Pharmaceutical companies in Ukraine aspire to develop their innovative medicinal products and successfully introduce them to the global market. However, along with the prospects of increased usage of these pharmaceuticals, there arises a challenge of heightened waste production, making them a part of the over twenty million tons of PPCPs produced annually. Consequently, one of the tasks in producing new pharmaceuticals is the development of methodologies and approaches not only for quality control but also for their determination in the environment matrices.
 The aim. Develop and validate GC-FID chromatographic method for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride, evaluate their applicability, and compare their "greenness" with the previously developed HPLC method.
 Materials and methods. The determination of the Tilorone dihydrochloride and Enisamium iodide was carried out by gas chromatography with a flame ionization detector using the Rxi-5 ms (30 m long, 0.25 mm outer diameter and 0.25 μm liquid stationary phase thickness)
 Results. Chromatographic GC-FID methods have been developed for the simultaneous determination of Enisamium iodide and Tilorone dihydrochloride. Optimal sample preparation conditions were established, and a validation process was conducted. A comparison with the previously developed HPLC method was made regarding "greenness."
 Conclusions. The developed GC-FID methodology is accurate and more environmentally friendly compared to the previously established methods. It can be recommended to determine Enisamium iodide and Tilorone dihydrochloride in the environmental matrices. It is considered environmentally friendly based on the overall GREENness (AGREE) scale, scoring 0.73 (>0.70), which demonstrates the environmentally favourable nature of the proposed analytical approach

One of the main steps in the pharmaceutical development of drugs is the choice of quality control methods. The correctness of the method must be confirmed by validation. In addition, manufacturers take into account various economic and environmental factors. It is especially important to determine the above aspects for domestic and promising drugs, such as the morpholinium thiazotate. The aim. During the development of methods for the routine analysis of medicinal products, attention should be paid to efficiency of analysis, budget, as well as their impact to the environment. Because of this reason, not only new methods for routine analysis should be developed. It is important this methods must be environmentally-friendly and cost-efficient. Materials and methods. The determination of the morpholinium thiazotate was carried out by HPLC using the SunFire C18 (150 mm × 4,6 mm, 5,0 μm) and gas chromatography with a flame ionization detector using the Rxi-5 ms (30 m long, 0.25 mm outer diameter and 0.25 μm liquid stationary phase thickness). Results. Various chromatographic methods for the routine quantitative analysis of morpholinium thiazotate were developed. The most suitable conditions for sample preparation were established. Proposed methods were compared to find the most ecological and economic. Conclusions. Proposed methods were accurate and reliable. However, an environmental impact assessment showed that GC-FID is a more environmentally friendly and economical method of analysis. Using 12 Principles of green analytical chemistry, the overall “analytical GREEnness (AGREE)” scale for the proposed analytical approach was computed 0.72, showing the good greener nature of the proposed analytical approach

