroendocrine tumours treatment.&#x0D; Methods: Tablets were prepared by using the wet granulation method and evaluated for weight variation, resistance to crushing, friability, disintegration time, content and content uniformity. Stability tests have been performed.&#x0D; Results: Characteristics of somatostatin analogue cyphetrylin powder have been investigated, and the wet granulation method has been chosen to prepare tablets. Technical pharmaceutical properties of formulations of different compositions and obtained tablets were estimated and the best formulation CF11 has been established with appropriate characteristics: disintegration time less than 15 min, resistance to crushing more than 30 kg·m·s−2, weight variation ˂7.5 %, active substance content closed to the nominal quantity and consistent cyphetrylin distribution in the batch. Performed examinations showed cyphetrylin tablets stability in the long-term study.&#x0D; Conclusion: Because of somatostatin analogue cyphetrylin powder properties tablets were prepared by the wet granulation method with starch, lactose, povidone, talc, microcrystalline cellulose, calcium stearate and magnesium stearate as excipients. The best formulation including povidone has characteristics corresponding to requirements of European Pharmacopoeia (PhEur) and State Pharmacopoeia of the Russian Federation (PhRu). Examination of cyphetrylin tablets showed its quality and stability within the time of observation–12 mo.

Development of the Composition and Manufacturing Technology of a New Combined Drug: Lavaflam.

Treatment of diseaes of the biliary system is one of the urgent problems of modern medicine. An original pharmaceutical drug "Lavaflam" in the form of combined tablets, which includes a composition of herbal components flamin (0.05 g) and lavender oil (0.02 g), was proposed for the complex treatment of diseases of the biliary system. Flamin is phytomedicine from the immortelle flowers [Helichrysum arenarium (L.) Moench, Asteraceae], which contains a complex of active substances from the flavonoids group (salipurposide, isosalipurposide, kaempferol, luteolin). It is applied as a choleretic and an anti-inflammatory agent in cholecystitis, cholangitis, and biliary dyskinesia. Tablets were prepared by pressing for separate granulation technology. The lavender oil granulate was prepared using the solid phase method, and β-cyclodextrin was used as an excipient substance. The flamin granulate was prepared by mixing with the spherical-shaped filler mannitol PARTECK M 200. On the basis of previous studies, the excipients of the designed composition tablet "Lavaflam" were β-cyclodextrin (0.27 g), mannitol PARTECK M 200 (0.20 g), croscarmellose sodium (0.03 g), potato starch (0.022 g), PEG 6000 (0.002 g), and magnesium stearate (0.006 g). The assay of the main components of lavender oil with the references linalol and linalyl acetate was performed using gas chromatography. The assay of the total flavonoids of flamin was performed using spectrophotometry with isosalipurposide as the reference. The new phytomedicine tablets "Lavaflam" meet European Pharmacopoeia requirements on the following parameters: appearance, geometric size, average weight, disintegration, friability, resistance of tablets to crushing, and quantification.

Question introduction to the State Pharmacopoeia of Ukraine monograph «Devil’s claw root»

The roots of Harpagophytum procumbens are the imported raw material on the territory of Ukraine, however a wide range of herbal drugs are based on it. Because of this conduct research works on verification of quality control techniques of the monograph «Devil’s claw root», which was included in the State Pharmacopoeia of Ukraine (SPhU) 2.0 for established of the relevance of the introduction of national requirements are need. Aim. To carry out experimental works on the verification of identification methods and quantitative determination of the active substances in the roots of Harpagophytum procumbens according to the requirements of the monograph of the European Pharmacopoeia (EPh) and SPhU 2.0. Materials and methods. Raw materials of Harpagophytum procumbens. The research was carried out by the way of determination the macroscopic, microscopic and quality indicators. Results. Studies showed that the roots of Harpagophytum procumbens, in the case determination of macroscopic, microscopic indicators, quality indicators such as: «Loss of drying», «Total ash», «Starch», qualitative and quantitative determination of biologically active substances (BAS) meet regulatory documentation – monographs EPh 8.0 and SPhU 2.0 «Devil’s claw Root». Conclusions. The data obtained indicate that the raw materials of Harpagophytum procumbens meet the requirements of EPh and SPhU 2.0. The results of the studies have been taken for the development of the monograph SPhU 2.0 «Devil’s claw Root», which is presented without the introduction of additional national parts.