The aim is to study the morphological and anatomical structure of the underground organs of R. confertus and to establish the type of medicinal plant raw materials, to determine a number of its indicators of quality and quantitative content of some groups of biologically active substances using modern methods of analysis.
 Materials and methods. It was used air-dry and freshly collected raw materials and a microscope Delta optic BioLight 300 (Poland) to study the macro- and microscopic characteristics of the plant raw materials. Determination of amount of hydroxycinnamic acids and total polyphenols was determined spectrophotometrically according to monograph «Nettle leaf» and the method 2.8.14 of the State Pharmacopoeia of Ukraine 2.0 (on a spectrophotometer Optizen POP (Korea)). The study of the component composition of hydroxycinnamic acids was performed by HPLC.
 Results. As a result of research of plant raw materials it was established that the underground organs of Rumex confertus are the roots of annual plants and the rhizomes and roots of two or three annual plants. Diagnostic features: morphological (for the root and rhizome - the nature of the surface and fracture) and anatomical (for the rhizome - aerenchyma in the cortex, the presence and location of scleroids and sclerenchyma in the plant raw materials of two or three annual plants, the presence in the cells of the cortex and pith parenchyma simple starch grains, druses and cells with yellow content in freshly harvested plant raw materials; for the root - the colour of peridermal cells, the degree of development of pith rays, the remainder of the primary xylem; a distinctive feature of the annual root from two or three annuals is the absence of scleroids). It was determined the borderline boundaries of indicators in the series of plant raw materials: loss on drying (not more than 13. 5 %), total ash (not more than 11 %), extractable matter (not less than 33 %) and the quantitative content of amount of hydroxycinnamic acids (not less than 1.3 %) and total polyphenols (not less than 3.5 %). It was identified chlorogenic and neochlorogenic acids.
 Conclusions. It was determined the type of underground organs of Rumex confertus: annuals had only roots (tap root systems), from the second to the third year of life the plants have both rhizomes and tap root system. It was established their morphological and anatomical diagnostic features and determined numerical indicators and the quantitative content of the amount of hydroxycinnamic acids and total polyphenols with using modern methods of analysis and chlorogenic and neochlorogenic acids were identified. The obtained data will be used in further research, including the development of the draft monograph of the State Pharmacopoeia of Ukraine 2.0 or the draft methods of quality control of medicinal plant raw materials Rumex confertus and the creation of herbal medicines

Refined sunflower oil is an auxiliary component of many dosage forms made in pharmacy. For their preparation, pharmacies purchase oils from different manufacturers and have the right to use it throughout the shelf life. However, the question arises of maintaining the stability of the fatty acid composition of the oil because of the influence of various environmental factors. In previous studies, the authors investigated the change in the composition of basic fatty acids (FA) of sunflower oil during heating or long-term storage, but there are no studies of the full FA composition of sunflower oil, as well as an assessment of its dependence on the period of the oil using.The aim. Detailed study of the FA composition of refined sunflower oil from different manufacturers, assessment of the effect of the shelf life of the oil on its FA composition and its compliance with the requirements of the European Pharmacopoeia (EP).Materials and methods. The determination of the FA composition of seven samples of refined sunflower oil was carried out by gas chromatography with a flame ionization detector using the Rt-2560 column (100 m×0.25 mm×0.20 µm).Results. The analysis of seven samples of refined sunflower oil showed the presence of 24 FA in most of them. Polyunsaturated FA predominate among them with a percentage of 52.45-60.86 %. The value of the fatty acids percentage in each groups (saturated, mono- and polyunsaturated fatty acids) are quite similar regardless of the oil shelf life at the time of the research. All test samples, except one, by the percentage content of the main FA correspond to EP requirements. The results of the quantitative content of the main FA of sunflower oil determining (palmitic, stearic, oleic, and linoleic) also indicate a similar FA composition of the studied samples.Conclusions. The studied samples of sunflower oil refined in terms of the content of basic FAs meet the requirements of the EP. The obtained results indicate the high stability of the oil and the absence of a significant effect of the storage period on the FA composition

The aim. To investigate both the profile of components and possible difference among herbal raw materials, semi-products and pharmaceuticals of Hedera helix for determination of main standartisation markers.Materials and methods. Investigation of components profile has been performed using the Shimadzu Nexera X2 chromatographic system coupled with a diode-area detector. The ACE C 18 column (250 mm × 4.6 mm with particle size 5 mm) was used for the separation of components. 0.1 % acetic acid and acetonitrile were used as mobile phase A and B, respectively. Studies have been performed on the leaves, dry extract and capsules of H. helix.Results. The determined profile had no significant variation among samples. It has been presented by 19 various components, such as phenolic acids, flavonoids and triterpene saponins. However, kaempferol, nicotiflorin and t-cinnamic acid were not found in the leaf raw material. Hederacoside C might be highlighted as the main marker of raw materials and products of H. helix due to its significant amount in comparison to other components. Its amount was in the range of 64,80 % up to 71,46 % of the total content of components. Moreover, according to some pharmacological studies, hederacoside C is responsible for pharmaceutical usage of H. helix pharmaceutics. Nevertheless, it is not recommended to standardize the plant-based medicines by one marker, since the pharmaceutical activity of such dosage forms is defined by synergism action of all constituents. Except for hederacoside C significant amounts in comparison to other components were found for chlorogenic acid and 4,5-dicaffeoylquinic acid about 5 % and 3 % respectively. Though the latter was found in small concentrations in leaves (0,058 %). This sample had a much higher amount of 3,5-dicaffeoylquinic acid, but in the case of extract and capsules, its content was lower 1,55 % and 0,66 % respectively. Thus, chlorogenic acid has been chosen as a second marker due to its high concentration in all samples and some pharmaceutical activities, such as antioxidant and anti-inflammatory effects.Conclusions. It was found, that standartisation of H. helix products is preferably to perform with determination both hederacoside C and chlorogenic acid. These components were dominant among all components; besides they possess a wide range of pharmaceutical effects. Hence, quantification of hederacoside C and chlorogenic acid is necessary to ensure the high quality of H. helix pharmaceuticals.

The aim. Development of optimal, high-precision and reproducible methods for quantitative determination of the main substance in the substance Epimidin - 1-(4-methoxyphenyl)-5-[2-[4-(4-methoxyphenyl)piperazin-1-yl]-2-oxo-ethyl]pyrazolo[3,4-d]pyrimidin-4-one by high performance liquid chromatography.Materials and methods. High performance liquid chromatography (HPLC) was performed using a ShimadzuNexeraX2 LC-30AD system (Shimadzu, Japan) equipped with a SPD-M20A diode array detector (DAD). ACE C18 column, size 250 x 4.6 mm, YMC with pre-column, particle size 5 μm, filled with octylsilyl silica gel for chromatography P. During the work acetonitrile and trifluoroacetic acid of HPLC class (Sigma-AldrichGmbH, Switzerland) were used, other chemicals and solvents were of analytical grade. In the study an analytical ware class A were used that meet the requirements of SPhU.Results. The following optimal conditions of chromatographic distribution are established: column C18 (250*4.6 mm); the speed of the mobile phase 1 ml / min; column thermostat temperature 35 °С; injection volume 10 μl; mobile phase A - 0.1 % trifluoroacetic acid; mobile phase B - acetonitrile P; the detection wavelength is 270 nm, the retention time of the test compound is 7.22 minutes. The performance of the column was determined for its main indicators, such as the number of theoretical plates (more than 25410) and the coefficient of symmetry (about 1.00). The technique was tested for the influence of various factors, such as flow rate, mobile phase composition and column thermostat temperature. It was established that the influence of these factors is insignificant and does not affect the results obtained by this method. The method was validated in accordance with the recommendations of SPhU on the parameters of specificity, linearity, correctness, precision, robustness (stability).Conclusions. For the first time, a high-precision and reproducible method for quantitative determination of the main substance in the substance Epimidin with anticonvulsant activity by high-performance liquid chromatography was developed. Conditions for chromatographic analysis (HPLC) were standardized. The requirements for the test “System suitability test criteria for chromatographic methods” are set. Statistical processing of the experimental results shows that the relative uncertainty of the average result is within acceptable limits. The correctness of the method was confirmed by validation studies. The developed technique will be used for pharmaceutical development and standardization of dosage form