Разработка отраслевого стандартного образца специфической активности эритропоэтина

Основываясь на рекомендациях ВОЗ и разработанных принципах стандартизации эритропоэтинов, в качестве материала для кандидата в стандартный образец выбрана одна из фармацевтических субстанций эпоэтина альфа, внесенных в Государственный реестр лекарственных средств. По результатам контроля производителя выбранная фармацевтическая субстанция соответствовала требованиям нормативной документации, дополнительный контроль подлинности методом капиллярного зонного электрофореза показал присутствие 8 изоформ, что соответствует требованиям Европейской фармакопеи. Разработаны условия получения отраслевого стандартного образца в лиофилизированной форме. Аттестацию отраслевого стандартного образца провели в межлабораторных исследованиях, значение аттестованной характеристики — специфической активности — составило 2400 ME в 1 ампуле, доверительный интервал от 94,5 до 105,8 %. Изучение стабильности разработанного отраслевого стандартного образца позволило установить срок годности 5 лет при температуре хранения –20 °C с возможностью его продления по результатам мониторинга стабильности.

Synthesis of [18F]-L-DOPA radiopharmaceutical on a modified GE TracerLAB Fx F-E platform

A procedure was developed for preparing [18F]-5-hydroxy-2-fluoro-L-thyrosine ([18F]-fluoro-L-dihydroxyphenylalanine, [18F]-L-DOPA) radiopharmaceutical on modified commercially available GE Tracer-LAB Fx F-E platform. This modification can be used for preparing both [18F]-L-DOPA and any other radiopharmaceuticals by electrophilic radiofluorination. The procedure allows preparation of the product meeting all the requirements of European Pharmacopoeia (8th ed.) for clinical use with the decay-corrected radiochemical yield of 20 ± 3% (n = 3).

Experimental Approaches to the Development of Inactivated Poliovirus Vaccine Based on Sabin Strains

The results of development of experimental series of inactivated poliovirus vaccine based on Sabin strains (s-IPV) using conventional modern techniques are presented. Comparative analysis of immunogenicity of the experimental s-IPV and commercial IPV prepared from «wild» poliovirus strains (Imovax, Sanofi Pasteur S.A., France) using several models of laboratory animals showed their identity regarding Sabin and «wild» strains. Original method (an ELISA using specific chicken egg yolk antibodies of class Y-IgY) for quantitative determination of D-antigen in elaboration of vaccine formulation has been developed and protected by patent; conventional control methods in accordance with the WHO and European Pharmacopoeia requirements have been adopted. Finally, at experimental level it was shown that basic characteristics of the s-IPV are comparable with those of commercial IPV prepared from «wild» poliovirus strains

Development of methods for identification of phenolic compounds in tansy flowers

Tanacetum vulgare L. is widespread herb in Ukraine, having a substantial resource base. The absence of national normative documentation for this type of herbal material points to the relevance of research in this area and development of modern methods for identification of phenolic compounds as a main group of biologically active substances.Aim. Development of harmonized with European Pharmacopoeia (PhEur) requirements method for identification of phenolic compounds in Tansy flowers.Methods. For the purpose mentioned above, a standardized TLC-method described in the State Pharmacopoeia of Ukraine (SPhU) monographs for identification of phenolic compounds in different types of herbal material was used. Validation parameters of the method for the studied herb were determined.Results. A TLC method for identification of phenolic compounds in Tansy flowers considering modern PhEur approaches to standardization of herbs was developed. Validation of the identification method was carried out for the following validation parameters: specificity, robustness, and precision. This method was offered for implementation to the SPhU monograph «Tansy flowers».Conclusion. TLC method for identification of phenolic compounds in Tansy flowers was developed for the first time. As a result of validation of the given method for identification of phenolic compounds, by validation parameters (specificity, robustness, and precision), following chromatography conditions were selected: Silica gel coated Aluminum-backed TLC plates, application volume for both test samples and reference samples was 10 μl, mobile phase: formic acid concentrated – water – methyl ethyl ketone – ethyl acetate (10:10:30:50 V/V/V/V), detection – spraying with solutions of aminoethyl diphenylborinate in methanol and macrogol in methanol and view under 365 nm UV. The method for quantitative determination of phenolic compounds in Tansy flowers was offered for implementation in «Tansy flowers» draft monograph

The efficacy of a multivalent calicivirus, herpesvirus and parvovirus vaccine and a rabies vaccine is not affected when administered in combination