Aim of this study was to analyze the influence of lifestyle (nutrition and physical activity) on constipation and evaluate the respondents’ attitude to this disorder.Methods. Pharmacy visitors who agreed to answer questions were included in the study. Data from respondents based on age (18-45 years old; 46-65 years old; above 65 years old) and body mass index (“normal,” “overweight,” “obese”) was analyzed. Nutrition and physical activity were analyzed for the purpose of identifying risk factors for constipation. Descriptive and comparative statistics were used – the respondents’ responses are presented in frequencies and percentages, Chi-square test was executed to measure association with knowledge, responses and gender, age, BMI.Results. The study identified a 12.8 % constant constipation rate in the study group with no significant differences between genders, however more males had no constipation problem (50.5 % vs 39.9 %). The age and body mass index had association with constipation (p<0.05). The consumption of coffee and/or tea was not related to constipation in the study group, however, the respondents’ motionless lifestyle was related to constant and occasional constipation (75.0 % and 41.1 %, respectively) (p<0.05).Conclusions. The consumption of carbohydrates, inadequate intake of fluids and motionless lifestyle were identified as risk factors for constipation in this study. Lifestyle modification recommendations might be included in the pharmacists’ consultation

Aim. The aim of the present study was to develop a method for the simultaneous determination of loratadine and auxiliary substances - methyl parahydroxybenzoate and propyl parahydroxybenzoate in the combined "Loratadine+" syrup in the presence of a bupleurum aurus grass extract.Materials and methods. Liquid chromatography separation was performed using a Shimadzu Nexera X2 LC-30AD HPLC system (Shimadzu, Japan) composed of a quaternary pump, an on-line degasser, a column temperature controller, the SIL-30AC autosampler (Shimadzu, Japan); the CTO-20AC thermostat (Shimadzu, Japan) as well as the SPD-M20A diode array detector (DAD).Results and discussion. Identification of the main component and impurities in the combined syrup was performed by determining the retention times of peaks of loratadine, methyl parahydroxybenzoate and propyl parahydroxybenzoate on the chromatogram of the test solution, obtained by quantifying them, which coincided with the retention times of the corresponding peaks on the chromatogram of the reference solution.When developing a quantitative determination method, it was found that using the gradient mode, the best separation between the compounds was observed, the separation coefficient between the peaks of methyl parahydroxybenzoate and the peaks closest to it became more than 2.5, in the case of propyl parahydroxybenzoate this index was more than 3.To confirm the correctness of the proposed method, validation studies were carried out in accordance with the requirements of SPHU. It was established that the uncertainty of sample preparation is 1.5 % for loratadine, 1.47 % for methyl parahydroxybenzoate, and 1.53 % for propyl parahydroxybenzoate, which does not exceed the acceptance criteria. The specificity of the technique was confirmed by comparing the chromatograms of the reference solution, the test solution and the chromatogram of the blank solution. Requirements for the linearity of the method were performed over the entire range of concentrations for loratadine and both excipients. The correlation coefficients were 0.9999, 0.9999 and 0.9995, respectively. The correctness of the technique was carried out according to two criteria - practical and statistical insignificance, which were determined in the course of experimental studies. The results of the assessment of intralaboratory precision showed that the obtained values of the confidence interval of the average result to the criteria of acceptability. Based on the results of the determination of robustness, it was found that for optimal chromatographic conditions, a freshly prepared reference solution can be used within 24 hours.Conclusions. A method was developed for the simultaneous quantitative determination of loratadine and auxiliary substances - methyl parahydroxybenzoate and propyl parahydroxybenzoate in the syrup of "Loratadine+". The conditions that allow to correctly determining all the components in the presence of a bupleurum aurus grass extract were determined. The correctness of the methodology is confirmed by validation studies

The aim. Development of methods of quantitative determination of Cardiazol substance using high-performance liquid chromatography.Materials and methods. The method of high-performance liquid chromato-graphy (HPLC) was to determine of quantitative determination of Cardiazol substance ([3-Allil-4-(41-methoxyphenyl)-3H-thiazole-2-ylidene]-(32-trifluoro-methylphenyl)amine hydrobromide) using Shimadzu Nexera X2 LC -30AD (Shimad-zu, Japan). The acetonitrile of the HPLC grade (Sigma-Aldrich GmbH, Switzerland) was used in the work and other chemicals and solvents were of analytical grade. The test substance was diluted in acetonitrile at a final concentration of 400 μg/ml.Results and discussion. The method of quantitative determination of Cardiazol substance with the help of highly effective liquid chromatography is developed. The developed conditions of testing are selected experimentally. The following optimal conditions for the chromatographic distribution were found: column C8 (250 *4.6 mm; speed of the mobile phase 1 ml/min; thermostat temperature of the column 35 ° C; detecting wavelength 300 nm; holding time of the test compound is 13.9 min. Suitability of determination methods.The following optimal conditions for chromatographic separation were revealed: a C8 column (250*4.6 mm, a mobile phase speed of 1 ml/min, a column thermostat temperature of 35 °C, a detection wavelength of 300 nm. Under the proposed conditions, the retention time of the tested component is 13.9 minutes. The performance of the column was determined for its main indicators, such as the theoretical number of plates (over 65000) and the coefficient of symmetry (about 1.00). The method of quantitative determination was tested in accordance with the recommendations of the Ukrainian and European Pharmacopoeia. The proposed method meets all requirements. The method has been tested for the effects of various factors such as flow rate, the composition of the mobile phase and the temperature of the column thermostat. It is established that the influence of these factors is insignificant and does not affect the results obtained by this method.Conclusions. An analytical method for quantitative determination of Сardiazole substance with cardioprotective action has been developed on the basis of the high-performance liquid chromatography method. The conditions for chromatographic analysis (HPLC) were standardized. Requirements for the test "Checking the suitability of the chromatographic system" were established. The statistical processing of the results of the experiment shows that the relative uncertainty of the average result was within the permissible limits. The developed method for the determination of Сardiazole will be used for further study of substance as a component of various dosage forms

Nowadays spices are known not only for their taste and flavour, but also for their medicinal value. Zingiber officinale Rosc. (Zingiberaceae family) contains a number of bioactive phenolic constituents, which in pure form or it‘s derivates might be potential antioxidants, in the most cases scientists discover 6-gingerol and 6-shogaol as a major constituents of ginger rhizomes.Methods. For this investigation we have chosen four different food supplements, containing ginger and one traditional herbal medicine. The presence of two major ginger constituents in the investigation objects were analysed through TLC analysis, which was performed using CAMAG TLC Visualizer and other method was HPLC analysis, which required a high performance liquid chromatographic system Waters 2695 with fotodiode detector Waters 966 PDA.Results. Our results suggest that ginger-containing food supplements and medicine contain two major constituents which leads to ginger biological active properties. Chromatographic analysis might be useful in providing information about quality of ginger rhizomes and commercial ginger products.Conclusions. Selected chromatography methods are suitable for qualitative and quantitative evaluation of 6-gingerol and 6-shogaol in dietary supplements and other products

Nowadays, combined medicines are becoming more and more widespread. The combination of active pharmaceutical ingredients is necessary to increase the therapeutic effect or shorten the treatment period, or prevent possible complications. The aim of the work is to develop a method for the quantitative determination of biologically active compounds of dry extract of Bupleurum aureum as part of a combined dosage form in the form of a syrup in a mixture with loratadine.Methods. Identification of flavonoids in the extract was carried out by HPLC. To determine the quantitative content of flavonoid substances, the method of absorption spectrophotometry in the visible spectral range was used, based on the formation of colored complexes of flavonoids with a solution of aluminum chloride in an acidic medium.Results of the research. As a result of the research, a spectrophotometric method was developed to quantify the amount of flavonoids in the combined syrup with loratadine and Bupleurum aureum dry extract. The HPLC method establishes the flavonoids contained in the extract.The resulting colored complexes of alcoholic extracts from the syrup after the reaction of interaction with a solution of aluminum chloride in an acetic acid medium were characterized by the presence of absorption maxima at a wavelength of 412 nm. The effect of background absorption is insignificant noise δ = 0.25 %, max δ = 0.51 %. The studied validation characteristics of the technique, which indicate a linear dependence of the amount of flavonoids in terms of rutin in the range of concentration of Bupleurum aureum extract in syrup from 80 % to 120 %, since the value of the correlation coefficient (r) is 0.9999 ³ 0.9981; the angular coefficient of the linear dependence (b) is 0.9947, the free term of the linear dependence (a) is 0.52 £ 1.60. The technique is precise, since the value of the relative confidence interval is less than the critical value for the convergence of the results: D % = 0.37 ≤ 2.60 and the criterion of insignificance of systematic error is fulfilled d = 0.01.Conclusions. The HPLC method established the presence in the dry extract of the aerial part of the Bupleurum aureum substance of the flavonoid structure, which became a prerequisite for the standardization of the active substance in the syrup in the sum of these biologically active compounds. A spectrophotometric method of quantitative determination in the visible region of the sum of flavonoids in terms of rutin in the combined dosage form in the form of a syrup in the presence of another active ingredient loratadine has been developed