The results of a serological study examining the antibody responses generated in cats following administration of a trivalent feline vaccine (feline calicivirus [FCV], feline herpesvirus [FHV] and feline panleucopaenia virus [FPV]; Versifel CVR) in combination with an inactivated rabies vaccine, in compliance with European Pharmacopoeia requirements to support new product registrations, are presented.Nine week old cats were allocated to one of three groups, 10 cats per group. Group 1 received the CVR vaccine on days 0 and 21, group 2 received the rabies vaccine on day 21 and group 3 received the CVR vaccine on day 0 and the CVR vaccine reconstituted with the rabies vaccine (i.e., administered simultaneously) on day 21. Blood samples were collected from each animal, on days 0, 21, 28, 35, 42 and 49; and antibody titres determined using haemagglutination inhibition assay or virus neutralisation test.FHV and FPV antibody responses in the group 3 combination administration were considered non-inferior to the responses in the group 1 CVR-only at all time points (days 28, 35, 42 and 49). However for FCV, group 3 was considered non-inferior to the responses in group 1 on days 28, 35 and 42; but not on day 49. For rabies, group 3 was only considered non-inferior to the responses in group 2 on day 49; an apparent inferiority was observed on days 28, 35 and 42. However, in all cases cats that received the combination administration seroconverted with antibody titres at a magnitude shown in other studies to be protective against virulent challenge, and also at a titre deemed adequate by the European Pharmacopeia monograph 04/2013:0451 (Rabies vaccine [inactivated] for Veterinary use).In conclusion, the data show that combining these two separate vaccines in one administration has limited impact on their ability to generate serological responses, and that these responses are still of a magnitude previously demonstrated to be protective.

Determination of (R)-timolol in (S)-timolol maleate active pharmaceutical ingredient: Validation of a new supercritical fluid chromatography method with an established normal phase liquid chromatography method

An enantioselective supercritical fluid chromatography (SFC) method was developed and validated to meet the current European Pharmacopoeia requirements of a limit test for the determination of S-timolol maleate enantiomeric purity in timolol maleate drug substance. The developed method is presented as an alternative to the current normal phase high performance liquid chromatography (NP-HPLC) method described in the European Pharmacopoeia (Timolol Maleate Monograph). Using a 4.6mm×250mm Chiralcel OD-H (dp: 5μm) column and a mobile phase of (93:7) CO2/0.1% (v/v) TEA in MeOH delivered at 4.0mLmin−1 resolution of 2.0 was achieved within 5min, representing a 3-fold reduction in run-time and an 11-fold reduction in solvent consumption relative to the NP-HPLC method. Method robustness was examined by the variation of flow rate (±0.5mLmin−1), column temperature (±5°C) and column back-pressure (±10bar) and resolution was maintained at ≥1.9 in all cases. R-timolol was resolved from all potential impurities and the limit of detection was improved by increasing the sample concentration threefold compared to the NP-HPLC method such that the method could detect the R-timolol enantiomer at 0.5% (w/w) with respect to S-timolol maleate. Additional validation parameters demonstrated that the potential of the method to be used for routine release testing of timolol maleate raw material for drug product manufacturing in which the quantitation of R-timolol impurity in S-timolol maleate drug substance would be a requirement.

Continuous direct tablet compression: effects of impeller rotation rate, total feed rate and drug content on the tablet properties and drug release

Context: Continuous processing is becoming popular in the pharmaceutical industry for its cost and quality advantages.Objective: This study evaluated the mechanical properties, uniformity of dosage units and drug release from the tablets prepared by continuous direct compression process.Materials and methods: The tablet formulations consisted of acetaminophen (3–30% (w/w)) pre-blended with 0.25% (w/w) colloidal silicon dioxide, microcrystalline cellulose (69–96% (w/w)) and magnesium stearate (1% (w/w)). The continuous tableting line consisted of three loss-in-weight feeders and a convective continuous mixer and a rotary tablet press. The process continued for 8 min and steady state was reached within 5 min. The effects of acetaminophen content, impeller rotation rate (39–254 rpm) and total feed rate (15 and 20 kg/h) on tablet properties were examined.Results and discussion: All the tablets complied with the friability requirements of European Pharmacopoeia and rapidly released acetaminophen. However, the relative standard deviation of acetaminophen content (10% (w/w)) increased with an increase in impeller rotation rate at a constant total feed rate (20 kg/h). A compression force of 12 kN tended to result in greater tablet hardness and subsequently a slower initial acetaminophen release from tablets when compared with those made with the compression force of about 8 kN.Conclusions: In conclusion, tablets could be successfully prepared by a continuous direct compression process and process conditions affected to some extent tablet properties.