Poisoning of benzodiazepines, particularly triazolam, estazolam and alprazolam usually is caused by consumption of the drug in bigger doses than prescribed. So, for the fast determination of material caused poisoning, selective and effective methods of analysis are requested.Methods. Benzodiazepines triazolam, estazolam and alprazolam, were chosen for investigation. Analysis was performed using chromatograph „Waters 2695” with a photodiode array detector (Waters 996, at wavelength 200-400 nm range), ACE C18 (2,1 mm x 5,0 cm, 5 μm) chromatographic column, gradient eluent flow (sulfuric acid buffer 0,1% and ACN), eluent flow rate 0,1 ml/min and injection volume of 10 µl.Results. Methodics for identification and quantification of triazolam, estazolam, alprazolam and their mixture was developed using reference solutions. Validated methodic was adapted for identification and quantification of triazolam, estazolam, alprazolam in medicinal products.Conclusions. Selected methodic is suitable for qualification and quantification of the medicinal preparations: ACE C18 (2,1 mm x 5,0 cm, 5 μm) chromatographic column, gradient eluent flow (sulfuric acid buffer 0,1% and ACN), eluent flow rate 0,1 ml/min, injection volume of 10 µl and diode array detector. Mixture of components has been examined and retention times have been stated as follows: alprazolam (13,216 min), estazolam (13,407 min) and triazolam (14,340 min). Retention time upon repetition of analysis have not exceeded the relative error of p <0,05 limitation.Limits of detection of alprazolam is 0,01 µg/ml, estazolam 0,012 µg/ml, triazolam 0,020 µg/ml. Limit of quantification of alprazolam is 0, 022 µg/ml, estazolam 0, 025 µg/ml, triazolam 0, 045 µg/ml

This study focused on bio-based succinic acid sample preparation and derivatization conditions optimization using GC-MS analytical method. Succinic acid, the precursor of a wide range bio-compounds, especially it is important in accumulation of mitochondrial metabolite succinate (citric acid cycle) and during ischemia controls reperfusion injury through mitochondrial reactive oxygen production. Accurate determination of analytes is the key in metabolomics to use as low molecular biomarkers in case to improve diagnostic methods.Methods. Gas chromatography-mass spectrometry (GC-MS) method. For the quantitative determination of the succinic acid applied derivatization process by silylation using -bis- (trimethylsilyl) -trifluoroacetamide (BSTFA).Results. The derivatization agent BSTFA, the derivatization time of 3-4 hours and derivatization temperature at 70 °C were selected as the optimal derivatization condition for quantification of succinic acid by GС/MS in biological samples. The results show that GC-MS SIM method with evaporation was the most effective to quantify succinate in biological samples after ischemia/reperfusion injury. Selected ion monitoring (SIM) allowed to monitor a subset of fragments with their related mass values in a certain retention time (RT) range for a set of targets.Conclusions. DC – MS has several advantages for measurements of succinate concentration in small kidney tissue samples (lyophilized mitochondria). The method can be applied in small pieces of tissue - biopy samples, tissues from various organs